Jorge A Ghiso, Ph.D.

Catabolism of Alzheimer's amyloid-b: Implications for brain clearance and plaque deposition
McIntee F.L.; Giannoni P.; Blais S.; Neubert T.; Mathews P.; Rostagno A.; Ghiso J.
2011 ;8:?-?, Neuro-degenerative diseases
Alzheimer's disease (AD) is the leading cause of dementia and the most common form of amyloidosis in humans. Extensive extracellular deposition of amyloid-beta (Abeta), a 40-42 amino acid degradation product of APP, is considered a hallmark feature of AD. Our attention is focused on the highly heterogeneous biochemical nature of the brain Abeta species, delving beyond Abeta40 and Abeta42, likely reflecting a complex balance between amyloidogenic and clearance pathways. We have fractionated water-soluble, detergent-soluble and formic acid soluble Abeta species from brains of transgenic mouse models of amyloid depostion and AD cases. Subsequently, we applied a combination of biochemical techniques including immunoprecipitation followed by identification of Abeta species with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Our biochemical data on the Abeta species present in sporadic AD cases and in transgenic mouse models highlight the presence of similar N-and C-terminally truncated fragments-likely reflecting the ability of multiple proteases to degrade Abeta in situ-and several post-translational modifications with still unclear roles in the amyloidogenesis mechanism. Notably, not all the brain Abeta peptides have identical solubility properties; whereas many of them are highly soluble in water-based physiologic solutions others require mild detergents or strong acids for extraction, suggesting their differential involvement in catabolic and fibrillogenic processes
— id: 136531, year: 2011, vol: 8, page: ?, stat: Journal Article,

Analysis of pyroglutamate-modified amyloid peptides in familial British and familial Danish dementia
Saul A.; Lashley T.; Revesz T.; Ghiso J.; Coomaraswamy J.; Bayer T.A.; Wirths O.
2011 ;8:?-?, Neuro-degenerative diseases
Mutations in the BRI2 gene cause rare neurodegenerative diseases referred to as familial British dementia (FBD) and familial Danish dementia (FDD). These disorders are both associated with neurodegeneration and extensive amyloid deposition in the CNS. FBD and FDD are distinguished from Alzheimer's disease (AD) and other dementing disorders by plaque deposition in the cerebellum and an accompanying cerebellar ataxia. In both lesions, 34-amino acid long peptides are generated (ABri/ADan), which share homology in their first 22 amino acids although they completely differ at their C-termini. Both peptides start with a glutamate at position 1, however, it has been demonstrated using massspectrometry that the majority of these peptides are N-terminally modified starting with a pyroglutamate (pGlu) residue at position 1. So far nothing is known about the immunohistochemical profile of pGlumodified ABri/ADan peptides in brain tissue. This type of post-translational modification is also known in AD where a large proportion of Abeta peptides are N-terminally truncated carrying the pGlu-modification at position 3 (Abeta3pGlu). In AD, this modification leads to a higher aggregation propensity, disturbed proteolytical degradation and increased toxicity of Abeta peptides. We have generated antibodies detecting the pGlu-modification at the N-terminus and characterized their specificity using western-and dot-blot analyses. Immunohistochemical stainings of human brain samples of FBD and FDD patients revealed the presence of pGlu-modified peptides in parenchymal and vascular deposits. In addition, an age-dependent increase in pGlu-modified ADan deposits was detected in the parenchyma and vessel walls of transgenic mice overexpressing the Danish mutant form of Bri2
— id: 136536, year: 2011, vol: 8, page: ?, stat: Journal Article,

Glycosylation of BRI2 on asparagine 170 is involved in its trafficking to the cell surface but not in its processing by furin or ADAM10
Tsachaki, Maria; Serlidaki, Despina; Fetani, Andriana; Zarkou, Vasiliki; Rozani, Ismini; Ghiso, Jorge; Efthimiopoulos, Spiros
2011 Oct;21(10):1382-1388, Glycobiology
Two different mutated forms of BRI2 protein are linked with familial British and Danish dementias, which present neuropathological similarities with Alzheimer's disease. BRI2 is a type II transmembrane protein that is trafficked through the secretory pathway to the cell surface and is processed by furin and ADAM10 (a disintegrin and metalloproteinase domain 10) to release secreted fragments of unknown function. Its apparent molecular mass (42-44 kDa) is significantly higher than that predicted by the number and composition of amino acids (30 kDa) suggesting that BRI2 is glycosylated. In support, bioinformatics analysis indicated that BRI2 bears the consensus sequence Asn-Thr-Ser (residues 170-173) and could be N-glycosylated at Asn170. Given that N-glycosylation is considered essential for protein folding, processing and trafficking, we examined whether BRI2 is N-glycosylated. Treatment of HEK293 (human embryonic kidney) cells expressing BRI2 with the N-glycosylation inhibitor tunicamycin or mutation of Asn170 to alanine reduced its molecular mass by approximately 2 kDa. These data indicate that BRI2 is N-glycosylated at Asn170. To examine the effect of N-glycosylation on BRI2 trafficking at the cell surface, we performed biotinylation and (35)S methionine pulse-chase experiments. These experiments showed that mutation of Asn170 to alanine reduced BRI2 trafficking at the cell surface and its steady state levels at the plasma membrane. Furthermore, we obtained data indicating that this mutation did not affect cleavage of BRI2 by furin or ADAM10. Our results confirm the theoretical predictions that BRI2 is N-glycosylated at Asn170 and show that this post-translational modification is essential for its expression at the cell surface but not for its proteolytic processing
— id: 137820, year: 2011, vol: 21, page: 1382, stat: Journal Article,

Aggregates of ABri and cell-secreted BRI2 containing ABri or ADan inhibit hippocampal LTP
Welzel A.; Freir D.; Herron C.; Ghiso G.; Sala Frigerio C.; Walsh D.
2011 ;8:?-?, Neuro-degenerative diseases
Introduction: Familial British Dementia (FBD) and Familial Danish Dementia (FDD) are rare heritable disorders that phenocopy Alzheimer's disease (AD). Both FBD and FDD are linked to mutations within coding regions of the BRI2 gene that result in the generation of longer BRI2 proteins which when processed by pro-protein convertases release 34 residue long peptides referred to as ABri and ADan. Aim: To assess if ABri and ADan have synaptotoxic roles equivalent to that proposed for the AD-associated amyloid beta-protein (Abeta). Methods: We addressed this issue by using synthetic ABri peptide and medium from cells that stably over-express mutant BRI2. Partially aggregated synthetic ABri and medium from F
— id: 136534, year: 2011, vol: 8, page: ?, stat: Journal Article,

Modeling familial Danish dementia in mice supports the concept of the amyloid hypothesis of Alzheimer's disease
Coomaraswamy, Janaky; Kilger, Ellen; Wolfing, Heidrun; Schafer, Claudia; Kaeser, Stephan A; Wegenast-Braun, Bettina M; Hefendehl, Jasmin K; Wolburg, Hartwig; Mazzella, Matthew; Ghiso, Jorge; Goedert, Michel; Akiyama, Haruhiko; Garcia-Sierra, Francisco; Wolfer, David P; Mathews, Paul M; Jucker, Mathias
2010 Apr 27;107(17):7969-7974, Proceedings of the National Academy of Sciences of the United States of America
Familial Danish dementia (FDD) is a progressive neurodegenerative disease with cerebral deposition of Dan-amyloid (ADan), neuroinflammation, and neurofibrillary tangles, hallmark characteristics remarkably similar to those in Alzheimer's disease (AD). We have generated transgenic (tg) mouse models of familial Danish dementia that exhibit the age-dependent deposition of ADan throughout the brain with associated amyloid angiopathy, microhemorrhage, neuritic dystrophy, and neuroinflammation. Tg mice are impaired in the Morris water maze and exhibit increased anxiety in the open field. When crossed with TauP301S tg mice, ADan accumulation promotes neurofibrillary lesions, in all aspects similar to the Tau lesions observed in crosses between beta-amyloid (Abeta)-depositing tg mice and TauP301S tg mice. Although these observations argue for shared mechanisms of downstream pathophysiology for the sequence-unrelated ADan and Abeta peptides, the lack of codeposition of the two peptides in crosses between ADan- and Abeta-depositing mice points also to distinguishing properties of the peptides. Our results support the concept of the amyloid hypothesis for AD and related dementias, and suggest that different proteins prone to amyloid formation can drive strikingly similar pathogenic pathways in the brain
— id: 137823, year: 2010, vol: 107, page: 7969, stat: Journal Article,

Differential activation of mitochondrial apoptotic pathways by vasculotropic amyloid-beta variants in cells composing the cerebral vessel walls
Fossati, S; Cam, J; Meyerson, J; Mezhericher, E; Romero, I A; Couraud, P O; Weksler, B B; Ghiso, J; Rostagno, A
2010 Jan;24(1):229-241, FASEB journal
Cerebral amyloid angiopathy (CAA) is an age-associated condition and a common finding in Alzheimer's disease in which amyloid-beta (Abeta) vascular deposits are featured in >80% of the cases. Familial Abeta variants bearing substitutions at positions 21-23 are primarily associated with CAA, although they manifest with strikingly different clinical phenotypes: cerebral hemorrhage or dementia. The recently reported Piedmont L34V Abeta mutant, located outside the hot spot 21-23, shows a similar hemorrhagic phenotype, albeit less aggressive than the widely studied Dutch E22Q variant. We monitored the apoptotic events occurring after stimulation of human brain microvascular endothelial and smooth muscle cells with nonfibrillar structures of both variants and wild-type Abeta40. Induction of analogous caspase-mediated mitochondrial pathways was elicited by all peptides, although within different time frames and intensity. Activated pathways were susceptible to pharmacological modulation either through direct inhibition of mitochondrial cytochrome c release or by the action of pan- and pathway-specific caspase inhibitors, giving a clear indication of the independent or synergistic engagement of both extrinsic and intrinsic mechanisms. Structural analyses of the Abeta peptides showed that apoptosis preceded fibril formation, correlating with the presence of oligomers and/or protofibrils. The data support the notion that rare genetic mutations constitute unique paradigms to understand the molecular pathogenesis of CAA
— id: 106090, year: 2010, vol: 24, page: 229, stat: Journal Article,

CEREBRAL AMYLOID ANGIOPATHY AND ALZHEIMER'S DISEASE
Ghiso J; Tomidokoro Y; Revesz T; Frangione B; Rostagno A
2010 Jul 8;61(Suppl):S111-S124, Hirosaki igaku = Hirosaki medical journal
Cerebral amyloid angiopathy (CAA) is increasingly recognized as a major contributor of Alzheimer's disease (AD) pathogenesis. To date, vascular deposits and not parenchymal plaques appear more sensitive predictors of dementia. Amyloid deposition in and around cerebral blood vessels plays a central role in a series of response mechanisms that lead to changes in the integrity of the blood-brain barrier, extravasations of plasma proteins, edema formation, release of inflammatory mediators and matrix metalloproteases which, in turn, produce partial degradation of the basal lamina with the potential to develop hemorrhagic complications. The progressive buildup of amyloid deposits in and around blood vessels chronically limits blood supply and causes focal deprivation of oxygen, triggering a secondary cascade of metabolic events several of which involve the generation of nitrogen and oxygen free radicals with consequent oxidative stress and cell toxicity. Many aspects of CAA in early- and late-onset AD -the special preference of Abeta40 to deposit in the vessel walls, the favored vascular compromise associated with many Abeta genetic variants, the puzzling observation that some of these vasculotropic variants solely manifest with recurrent hemorrhagic episodes while others are mainly associated with dementia- await clarification. Non-Abeta cerebral amyloidoses reinforce the viewpoint that plaque burden is not indicative of dementia while highlighting the relevance of nonfibrillar lesions and vascular involvement in the disease pathogenesis. The lessons learned from the comparative study of Abeta and non-Abeta cerebral amyloidosis provide new avenues and alternative models to study the role of amyloid in the molecular basis of neurodegeneration
— id: 138253, year: 2010, vol: 61, page: S111, stat: Journal Article,

Matrix metalloproteinase 2 (MMP-2) degrades soluble vasculotropic amyloid-beta E22Q and L34V mutants, delaying their toxicity for human brain microvascular endothelial cells
Hernandez-Guillamon, Mar; Mawhirt, Stephanie; Fossati, Silvia; Blais, Steven; Pares, Mireia; Penalba, Anna; Boada, Merce; Couraud, Pierre-Olivier; Neubert, Thomas A; Montaner, Joan; Ghiso, Jorge; Rostagno, Agueda
2010 Aug 27;285(35):27144-27158, Journal of biological chemistry
Patients carrying mutations within the amyloid-beta (Abeta) sequence develop severe early-onset cerebral amyloid angiopathy with some of the related variants manifesting primarily with hemorrhagic phenotypes. Matrix metalloproteases (MMPs) are typically associated with blood brain barrier disruption and hemorrhagic transformations after ischemic stroke. However, their contribution to cerebral amyloid angiopathy-related hemorrhage remains unclear. Human brain endothelial cells challenged with Abeta synthetic homologues containing mutations known to be associated in vivo with hemorrhagic manifestations (AbetaE22Q and AbetaL34V) showed enhanced production and activation of MMP-2, evaluated via Multiplex MMP antibody arrays, gel zymography, and Western blot, which in turn proteolytically cleaved in situ the Abeta peptides. Immunoprecipitation followed by mass spectrometry analysis highlighted the generation of specific C-terminal proteolytic fragments, in particular the accumulation of Abeta-(1-16), a result validated in vitro with recombinant MMP-2 and quantitatively evaluated using deuterium-labeled internal standards. Silencing MMP-2 gene expression resulted in reduced Abeta degradation and enhanced apoptosis. Secretion and activation of MMP-2 as well as susceptibility of the Abeta peptides to MMP-2 degradation were dependent on the peptide conformation, with fibrillar elements of AbetaE22Q exhibiting negligible effects. Our results indicate that MMP-2 release and activation differentially degrades Abeta species, delaying their toxicity for endothelial cells. However, taking into consideration MMP ability to degrade basement membrane components, these protective effects might also undesirably compromise blood brain barrier integrity and precipitate a hemorrhagic phenotype
— id: 112035, year: 2010, vol: 285, page: 27144, stat: Journal Article,

A strategy for designing a peptide probe for detection of beta-amyloid oligomers
Hu, Yang; Su, Baihao; Kim, Chung-Sei; Hernandez, Michael; Rostagno, Agueda; Ghiso, Jorge; Kim, Jin Ryoun
2010 Nov 22;11(17):2409-2418, Chembiochem : a European journal of chemical biology
Aggregation of beta-amyloid (Abeta) is implicated in the pathology of Alzheimer's disease. Development of a robust strategy to detect Abeta oligomeric intermediates, which have been identified as significant toxic agents, would be highly beneficial in the screening of drug candidates as well as enhancing our understanding of Abeta oligomerization. Rapid, specific and quantitative detection, currently unavailable, would be highly preferred for accurate and reliable probing of transient Abeta oligomers. Here, we report the development of a novel peptide probe, PG46, based on the nature of Abeta self-assembly and the conformation-sensitive fluorescence of the biarsenical dye, FlAsH. PG46 was found to bind to Abeta oligomers and displayed an increase in FlAsH fluorescence upon binding. No such event was observed when PG46 was co-incubated with Abeta low-molecular-weight species or Abeta fibrils. Abeta oligomer detection was fast, and occurred within one hour without any additional sample incubation or preparation. We anticipate that the development of a strategy for detection of amyloid oligomers described in this study will be directly relevant to a host of other amyloidogenic proteins
— id: 133836, year: 2010, vol: 11, page: 2409, stat: Journal Article,

Catabolism of Alzheimer's amyloid-beta: Implications for brain clearance and plaque deposition
McIntee F.L.; Neubert T.; Blais S.; Ghiso J.
2010 ;3(2):S47-S47, Clinical & Translational Science
OBJECTIVES: Alzheimer's disease (AD) is the leading cause of dementia and the most common form of amyloidosis in humans. Extensive extracellular deposition of amyloid-beta (Abeta), a 40-42 amino acid degradation product of APP, is considered a hallmark feature of AD. Our attention is focused on the highly heterogeneous biochemical nature of the brain Abeta species. METHODS AND POPULATION: We have fractionated water-soluble, detergent-soluble and formic acid-solube Abeta species from transgenic mouse models of amyloid deposition and AD cases. Subsequently, we applied a combination of biochemical techniques including immunoprecipitation followed by identification of Abeta fragments with a novel highly sensitive matrixassisted laser desorption/ionization time-of-flight mass spectrometry technique. RESULTS: Our biochemical data on the Abeta species present in sporadic and familial AD cases and in transgenic mouse models highlight the presence of similar N- and C-terminally truncated fragments-likely reflecting the ability of multiple proteases to degrade Abeta in situ- and several post-translational modifications with still unclear roles in the amyloidogenesis mechanism. Notably, not all the brain Abeta peptides have identical solubility properties. SIGNIFICANCE OF STUDY: In view of these findings and the growing evidence that an imbalance between Abeta production and clearance plays a major role in the process of Abeta deposition, we hypothesize that certain truncated and post-translationally modified Abeta fragments have a distinct and opposite role in clearance and fibrillization mechanisms and that the spectrum and abundance of these species vary according to the magnitude of the amyloid load
— id: 111407, year: 2010, vol: 3, page: S47, stat: Journal Article,

Deficient high-affinity binding of Pittsburgh compound B in a case of Alzheimer's disease
Rosen, Rebecca F; Ciliax, Brian J; Wingo, Thomas S; Gearing, Marla; Dooyema, Jeromy; Lah, James J; Ghiso, Jorge A; LeVine, Harry 3rd; Walker, Lary C
2010 Feb;119(2):221-233, Acta neuropathologica
Radiolabeled Pittsburgh compound B (PIB) is a benzothiazole imaging agent that usually binds with high affinity, specificity, and stoichiometry to cerebral beta-amyloid (Abeta) in patients with Alzheimer's disease. Among a cohort of ten AD subjects examined postmortem, we describe a case of idiopathic, end-stage Alzheimer's disease with heavy Abeta deposition yet substantially diminished high-affinity binding of (3)H-PIB to cortical homogenates and unfixed cryosections. Cortical tissue samples were analyzed by immunohistochemistry, electron microscopy, ELISA, immunoblotting, MALDI-TOF mass spectrometry, in vitro (3)H-PIB binding and (3)H-PIB autoradiography. The PIB-refractory subject met the histopathological criteria for AD. However, cortical tissue from this case contained more vascular beta-amyloidosis, higher levels of insoluble Abeta40 and Abeta42, and a higher ratio of Abeta40:Abeta42 than did tissue from the nine comparison AD cases. Furthermore, cerebral Abeta from the PIB-refractory subject displayed an unusual distribution of low- and high-molecular weight Abeta oligomers, as well as a distinct pattern of N- and C-terminally truncated Abeta peptides in both the soluble and insoluble cortical extracts. Genetically, the patient was apolipoprotein-E3/4 heterozygous, and exhibited no known AD-associated mutations in the genes for the beta-amyloid precursor protein, presenilin1 or presenilin2. Our findings suggest that PIB may differentially recognize polymorphic forms of multimeric Abeta in humans with Alzheimer's disease. In addition, while the prevalence of PIB-refractory cases in the general AD population remains to be determined, the paucity of high-affinity binding sites in this AD case cautions that minimal PIB retention in positron-emission tomography scans of demented patients may not always rule out the presence of Alzheimer-type Abeta pathology
— id: 101719, year: 2010, vol: 119, page: 221, stat: Journal Article,

SDS-PAGE/immunoblot detection of Abeta multimers in human cortical tissue homogenates using antigen-epitope retrieval
Rosen, Rebecca F; Tomidokoro, Yasushi; Ghiso, Jorge A; Walker, Lary C
2010 ;(38):?-?, Journal of visualized experiments : JoVE
The anomalous folding and polymerization of the beta-amyloid (Abeta) peptide is thought to initiate the neurodegenerative cascade in Alzheimer's disease pathogenesis(1). Abeta is predominantly a 40- or 42-amino acid peptide that is prone to self-aggregation into beta-sheet-rich amyloid fibrils that are found in the cores of cerebral senile plaques in Alzheimer's disease. Increasing evidence suggests that low molecular weight, soluble Abeta multimers are more toxic than fibrillar Abeta amyloid(2). The identification and quantification of low- and high-molecular weight multimeric Abeta species in brain tissue is an essential objective in Alzheimer's disease research, and the methods employed also can be applied to the identification and characterization of toxic multimers in other proteopathies(3). Naturally occurring Abeta multimers can be detected by SDS-polyacrylamide gel electrophoresis followed by immunoblotting with Abeta-specific antibodies. However, the separation and detection of multimeric Abeta requires the use of highly concentrated cortical homogenates and antigen retrieval in small pore-size nitrocellulose membranes. Here we describe a technique for the preparation of clarified human cortical homogenates, separation of proteins by SDS-PAGE, and antigen-epitope retrieval/Western blotting with antibody 6E10 to the N-terminal region of the Abeta peptide. Using this protocol, we consistently detect Abeta monomers, dimers, trimers, tetramers, and higher molecular weight multimers in cortical tissue from humans with Alzheimer's pathology
— id: 137822, year: 2010, vol: , page: ?, stat: Journal Article,

Cerebral amyloidosis: amyloid subunits, mutants and phenotypes
Rostagno, A; Holton, J L; Lashley, T; Revesz, T; Ghiso, Jorge
2010 Feb;67(4):581-600, Cellular & molecular life sciences: CMLS
Cerebral amyloid diseases are part of a complex group of chronic and progressive entities bracketed together under the common denomination of protein folding disorders and characterized by the intra- and extracellular accumulation of fibrillar aggregates. Of the more than 25 unrelated proteins known to produce amyloidosis in humans only about a third of them are associated with cerebral deposits translating in cognitive deficits, dementia, stroke, cerebellar and extrapyramidal signs, or a combination thereof. The familial forms reviewed herein, although infrequent, provide unique paradigms to examine the role of amyloid in the mechanism of disease pathogenesis and to dissect the link between vascular and parenchymal amyloid deposition and their differential contribution to neurodegeneration
— id: 106366, year: 2010, vol: 67, page: 581, stat: Journal Article,

APOE genotype results in differential effects on the peripheral clearance of amyloid-beta42 in APOE knock-in and knock-out mice
Sharman, Matthew J; Morici, Michael; Hone, Eugene; Berger, Tamar; Taddei, Kevin; Martins, Ian J; Lim, Wei Ling F; Singh, Sajla; Wenk, Markus R; Ghiso, Jorge; Buxbaum, Joseph D; Gandy, Sam; Martins, Ralph N
2010 ;21(2):403-409, Journal of Alzheimer's Disease
The epsilon4 allele of apolipoprotein E (APOE) is currently the major genetic risk factor identified for Alzheimer's disease (AD). Previous in vivo data from our laboratory has demonstrated that amyloid-beta (Abeta) is rapidly removed from the plasma by the liver and kidney and that the rate of its clearance is affected by ApoE in C57BL/6J and APOE-/- mice. To expand upon these findings, we assessed the peripheral clearance of human synthetic Abeta42 in APOE epsilon2, epsilon3, and epsilon4 knock-in and APOE knock-out mice injected with lipidated recombinant apoE2, E3, and E4 protein. Our results show that APOE does influence the rate at which the mice are able to clear Abeta42 from their bloodstream. Both APOE epsilon4 mice and APOE knock-out mice treated with lipidated recombinant apoE4 demonstrated increased retention of plasma Abeta42 over time compared to APOE epsilon2/APOE knock-out rE2 and APOE epsilon3/APOE knock-out rE3 mice. These findings suggest that the peripheral clearance of Abeta42 is significantly altered by APOE genotype. Given that APOE epsilon4 is a risk factor for AD, then these novel findings provide some insight into the role of ApoE isoforms on the peripheral clearance of Abeta which may impact on clearance from the brain
— id: 137821, year: 2010, vol: 21, page: 403, stat: Journal Article,

Profiling brain and plasma lipids in human APOE epsilon2, epsilon3, and epsilon4 knock-in mice using electrospray ionization mass spectrometry
Sharman, Matthew J; Shui, Guanghou; Fernandis, Aaron Z; Lim, Wei Ling F; Berger, Tamar; Hone, Eugene; Taddei, Kevin; Martins, Ian J; Ghiso, Jorge; Buxbaum, Joseph D; Gandy, Sam; Wenk, Markus R; Martins, Ralph N
2010 ;20(1):105-111, Journal of Alzheimer's Disease
It is known that apolipoprotein E (ApoE) is essential for normal lipid metabolism. ApoE is the major apolipoprotein in the central nervous system and plays a key role in neurobiology by mediating the transport of cholesterol, phospholipids, and sulfatides. We therefore examined APOE epsilon2, epsilon3, and epsilon4 knock-in mice, using electrospray ionization mass spectrometry to determine if APOE genotype or age leads to altered levels in the brain of a number of glycerophospholipids (phosphatidylinositol, PI; phosphatidylethanolamine, PE; phosphatidic acid, PA, phosphatidylserine, PS; phosphatidylcholine, PC), sphingolipids (sphingomyelin, SM; ceramide, Cer), cholesterol, and triacylglycerols. We observed slight changes within individual PI, PE, PC, Cer, and SM lipid levels in APOE epsilon2 and epsilon4 mice compared to APOE epsilon3 mice. However, overall, we did not observe any major effects in APOE epsilon4 knock-in mice for the levels of the glycerophospholipids measured, as compared to APOE epsilon2 and epsilon3 mice. Our findings indicate that variations in ApoE isoforms do not per se affect bulk lipid homeostasis in the brain. These findings indicate that APOE epsilon4 is not associated with disturbances in brain sterol or sphingolipids in the absence of environmental factors
— id: 133514, year: 2010, vol: 20, page: 105, stat: Journal Article,

PYROGLUTAMATE FORMATION AT THE N-TERMINI OF ABRI MOLECULES IN FAMILIAL BRITISH DEMENTIA IS NOT RESTRICTED TO THE CENTRAL NERVOUS SYSTEM
Tomidokoro Y; Tamaoka A; Holton JL; Lashley T; Frangione B; Revesz T; Rostagno A; Ghiso J
2010 Jul 8;61(Suppl):S262-S269, Hirosaki igaku = Hirosaki medical journal
Amyloid molecules harboring pyroglutamate (pGlu) residue at the N-termini are considered to be important for the development of cerebral amyloidosis such as Alzheimer's disease and thought to be either spontaneously generated or being catalyzed by glutaminyl cyclase. Familial British dementia (FBD) is an autosomal dominant form of dementia neuropathologically characterized by parenchymal amyloid and preamyloid deposits, extensive cerebral amyloid angiopathy, and neurofibrillary tangles. FBD is caused by a stop to Arg mutation in the BRI2 gene, generating de novo created amyloid molecule ABri which accumulates in FBD brains but is not present in the normal population. Soluble ABri molecules present in the circulation of carriers of the BRI2 mutation are 34 amino acids long exclusively harboring Glu residue at the N-termini (ABri1-34E), whereas water- and formic acid-soluble ABri molecules extracted from FBD brains have abundant ABri species bearing pGlu residue (ABri1-34pE), suggesting that pyroglutamate formation occurs at the site of deposition. In order to further clarify the mechanism (s) of ABri deposition, we studied whether pyroglutamate formation indeed occurs outside the central nervous system taking advantage that FBD is also a systemic amyloidosis. Soluble and fibrillar ABri molecules extracted from systemic organs and analyzed biochemically using a combination of immunoprecipitation, mass spectrometry, and western blot analysis were oligomeric in size and contained a large proportion of ABri1-34pE. The data indicate that pyroglutamate formation at the N-termini of ABri molecules is an early step in the process of FBD amyloid deposition, and its formation is not restricted to the central nervous system
— id: 138254, year: 2010, vol: 61, page: S262, stat: Journal Article,

Iowa variant of familial Alzheimer's disease: accumulation of posttranslationally modified AbetaD23N in parenchymal and cerebrovascular amyloid deposits
Tomidokoro, Yasushi; Rostagno, Agueda; Neubert, Thomas A; Lu, Yun; Rebeck, G William; Frangione, Blas; Greenberg, Steven M; Ghiso, Jorge
2010 Apr;176(4):1841-1854, American journal of pathology
Mutations within the amyloid-beta (Abeta) sequence, especially those clustered at residues 21-23, which are linked to early onset familial Alzheimer's disease (AD), are primarily associated with cerebral amyloid angiopathy (CAA). The basis for this predominant vascular amyloid burden and the differential clinical phenotypes of cerebral hemorrhage/stroke in some patients and dementia in others remain unknown. The AbetaD23N Iowa mutation is associated with progressive AD-like dementia, often without clinically manifested intracerebral hemorrhage. Neuropathologically, the disease is characterized by predominant preamyloid deposits, severe CAA, and abundant neurofibrillary tangles in the presence of remarkably few mature plaques. Biochemical analyses using a combination of immunoprecipitation, mass spectrometry, amino acid sequence, and Western blot analysis performed after sequential tissue extractions to separately isolate soluble components, preamyloid, and fibrillar amyloid species indicated that the Iowa deposits are complex mixtures of mutated and nonmutated Abeta molecules. These molecules exhibited various degrees of solubility, were highly heterogeneous at both the N- and C-termini, and showed partial aspartate isomerization at positions 1, 7, and 23. This collection of Abeta species-the Iowa brain Abeta peptidome-contained clear imprints of amyloid clearance mechanisms yet highlighted the unique neuropathological features shared by a non-Abeta cerebral amyloidosis, familial Danish dementia, in which neurofibrillary tangles coexist with extensive pre-amyloid deposition in the virtual absence of fibrillar lesions. These data therefore challenge the importance of neuritic plaques as the sole contributors for the development of dementia
— id: 108922, year: 2010, vol: 176, page: 1841, stat: Journal Article,

BRI2 homodimerizes with the involvement of intermolecular disulfide bonds
Tsachaki, Maria; Ghiso, Jorge; Rostagno, Agueda; Efthimiopoulos, Spiros
2010 Jan;31(1):88-98, Neurobiology of aging
Familial British and Familial Danish Dementia (FBD and FDD) are two dominantly inherited neurodegenerative diseases that present striking similarities with Alzheimer's disease. The genetic defects underlying those dementias are mutations in the gene that encodes for BRI2 protein. Cleavage of mutated BRI2 by furin releases the peptides ABri or ADan, which accumulate in the brains of patients. BRI2 normal function is yet unknown. To unwind aspects of its cellular role, we investigated the possibility that BRI2 forms dimers, based on structural elements of the protein, the GXXXG motif within its transmembrane domain and the odd number of cysteine residues. We found that BRI2 dimerizes in cells and that dimers are held via non-covalent interactions and via disulfide bridges between the cysteines at position 89. Additionally, we showed that BRI2 dimers are formed in the ER and appear at the cell surface. Finally, BRI2 dimers were found to exist in mouse brain. Revealing the physiological properties of BRI2 is critical in the elucidation of the deviations that lead to neurodegeneration
— id: 81089, year: 2010, vol: 31, page: 88, stat: Journal Article,

Neuroprotective natural antibodies to assemblies of amyloidogenic peptides decrease with normal aging and advancing Alzheimer's disease
Britschgi, M; Olin, C E; Johns, H T; Takeda-Uchimura, Y; LeMieux, M C; Rufibach, K; Rajadas, J; Zhang, H; Tomooka, B; Robinson, W H; Clark, C M; Fagan, A M; Galasko, D R; Holtzman, D M; Jutel, M; Kaye, J A; Lemere, C A; Leszek, J; Li, G; Peskind, E R; Quinn, J F; Yesavage, J A; Ghiso, J A; Wyss-Coray, T
2009 Jul 21;106(29):12145-12150, Proceedings of the National Academy of Sciences of the United States of America
A number of distinct beta-amyloid (Abeta) variants or multimers have been implicated in Alzheimer's disease (AD), and antibodies recognizing such peptides are in clinical trials. Humans have natural Abeta-specific antibodies, but their diversity, abundance, and function in the general population remain largely unknown. Here, we demonstrate with peptide microarrays the presence of natural antibodies against known toxic Abeta and amyloidogenic non-Abeta species in plasma samples and cerebrospinal fluid of AD patients and healthy controls aged 21-89 years. Antibody reactivity was most prominent against oligomeric assemblies of Abeta and pyroglutamate or oxidized residues, and IgGs specific for oligomeric preparations of Abeta1-42 in particular declined with age and advancing AD. Most individuals showed unexpected antibody reactivities against peptides unique to autosomal dominant forms of dementia (mutant Abeta, ABri, ADan) and IgGs isolated from plasma of AD patients or healthy controls protected primary neurons from Abeta toxicity. Aged vervets showed similar patterns of plasma IgG antibodies against amyloid peptides, and after immunization with Abeta the monkeys developed high titers not only against Abeta peptides but also against ABri and ADan peptides. Our findings support the concept of conformation-specific, cross-reactive antibodies that may protect against amyloidogenic toxic peptides. If a therapeutic benefit of Abeta antibodies can be confirmed in AD patients, stimulating the production of such neuroprotective antibodies or passively administering them to the elderly population may provide a preventive measure toward AD
— id: 127730, year: 2009, vol: 106, page: 12145, stat: Journal Article,

Erratum: Genetics and molecular pathogenesis of sporadic and hereditary cerebral amyloid angiopathies (Acta Neuropathologica (2009) vol. 118 (115-130) 10.1007/s00401-009-0501-8)
Revesz T.; Holton J.L.; Lashley T.; Plant G.; Frangione B.; Rostagno A.; Ghiso J.
2009 ;118(2):321-, Acta neuropathologica
— id: 101651, year: 2009, vol: 118, page: 321, stat: Journal Article,

Genetics and molecular pathogenesis of sporadic and hereditary cerebral amyloid angiopathies
Revesz, Tamas; Holton, Janice L; Lashley, Tammaryn; Plant, Gordon; Frangione, Blas; Rostagno, Agueda; Ghiso, Jorge
2009 Jul;118(1):115-130, Acta neuropathologica
In cerebral amyloid angiopathy (CAA), amyloid fibrils deposit in walls of arteries, arterioles and less frequently in veins and capillaries of the central nervous system, often resulting in secondary degenerative vascular changes. Although the amyloid-beta peptide is by far the commonest amyloid subunit implicated in sporadic and rarely in hereditary forms of CAA, a number of other proteins may also be involved in rare familial diseases in which CAA is also a characteristic morphological feature. These latter proteins include the ABri and ADan subunits in familial British dementia and familial Danish dementia, respectively, which are also known under the umbrella term BRI2 gene-related dementias, variant cystatin C in hereditary cerebral haemorrhage with amyloidosis of Icelandic-type, variant transthyretins in meningo-vascular amyloidosis, disease-associated prion protein (PrP(Sc)) in hereditary prion disease with premature stop codon mutations and mutated gelsolin (AGel) in familial amyloidosis of Finnish type. In this review, the characteristic morphological features of the different CAAs is described and the implication of the biochemical, genetic and transgenic animal data for the pathogenesis of CAA is discussed
— id: 92856, year: 2009, vol: 118, page: 115, stat: Journal Article,

Hereditary forms of cerebrovascular amyloidosis
Rostagno A; Ghiso J
Vascular cognitive impairment in clinical practice New York : Cambridge University Press, 2009,
— id: 5117, year: 2009, vol: , page: 139, stat: Chapter,

Isolation and biochemical characterization of amyloid plaques and paired helical filaments
Rostagno, Agueda; Ghiso, Jorge
2009 Sep;Chapter 3:Unit 3.33 3.33.1-Unit 3.33 3.3333, Current protocols in cell biology
Extracellular deposits of amyloid fibrils in the form of parenchymal plaques and cerebrovascular lesions, as well as intracellular accumulation of paired-helical filaments in the form of neurofibrillary tangles (NFT) in selected neuronal populations are the main neuropathologic hallmarks of Alzheimer's disease. Amyloid fibrils composed of polymeric structures of the amyloid-beta (Abeta) concentrate at the center of senile plaques and accumulate in the walls of cerebral blood vessels, exhibiting extensive Congo red/thioflavin S staining. Intraneuronal NFT are composed of building blocks of aberrantly hyperphosphorylated species of the microtubule-associated protein tau, which accumulate in the perinuclear cytoplasm of vulnerable neurons in the form of paired helical filaments (PHF). This unit presents a variety of protocols for the isolation, biochemical analysis, and characterization of amyloid fibrils and neurofibrillary tangles
— id: 137824, year: 2009, vol: Chapter 3, page: Unit 3.33 3.33.1, stat: Journal Article,

Dutch and Arctic mutant peptides of beta amyloid(1-40) differentially affect the FGF-2 pathway in brain endothelium
Solito, Raffaella; Corti, Federico; Fossati, Silvia; Mezhericher, Emiliya; Donnini, Sandra; Ghiso, Jorge; Giachetti, Antonio; Rostagno, Agueda; Ziche, Marina
2009 Feb 1;315(3):385-395, Experimental cell research
Single point mutations of the amyloid precursor protein generate Abeta variants bearing amino acid substitutions at positions 21-23. These mutants are associated with distinct hereditary phenotypes of cerebral amyloid angiopathy, manifesting varying degrees of tropism for brain vessels, and impaired microvessel remodeling and angiogenesis. We examined the differential effects of E22Q (Dutch), and E22G (Arctic) variants in comparison to WT Abeta on brain endothelial cell proliferation, angiogenic phenotype expression triggered by fibroblast growth factor (FGF-2), pseudo-capillary sprouting, and induction of apoptosis. E22Q exhibited a potent anti-angiogenic profile in contrast to E22G, which had a much weaker effect. Investigations on the FGF-2 signaling pathway revealed the greatest differences among the peptides: E22Q and WT peptides suppressed FGF-2 expression while E22G had barely any effect. Phosphorylation of the FGF-2 receptor, FGFR-1, and the survival signal Akt were abolished by E22Q and WT peptides, but not by E22G. The biological dissimilar effect of the mutant and WT peptides on cerebral EC cannot be assigned to a particular Abeta structure, suggesting that the toxic effect of the Abeta assemblies goes beyond mere multimerization
— id: 96457, year: 2009, vol: 315, page: 385, stat: Journal Article,

Tauroursodeoxycholic acid prevents E22Q Alzheimer's Abeta toxicity in human cerebral endothelial cells
Viana, R J S; Nunes, A F; Castro, R E; Ramalho, R M; Meyerson, J; Fossati, S; Ghiso, J; Rostagno, A; Rodrigues, C M P
2009 Mar;66(6):1094-1104, Cellular & molecular life sciences: CMLS
The vasculotropic E22Q mutant of the amyloid-beta (Abeta) peptide is associated with hereditary cerebral hemorrhage with amyloidosis Dutch type. The cellular mechanism(s) of toxicity and nature of the AbetaE22Q toxic assemblies are not completely understood. Comparative assessment of structural parameters and cell death mechanisms elicited in primary human cerebral endothelial cells by AbetaE22Q and wild-type Abeta revealed that only AbetaE22Q triggered the Bax mitochondrial pathway of apoptosis. AbetaE22Q neither matched the fast oligomerization kinetics of Abeta42 nor reached its predominant beta-sheet structure, achieving a modest degree of oligomerization with a secondary structure that remained a mixture of beta and random conformations. The endogenous molecule tauroursodeoxycholic acid (TUDCA) was a strong modulator of AbetaE22Q-triggered apoptosis but did not significantly change the secondary structures and fibrillogenic propensities of Abeta peptides. These data dissociate the pro-apoptotic properties of Abeta peptides from their distinct mechanisms of aggregation/fibrillization in vitro, providing new perspectives for modulation of amyloid toxicity
— id: 101612, year: 2009, vol: 66, page: 1094, stat: Journal Article,

Human chorionic gonadotropin (a luteinizing hormone homologue) decreases spatial memory and increases brain amyloid-beta levels in female rats
Berry, Anne; Tomidokoro, Yasushi; Ghiso, Jorge; Thornton, Jan
2008 Jun;54(1):143-152, Hormones & behavior
Numerous studies have suggested that estradiol (E) improves spatial memory as female rats with E perform better than those without E. However there is an inverse relationship between E and luteinizing hormone (LH) levels and LH could play a role. We examined whether treatment with the LH homologue human chorionic gonadotropin (hCG), would impair spatial memory of adult E-treated female rats. In the object location memory task, ovariectomized (ovxed) rats treated with E and either a single high dose (400 IU/kg) or a lower repeated dose of hCG (75 IU/kg hourly for 8 h) showed spatial memory disruption compared to ovxed rats treated with estradiol alone. Impairment was attributed to memory disruption as performance improved with shortened delay between task exposure and testing. Tests on another spatial memory task, the Barnes maze, confirmed that hCG (400 IU/kg) can impair memory: although E+veh treated animals made significantly fewer hole errors across time, E+hCG-treated did not. In humans, high LH levels have been correlated with Alzheimer's disease (AD). Because brain amyloid-beta (Abeta) species have been implicated as a toxic factor thought to cause memory loss in AD, we analyzed whether hCG-treated animals had increased Abeta levels. Levels of Abeta from whole brains or hippocampi were assessed by Western blot. hCG treatment to E-implanted females significantly increased soluble Abeta40 and Abeta42 levels. These results indicate that high levels of LH/hCG can impair spatial memory, and an increase in brain Abeta species may account for the memory impairment in hCG-treated rats
— id: 81090, year: 2008, vol: 54, page: 143, stat: Journal Article,

Differential apoptotic response of primary human cerebral endothelial cells to oligomeric assemblies of amyloid beta genetic variants
Cam J; Meyerson J; Mezhericher E; Frangione B; Ghiso J; Rostagno A
New trends in Alzheimer and Parkinson related disorders : ADPD 2007 Bologna : Medimond International Proceedings, 2008,
— id: 5118, year: 2008, vol: , page: 141, stat: Chapter,

Assembly of a membrane receptor complex: roles of the uroplakin II prosequence in regulating uroplakin bacterial receptor oligomerization
Hu, Chih-Chi Andrew; Bachmann, Thomas; Zhou, Ge; Liang, Feng-Xia; Ghiso, Jorge; Kreibich, Gert; Sun, Tung-Tien
2008 Sep 1;414(2):195-203, Biochemical journal
The apical surface of the mammalian urothelium is almost completely covered by two-dimensional protein crystals (known as urothelial plaques) of hexagonally packed 16 nm particles consisting of two UP (uroplakin) heterodimers, i.e. UPs Ia/II and Ib/III pairs. UPs are functionally important as they contribute to the urothelial permeability barrier function, and UPIa may serve as the receptor for the uropathogenic Escherichia coli that causes over 90% of urinary tract infections. We study here how the UP proteins are assembled and targeted to the urothelial apical surface, paying special attention to the roles of the prosequence of UPII in UP oligomerization. We show that (i) the formation of the UPIa/UPII heterodimer, necessary for ER (endoplasmic reticulum) exit, requires disulfide formation in the prosequence domain of proUPII (the immature form of UPII still containing its prosequence); (ii) differentiation-dependent N-glycosylation of the prosequence leads to UP stabilization; (iii) a failure to form tetramers in cultured urothelial cells, in part due to altered glycosylation of the prosequence, may block two-dimensional crystal formation; and (iv) the prosequence of UPII remains attached to the mature protein complex on the urothelial apical surface even after it has been cleaved by the trans-Golgi-network-associated furin. Our results indicate that proper secondary modifications of the prosequence of UPII play important roles in regulating the oligomerization and function of the UP protein complex
— id: 81088, year: 2008, vol: 414, page: 195, stat: Journal Article,

Expression of BRI2 mRNA and protein in normal human brain and familial British dementia: its relevance to the pathogenesis of disease
Lashley, T; Revesz, T; Plant, G; Bandopadhyay, R; Lees, A J; Frangione, B; Wood, N W; de Silva, R; Ghiso, J; Rostagno, A; Holton, J L
2008 Oct;34(5):492-505, Neuropathology & applied neurobiology
INTRODUCTION: Two different disease-specific mutations in the BRI2 gene, situated on chromosome 13, have been identified as giving rise to familial British dementia (FBD) and familial Danish dementia (FDD). Each mutation results in extension of the open reading frame generating the disease-specific precursor proteins which are cleaved by furin-like proteolysis releasing the amyloidogenic C-terminal peptides ABri and ADan in FBD and FDD, respectively. MATERIAL AND METHODS: To understand the mechanism of the formation of amyloid lesions in FBD, we studied the origin of the precursor proteins and furin in the human brain. We used control brains, cases of sporadic Alzheimer's disease (AD), variant AD with cotton wool plaques and FBD to study BRI2 mRNA expression using in situ hybridization. Furin and BRI2 protein expression was investigated using Western blotting and immunohistochemistry. RESULTS: BRI2 mRNA and BRI2 protein are widely expressed primarily by neurones and glia and are deposited in the amyloid lesions in FBD. They were, however, not expressed by cerebrovascular components. Furin expression showed a similar pattern except that it was also present in cerebrovascular smooth muscle cells. CONCLUSIONS: These findings suggest that neurones and glia and are a major source of BRI2 protein and that in FBD, the mutated precursor protein may undergo furin cleavage within neurones to produce the amyloid peptide ABri. The failure to demonstrate BRI2 in blood vessels under the conditions tested suggests that vascular amyloid peptide production does not contribute significantly to cerebral amyloid angiopathy (CAA) in FBD and FDD, lending indirect support to the drainage hypothesis of CAA
— id: 101670, year: 2008, vol: 34, page: 492, stat: Journal Article,

Preamyloid lesions and cerebrovascular deposits in the mechanism of dementia: lessons from non-beta-amyloid cerebral amyloidosis
Rostagno, Agueda; Ghiso, Jorge
2008 ;5(3-4):173-175, Neuro-degenerative diseases
The importance of amyloid plaques in the pathogenesis of dementia is usually centered on beta-amyloid (Abeta) and its role in Alzheimer's disease (AD). However, since fibrillar plaques correlate poorly with neurodegeneration, challenging their importance in the mechanism(s) of dementia, investigators turned their focus to the importance of soluble oligomers and the role of preamyloid and cerebrovascular deposits. Two non-Abeta cerebral amyloidoses, familial British and Danish dementias (FBD and FDD), share many aspects of AD, including cognitive impairment and the presence of neurofibrillary tangles in limbic areas. The lack of compact plaques in FDD and in many areas in FBD further questions the importance of these lesions in the mechanism of dementia. The main components of the deposits--ABri and ADan--are structurally unrelated to Abeta and yet they all have a high tendency to oligomerize and assemble into amyloid fibrils in vitro and form ion-like channels in lipid membranes. Thus, different amyloid species have the capability to induce similar neuropathological changes, which are neither exclusive for Abeta nor dependent on the presence of compact plaques. These findings reaffirm the notion that non-Abeta amyloidoses constitute alternative models to study the role of preassembled amyloid subunits in neuronal death
— id: 78738, year: 2008, vol: 5, page: 173, stat: Journal Article,

Regional brain differences in the degradation of deposited ADan species in familial Danish dementia
Tomidokoro, Y; Tamaoka, A; Blas, F; Ghiso, J
2008 MAR ;61(2):S201-S201, Neuroscience research
— id: 98112, year: 2008, vol: 61, page: S201, stat: Journal Article,

BRI2 as a central protein involved in neurodegeneration
Tsachaki, Maria; Ghiso, Jorge; Efthimiopoulos, Spiros
2008 Dec;3(12):1548-1554, Biotechnology journal
BRI2 is a protein that when mutated causes familial British and familial Danish dementias. Upon cleavage, the mutated BRI2 proteins release the peptides ABri and ADan, which are amyloidogenic and accumulate in the brains of patients. Although BRI2 has an unknown function, several reports indicate that it could play multiple roles. For example, the fact that it exists at the cell surface as a homodimer indicates that it could be involved in cell signaling events by acting as a receptor. BRI2 also interacts with amyloid precursor protein (APP), involved in Alzheimer's disease (AD). In cell cultures and mouse models of AD, BRI2 inhibits APP processing and reduces amyloid beta peptide deposition. The interaction between the two proteins could be responsible for the neuropathological similarities between familial British/Danish dementias and AD. The study of BRI2, which is central in familial British and Danish dementia, could unravel underlying molecular mechanisms of neurodegeneration
— id: 96456, year: 2008, vol: 3, page: 1548, stat: Journal Article,

Differential apoptotic response of primary human cerebral endothelial cells to oligomeric assemblies of amyloid beta genetic variants
Cam J; Meyerson J; Lin H; Frangione B; Ghiso J; Rostagno A
2007 ;4:66-66 #S1, Neuro-degenerative diseases
— id: 73970, year: 2007, vol: 4, page: 66, stat: Journal Article,

Proteomic analysis of exfoliation deposits
Ovodenko, Boris; Rostagno, Agueda; Neubert, Thomas A; Shetty, Vivekananda; Thomas, Stefani; Yang, Austin; Liebmann, Jeffrey; Ghiso, Jorge; Ritch, Robert
2007 Apr;48(4):1447-1457, Investigative ophthalmology & visual science. IOVS
PURPOSE: To increase knowledge of the biochemical composition of lenticular exfoliation material (XFM) by using proteomic approaches. METHODS: Anterior lens capsules from patients with and without exfoliation syndrome (XFS) were homogenized in formic acid and subjected to cyanogen bromide (CNBr) cleavage, and the pattern of chemically generated fragments was compared by SDS-PAGE after silver staining. Unique XFS bands not present in control cases were excised, digested with TPCK-trypsin, and the resultant peptides sequenced with quadrupole time-of-flight mass spectrometry (MS). In parallel experiments, CNBr-fragmented XFM was separately digested in solution with trypsin and elastase, and the resultant peptide mixture was analyzed by liquid chromatography coupled to tandem MS followed by identification through homology searches at nonredundant protein databases. Immunolocalization of the MS-identified components were performed in XFS versus control samples by using conventional immunohistochemical methods and light microscopy. RESULTS: In addition to fibrillin-1, fibronectin, vitronectin, laminin, and amyloid P-component, which are well-known extracellular matrix and basement membrane components of XFM, the proteomic approaches identified the multifunctional protein clusterin and tissue inhibitor of metalloprotease (TIMP)-3 as well as novel molecules, among them fibulin-2, desmocollin-2, the glycosaminoglycans syndecan-3, and versican, membrane metalloproteases of the ADAM family (a disintegrin and metalloprotease), and the initiation component of the classic complement activation pathway C1q. In all cases, classic immunohistochemistry confirmed their location in XFM. CONCLUSIONS: A novel solubilization strategy combined with sensitive proteomic analysis emphasizes the complexity of the XFS deposits and opens new avenues to study the molecular mechanisms involved in the pathogenesis and progression of XFS
— id: 71391, year: 2007, vol: 48, page: 1447, stat: Journal Article,

Protein misfolding, aggregation, and fibril formation : common features of cerebral and non-cerebral amyloid disease
Rostagno A; Lal R; Ghiso J
Neurobiology of Alzheimer's disease New York : Oxford University Press, 2007,
— id: 5115, year: 2007, vol: , page: 133, stat: Chapter,

Preferential association of serum amyloid P component with fibrillar deposits in familial British and Danish dementias: similarities with Alzheimer's disease
Rostagno, Agueda; Lashley, Tammaryn; Ng, Douglas; Meyerson, Jordana; Braendgaard, Hans; Plant, Gordon; Bojsen-Moller, Marie; Holton, Janice; Frangione, Blas; Revesz, Tamas; Ghiso, Jorge
2007 Jun 15;257(1-2):88-96, Journal of the neurological sciences
Two hereditary forms of cerebrovascular amyloidosis, familial British and Danish dementias (FBD and FDD), share striking similarities with Alzheimer's disease (AD) despite structural differences among their amyloid subunits (ABri in FBD, ADan in FDD, and Abeta in AD). Neuropathological lesions in these disorders include neurofibrillary tangles, parenchymal amyloid and pre-amyloid deposits and overwhelming cerebral amyloid angiopathy co-localizing with reactive microglia and multiple amyloid associated proteins including activation products of the complement cascade. Immunohistochemical analysis of FBD and FDD brain lesions unveiled the presence of serum amyloid P-component (SAP) primarily associated with thioflavin positive amyloid deposits in spite of the significant pre-amyloid burden existing in both disorders. Using affinity chromatography and ELISA binding assays we demonstrated specific, calcium-dependent, saturable, high affinity binding interactions between SAP and ABri/ADan peptides, with dissociation constant values in the sub-nanomolar range and within the same order of magnitude as those resulting from the interaction of SAP with Alzheimer's Abeta1-40 and Abeta1-42. The preferential association of SAP with fibrillar amyloid lesions and not with non-fibrillar pre-amyloid deposits is puzzling, suggesting that SAP modulates the assembly and stability of the final fibril rather than participating in the early steps of protein misfolding and oligomerization
— id: 73958, year: 2007, vol: 257, page: 88, stat: Journal Article,

Oligomeric assemblies of the Abeta Dutch mutant induce the formation of nucleosomes in primary cerebral endothelial cells
Cam J; Meyerson JL; Frangione B; Ghiso J; Rostagno A
Alzheimer's disease : new advances Bologna : Medimond International Proceedings, 2006,
— id: 5116, year: 2006, vol: , page: 397, stat: Chapter,

Oligomeric assemblies of the Abeta Dutch mutant induce formation of nucleosomes in primary cerebral endothelial cells
Cam J; Meyerson JL; Ng D; Frangione B; Ghiso J; Rostagno A
2006 ;2:S531-S531, Alzheimer's & Dementia
— id: 101631, year: 2006, vol: 2, page: S531, stat: Journal Article,

Oligomeric assemblies of the A-beta Dutch mutant induces the formation of nucleosomes in primary cerebral endothelial cells
Cam J; Meyerson JL; Ng D; Frangione B; Ghiso J; Rostango A
2006 ;2(3 Suppl 1):S531-S531, Alzheimer's & Dementia
— id: 73968, year: 2006, vol: 2, page: S531, stat: Journal Article,

Brain neprilysin activity and susceptibility to transgene-induced Alzheimer amyloidosis
Carter, Troy L; Pedrini, Steve; Ghiso, Jorge; Ehrlich, Michelle E; Gandy, Sam
2006 Jan 16;392(3):235-239, Neuroscience letters
Neprilysin (NEP) is a zinc metalloproteinase that degrades enkephalins, endothelins, and the Alzheimer's disease amyloid beta (Abeta) peptides. NEP-deficient mice possess increased levels of brain Abeta(1-40) and Abeta(1-42). The objective of this study was to determine whether tissue NEP specific activity differs according to age and/or across mouse strains, especially those strains predisposed toward formation of Abeta-amyloid plaques following overexpression of the human Alzheimer amyloid precursor protein (APP). The C57Bl/6J mouse strain appears to be relatively susceptible to cerebral amyloidosis, whereas the Swiss Webster (SW) strain appears more resistant. We investigated whether NEP specific activity in brain and kidney homogenates from SW and C57 mice of 6, 40, and 80 weeks old varied according to mouse strain, age, and gender. Among the variables tested, NEP specific activity varied most dramatically across mouse strain, with the kidney and brain of SW mice displaying the highest activities. Aging was associated with a reduction in brain NEP specific activity in both strains. Gender-specific differences were identified in kidney but not in brain. We conclude that aging- and strain-dependent differences in NEP specific activity may play a role in the differential susceptibility of some mouse strains for developing cerebral amyloidosis following human APP overexpression
— id: 81093, year: 2006, vol: 392, page: 235, stat: Journal Article,

Post-translational modifications in A-beta and non-A-beta amyloidosis
Ghiso J; Tomidokoro Y; Lashley T; Holton J; Revesz T; Rostagno A; Frangione B
2006 ;2(3 Suppl 1):S479-S479, Alzheimer's & Dementia
— id: 73967, year: 2006, vol: 2, page: S479, stat: Journal Article,

Genetic alterations of the BRI2 gene: familial British and Danish dementias
Ghiso, J; Rostagno, A; Tomidokoro, Y; Lashley, T; Bojsen-Moller, M; Braendgaard, H; Plant, G; Holton, J; Lal, R; Revesz, T; Frangione, B
2006 Jan;16(1):71-79, Brain pathology
Classic arguments sustaining the importance of amyloid in the pathogenesis of dementia are usually centered on amyloid beta (Abeta) and its role in neuronal loss characteristic of Alzheimer disease, the most common form of human cerebral amyloidosis. Two non-Abeta cerebral amyloidoses, familial British and Danish dementias, share many aspects of Alzheimer disease, including the presence of neurofibrillary tangles, parenchymal pre-amyloid and amyloid deposits, cerebral amyloid angiopathy, and a widespread inflammatory response. Both early-onset conditions are linked to specific mutations in the BRI2 gene, causing the generation of longer-than-normal protein products and the release of 2 de novo created peptides ABri and ADan, the main components of amyloid fibrils in these inherited dementias. Although the molecular mechanisms and signal transduction pathways elicited by the amyloid deposits and their relation to cognitive impairment remain to be clarified, new evidence indicates that, independent of the differences in their primary structures, Abeta, ABri, and ADan subunits are able to form morphologically compatible ion-channel-like structures and elicit single ion-channel currents in reconstituted lipid membranes. These findings reaffirm the notion that non-Abeta amyloidosis constitute suitable alternative models to study the role of amyloid deposition in the mechanism of neuronal cell death
— id: 64171, year: 2006, vol: 16, page: 71, stat: Journal Article,

Molecular chaperons, amyloid and preamyloid lesions in the BRI2 gene-related dementias: a morphological study
Lashley, T; Holton, J L; Verbeek, M M; Rostagno, A; Bojsen-Moller, M; David, G; van Horssen, J; Braendgaard, H; Plant, G; Frangione, B; Ghiso, J; Revesz, T
2006 Oct;32(5):492-504, Neuropathology & applied neurobiology
Molecular chaperons or amyloid-associated proteins (AAPs) are deposited in vascular and parenchymal amyloid lesions in Alzheimer's disease (AD) and other amyloidoses. AAPs, such as apolipoprotein E (ApoE) or apolipoprotein J (ApoJ) have been strongly implicated in the pathogenesis of AD in vitro and in vivo. Furthermore the possession of the ApoE in4 allele is a well-studied risk factor for AD. In view of the similarities between AD and both familial British dementia (FBD) and familial Danish dementia (FDD), we investigated the presence of AAPs in these two diseases to understand better their role in the general process of amyloidogenesis. Immunohistochemistry for ApoE, ApoJ, serum amyloid P (SAP), alpha-1-antichymotrypsin, cystatin C, heparan sulphate proteoglycans, such as agrin, perlecan, syndecans, glypican-1 and for heparan sulphate glycosaminoglycan (HS GAG) side chains was carried out together with immunohistochemical preparations specific to the amyloid subunits. Significant or extensive staining for ApoE, ApoJ, agrin, glypican-1 and HS GAG side chains was found in both amyloid (fibrillar) and preamyloid (nonfibrillar) deposits in FBD and FDD. The remaining AAPs, including SAP, were predominantly found in amyloid lesions. Only very weak staining was present in a small proportion of the amyloid lesions using perlecan immunohistochemistry. These findings suggest that the deposition patterns of AAPs in FBD and FDD are mostly similar to those in AD. The presence of AAPs in the preamyloid lesions supports the notion that chaperon molecules may play a role in the early steps of fibrillogenesis
— id: 73959, year: 2006, vol: 32, page: 492, stat: Journal Article,

Exogenous induction of cerebral beta-amyloidogenesis is governed by agent and host
Meyer-Luehmann, Melanie; Coomaraswamy, Janaky; Bolmont, Tristan; Kaeser, Stephan; Schaefer, Claudia; Kilger, Ellen; Neuenschwander, Anton; Abramowski, Dorothee; Frey, Peter; Jaton, Anneliese L; Vigouret, Jean-Marie; Paganetti, Paolo; Walsh, Dominic M; Mathews, Paul M; Ghiso, Jorge; Staufenbiel, Matthias; Walker, Lary C; Jucker, Mathias
2006 Sep 22;313(5794):1781-1784, Science
Protein aggregation is an established pathogenic mechanism in Alzheimer's disease, but little is known about the initiation of this process in vivo. Intracerebral injection of dilute, amyloid-beta (Abeta)-containing brain extracts from humans with Alzheimer's disease or beta-amyloid precursor protein (APP) transgenic mice induced cerebral beta-amyloidosis and associated pathology in APP transgenic mice in a time- and concentration-dependent manner. The seeding activity of brain extracts was reduced or abolished by Abeta immunodepletion, protein denaturation, or by Abeta immunization of the host. The phenotype of the exogenously induced amyloidosis depended on both the host and the source of the agent, suggesting the existence of polymorphic Abeta strains with varying biological activities reminiscent of prion strains
— id: 81091, year: 2006, vol: 313, page: 1781, stat: Journal Article,

Matrix metalloproteases and A-beta clearance
Tomidokoro Y; Lashley T; Revesz T; Greenberg S; Frangione B; Rostagno A; Ghiso J
2006 ;2(3 Suppl 1):S536-S537, Alzheimer's & Dementia
— id: 73969, year: 2006, vol: 2, page: S536, stat: Journal Article,

BRI2 modulates amyloid precursor protein processing and inhibits A-beta generation
Tsachaki M; Fotinopoulou A; Vlavaki M; Poulopoulos A; Rostagno A; Fragione B; Ghiso J; Efthimiopoulos S
2006 ;2(3 Suppl 1):S36-S36, Alzheimer's & Dementia
— id: 73966, year: 2006, vol: 2, page: S36, stat: Journal Article,

Radiolabeling of amyloid-beta peptides
Calero, Miguel; Ghiso, Jorge
2005 ;299:325-348, Methods in molecular biology
Nowadays, a wide variety of protocols for labeling proteins is available. However, radiolabeling remains one of the most powerful, sensitive and accurate methods to trace and quantitate proteins. Additionally, radiolabeling techniques are steadily gaining importance for diagnosis and treatment in nuclear medicine. There is a considerable number of radioisotopes, but only some are commonly used for basic biomedical research. Among them, the iodine radioisotopes (gamma-emitters) have several advantages for the labeling of proteins. This chapter focuses on radioiodination protocols for amyloidogenic peptides, using the Abeta peptides as a paradigm. The chloramine T, Iodo-Gen, and lactoperoxidase methods can be successfully applied to radioiodination of different amyloid peptides as long as free tyrosyl (or histidyl) groups are avail-able. However, these methods differ in their yield and the degree of oxidative damage conferred to labile peptides. When no tyrosines are available, the Bolton-Hunter methodology can be used. The labeling by the tyramine-cellobiose ligand trapping method is applicable to the study of cellular uptake and catabolism of amyloid peptides
— id: 81096, year: 2005, vol: 299, page: 325, stat: Journal Article,

Clusterin and Alzheimer's disease
Calero, Miguel; Rostagno, Agueda; Frangione, Blas; Ghiso, Jorge
2005 ;38:273-298, Sub-cellular biochemistry
Clusterin (apolipoprotein J) is a ubiquitous multifunctional glycoprotein with the capability to interact with a broad spectrum of molecules, among them the Alzheimer's Abeta peptide. Due to its co-localization with fibrillar deposits in systemic and cerebral amyloid disorders, clusterin is also considered an amyloid-associated protein. Although no genuine function has been attributed to this protein so far, it has been implicated in a wide variety of physiological and pathological processes, a role that may vary according to the protein maturation, sub-cellular localization, and the presence of certain tissue- or cell-specific factors. This review focuses on the importance of clusterin in health and disease conditions, with particular emphasis in its role in Abeta amyloidosis and other disorders of protein folding
— id: 56305, year: 2005, vol: 38, page: 273, stat: Journal Article,

BRI2 interacts with APP and regulates Abeta production
Fotinopoulou, Angeliki; Tsachaki, Maria; Vlavaki, Maria; Poulopoulos, Alexandros; Rostagno, Agueda; Frangione, Blas; Ghiso, Jorge; Efthimiopoulos, Spiros
2005 Sep;280(35):30768-30772, Journal of biological chemistry
Transmembrane proteins BRI2 and APP co-localize with Ass amyloid lesions in sporadic Alzheimer's disease and mutations in both precursor proteins are linked to early-onset familial cases of cerebral amyloidosis associated with dementia and/or cerebral haemorrhage. A specific interaction between BRI2 and APP was unveiled by immunoprecipitation experiments using transfected and non-transfected cells. The use of deletion mutants further revealed that stretches 648-719 of APP751 and 46-106 of BRI2, both inclusive of the full transmembrane domains, are sufficient for the interaction. Removal of most of the APP and BRI2 extracellular domains without affecting the interaction implies that both proteins interact when are expressed on the same cell membrane (cis) rather than on adjacent cells (trans). The presence of BRI2 had a modulatory effect on APP processing, specifically increasing the levels of cellular APP as well as ss-secretase-generated C-terminal fragments while decreasing the levels of a-secretase-generated C-terminal fragments as well as the secretion of total APP and Ass peptides. Determining the precise molecular pathways affected by the specific binding between APP and BRI2 could result in the identification of common therapeutic targets for these sporadic and familial neurodegenerative disorders
— id: 56303, year: 2005, vol: 280, page: 30768, stat: Journal Article,

Familial British and Danish dementias
Ghiso J; Rostagno A; Tomidokoro Y; Lashley T; Holton J; Plant G; Revesz T; Frangione B
Amyloid proteins : the beta sheet conformation and disease Weinheim : Wiley-VCH, 2005,
— id: 5113, year: 2005, vol: , page: 515, stat: Chapter,

Biochemistry and genetics of cerebral amyloid diseases
Ghiso, J
2005 APR ;31(2):216-216, Neuropathology & applied neurobiology
— id: 50152, year: 2005, vol: 31, page: 216, stat: Journal Article,

Amyloid-associated proteins (AAPs) in familial British dementia (FBD) and familial Danish dementia (FDD)
Lashley, T; Holton, JL; Frangione, B; van Horssen, J; Rostagno, A; Verbeek, MM; Ghiso, J; Revesz, T
2005 APR ;31(2):244-244, Neuropathology & applied neurobiology
— id: 73960, year: 2005, vol: 31, page: 244, stat: Journal Article,

The familial dementia BRI2 gene binds the Alzheimer gene amyloid-beta precursor protein and inhibits amyloid-beta production
Matsuda, Shuji; Giliberto, Luca; Matsuda, Yukiko; Davies, Peter; McGowan, Eileen; Pickford, Fiona; Ghiso, Jorge; Frangione, Blas; D'Adamio, Luciano
2005 Aug 12;280(32):28912-28916, Journal of biological chemistry
Alzheimer disease (AD), the most common senile dementia, is characterized by amyloid plaques, vascular amyloid, neurofibrillary tangles, and progressive neurodegeneration. Amyloid is mainly composed by amyloid-beta (A(beta)) peptides, which are derive from processing of the beta-amyloid precursor protein (APP), better named amyloid-beta precursor protein (A(beta)PP), by secretases. The A(beta)PP intracellular domain (AID), which is released together with A(beta), has signaling function, since it modulates apoptosis and transcription. Despite its biological and pathological importance, the mechanisms regulating A(beta)PP processing are poorly understood. As cleavage of other gamma-secretase substrates is regulated by membrane bound proteins, we have postulated the existence of integral membrane proteins that bind A(beta)PP and regulate its processing. Here, we show that BRI2, a type II membrane protein, interacts with A(beta)PP. Interestingly, 17 amino acids corresponding to the NH2-terminal portion of A(beta) are necessary for this interaction. Moreover, BRI2 expression regulates A(beta)PP processing resulting in reduced A(beta) and AID levels. Altogether, these findings characterize the BRI2-A(beta)PP interaction as a regulatory mechanism of A(beta)PP processing that inhibits A(beta) production. Notably, BRI2 mutations cause familial British (FBD) and Danish dementias (FDD) that are clinically and pathologically similar to AD. Finding that BRI2 pathogenic mutations alter the regulatory function of BRI2 on A(beta)PP processing would define dysregulation of A(beta)PP cleavage as a pathogenic mechanism common to AD, FDD, and FBD
— id: 81095, year: 2005, vol: 280, page: 28912, stat: Journal Article,

Insulin-degrading enzyme degrades amyloid peptides associated with British and Danish familial dementia
Morelli, Laura; Llovera, Ramiro E; Alonso, Leonardo G; Frangione, Blas; de Prat-Gay, Gonzalo; Ghiso, Jorge; Castano, Eduardo M
2005 Jul 8;332(3):808-816, Biochemical & biophysical research communications
Familial British dementia (FBD) and familial Danish dementia (FDD) are autosomal dominant disorders characterized by cerebrovascular and parenchymal amyloid deposition and neurofibrillary degeneration. In both conditions, the genetic defects cause the loss of the normal stop codon in the precursor BRI, generating novel 34-residue peptides named ABri and ADan in FBD and FDD, respectively. ABri and ADan show a strong tendency to aggregate into non-fibrillar and fibrillar structures at neutral pH and this property seems to be directly related to neurotoxicity. Here we report that a recombinant insulin-degrading enzyme (rIDE) was capable of degrading monomeric ABri and ADan in vitro more efficiently than oligomeric species. These peptides showed high beta-structure content and were more resistant to proteolysis as compared to the BRI wild-type product of 23 amino acids. Specific sites of cleavage within the C-terminal pathogenic extensions raise the possibility that proteolysis of monomeric soluble precursors by IDE may delay ABri and ADan aggregation in vivo
— id: 81097, year: 2005, vol: 332, page: 808, stat: Journal Article,

Complement activation in exfoliation syndrome
Ovodenko, B.; Rostagno, A.; Liebmann, J. M.; Bley, L. M.; Jofe, M. A.; Smolyak, R. M.; Pinhas, D.; Ghiso, J. A.; Ritch, R.
2005 ;46(Suppl. S):3763-3763, Investigative ophthalmology & visual science. IOVS
Purpose: To identify components of exfoliation material (XFM) in eyes with XFS. Methods: Anterior lens capsules from patients with and without XFS were divided into two groups, homogenized in formic acid, and subjected to cyanogen bromide (CNBr). Aliquots of resultant peptides were separated on SDS-PAGE and silver stained. Differentially stained bands were excised from the gel, digested with trypsin, and sequenced using Quadrupole Time-of-Flight mass spectrometry (Q-TOF MS). The resultant deconvoluted MS/MS spectra were directly used to search the NCBI nonredundant protein database using the Mascot search program (Matrix Science, UK). The remainder of CNBr-fragmented material was digested witheither trypsin or elastase and analyzed with liquid chromatography coupled totandem mass spectrometry (LC-MS/MS). The resultant MS/MS spectra were analyzed by Bioworks 3.1 (Thermo Finnigan, USA) utilizing the SEQUEST database. Immunohistochemical analysis for the components of the complement pathways was performed in XFS vs. control samples using conventional horseradish peroxidase-based immunohistochemistry. Results: Both biochemical approaches yielded similar Results: Specifically, components involved in inhibition of complement activation (e. g., clusterin and vitronectin) were identified. Immunohistochemistry revealed very strong staining of XFM with anti-clusterin antibody and moderate staining with antibodies against normal components and activation products of the classical complement pathway: C1q, C3c, and C4c. In the samples examined, there was no staining of XFM with anti-apolipoprotein- E, anti-Bb, and anti-C5b-9 antibodies. Conclusions: Deposition of normal components and activation products of the classical complement pathway in XFM could be due to ocular ischemia and/or inflammatory process. Clusterin, a ubiquitous extracellular protein, is found in a wide variety of lesions and has multiple functions, such as protection against apoptosis and oxidative stress, inhibition of the membrane attack complex of complement proteins, and inhibition of amyloid aggregation. The apparently great abundance of clusterin in XFM, along with the presence of complement activation products is intriguing and warrants further study. Complement inhibition should be investigated as a potential therapy for mitigating cytotoxicity in the anterior segment in XFS
— id: 101615, year: 2005, vol: 46, page: 3763, stat: Journal Article,

Amyloid ion channels: a common structural link for protein-misfolding disease
Quist, Arjan; Doudevski, Ivo; Lin, Hai; Azimova, Rushana; Ng, Douglas; Frangione, Blas; Kagan, Bruce; Ghiso, Jorge; Lal, Ratnesh
2005 Jul 26;102(30):10427-10432, Proceedings of the National Academy of Sciences of the United States of America
Protein conformational diseases, including Alzheimer's, Huntington's, and Parkinson's diseases, result from protein misfolding, giving a distinct fibrillar feature termed amyloid. Recent studies show that only the globular (not fibrillar) conformation of amyloid proteins is sufficient to induce cellular pathophysiology. However, the 3D structural conformations of these globular structures, a key missing link in designing effective prevention and treatment, remain undefined as of yet. By using atomic force microscopy, circular dichroism, gel electrophoresis, and electrophysiological recordings, we show here that an array of amyloid molecules, including amyloid-beta(1-40), alpha-synuclein, ABri, ADan, serum amyloid A, and amylin undergo supramolecular conformational change. In reconstituted membranes, they form morphologically compatible ion-channel-like structures and elicit single ion-channel currents. These ion channels would destabilize cellular ionic homeostasis and hence induce cell pathophysiology and degeneration in amyloid diseases
— id: 81094, year: 2005, vol: 102, page: 10427, stat: Journal Article,

Diversity of senile plaques in Alzheimer's disease as revealed by a new monoclonal antibody that recognizes an internal sequence of the Abeta peptide
Rabano, Alberto; Jimenez-Huete, Adolfo; Acevedo, Boris; Calero, Miguel; Ghiso, Jorge; Valdes, Israel; Gavilondo, Jorge; Frangione, Blas; Mendez, Enrique
2005 Oct;2(4):409-417, Current Alzheimer research
In order to have more specific tools available to approach amyloidogenesis in Alzheimer's disease (AD), we have produced several polyclonal and monoclonal antibodies that recognize specific sequences of the amyloid beta (Abeta) peptide. Here we present results that demonstrate that our monoclonal antibody EM5 recognizes an internal sequence (residues 11-16) of the Abeta peptide. This strategic localization of the epitope allowed us to employ this antibody, together with two previously reported polyclonal antibodies (EM2 and EM3, specific for AbetaX-40 and AbetaX-42, respectively), in an immunohistochemical study aimed at exploring the differential distribution of longer (AbetaX-40/42) and shorter (Abeta17-X) peptides along the various types of amyloid deposits of AD. This antibody panel was used in six AD brains, on sections from associative neocortex, striatum and cerebellar cortex. Single and double immunostaining revealed specific staining of vascular amyloid deposits and neuritic plaques by EM5 antibody, with high co-localization of EM2. Our results suggest that EM5 antibody recognizes pathogenic forms of Abeta deposits (amyloid angiopathy and neuritic plaques) and reveals the existence of a subset of plaques with a profile similar to vascular deposits. Additionally, our results show that diffuse plaques in AD brains may contain Abeta17-X peptides as its principal component. EM5 may be a useful tool in research both on human and transgenic mice tissue that may aid in the study of molecular heterogeneity of plaques in AD
— id: 81092, year: 2005, vol: 2, page: 409, stat: Journal Article,

Cerebral amyloid angiopathy
Revesz T; Ghiso J; Plant G; Lashley T; Rostagno A; Frangione B; Holton JL
Cerebrovascular diseases Basel, Switzerland : ISN Neuropath Press, 2005,
— id: 5114, year: 2005, vol: , page: 94, stat: Chapter,

Chromosome 13 dementias
Rostagno, A; Tomidokoro, Y; Lashley, T; Ng, D; Plant, G; Holton, J; Frangione, B; Revesz, T; Ghiso, J
2005 Aug;62(16):1814-1825, Cellular & molecular life sciences: CMLS
The importance of cerebral amyloid deposition in the mechanism of neurodegeneration is still debatable. Classic arguments are usually centered on amyloid beta(Abeta) and its role in the neuronal loss characteristic of Alzheimer's disease, the most common form of human cerebral amyloidosis. Two non-Abeta cerebral amyloidoses, familial British and Danish dementias (FBD and FDD), share many aspects of Alzheimer's disease, including the presence of neurofibrillary tangles, parenchymal preamyloid and amyloid deposits, cerebral amyloid angiopathy and a variety of amyloid-associated proteins and inflammatory components. Both early-onset conditions are linked to specific mutations at or near the stop codon of the chromosome 13 gene BRI2 that cause generation of longer-than-normal protein products. Furin-like processing of these longer precursors releases two de novo-created peptides, ABri and ADan, which deposit as amyloid fibrils in FBD and FDD, respectively. Due to the similar pathology generated by completely unrelated amyloid subunits, FBD and FDD, collectively referred to as chromosome 13 dementias, constitute alternative models for studying the role of amyloid deposition in the mechanism of neuronal cell death
— id: 56304, year: 2005, vol: 62, page: 1814, stat: Journal Article,

Familial Danish dementia: co-existence of Danish and Alzheimer amyloid subunits (ADan AND A{beta}) in the absence of compact plaques
Tomidokoro, Yasushi; Lashley, Tammaryn; Rostagno, Agueda; Neubert, Thomas A; Bojsen-Moller, Marie; Braendgaard, Hans; Plant, Gordon; Holton, Janice; Frangione, Blas; Revesz, Tamas; Ghiso, Jorge
2005 Nov 4;280(44):36883-36894, Journal of biological chemistry
Familial Danish dementia is an early onset autosomal dominant neurodegenerative disorder linked to a genetic defect in the BRI2 gene and clinically characterized by dementia and ataxia. Cerebral amyloid and preamyloid deposits of two unrelated molecules (Danish amyloid (ADan) and beta-amyloid (Abeta)), the absence of compact plaques, and neurofibrillary degeneration indistinguishable from that observed in Alzheimer disease (AD) are the main neuropathological features of the disease. Biochemical analysis of extracted amyloid and preamyloid species indicates that as the solubility of the deposits decreases, the heterogeneity and complexity of the extracted peptides exponentially increase. Nonfibrillar deposits were mainly composed of intact ADan-(1-34) and its N-terminally modified (pyroglutamate) counterpart together with Abeta-(1-42) and Abeta-(4-42) in approximately 1:1 mixture. The post-translational modification, glutamate to pyroglutamate, was not present in soluble circulating ADan. In the amyloid fractions, ADan was heavily oligomerized and highly heterogeneous at the N and C terminus, and, when intact, its N terminus was post-translationally modified (pyroglutamate), whereas Abeta was mainly Abeta-(4-42). In all cases, the presence of Abeta-(X-40) was negligible, a surprising finding in view of the prevalence of Abeta40 in vascular deposits observed in sporadic and familial AD, Down syndrome, and normal aging. Whether the presence of the two amyloid subunits is imperative for the disease phenotype or just reflects a conformational mimicry remains to be elucidated; nonetheless, a specific interaction between ADan oligomers and Abeta molecules was demonstrated in vitro by ligand blot analysis using synthetic peptides. The absence of compact plaques in the presence of extensive neuro fibrillar degeneration strongly suggests that compact plaques, fundamental lesions for the diagnosis of AD, are not essential for the mechanism of dementia
— id: 61252, year: 2005, vol: 280, page: 36883, stat: Journal Article,

Molecular pathology of non-A beta cerebral amyloidosis
Ghiso, J
2004 JUL ;25(10):S89-S90, Neurobiology of aging
— id: 47717, year: 2004, vol: 25, page: S89, stat: Journal Article,

Systemic catabolism of Alzheimer's Abeta40 and Abeta42
Ghiso, Jorge; Shayo, Marcos; Calero, Miguel; Ng, Douglas; Tomidokoro, Yasushi; Gandy, Samuel; Rostagno, Agueda; Frangione, Blas
2004 Oct 29;279(44):45897-45908, Journal of biological chemistry
To better understand the physiologic excretion and/or catabolism of circulating peripheral amyloid beta (Abeta), we labeled human Abeta40 (monomeric, with predominant unordered structure) and Abeta42 (mixture of monomers and oligomers in approximately 50:50 ratio, rich in beta-sheet conformation) with either Na(125)I or (125)I-tyramine cellobiose, also known as the cell-trapping ligand procedure, testing their blood clearance and organ uptake in B6SJLF1/J mice. Irrespective of the labeling protocol, the peptide conformation, and the degree of oligomerization, both Abeta40 and Abeta42 showed a short half-life of 2.5-3.0 min. The liver was the major organ responsible for plasma clearance, accounting for >60% of the peptide uptake, followed by the kidney. In vivo, hepatocytes captured >90% of the radiolabeled peptides which, after endocytosis, were preferentially catabolized and excreted into the bile. Biliary excretion of intact as well as partially degraded Abeta species became obviously relevant at doses above 10 microg. The use of biotin-labeled Abeta allowed the visualization of the interaction with HepG2 cells in culture, whereas competitive inhibition experiments with unlabeled Abeta demonstrated the specificity of the binding. The capability of the liver to uptake, catabolize, and excrete large doses of Abeta, several orders of magnitude above its physiologic concentration, may explain not only the femtomolar plasma levels of Abeta but the little fluctuation observed with age and disease stages
— id: 47833, year: 2004, vol: 279, page: 45897, stat: Journal Article,

An animal model of vascular amyloidosis
Ghiso, Jorge; Wisniewski, Thomas
2004 Sep;7(9):902-904, Nature neuroscience
— id: 44510, year: 2004, vol: 7, page: 902, stat: Journal Article,

The possible origin of the amyloid peptides in the BRI2 gene-related dementias
Lashley, T; Holton, JL; Frangione, B; Bandopadhyay, R; Ghiso, J; Rostagno, A; Revesz, T
2004 JUL ;25(10):S171-S171, Neurobiology of aging
— id: 47722, year: 2004, vol: 25, page: S171, stat: Journal Article,

Soluble Abeta homeostasis in AD and DS: impairment of anti-amyloidogenic protection by lipoproteins
Matsubara, Etsuro; Sekijima, Yoshiki; Tokuda, Takahiko; Urakami, Katsuya; Amari, Masakuni; Shizuka-Ikeda, Masami; Tomidokoro, Yasushi; Ikeda, Masaki; Kawarabayashi, Takeshi; Harigaya, Yasuo; Ikeda, Shu-ichi; Murakami, Tetsuro; Abe, Koji; Otomo, Eiichi; Hirai, Shunsaku; Frangione, Blas; Ghiso, Jorge; Shoji, Mikio
2004 Aug;25(7):833-841, Neurobiology of aging
In order to assess whether lipoproteins are physiologically able to balance and modulate the sAbeta homeostasis in vivo, soluble Abeta levels in lipoprotein-depleted plasma were measured as a function of age in normal controls, Alzheimer's disease (AD) patients, and Down's syndrome (DS) cases. The reshaping of sAbeta homeostasis, in particular the sAbeta42-lipoprotein interaction, takes place over normal-60's, whereas mild AD patients appear to have impaired this anti-amyloidogenic mechanism resulting in a significant increase of lipoprotein-free sAbeta42. Similar loss of function takes place in Down's syndrome patients. Lipoprotein-free sAbeta remains significantly elevated from the pre-symptomatic through the symptomatic stages of the disease, and declines with the progression of the AD-like pathology. The dissociation of sAbeta from lipoprotein-particles also occurs in brain parenchyma and the presence of soluble dimeric lipoprotein-free Abeta prior to its parenchymal deposition in AD brains would support the hypothesis that functionally declined lipoproteins may be major determinants in the production of metabolic conditions leading to higher levels of sAbeta species and cerebral amyloidosis
— id: 81099, year: 2004, vol: 25, page: 833, stat: Journal Article,

Insulin degrading enzyme from human brain microvessels degrades amyloid beta and its Flemish, Dutch and Italian vasculotropic variants
Morelli, L; Llovera, RE; Frangione, B; Ghiso, J; Castano, EM
2004 JUL ;25(10):S146-S146, Neurobiology of aging
— id: 47720, year: 2004, vol: 25, page: S146, stat: Journal Article,

Insulin-degrading enzyme in brain microvessels: proteolysis of amyloid {beta} vasculotropic variants and reduced activity in cerebral amyloid angiopathy
Morelli, Laura; Llovera, Ramiro E; Mathov, Irina; Lue, Lih-Fen; Frangione, Blas; Ghiso, Jorge; Castano, Eduardo M
2004 Dec 31;279(53):56004-56013, Journal of biological chemistry
The accumulation of amyloid beta (Abeta) in the walls of small vessels in the cerebral cortex is associated with diseases characterized by dementia or stroke. These include Alzheimer's disease, Down syndrome, and sporadic and hereditary cerebral amyloid angiopathies (CAAs) related to mutations within the Abeta sequence. A higher tendency of Abeta to aggregate, a defective clearance to the systemic circulation, and insufficient proteolytic removal have been proposed as mechanisms that lead to Abeta accumulation in the brain. By using immunoprecipitation and mass spectrometry, we show that insulin-degrading enzyme (IDE) from isolated human brain microvessels was capable of degrading (125)I-insulin and cleaved Abeta-(1-40) wild type and the genetic variants Abeta A21G (Flemish), Abeta E22Q (Dutch), and Abeta E22K (Italian) at the predicted sites. In microvessels from Alzheimer's disease cases with CAA, IDE protein levels showed a 44% increase as determined by sandwich enzyme-linked immunosorbent assay and Western blot. However, the activity of IDE upon radiolabeled insulin was significantly reduced in CAA as compared with age-matched controls. These results support the notion that a defect in Abeta proteolysis by IDE contributes to the accumulation of this peptide in the cortical microvasculature. Moreover they raise the possibility that IDE inhibition or inactivation is a pathogenic mechanism that may open novel strategies for the treatment of cerebrovascular Abeta amyloidoses
— id: 81098, year: 2004, vol: 279, page: 56004, stat: Journal Article,

Familial and sporadic cerebral amyloid angiopathies associated with dementia and the BRI dementias
Plant, Gordon T; Revesz, Tamas; Holton, Janice L; Ghiso, Jorge; Frangione, Blas
The neuropathology of dementia Cambridge UK : Cambridge University Press, 2004,
— id: 4822, year: 2004, vol: , page: ?, stat: Chapter,

Cholesterol in neurologic disorders of the elderly: stroke and Alzheimer's disease
Reiss, Allison B; Siller, Keith A; Rahman, Mohammad M; Chan, Edwin S L; Ghiso, Jorge; de Leon, Mony J
2004 Oct;25(8):977-989, Neurobiology of aging
Mechanisms for the regulation of intracellular cholesterol levels in various types of brain and vascular cells are of considerable importance in our understanding of the pathogenesis of a variety of diseases, particularly atherosclerosis and Alzheimer's disease (AD). It is increasingly clear that conversion of brain cholesterol into 24-hydroxycholesterol and its subsequent release into the periphery is important for the maintenance of brain cholesterol homeostasis. Recent studies have shown elevated plasma concentrations of 24-hydroxycholesterol in patients with AD and vascular dementia, suggesting increased brain cholesterol turnover during neurodegeneration. The oxygenases involved in the degradation and excretion of cholesterol, including the cholesterol 24-hydroxylase and the 27-hydroxylase, are enzymes of the cytochrome P-450 family. This review focuses on the newly recognized importance of cholesterol and its oxygenated metabolites in the pathogenesis of ischemic stroke and AD. The reduction in stroke and AD risk in patients treated with cholesterol-lowering statins is also discussed
— id: 45967, year: 2004, vol: 25, page: 977, stat: Journal Article,

Familial British and Danish dementias: BRI2 gene and protein expression by human cerebral cells
Rostagno, A; Zhao, ZH; Ng, D; Lashley, T; Holton, J; Frangione, B; Revesz, T; Ghiso, J
2004 JUL ;25(10):S170-S171, Neurobiology of aging
— id: 47721, year: 2004, vol: 25, page: S170, stat: Journal Article,

Biochemical analysis of A beta amyloid deposits in the Iowa variant of Alzheimer's disease
Tomidokoro, Y; Rostagno, A; Greenberg, SM; Frangione, B; Rebeck, WG; Ghiso, J
2004 JUL ;25(10):S38-S38, Neurobiology of aging
— id: 47713, year: 2004, vol: 25, page: S38, stat: Journal Article,

SiRNA-mediated BRI2 gene silencing in human neuronal cells
Zhao, ZH; Rostagno, A; Revesz, T; Frangione, B; Ghiso, J
2004 JUL ;25(10):S470-S471, Neurobiology of aging
— id: 47742, year: 2004, vol: 25, page: S470, stat: Journal Article,

RAGE mediates amyloid-beta peptide transport across the blood-brain barrier and accumulation in brain
Deane, Rashid; Du Yan, Shi; Submamaryan, Ram Kumar; LaRue, Barbara; Jovanovic, Suzana; Hogg, Elizabeth; Welch, Deborah; Manness, Lawrence; Lin, Chang; Yu, Jin; Zhu, Hong; Ghiso, Jorge; Frangione, Blas; Stern, Alan; Schmidt, Ann Marie; Armstrong, Don L; Arnold, Bernd; Liliensiek, Birgit; Nawroth, Peter; Hofman, Florence; Kindy, Mark; Stern, David; Zlokovic, Berislav
2003 Jul;9(7):907-913, Nature medicine
Amyloid-beta peptide (Abeta) interacts with the vasculature to influence Abeta levels in the brain and cerebral blood flow, providing a means of amplifying the Abeta-induced cellular stress underlying neuronal dysfunction and dementia. Systemic Abeta infusion and studies in genetically manipulated mice show that Abeta interaction with receptor for advanced glycation end products (RAGE)-bearing cells in the vessel wall results in transport of Abeta across the blood-brain barrier (BBB) and expression of proinflammatory cytokines and endothelin-1 (ET-1), the latter mediating Abeta-induced vasoconstriction. Inhibition of RAGE-ligand interaction suppresses accumulation of Abeta in brain parenchyma in a mouse transgenic model. These findings suggest that vascular RAGE is a target for inhibiting pathogenic consequences of Abeta-vascular interactions, including development of cerebral amyloidosis
— id: 42003, year: 2003, vol: 9, page: 907, stat: Journal Article,

Familial and sporadic cerebral amyloid angiopathies
Fragione B; Prelli F; Ghiso J; Revesz T
Third International Congress on Vascular Dementia : Prague, Czech Republic, October 23-26, 2003 Bologna : Monduzzi Editore, 2003,
— id: 5153, year: 2003, vol: , page: 35, stat: Chapter,

Melatonin increases survival and inhibits oxidative and amyloid pathology in a transgenic model of Alzheimer's disease
Matsubara, Etsuro; Bryant-Thomas, Tara; Pacheco Quinto, Javier; Henry, Tracey L; Poeggeler, Burkhard; Herbert, Donald; Cruz-Sanchez, Felix; Chyan, Yau-Jan; Smith, Mark A; Perry, George; Shoji, Mikio; Abe, Koji; Leone, Anna; Grundke-Ikbal, Inge; Wilson, Glen L; Ghiso, Jorge; Williams, Christina; Refolo, Lorenzo M; Pappolla, Miguel A; Chain, Daniel G; Neria, Eyal
2003 Jun;85(5):1101-1108, Journal of neurochemistry
Increased levels of a 40-42 amino-acid peptide called the amyloid beta protein (A beta) and evidence of oxidative damage are early neuropathological markers of Alzheimer's disease (AD). Previous investigations have demonstrated that melatonin is decreased during the aging process and that patients with AD have more profound reductions of this hormone. It has also been recently shown that melatonin protects neuronal cells from A beta-mediated oxidative damage and inhibits the formation of amyloid fibrils in vitro. However, a direct relationship between melatonin and the biochemical pathology of AD had not been demonstrated. We used a transgenic mouse model of Alzheimer's amyloidosis and monitored over time the effects of administering melatonin on brain levels of A beta, abnormal protein nitration, and survival of the mice. We report here that administration of melatonin partially inhibited the expected time-dependent elevation of beta-amyloid, reduced abnormal nitration of proteins, and increased survival in the treated transgenic mice. These findings may bear relevance to the pathogenesis and therapy of AD
— id: 81101, year: 2003, vol: 85, page: 1101, stat: Journal Article,

Differential degradation of amyloid beta genetic variants associated with hereditary dementia or stroke by insulin-degrading enzyme
Morelli, Laura; Llovera, Ramiro; Gonzalez, Silvia A; Affranchino, Jose L; Prelli, Frances; Frangione, Blas; Ghiso, Jorge; Castano, Eduardo M
2003 Jun 27;278(26):23221-23226, Journal of biological chemistry
Inherited amino acid substitutions at position 21, 22, or 23 of amyloid beta (Abeta) lead to presenile dementia or stroke. Insulin-degrading enzyme (IDE) can hydrolyze Abeta wild type, yet whether IDE is capable of degrading Abeta bearing pathogenic substitutions is not known. We studied the degradation of all of the published Abeta genetic variants by recombinant rat IDE (rIDE). Monomeric Abeta wild type, Flemish (A21G), Italian (E22K), and Iowa (D23N) variants were readily degraded by rIDE with a similar efficiency. However, proteolysis of Abeta Dutch (E22Q) and Arctic (E22G) was significantly lower as compared with Abeta wild type and the rest of the mutant peptides. In the case of Abeta Dutch, inefficient proteolysis was related to a high content of beta structure as assessed by circular dichroism. All of the Abeta variants were cleaved at Glu3-Phe4 and Phe4-Arg5 in addition to the previously described major sites within positions 13-15 and 18-21. SDS-stable Abeta dimers were highly resistant to proteolysis by rIDE regardless of the variant, suggesting that IDE recognizes a conformation that is available for interaction only in monomeric Abeta. These results raise the possibility that upregulation of IDE may promote the clearance of soluble Abeta in hereditary forms of Abeta diseases
— id: 42004, year: 2003, vol: 278, page: 23221, stat: Journal Article,

Inherited amyloidoses and neurodegeneration : familial British dementia and familial Danish dementia
Revesz T; Ghiso J; Holton J; Frangione B
Neurodegeneration : the molecular pathology of dementia and movement disorders Basel Switzerland : International Society of Neuropathology, 2003,
— id: 5152, year: 2003, vol: , page: 380, stat: Chapter,

Cerebral amyloid angiopathies: a pathologic, biochemical, and genetic view
Revesz, Tamas; Ghiso, Jorge; Lashley, Tammaryn; Plant, Gordon; Rostagno, Agueda; Frangione, Blas; Holton, Janice L
2003 Sep;62(9):885-898, Journal of neuropathology & experimental neurology
Amyloid deposition can take place in the walls of arteries, arterioles, and, less often, capillaries and veins of the central nervous system, a phenomenon known as cerebral amyloid angiopathy (CAA). The major clinicopathological manifestations of CAA include cerebral hemorrhage, ischemic lesions, and dementia. CAA may be classified according to the amyloid protein deposited. In the most common form, sporadic CAA, and in CAA related to sporadic Alzheimer disease (AD). A beta deposition is characteristic. CAA can also be severe in variants of familial AD caused by mutations of the amyloid-beta precursor protein or presenilin-1 genes in which deposition of A beta variants and/or wild-type A beta occurs. Other amyloid proteins involved in familial CAAs include 1) the mutant cystatin C (ACys) in hereditary cerebral hemorrhage with amyloidosis of Icelandic type, 2) variant transthyretins (ATTR) in meningo-vascular amyloidoses, 3) mutated gelsolin (AGel) in familial amyloidosis of Finnish type, 4) disease-associated prion protein (PrP(Sc)) in a variant of the Gerstmann-Straussler-Scheinker syndrome, and 5) ABri and ADan in CAAs observed in the recently described BRI2 gene-related dementias, familial British dementia and familial Danish dementia, respectively. This review addresses issues related to the correlation between morphology, biochemistry, and genetics, and briefly discusses both the pathogenesis and animal models of CAAs
— id: 42002, year: 2003, vol: 62, page: 885, stat: Journal Article,

Amyloidosis
Rostagno A; Ghiso J
Encyclopedia of the neurological sciences Boston MA : Academic Press, 2003,
— id: 5112, year: 2003, vol: , page: 129, stat: Chapter,

P - component in familial british and danish dementias
Rostagno, A. A.; McGinty, R.; Ng, D.; Lashley, T.; Holton, J.; Frangione, B.; Revesz, T.; Ghiso, J.
2003 ;2003(9):Abstract No. 203.3-Abstract No. 203.3, Society for Neuroscience Abstract Viewer & Itinerary Planner
P-component is a member of the pentraxin family of proteins that has been found associated with amyloid deposits in vivo in both systemic and localized forms of amyloidosis including AD. Recently, two novel familial forms of cerebrovascular amyloidosis have been described. These hereditary conditions, familial British dementia (FBD) and familial Danish dementia (FDD), both result from genetic alterations in the BRI2 gene and show striking clinical and neuropathological similarities with AD. Despite structural differences among the amyloid subunits (ABri in FBD, ADan in FDD, and Abeta in AD) all these disorders are characterized by the presence of neurofibrillary tangles and parenchymal and vascular amyloid deposits co-localizing with reactive microglia and activation products of the complement cascade. Immunohistochemical studies of FBD and FDD brain tissue identified P-component associated with the amyloid lesions. Using affinity chromatography, ELISA binding assays and electrophoretic techniques we were able to demonstrate a specific binding interaction between P-component and ABri/ADan peptides, study the biochemical parameters of the interaction, and compare them with that of Abeta amyloid species. Our studies revealed a high affinity, Calcium dependent, saturable binding between P-component and ABri/ADan peptides with Kd values in the low nanomolar range and in the same order of magnitude to those resulting from the interaction of P-component with Abeta1-40 and Abeta1-42. The high affinity interaction of P-component with ABri and ADan peptides in vitro suggests a likely mechanism for their in vivo co-localization in FBD and FDD lesions. Whether the presence of P-component protects the ABri/ADan amyloid deposits from enzymatic degradation as demonstrated for other amyloids is being investigated
— id: 101616, year: 2003, vol: 2003, page: Abstract No. 203.3, stat: Journal Article,

pH-dependent amyloid and protofibril formation by the ABri peptide of familial British dementia
Srinivasan, Rekha; Jones, Eric M; Liu, Keqian; Ghiso, Jorge; Marchant, Roger E; Zagorski, Michael G
2003 Nov 7;333(5):1003-1023, Journal of molecular biology
The ABri is a 34 residue peptide that is the major component of amyloid deposits in familial British dementia. In the amyloid deposits, the ABri peptide adopts aggregated beta-pleated sheet structures, similar to those formed by the Abeta peptide of Alzheimer's disease and other amyloid forming proteins. As a first step toward elucidating the molecular mechanisms of the beta-amyloidosis, we explored the ability of the environmental variables (pH and peptide concentration) to promote beta-sheet fibril structures for synthetic ABri peptides. The secondary structures and fibril morphology were characterized in parallel using circular dichroism, atomic force microscopy, negative stain electron microscopy, Congo red, and thioflavin-T fluorescence spectroscopic techniques. As seen with other amyloid proteins, the ABri fibrils had characteristic binding with Congo red and thioflavin-T, and the relative amounts of beta-sheet and amyloid fibril-like structures are influenced strongly by pH. In the acidic pH range 3.1-4.3, the ABri peptide adopts almost exclusively random structure and a predominantly monomeric aggregation state, on the basis of analytical ultracentrifugation measurements. At neutral pH, 7.1-7.3, the ABri peptide had limited solubility and produced spherical and amorphous aggregates with predominantly beta-sheet secondary structure, whereas at slightly acidic pH, 4.9, spherical aggregates, intermediate-sized protofibrils, and larger-sized mature amyloid fibrils were detected by atomic force microscopy. With aging at pH 4.9, the protofibrils underwent further association and eventually formed mature fibrils. The presence of small amounts of aggregated peptide material or seeds encourage fibril formation at neutral pH, suggesting that generation of such seeds in vivo could promote amyloid formation. At slightly basic pH, 9.0, scrambling of the Cys5-Cys22 disulfide bond occurred, which could lead to the formation of covalently linked aggregates. The presence of the protofibrils and the enhanced aggregation at slightly acidic pH is consistent with the behavior of other amyloid-forming proteins, which supports the premise that a common mechanism may be involved in protein misfolding and beta-amyloidosis
— id: 81100, year: 2003, vol: 333, page: 1003, stat: Journal Article,

Biochemical analysis of Abeta amyloid deposits in the Iowa variant of Alzheimer's disease
Tomidokoro, Y.; Rostagno, A.; Greenberg, S.; Frangione, B.; Rebeck, W.; Ghiso, J.
2003 ;2003(9):Abstract No. 772.2-Abstract No. 772.2, Society for Neuroscience Abstract Viewer & Itinerary Planner
Several mutations within the Abeta sequence of the APP gene are associated with autosomal dominant cerebral amyloid angiopathy. An Asp to Asn mutation at position 23 of Abeta due to a single nucleotide change at codon 694 of the APP gene causes early onset dementia with leukoencephalopathy and cortical calcification in the Iowa kindred. Severe cerebral amyloid angiopathy and cortical pre-amyloid deposits as well as widespread neurofibrillary tangles are the main neuropathological features of the disease. We have extracted the deposited Abeta species from affected brain areas and biochemically analyzed them using a combination of immunoprecipitation, mass spectrometry, amino acid sequence and western blot analysis. Corroborating previous histological data, western blots with C-terminal specific antibodies revealed an extensive accumulation of Abetax-40 in cortical lesions whereas Abetax-42 was only a minor component. Amino acid sequence of the formic-acid soluble extracts showed high degree of N-terminal heterogeneity, being Abeta starting at position 2 (Ala) the predominant species followed by Abeta4 (Phe) and Abeta1 (Asp) in a 4:2:1 ratio, respectively. IP/mass spect confirmed the presence of Abeta1-40, Abeta2-40 and Abeta4-40 as well as minor components starting at Asp1 and Ala2 but ending at Gly38. Interestingly, amino acid sequence analysis demonstrated the presence of both Asp and Asn at position 23 of the Abeta sequence at a 1:4 ratio, respectively, indicating that the deposited amyloid is a mixture of mutant and wild-type Abeta. Whether one of them is an innocent bystander being recruited by the other (conformational mimicry) or both mutated and non-mutated Abeta peptides are important for the amyloidogenesis process in the Iowa family is being currently investigated
— id: 101617, year: 2003, vol: 2003, page: Abstract No. 772.2, stat: Journal Article,

Brain clearance of Alzheimer's amyloid-beta40 in the squirrel monkey: a SPECT study in a primate model of cerebral amyloid angiopathy
Bading, James R; Yamada, Shinya; Mackic, Jasmina B; Kirkman, Linda; Miller, Carol; Calero, Miguel; Ghiso, Jorge; Frangione, Blas; Zlokovic, Berislav V
2002 Jun;10(4):359-368, Journal of drug targeting
Squirrel monkey is a valuable model to study pathogenesis of cerebrovascular amyloid angiopathy (CAA). Previous studies suggested that circulating amyloid-beta40 peptide (Abeta40) crosses the blood-brain barrier (BBB) and may therefore enhance cerebrovascular amyloidosis in aged squirrel monkeys. In the present study, we used single photon emission computed tomography (SPECT) to determine elimination of 123I-Abeta40 and 99mTc-DTPA, an extracellular marker, from the brain in squirrel monkeys at different age. Following intracerebral microinfusions, the time-activity brain clearance curves indicated bi-exponential removal of 123I-Abeta40 with an initial rapid washout (1.1 < or = t 1/2 < or = 2.7 h). This, plus the observed appearance of 123I-radioactivity in plasma suggest significant brain-to-blood transport. In contrast, 99mTc-DTPA was removed slowly by brain interstitial fluid bulk flow (monoexponential decay with 6.8 < or = t 1/2 < or = 16.8 h). A comparison of three middle aged (11-16 years old) vs. two old (22 yrs old) monkeys was consistent with an age-related decline in the BBB capacity to remove 123I-Abeta from the brain. This correlated with an age-dependent increase in A1beta40/42 cerebrovascular immunoreactivity and amyloid deposition. Thus, vascular clearance plays an important role in reducing Abeta levels in the squirrel monkey brain and impaired Abeta40 elimination across the BBB may contribute to the development of CAA
— id: 42008, year: 2002, vol: 10, page: 359, stat: Journal Article,

CATABOLISM OF HUMAN ABETA IN WILD - TYPE MICE
Calero, M.; Shayo, M.; Fleire, S.; Rostagno, A.; Frangione, B.; Ghiso, J.
2002 ;2002(9):Abstract No. 19.10-Abstract No. 19.10, Society for Neuroscience Abstract Viewer & Itinerary Planner
ABETA is the main component of amyloid deposits in Alzheimers disease. A soluble form (sABETA) identical to the deposited ABETA is found in biological fluids and has the ability to cross the blood-brain barrier. The primary structure of ABETA and sABETA are indistinguishable and it is not clear whether sABETA reflects systemic production, brain clearance, or both. The pharmacokinetic parameters of synthetic soluble ABETA40 and ABETA42 species labeled with 125I or 125I-tyramine cellobiose were studied. Blood was drawn at different time points after a single dose of intravenous bolus-injection of the ABETA peptides to assess the distribution and elimination phases. Tissues were harvested at the end the experiment to determine organ uptake. The liver was found to be the major organ responsible for the clearance (>60%). Liver perfusion with collagenase followed by cell fractionation demonstrated that hepatocytes uptake >87% of the peptides while only 2% was found associated with Kupffer cells. (125I)-tyramine cellobiose ABETAwas also present in the gallbladder and small intestine, pointing to the biliary excretion as one of the clearance mechanisms. TCA-precipitable counts in plasma decreased exponentially shortly after the i.v. injection, suggesting that hydrolysis/enzymatic degradation are also important. Less than 15% biotransformation occurs via conjugation to plasma proteins under the conditions tested. The findings indicate that the liver plays a major role in the catabolism of sABETA, an issue to be considered in view of the current attempts to use synthetic homologues for therapeutic use
— id: 101618, year: 2002, vol: 2002, page: Abstract No. 19.10, stat: Journal Article,

Catabolism of human amyloid beta in wild-type B6SJFL1/J mice
Calero, M; Shayo, M; Fleire, S; Frangione, B; Ghiso, J
2002 Jul-Aug;23(1):925-, Neurobiology of aging
— id: 32423, year: 2002, vol: 23, page: 925, stat: Journal Article,

Transport of the precursor protein of familiar British and Danish dementias into secretory vesicles in neuronal cells
Choi, S; Ghiso, J; Frangione, B; Levy, E
2002 Jul-Aug;23(1):66-, Neurobiology of aging
— id: 32407, year: 2002, vol: 23, page: 66, stat: Journal Article,

Involvement of apolipoprotein J (apo J) in brain amyloidosis
Ghiso, J; Calero, M; Magnotti, L; Ng, D; Rostagno, A; Frangione, B
2002 Jul-Aug;23(1):1475-, Neurobiology of aging
— id: 32427, year: 2002, vol: 23, page: 1475, stat: Journal Article,

Inhibition of FAK signaling activated by urokinase receptor induces dormancy in human carcinoma cells in vivo
Ghiso, JAA
2002 APR 11 ;21(16):2513-2524, Oncogene
Activation of focal adhesion kinase (FAK), overexpressed in several human cancers, induces survival, proliferation and motility of cells in culture, but its functional importance in human tumor growth in vivo has not been elucidated. I explored the role of FAK in regulating tumorigenicity of human carcinoma cells, HEp3. These cells overexpress urokinase receptor (uPAR) which, by activating x5beta1 integrin, initiates an intracellular signal through FAK and Src leading to ERK activation and tumorigenicity in vivo. Down regulation of uPAR in these cells led to a similar to3-5-fold reduction in FAK phosphorylation and association with Src and dormancy in vivo. Both FAK phosphorylation and ability to grow in vivo were restored by re-expression of uPAR. The FAK signaling pathway in T-HEp3 cells, measured by FAK phosphorylation, GTP-loaded Ras and ERK activation, was inhibited by transient or stable transfection of FAK related non-kinase (FRNK), known to have a dominant negative function, but not by a FRNK mutant version (S1034-FRNK). Most importantly, while vector- and mutant-S1034-FRNK transfected cells inoculated onto chicken embryo CAMs formed progressively growing tumors, the HA-FRNK-expressing T-HEp3 cells did not proliferate in vivo and remained dormant for at least 6 weeks. Both cell types had similar rate of apoptosis in vivo and the p38(SAPK) or PI3K-Akt signaling pathways were unaffected by FRNK. FRNK induced dormancy could be reverted by expression of an active-R4F-Mek1 mutant. These results show that active FAK is an important mediator of uPAR-regulated tumorigenicity of HEp3 cells and that interruption of FAK mitogenic signaling either through down-regulation of uPAR or by expression of FRNK can force human carcinoma cells into dormancy
— id: 55316, year: 2002, vol: 21, page: 2513, stat: Journal Article,

Amyloidosis and Alzheimer's disease
Ghiso, Jorge; Frangione, Blas
2002 Dec 7;54(12):1539-1551, Advanced drug delivery reviews
Alzheimer's disease (AD) is the most frequent type of amyloidosis in humans and the commonest form of dementia. Extracellular Abeta amyloid deposits in the form of amyloid plaques and cerebral amyloid angiopathy as well as intraneuronal neurofibrillary tangles co-exist in the brain parenchyma of AD patients, the cognitive areas being the most severely affected. This review focuses on the potential role of amyloid in the development of neurodegeneration and presents studies of AD and other unrelated inherited dementia syndromes associated with neuronal loss and amyloid deposition in the brain
— id: 39364, year: 2002, vol: 54, page: 1539, stat: Journal Article,

Tau in familial Danish dementia brain is similar, but not identical, to that found in familial British dementia and PHF- tau in Alzheimer's disease
Hanger, D; Gibb, G; Anderton, B; Ghiso, J; Rostagno, A; Frangione, B; Holton, J; Revesz, T
2002 Jul-Aug;23(1):1845-, Neurobiology of aging
— id: 32437, year: 2002, vol: 23, page: 1845, stat: Journal Article,

Familial British Dementia (FBD): a cerebral amyloidosis with systemic amyloid deposition
Holton J; Ghiso J; Lashley T; Ganguly M; Strand K; Rostagno A; Plant G; Frangione B; Revesz T
2002 ;28(2):148-148, Neuropathology & applied neurobiology
— id: 73971, year: 2002, vol: 28, page: 148, stat: Journal Article,

Morphological evidence of the activation of the classical complement pathway in the BRI gene-related dementias
Holton, J; Lashley, T; Revesz, T; Rostagno, A; Frangione, B; Ghiso, J
2002 Jul-Aug;23(1):774-, Neurobiology of aging
— id: 32416, year: 2002, vol: 23, page: 774, stat: Journal Article,

Familial Danish dementia: a novel form of cerebral amyloidosis associated with deposition of both amyloid-Dan and amyloid-beta
Holton, Janice L; Lashley, Tammaryn; Ghiso, Jorge; Braendgaard, Hans; Vidal, Ruben; Guerin, Christopher J; Gibb, Graham; Hanger, Diane P; Rostagno, Agueda; Anderton, Brian H; Strand, Catherine; Ayling, Hilary; Plant, Gordon; Frangione, Blas; Bojsen-Moller, Marie; Revesz, Tamas
2002 Mar;61(3):254-267, Journal of neuropathology & experimental neurology
Familial Danish dementia (FDD) is pathologically characterized by widespread cerebral amyloid angiopathy (CAA), parenchymal protein deposits, and neurofibrillary degeneration. FDD is associated with a mutation of the BRI2 gene located on chromosome 13. In FDD there is a decamer duplication, which abolishes the normal stop codon, resulting in an extended precursor protein and the release of an amyloidogenic fragment, ADan. The aim of this study was to describe the major neuropathological changes in FDD and to assess the distribution of ADan lesions, neurofibrillary pathology, glial, and microglial response using conventional techniques, immunohistochemistry, confocal microscopy, and immunoelectron microscopy. We showed that ADan is widely distributed in the central nervous system (CNS) in the leptomeninges, blood vessels, and parenchyma. A predominance of parenchymal pre-amyloid (non-fibrillary) lesions was found. Abeta was also present in a proportion of both vascular and parenchymal lesions. There was severe neurofibrillary pathology, and tau immunoblotting revealed a triplet electrophoretic migration pattern comparable with PHF-tau. FDD is a novel form of CNS amyloidosis with extensive neurofibrillary degeneration occurring with parenchymal, predominantly pre-amyloid rather than amyloid, deposition. These findings support the notion that parenchymal amyloid fibril formation is not a prerequisite for the development of neurofibrillary tangles. The significance of concurrent ADan and Abeta deposition in FDD is under further investigation
— id: 42011, year: 2002, vol: 61, page: 254, stat: Journal Article,

Circulating amyloid-beta peptide crosses the blood-brain barrier in aged monkeys and contributes to Alzheimer's disease lesions
Mackic, Jasmina B; Bading, James; Ghiso, Jorge; Walker, Larry; Wisniewski, Thomas; Frangione, Blas; Zlokovic, Berislav V
2002 Jun;38(6):303-313, Vascular pharmacology
1. We studied cerebrovascular sequestration and blood-brain barrier (BBB) permeability to [125I]- or [123I]-labeled amyloid-beta peptides (A beta) in aged rhesus and aged squirrel monkey, the nonhuman primate models of cerebral beta-amyloidosis and cerebrovascular amyloid angiopathy (CAA), respectively. 2. In aged rhesus, the half-time of elimination of [125I]A beta 1-40, t1/2e, was faster by 1.34 h, the systemic clearance, Clss, increased by 4.21 ml/min/kg and the mean residence time of intact peptide in the circulation shortened by 2 h. 3. Cerebrovascular sequestration of [125I]A beta 1-40 was significant in aged squirrel monkey (20.8 ml/g x 10(2)), but undetectable in the rhesus. 4. The permeability surface area product, PS, for [14C]inulin was low in both species (0.11-0.18 ml/g/s x 10(6)) indicating an intact barrier. 5. The BBB permeability to A beta 1-40 was 34.8- and 13.7-fold higher than for [14C]inulin in aged squirrel and rhesus, respectively, suggesting a specialized A beta transport across the BBB. 6. The single photon computed emission tomography studies confirmed a saturable [123I]A beta 1-40 transport at the BBB in primates (Km = 40 nM). 7. Brain autoradiographic analysis of [125I]A beta 1-42 or [125I]A beta 1-40 after intracarotid infusions of radiotracers confirmed co-localization of the signal with A beta-immunoreactive plaques in rhesus monkeys. 8. Metabolism of [125I]A beta 1-40 in brain and plasma was slower in aged squirrel compared to aged rhesus, by 2.9- and 2.6-fold, respectively. 9. Thus, transport of circulating A beta across the BBB contributes to brain parenchymal accumulation of amyloid in aged nonhuman primates. Negligible capillary binding, rapid systemic and brain degradation, and accelerated body elimination of blood-borne A beta, may prevent the development of CAA in rhesus in contrast to squirrel monkeys
— id: 42006, year: 2002, vol: 38, page: 303, stat: Journal Article,

Vascular Amyloidosis in Neurodegenerative Conditions
Matsubara E; Shoji M; Abe K; Frangione B; Ghiso J
2002 Sep;15(7):439-444, Drug news & perspectives
Cerebral amyloid angiopathy defines the deposition of amyloid fibrils in the walls of medium- and small-size leptomeningeal and cortical arteries and arterioles. This condition is an important cause of cerebral hemorrhages and is also associated with cerebral infarctions and diffused white matter changes. In many instances, vascular amyloid deposits co-exist with intraneuronal neurofibrillary tangles, being the cognitive areas the most severely affected. However, the importance of cerebral amyloid angiopathy as a causative element in the process of neurodegeneration is still debatable. This review discusses inherited dementia syndromes associated with neuronal loss and amyloid deposition in the brain, with particular focus on familial Alzheimer's disease and chromosome 13 dementia. (c) 2002 Prous Science. All rights reserved
— id: 42005, year: 2002, vol: 15, page: 439, stat: Journal Article,

Platelet microparticles as carriers of soluble Alzheimer's amyloid beta (sAbeta)
Matsubara, E; Shoji, M; Murakami, T; Abe, K; Frangione, B; Ghiso, J
2002 Nov;977(6):340-348, Annals of the New York Academy of Sciences
— id: 42007, year: 2002, vol: 977, page: 340, stat: Journal Article,

Substitution at codon 22 reduces clearance of Alzheimer's amyloid-beta peptide from the cerebrospinal fluid and prevents its transport from the central nervous system into blood
Monro, O R; Mackic, J B; Yamada, S; Segal, M B; Ghiso, J; Maurer, C; Calero, M; Frangione, B; Zlokovic, B V
2002 May-Jun;23(3):405-412, Neurobiology of aging
A point mutation of G to C at codon 693 of the amyloid-beta (Abeta) precursor protein gene results in Glu to Gln substitution at position 22 of the Abeta (AbetaQ22) associated with hereditary cerebrovascular amyloidosis with hemorrhage Dutch type. Factors that regulate AbetaQ22 levels in the central nervous system (CNS) are largely unknown. By using ventriculo-cisternal perfusion technique in guinea pigs, we demonstrated that clearance from the cerebrospinal fluid and transport from the CNS to blood of [(125)I]-AbetaQ22 (1 nM) were reduced by 36% and 52%, respectively, in comparison to the wild type Abeta(1-40) peptide. In contrast to significant uptake and transport of Abeta(1-40) across the brain capillaries and leptomeningeal vessels, AbetaQ22 was not taken up at these CNS vascular transport sites, which was associated with its 53% greater accumulation in the brain. The CNS clearance of Abeta(1-40) was half-saturated at 23.6 nM; AbetaQ22 had about 6.8-fold less affinity for the CNS efflux transporters and its elimination relied mainly on transport across the choroid plexus. Thus, the Dutch mutation impairs elimination of Abeta from brain by reducing its rapid transport across the blood-brain barrier and the vascular drainage pathways, which in turn may result in accumulation of the peptide around the blood vessels and in brain
— id: 42010, year: 2002, vol: 23, page: 405, stat: Journal Article,

The British and Danish forms of familial BRI dementia
Plant, GT; Braendgaard, H; Revesz, T; Vidal, R; Ghiso, J; Frangione, B
2002 Aug;73(2):223-223, Journal of neurology neurosurgery & psychiatry
— id: 32382, year: 2002, vol: 73, page: 223, stat: Journal Article,

Familial British dementia (FBD), a form of cerebral amyloidosis with systemic amyloid deposition
Revesz, T; Lashley, T; Ganguly, M; Strand, C; Plant, G; Holton, J; Ghiso, J; Rostagno, A; Frangione, B
2002 Jul-Aug;23(1):1759-, Neurobiology of aging
— id: 32434, year: 2002, vol: 23, page: 1759, stat: Journal Article,

Sporadic and familial cerebral amyloid angiopathies
Revesz, Tamas; Holton, Janice L; Lashley, Tammaryn; Plant, Gordon; Rostagno, Agueda; Ghiso, Jorge; Frangione, Blas
2002 Jul;12(3):343-357, Brain pathology
Cerebral amyloid angiopathy (CAA) is the term used to describe deposition of amyloid in the walls of arteries, arterioles and, less often, capillaries and veins of the central nervous system. CAAs are an important cause of cerebral hemorrhage and may also result in ischemic lesions and dementia. A number of amyloid proteins are known to cause CAA. The most common sporadic CAA, caused by A beta deposition, is associated with aging and is a common feature of Alzheimer disease (AD). CAA occurs in several familial conditions, including hereditary cerebral hemorrhage with amyloidosis of Icelandic type caused by deposition of mutant cystatin C, hereditary cerebral hemorrhage with amyloidosis Dutch type and familial AD with deposition of either A beta variants or wild-type A beta, the transthyretin-related meningo-vascular amyloidoses, gelsolin as well as familial prion disease-related CAAs and the recently described BRI2 gene-related CAAs in familial British dementia and familial Danish dementia. This review focuses on the morphological, biochemical, and genetic aspects as well as the clinical significance of CAAs with special emphasis on the BRI2 gene-related cerebrovascular amyloidoses. We also discuss data relevant to the pathomechanism of the different forms of CAA with an emphasis on the most common A beta-related types
— id: 42009, year: 2002, vol: 12, page: 343, stat: Journal Article,

Complement activation in chromosome 13 dementias
Rostagno, A; Magnotti, L; Tomidokoro, Y; Frangione, B; Ghiso, J; Revesz, T; Lashley, T; Holton, J
2002 Jul-Aug;23(1):1661-, Neurobiology of aging
— id: 32431, year: 2002, vol: 23, page: 1661, stat: Journal Article,

Tumoral non-amyloidotic monoclonal immunoglobulin light chain deposits ('aggregoma'): presenting feature of B-cell dyscrasia in three cases with immunohistochemical and biochemical analyses
Rostagno, Agueda; Frizzera, Glauco; Ylagan, Lourdes; Kumar, Asok; Ghiso, Jorge; Gallo, Gloria
2002 Oct;119(1):62-69, British journal of haematology
Tumoral monoclonal immunoglobulin (Ig) light chain non-fibrillar deposits ('aggregomas'), which can be considered analogous to solitary light chain amyloidomas, are a rare presenting feature of B-cell dyscrasias. It is not certain if they are truly localized or if in reality they represent an initial expression of a silent systemic non-amyloid light chain deposition disease (LCDD). This report describes three patients, two of whom presented with cervical masses and the third with a solitary lung nodule, each comprising granular aggregates of monoclonal kappa light chain. Extracted deposits from the lymph node of one patient were shown by N-terminal amino acid sequence analysis to belong to the variable-region kappa I (Vkappa I) light chain subgroup, the first reported kappa-LCDD protein encoded by the L9 gene and the first report of an expressed protein related to this gene. Extracted deposits from the lung nodule of the second patient belonged to the Vkappa IV light chain subgroup encoded by the B3 germ line gene. The N-terminal amino acid sequences of the light chains from the aggregomas were compared with the related germ line sequences and to the N-terminal amino acid sequences of the nine other known kappa-LCDD light chains reported thus far from patients with systemic LCDD
— id: 39580, year: 2002, vol: 119, page: 62, stat: Journal Article,

Complement activation in chromosome 13 dementias. Similarities with Alzheimer's disease
Rostagno, Agueda; Revesz, Tamas; Lashley, Tammaryn; Tomidokoro, Yasushi; Magnotti, Laura; Braendgaard, Hans; Plant, Gordon; Bojsen-Moller, Marie; Holton, Janice; Frangione, Blas; Ghiso, Jorge
2002 Dec 20;277(51):49782-49790, Journal of biological chemistry
Chromosome 13 dementias, familial British dementia (FBD) and familial Danish dementia (FDD), are associated with neurodegeneration and cerebrovascular amyloidosis, with striking neuropathological similarities to Alzheimer's disease (AD). Despite the structural differences among the amyloid subunits (ABri in FBD, ADan in FDD, and Abeta in AD), these disorders are all characterized by the presence of neurofibrillary tangles and parenchymal and vascular amyloid deposits co-localizing with markers of glial activation, suggestive of local inflammation. Proteins of the complement system and their pro-inflammatory activation products are among the inflammation markers associated with AD lesions. Immunohistochemistry of FBD and FDD brain sections demonstrated the presence of complement activation components of the classical and alternative pathways as well as the neo-epitope of the membrane attack complex. Hemolytic experiments and enzyme-linked immunosorbent assays specific for the activation products iC3b, C4d, Bb, and C5b-9 indicated that ABri and ADan are able to fully activate the complement cascade at levels comparable to those generated by Abeta1-42. ABri and ADan specifically bound C1q with high affinity and formed stable complexes in physiological conditions. Activation proceeds approximately 70-75% through the classical pathway while only approximately 25-30% seems to occur through the alternative pathway. The data suggest that the chronic inflammatory response generated by the amyloid peptides in vivo might be a contributing factor for the pathogenesis of FBD and FDD and, in more general terms, to other neurodegenerative conditions
— id: 39388, year: 2002, vol: 277, page: 49782, stat: Journal Article,

FAMILIAL DANISH DEMENT
Tomidokoro, Y.; Frangione, B.; Lashley, T.; Fleire, S.; Rostagno, A.; Holton, J.; Houlden, H.; Bojsen-Moller, M.; Braendgaard, H.; Plant, G.; Revesz, T.; Ghiso, J.
2002 ;2002(1):Abstract No. 328.9-Abstract No. 328.9, Society for Neuroscience Abstract Viewer & Itinerary Planner
Familial Danish Dementia (FDD) is an early onset autosomal dominant disorder characterized by cataracts, deafness, progressive ataxia and dementia. Cerebral ADan amyloid and pre-amyloid deposits as well as NFTs in the hippocampus are the main neuropathological features of the disease. Surprisingly, Abeta was also found co-deposited in the amyloid lesions. We have biochemically characterized by immunoprecipitation, mass spectrometry, amino acid sequence and/or immunoblot analysis the ADan/Abeta species found in amyloid and pre-amyloid lesions in FDD brain and investigated the presence of soluble ADan and Abeta in plasma from carriers of the disease. ADan was composed by a mixture of full-length and C-terminal truncated species bearing N-terminal pyroglutamate (4044.6 Da). Abeta was mainly 1-42 and 4-42. Plasma soluble ADan contained N-terminal glutamate (4062.6 Da). The difference of 18 mass units accounts for the loss of one molecule of water. The conversion glutamate-pyroglutamate is chemically irreversible, indicating that sADan does not represent ADan being cleared from the brain amyloid but rather from soluble unmodified sADan species systemically produced and/or transported from the brain soluble pool. Thus, patients with FDD have two different types of amyloid molecules deposited in the brain and their respective soluble precursors are present in biological fluids. The significance of these unusual findings will be discussed
— id: 101619, year: 2002, vol: 2002, page: Abstract No. 328.9, stat: Journal Article,

Co-existence of amyloid ADan and amyloid-beta in familial Danish dementia
Tomidokoro, Y; Fleire, S; Rostagno, A; Frangione, B; Ghiso, J; Lashley, T; Holton, J; Houlden, H; Revesz, T; Bojsen-Moller, M; Braendgaard, H; Plant, G
2002 Jul-Aug;23(1):1663-, Neurobiology of aging
— id: 32432, year: 2002, vol: 23, page: 1663, stat: Journal Article,

Human cerebrospinal fluid apolipoprotein E isoforms are apparently inefficient at complexing with synthetic Alzheimer's amyloid-[beta] peptide (A[beta] 1-40 ) in vitro
Zhou, Zhongmin; Relkin, Norman; Ghiso, Jorge; Smith, Jonathan D; Gandy, Sam
2002 Jul;8(7):376-381, Molecular medicine
BACKGROUND: Variation at the apolipoprotein E locus on chromosome 19 plays a role in more cases of Alzheimer's disease than does any other identified genetic determinant. We have previously reported the isoform-specific interaction of native human apolipoprotein E (APOE, gene; apoE, protein) epsilon 3 with the amyloid-ss peptide, Ass(1-40), the major component of the cerebral amyloid deposits that appear to cause Alzheimer's disease. MATERIALS AND METHODS: In order to investigate the apoE: A beta interaction further, a modified assay was developed based on co-immunoprecipitation of the complex using an anti-apoE antibody (anti-apoE IP assay). RESULTS: Application of this assay demonstrated that the interaction of Ass(1-40) and apoE can be distinguished into two types: sodium dodecyl sulfate (SDS) -resistant and SDS-releasable. The SDS-resistant interaction between epsilon;3 and Ass(1-40) is apparently maximal at an Ass(1-40) concentration of approximately 75 micro M, and an Ass(1-40) /epsilon 3 molar ratio of about 250:1. The major apoE-isoform-specific difference in interaction with Ass(1-40) is the ability of Ass(1-40) to form SDS- resistant complexes with epsilon 3 but not with epsilon 4. Using the anti-apoE co-IP assay, we found that human cerebrospinal fluid (CSF) epsilon 3 can also form an SDS-resistant complex with Ass(1-40) but human CSF epsilon;4 cannot. However, when compared with apoE epsilon;3 collected from the conditioned medium of APOE epsilon 3-transfected cells, the competence of equal concentrations of CSF apoE epsilon 3 to form SDS-resistant complexes with Ass(1-40) is apparently diminished. A 1:1 mixture of CSF plus apoE epsilon 3-containing conditioned medium is associated with diminished Ass(1-40) /epsilon;3 complex formation to a greater extent than that observed when an identical volume of phosphate-buffered saline is added to apoE epsilon;3 medium. CONCLUSIONS: These results suggest the presence in CSF of factors that interfere with the formation of complexes between synthetic Ass(1-40) and apoE epsilon 3
— id: 81102, year: 2002, vol: 8, page: 376, stat: Journal Article,

Familial cerebral amyloid angiopathy related to stroke and dementia
Frangione B; Revesz T; Vidal R; Holton J; Lashley T; Houlden H; Wood N; Rostagno A; Plant G; Ghiso J
2001 Jul;8 Suppl 1(1):36-42, Amyloid
The term cerebral amyloid angiopathy (CAA) refers to the specific deposition of amyloid fibrils in the walls of leptomeningeal and cortical arteries, arterioles and, although less frequently in capillaries and veins. It is commonly associated with Alzheimers disease, Down's syndrome and normal aging, as well as with a variety of familial conditions related to stroke and/or dementia: hereditary cerebral hemorrhage with amyloidosis of Icelandic type (HCHWA-I), various inherited disorders linked to Abeta mutants (including the Dutch variant of HCHWA), and the recently described chromosome 13 familial dementia in British and Danish kindreds. This review focuses on four different types of hereditary CAA, emphasizing the notion that CAA is not only related to stroke but also to neurodegeneration and dementia of the Alzheimer's type
— id: 39471, year: 2001, vol: 8 Suppl 1, page: 36, stat: Journal Article,

Cerebral amyloidosis, amyloid angiopathy, and their relationship to stroke and dementia
Ghiso J; Frangione B
2001 Feb;3(1):65-73, Journal of Alzheimer's Disease
Cerebral amyloid angiopathy (CAA) is the common term used to define the deposition of amyloid in the walls of medium- and small-size leptomeningeal and cortical arteries, arterioles and, less frequently, capillaries and veins. CAA is an important cause of cerebral hemorrhages although it may also lead to ischemic infarction and dementia. It is a feature commonly associated with normal aging, Alzheimer disease (AD), Down syndrome (DS), and Sporadic Cerebral Amyloid Angiopathy. Familial conditions in which amyloid is chiefly deposited as CAA include hereditary cerebral hemorrhage with amyloidosis of Icelandic type (HCHWA-I), familial CAA related to Abeta variants, including hereditary cerebral hemorrhage with amyloidosis of Dutch origin (HCHWA-D), the transthyretin-related meningocerebrovascular amyloidosis of Hungarian and Ohio kindreds, the gelsolin-related spinal and cerebral amyloid angiopathy, familial PrP-CAA, and the recently described chromosome 13 familial dementia in British and Danish kindreds. This review focuses on the various molecules and genetic variants that target the cerebral vessel walls producing clinical features related to stroke and/or dementia, and discusses the potential role of amyloid in the mechanism of neurodegeneration
— id: 39451, year: 2001, vol: 3, page: 65, stat: Journal Article,

Chromosome 13 dementia syndromes as models of neurodegeneration
Ghiso J; Revesz T; Holton J; Rostagno A; Lashley T; Houlden H; Gibb G; Anderton B; Bek T; Bojsen-Moller M; Wood N; Vidal R; Braendgaard H; Plant G; Frangione B
2001 Dec;8(4):277-284, Amyloid
Two hereditary conditions, familial British dementia (FBD) and familial Danish dementia (FDD), are associated with amyloid deposition in the central nervous system and neurodegeneration. The two amyloid proteins, ABri and ADan, are degradation products of the same precursor molecule BriPP bearing different genetic defects, namely a Stop-to-Arg mutation in FBD and a ten-nucleotide duplication-insertion immediately before the stop codon in FDD. Both de novo created amyloid peptides have the same length (34 amino acids) and the same post-translational modification (pyroglutamate) at their N-terminus. Neurofibrillary tangles containing the classical paired helical filaments as well as neuritic components in many instances co-localize with the amyloid deposits. In both disorders, the pattern of hyperphosphorylated tau immunoreactivity is almost indistinguishable from that seen in Alzheimer's disease. These issues argue for the primary importance of the amyloid deposits in the mechanism(s) of neuronal cell loss. We propose FBD and FDD, the chromosome 13 dementia syndromes, as models to study the molecular basis of neurofibrillary degeneration, cell death and amyloid formation in the brain
— id: 32472, year: 2001, vol: 8, page: 277, stat: Journal Article,

Famililal British dementia
Ghiso J; Revesz T; Rostagno A; Vidal R; Plant G; Frangione B
Alzheimer's disease : advances in etiology, pathogenesis and therapeutics New York : Wiley, 2001,
— id: 5111, year: 2001, vol: , page: 487, stat: Chapter,

Familial British dementia
Ghiso J; Revesz T; Rostango A; Vidal R; Plant G; Frangione B
Alzheimer's Disease : advances in etiology, pathogenesis and therapeutics Chichester : John Wiley, 2001,
— id: 3843, year: 2001, vol: , page: 487, stat: Chapter,

A newly formed amyloidogenic fragment due to a stop codon mutation causes familial British dementia
Ghiso J; Vidal R; Rostagno A; Mead S; Revesz T; Plant G; Frangione B
2001 ;3:12-13, Dementia reviews
— id: 73974, year: 2001, vol: 3, page: 12, stat: Journal Article,

Systemic Amyloid Deposits in Familial British Dementia
Ghiso JA; Holton J; Miravalle L; Calero M; Lashley T; Vidal R; Houlden H; Wood N; Neubert TA; Rostagno A; Plant G; Revesz T; Frangione B
2001 Nov 23;276(47):43909-43914, Journal of biological chemistry
Familial British dementia (FBD) is an early onset inherited disorder that, like familial Alzheimer's disease (FAD), is characterized by progressive dementia, amyloid deposition in the brain, and neurofibrillary degeneration of limbic neurons. The primary structure of the amyloid subunit (ABri) extracted from FBD brain tissues (Vidal, R., Frangione, B., Rostagno, A., Mead, S., Revesz, T., Plant, G., and Ghiso, J. (1999) Nature 399, 776-781) is entirely different and unrelated to any previously known amyloid protein. Patients with FBD have a single nucleotide substitution at codon 267 in the BRI2 gene, resulting in an arginine replacing the stop codon and a longer open reading frame of 277 amino acids instead of 266. The ABri peptide comprises the 34 C-terminal residues of the mutated precursor ABriPP-277 and is generated via furin-like proteolytic processing. Here we report that carriers of the Stop-to-Arg mutation have a soluble form of the amyloid peptide (sABri) in the circulation with an estimated concentration in the range of 20 ng/ml, several fold higher than that of soluble Abeta. In addition, ABri species identical to those identified in the brain were also found as fibrillar components of amyloid deposits predominantly in the blood vessels of several peripheral tissues, including pancreas and myocardium. We hypothesize that the high concentration of the soluble de novo created amyloidogenic peptide and/or the insufficient tissue clearance are the main causative factors for the formation of amyloid deposits outside the brain. Thus, FBD constitutes the first documented cerebral amyloidosis associated with neurodegeneration and dementia in which the amyloid deposition is also systemic
— id: 23926, year: 2001, vol: 276, page: 43909, stat: Journal Article,

Regional distribution of amyloid-Bri deposition and its association with neurofibrillary degeneration in familial British dementia
Holton JL; Ghiso J; Lashley T; Rostagno A; Guerin CJ; Gibb G; Houlden H; Ayling H; Martinian L; Anderton BH; Wood NW; Vidal R; Plant G; Frangione B; Revesz T
2001 Feb;158(2):515-526, American journal of pathology
Familial British dementia (FBD), pathologically characterized by cerebral amyloid angiopathy (CAA), amyloid plaques, and neurofibrillary degeneration, is associated with a stop codon mutation in the BRI gene resulting in the production of an amyloidogenic fragment, amyloid-Bri (ABri). The aim of this study was to assess the distribution of ABri fibrillar and nonfibrillar lesions and their relationship to neurofibrillary pathology, astroglial and microglial response using immunohistochemistry, confocal microscopy, and immunoelectron microscopy in five cases of FBD. Abnormal tau was studied with immunoblotting. We present evidence that ABri is deposited throughout the central nervous system in blood vessels and parenchyma where both amyloid (fibrillar) and pre-amyloid (nonfibrillar) lesions are formed. Ultrastructurally amyloid lesions appear as bundles of fibrils recognized by an antibody raised against ABri, whereas Thioflavin S-negative diffuse deposits consist of amorphous electron-dense material with sparse, dispersed fibrils. In contrast to nonfibrillar lesions, fibrillar ABri is associated with a marked astrocytic and microglial response. Neurofibrillary tangles and neuropil threads occurring mainly in limbic structures, are found in areas affected by all types of ABri lesions whereas abnormal neurites are present around amyloid lesions. Immunoblotting for tau revealed a triplet electrophoretic migration pattern. Our observations confirm a close link between ABri deposition and neurodegeneration in FBD
— id: 42013, year: 2001, vol: 158, page: 515, stat: Journal Article,

Familial Danish dementia (FDD); A novel form of cerebral amyloidosis associated with deposition of two amyloidogenic peptides
Holton, JL; Lashley, T; Vidal, R; Rostagno, A; Gibb, G; Anderton, BH; Braendgaard, H; Plant, GT; Bojsen-Moller, M; Ghiso, J; Frangione, B; Revesz, T
2001 MAY ;60(5):536-536, Journal of neuropathology & experimental neurology
— id: 55068, year: 2001, vol: 60, page: 536, stat: Journal Article,

Induction of NADPH cytochrome P450 reductase by the Alzheimer beta-protein. Amyloid as a "foreign body"
Pappolla, M A; Omar, R A; Chyan, Y J; Ghiso, J; Hsiao, K; Bozner, P; Perry, G; Smith, M A; Cruz-Sanchez, F
2001 Jul;78(1):121-128, Journal of neurochemistry
A large body of data suggests that the Alzheimer's amyloid peptide (Abeta) causes degeneration and death of neurons by mechanisms that involve reactive oxygen species. The pathways involved in Abeta-mediated oxidative injury are only partially understood. We theorized that abnormal microaggregates and/or pathological conformations of Abeta peptides may behave as xenobiotics and trigger the induction of NADPH cytochrome P450 reductase (CP450r), an enzyme which, if induced by non-physiological substrates (such as xenobiotics like drugs or other 'foreign molecules'), is known to cause oxidative stress. In order to test this hypothesis, i.e. that Abeta can increase the expression of CP450r, SK-N-SH human neuroblastoma cells were exposed to Abeta25-35 and Abeta1-42 and then examined for induction of this enzyme in immunoblots, using specific antibodies. Following exposure to Abeta peptides, neuroblastoma cells showed a clear-cut induction of CP450r. To determine whether this mechanism is operational in vivo, we investigated the expression of CP450r in a transgenic mouse model of Alzheimer's disease (AD) and in brains from patients afflicted with AD, using an immunocytochemical approach. Tissue sections from brains of transgenic mice exhibited strong immunoreactivity for CP450r, surrounding amyloid deposits. The pattern of expression of CP450r was similar to that exhibited by neuritic and oxidative stress markers. Sections from non-transgenic mice showed no detectable immunoreactivity. Immunostaining of sections from four brains with neuropathologically confirmed AD showed a pattern of abnormality different from transgenic mice that was characterized by abnormal immunoreactivity for CP450r within the cytoplasm of cortical neurons. No labeling was seen in sections from aged-matched control brains. The data showed that CP450r is induced by Alzheimer amyloid peptide and that such a response must be considered as one possible mechanism whereby Abeta causes oxidative stress
— id: 102280, year: 2001, vol: 78, page: 121, stat: Journal Article,

Retinal angiopathy in the British and Danish forms of Familial BRI Dementia
Plant, GT; Guerin, CJ; Holton, J; Houlden, H; Braendgaard, H; Bek, T; Ghiso, J; Frangione, B; Revesz, T
2001 MAR 15 ;42(4):S648-S648, Investigative ophthalmology & visual science. IOVS
— id: 54980, year: 2001, vol: 42, page: S648, stat: Journal Article,

Melatonin reverses the profibrillogenic activity of apolipoprotein E4 on the Alzheimer amyloid Abeta peptide
Poeggeler B; Miravalle L; Zagorski MG; Wisniewski T; Chyan YJ; Zhang Y; Shao H; Bryant-Thomas T; Vidal R; Frangione B; Ghiso J; Pappolla MA
2001 Dec 11;40(49):14995-15001, Biochemistry
Inheritance of apoE4 is a strong risk factor for the development of late-onset sporadic Alzheimer's disease (AD). Several lines of evidence suggest that apoE4 binds to the Alzheimer Abeta protein and, under certain experimental conditions, promotes formation of beta-sheet structures and amyloid fibrils. Deposition of amyloid fibrils is a critical step in the development of AD. We report here that addition of melatonin to Abeta in the presence of apoE resulted in a potent isoform-specific inhibition of fibril formation, the extent of which was far greater than that of the inhibition produced by melatonin alone. This effect was structure-dependent and unrelated to the antioxidant properties of melatonin, since it could be reproduced neither with the structurally related indole N-acetyl-5-hydroxytryptamine nor with the antioxidants ascorbate, alpha-tocophenol, and PBN. The enhanced inhibitory effects of melatonin and apoE were lost when bovine serum albumin was substituted for apoE. In addition, Abeta in combination with apoE was highly neurotoxic (apoE4 > apoE3) to neuronal cells in culture, and this activity was also prevented by melatonin. These findings suggest that reductions in brain melatonin, which occur during aging, may contribute to a proamyloidogenic microenvironment in the aging brain
— id: 42012, year: 2001, vol: 40, page: 14995, stat: Journal Article,

Deposition of amyloid-BRI (ABri) is associated with neurofibrillary degeneration in familial British dementia (FBD)
Revesz, T; Lashley, T; Vidal, R; Rostagno, K; Gibb, G; Anderton, BH; Plant, G; Frangione, B; Ghiso, J; Holton, JL
2001 MAY ;60(5):536-536, Journal of neuropathology & experimental neurology
— id: 55069, year: 2001, vol: 60, page: 536, stat: Journal Article,

Complement activation in Bri dementias and Alzheimer's disease
Rostagno, A.; Revesz, T.; Holton, J.; Lashley, T.; Frangione, B.; Ghiso, J.
2001 ;27(2):2562-2562, Abstracts (Society for Neuroscience)
Familial British dementia (FBD) and familial Danish dementia (FDD), are associated with amyloid deposition in the CNS and neurodegeneration. Amyloids ABri and ADan are C-terminal degradation products of the same precursor BriPP codified by the chromosome 13 BRI2 gene bearing different genetic defects, namely a Stop-to-Arg mutation in FBD and a ten-nucleotide insertion before the stop codon in FDD. Both de novo created amyloid peptides are 34 amino acids long, share 100% identity of the first 22 residues and pyroglutamate at their N-terminus. Neuritic components and NFTs containing PHF co-localize with the amyloid deposits in both disorders and the pattern of hyperphosphorylated tau immunoreactivity is almost indistinguishable from that seen in AD. To explore the role of inflammatory factors in these familial disorders, complement activation was assessed via immunohistochemistry, hemolytic assays and ELISA. Components and activation products (i.e. C1q, iC3b, C3d, C4d, C5b-9) were found to co-localize with plaques and vascular deposits in both diseases, suggesting in situ activation. ABri and ADan synthetic peptides activated the classical pathway in vitro and resulted in the formation of the activation products iC3b, C4d and SC5b-9 at levels comparable to those generated by Abeta42. The ability of ABri and ADan to trigger the complement cascade in vitro together with the presence of complement proteins and activation products as integral components of parenchymal and vascular amyloid deposits suggest that, as indicated in AD, the complement system may contribute to the mechanism of neurodegeneration leading to dementia
— id: 101620, year: 2001, vol: 27, page: 2562, stat: Journal Article,

Amyloid and nonfibrillar deposits in mice transgenic for wild-type human transthyretin: a possible model for senile systemic amyloidosis
Teng MH; Yin JY; Vidal R; Ghiso J; Kumar A; Rabenou R; Shah A; Jacobson DR; Tagoe C; Gallo G; Buxbaum J
2001 Mar;81(3):385-396, Laboratory investigation
The human serum protein transthyretin (TTR) is highly fibrillogenic in vitro and is the fibril precursor in both autosomal dominant (familial amyloidotic polyneuropathy [FAP] and familial amyloidotic cardiomyopathy [FAC]) and sporadic (senile systemic amyloidosis [SSA]) forms of human cardiac amyloidosis. We have produced mouse strains transgenic for either wild-type or mutant (TTRLeu55Pro) human TTR genes. Eighty-four percent of C57BI/6xDBA/2 mice older than 18 months, transgenic for the wild-type human TTR gene, develop TTR deposits that occur primarily in heart and kidney. In most of the animals, the deposits are nonfibrillar and non-Congophilic, but 20% of animals older than 18 months that bear the transgene have human TTR cardiac amyloid deposits identical to the lesions seen in SSA. Amino terminal amino acid sequence analysis and mass spectrometry of the major component extracted from amyloid and nonamyloid deposits revealed that both were intact human TTR monomers with no evidence of proteolysis or codeposition of murine TTR. This is the first instance in which the proteins from amyloid and nonfibrillar deposits in the same or syngeneic animals have been shown to be identical by sequence analysis. It is also the first time in any form of amyloidosis that nonfibrillar deposits have been shown to systematically occur temporally before the appearance of fibrils derived from the same precursor in the same tissues. These findings suggest, but do not prove, that the nonamyloid deposits represent a precursor of the fibril. The differences in the ultrastructure and binding properties of the deposits, despite the identical sizes and amino terminal amino acid sequences of the TTR and the dissociation of deposition and fibril formation, provide evidence that in vivo factors, perhaps associated with aging, impact on both systemic precursor deposition and amyloid fibril formation
— id: 18579, year: 2001, vol: 81, page: 385, stat: Journal Article,

Sequence, genomic structure and tissue expression of Human BRI3, a member of the BRI gene family
Vidal R; Calero M; Revesz T; Plant G; Ghiso J; Frangione B
2001 Mar 21;266(1-2):95-102, Gene
The BRI3 gene is a member of the BRI gene family, made up of at least three different genes (BRI1-3). Previous studies established the cDNA sequence and structure of the human and mouse BRI1 and BRI2 genes and we recently reported that mutations in the BRI2 isoform, located on chromosome 13, are associated with dementia in humans. In the present work, we determine the complete cDNA sequence and genomic organization of the human BRI3 gene. BRI3 codes for a polypeptide of 267 amino acids, with a Mr of 30 KDa and a pI of 8.47. The amino acid sequence is 43.7% identical to the sequence of the human BRI2, and 38.3% identical to that of human BRI1, with the highest percentage of amino acid identity being concentrated on the C-terminal half of the molecules. In Northern blots, BRI3 cDNA hybridizes only one message of approximately 2.1 kilobases, which is predominantly present in the human brain. The BRI3 gene is localized on chromosome 2 and consists of six exons spanning more than 20 kb. Homology search of EST data banks retrieved a Caenorhabditis briggsae homolog of BRI, indicating that the BRI gene belongs to a strongly conserved gene family. These studies, aimed at characterizing the members of the BRI gene family, may provide valuable clues to the understanding of their normal function and how mutations in BRI2 can cause neurodegeneration and dementia similar to Alzheimer's disease
— id: 21212, year: 2001, vol: 266, page: 95, stat: Journal Article,

A dacamer duplication in the BRI gene originates a de novo amyloid peptide that causes dementia in a Danish kindred
Vidal R; Revesz T; Rostango A; Bek T; Braendgaard H; Plant G; Ghiso J; Frangione B
Alzheimer's Disease : advances in etiology, pathogenesis and therapeutics Chichester : John Wiley, 2001,
— id: 3844, year: 2001, vol: , page: 507, stat: Chapter,

Inclusion body myositis, muscle blood vessel and cardiac amyloidosis, and transthyretin Val122Ile allele
Askanas V; Engel WK; Alvarez RB; Frangione B; Ghiso J; Vidal R
2000 Apr;47(4):544-549, Annals of neurology
Typical of sporadic inclusion body myositis muscle biopsies are vacuolated muscle fibers containing intracellular amyloid deposits and accumulations of 'Alzheimer-characteristic' proteins. There is no muscle blood vessel or cardiac amyloidosis. We report on a 70-year-old African-American man homozygous for the transthyretin Val122Ile allele who has both sporadic inclusion body myositis and cardiac amyloidosis. His unique pathological features included transthyretin immunoreactivity in prominent muscle blood vessel amyloid and congophilic amyloid deposits within vacuolated muscle fibers
— id: 9381, year: 2000, vol: 47, page: 544, stat: Journal Article,

Apolipoprotein J (clusterin) and Alzheimer's disease [In Process Citation]
Calero M; Rostagno A; Matsubara E; Zlokovic B; Frangione B; Ghiso J
2000 Aug 15;50(4):305-315, Microscopy research & technique
Apolipoprotein J (clusterin) is a ubiquitous multifunctional glycoprotein capable of interacting with a broad spectrum of molecules. In pathological conditions, it is an amyloid associated protein, co-localizing with fibrillar deposits in systemic and localized amyloid disorders. In Alzheimer's disease, the most frequent form of amyloidosis in humans and the major cause of dementia in the elderly, apoJ is present in amyloid plaques and cerebrovascular deposits but is rarely seen in NFT-containing neurons. ApoJ expression is up-regulated in a wide variety of insults and may represent a defense response against local damage to neurons. Four different mechanisms of action could be postulated to explain the role of apoJ as a neuroprotectant during cellular stress: (1) function as an anti-apoptotic signal, (2) protection against oxidative stress, (3) inhibition of the membrane attack complex of complement proteins locally activated as a result of inflammation, and (4) binding to hydrophobic regions of partially unfolded, stressed proteins, and therefore avoiding aggregation in a chaperone-like manner. This review focuses on the association of apoJ in biological fluids with Alzheimer's soluble Abeta. This interaction prevents Abeta aggregation and fibrillization and modulates its blood-brain barrier transport at the cerebrovascular endothelium.
— id: 9372, year: 2000, vol: 50, page: 305, stat: Journal Article,

Carboxyl-Terminal Fragments of Alzheimer {beta}-Amyloid Precursor Protein Accumulate in Restricted and Unpredicted Intracellular Compartments in Presenilin 1 Deficient Cells
Chen F; Yang DS; Petanceska S; Yang A; Tandon A; Yu G; Rozmahel R; Ghiso J; Nishimura M; Zhang DM; Kawarai T; Levesque G; Mills J; Levesque L; Song YQ; Rogaeva E; Westaway D; Mount H; Gandy S; St George-Hyslop P; Fraser PE
2000 Nov;275(47):36794-36802, Journal of biological chemistry
Absence of functional presenilin 1 (PS1) protein leads to loss of gamma-secretase cleavage of the amyloid precursor protein (betaAPP), resulting in a dramatic reduction in amyloid beta peptide (Abeta) production and accumulation of alpha- or beta-secretase-cleaved C-terminal fragments of betaAPP (alpha- or beta-CTFs). The major C-terminal fragment (CTF) in brain was identified as Abeta[11-98], which is consistent with the observation that cultured neurons generate primarily Abeta[11-40]. In PS1-/- murine neurons and fibroblasts expressing the loss-of-function PS1Asp385Ala mutant, CTFs accumulated in the endoplasmic reticulum (ER), Golgi, and lysosomes, but not late endosomes. There were some subtle differences in the subcellular distribution of CTFs in PS1-/- neurons as compared to PS1Asp385Ala mutant fibroblasts. However, there was no obvious redistribution of full-length betaAPP or of markers of other organelles in either mutant. Blockade of ER-to-Golgi trafficking indicated that in PS1-/- neurons (as in normal cells) trafficking of betaAPP to the Golgi compartment is necessary before alpha- and beta-secretase cleavages occur. Thus, while we cannot exclude a specific role for PS1 in trafficking of CTFs, these data argue against a major role in general protein trafficking. These results are more compatible with a role for PS1 either as the actual gamma-secretase catalytic activity or in other functions indirectly related to gamma-secretase catalysis (e.g., an activator of gamma-secretase, a substrate-adaptor for gamma-secretase, or delivery of gamma-secretase to betaAPP-containing compartments)
— id: 9785, year: 2000, vol: 275, page: 36794, stat: Journal Article,

Familial cerebral amyloid angiopathies and dementia [In Process Citation]
Frangione B; Vidal R; Rostagno A; Ghiso J
2000 ;14 Suppl 1:S25-S30, Alzheimer disease & associated disorders
Amyloidosis is a disorder of protein conformation leading to aggregation. The term defines a diverse group of proteins normally present in body fluids as soluble precursors that can be deposited as insoluble amyloid fibrils in different tissues producing organ dysfunction and cell death. These fibrils are composed of self-assembled, low molecular weight mass peptides adopting beta-pleated sheet structure, the conformation responsible for their physicochemical properties and tinctoreal characteristics. So far, 20 different proteins have been identified as subunits of amyloid fibrils (Westermark et al., 1999). Collectively, they are products of normal genes; however, several amyloid precursors contain abnormal amino acid substitutions that can impose an unusual potential for self-aggregation. The molecular mass of the amyloid peptides is within the 4 to 30-kDa range, with heterogeneity at the amino- and carboxyl-terminal portions found in most amyloid proteins. Increased levels of amyloid precursors, either in the circulation or locally in sites of deposition, are usually the result of overexpression or defective clearance, or both. Of the 20 amyloid proteins identified, few of them are known to cause amyloid deposition in the central nervous system, which in turn results in cognitive deficits, dementia, stroke, cerebellar and extrapyramidal signs, or a combination of them
— id: 9374, year: 2000, vol: 14 Suppl 1, page: S25, stat: Journal Article,

Neuropathology and genetics of Prion protein and British cerebral amyloid angiopathies
Ghetti B; Piccardo P; Frangione B; Vidal R; Ghiso J
Cerebral amyloid angiopathy in Alzheimer's disease and related disorders Boston : Kluwer, 2000,
— id: 5150, year: 2000, vol: , page: 237, stat: Chapter,

A newly formed amyloidogenic fragment due to a stop codon mutation causes familial British dementia
Ghiso J; Vidal R; Rostagno A; Mead S; Revesz T; Plant G; Frangione B
2000 Apr;903:129-137, Annals of the New York Academy of Sciences
Familial British dementia (FBD) is an early-onset autosomal dominant disorder characterized by progressive cognitive impairment, spasticity, and cerebellar ataxia. Hippocampal neurofibrillar degeneration and widespread parenchymal and vascular amyloid deposits are the main neuropathological lesions. Amyloid fibrils are composed of a novel 34 amino acid subunit (ABri) with no sequence identity to any known amyloid molecule. The peptide derives from a larger precursor protein codified by a single gene BRI on chromosome 13. Affected family members have a single base substitution at the stop codon of the BRI gene that generates a longer open-reading frame resulting in a larger precursor protein. The release of the 34 C-terminal amino acids from the mutated precursor originates the ABri amyloid subunit. Our discovery of a new amyloid associated with the development of dementia supports the concept that amyloid peptides may be of primary importance in the initiation of neurodegeneration
— id: 9377, year: 2000, vol: 903, page: 129, stat: Journal Article,

Amyloidogenesis in familial British dementia is associated with a genetic defect on chromosome 13
Ghiso J; Vidal R; Rostagno A; Miravalle L; Holton JL; Mead S; Revesz T; Plant G; Frangione B
2000 ;920(1):84-92, Annals of the New York Academy of Sciences
Familial British dementia (FBD) is a disorder characterized by the presence of amyloid deposits in cerebral blood vessels and brain parenchyma coexisting with neurofibrillary tangles in limbic areas. The amyloid subunit (ABri) is a 4 kDa fragment of a 266 amino acid type II single-spanning transmembrane precursor protein encoded by the BRI gene located on chromosome 13. In FBD patients, a single base substitution at the stop codon of this gene generates a larger 277-residue precursor (ABriPP-277). Proteolytic processing by a furin-like enzyme at the C-terminus of the elongated precursor generates the 34 amino acid ABri that undergoes rapid aggregation and fibrillization. ABri is structually unrelated to all known amyloids including A beta, the main component of the amyloid lesions in Alzheimer's disease (AD), indicating that cerebral deposition of amyloid molecules other than A beta can trigger similar neuropathological changes leading to neuronal loss and dementia. These data support the concept that amyloid deposition in the vascular wall and brain parenchyma is of primary importance in the initiation of neurogeneration
— id: 39490, year: 2000, vol: 920, page: 84, stat: Journal Article,

Familial British dementia is a systemic amyloidosis
Ghiso, J.; Miravalle, L.; Calero, M.; Vidal, R.; Holden, H.; Holton, J.; Lashley, T.; Rostagno, A.; Wood, N.; Revesz, T.; Plant, G.; Frangione, B.
2000 ;26(1-2):Abstract No.-201.11, Abstracts (Society for Neuroscience)
Familial British dementia (FBD) is an autosomal dominant neurodegenerative disorder clinically characterized by progressive dementia, spastic tetraparesis and cerebellar ataxia, with an age of onset in the fourth decade. The neuropathology of FBD (amyloid angiopathy, parenchymal plaques and neurofibrillary tangles) is similar to that of AD. Amyloid deposits in cerebral blood vessels and parenchymal plaques are mainly composed of a 4 kDa subunit, ^ABri, derived from a type II transmembrane precursor molecule mutated at the stop codon. The mutation produces a longer open reading frame that generates a larger 277 aa precursor. ^ABri is a 34 amino acids peptide released by proteolytic processing of the C-terminus of the mutated precursor protein (ABriPP277). We have identified soluble ABri (sABri) in serum and CSF of patients with FBD using a combination of immunoprecipitation, mass spectrometry and western blot analysis. The 4 kDa component was present in all tested carriers of the Stop-to-Arg mutation and consistently absent in non-carrier family members and normal controls. In contrast to the CNS-deposited ^ABri, sABri was monomeric and devoided of N-terminal pyroglutamate. In view of these findings, we tested systemic organs in an autopsy case of FBD for amyloid deposits. Antibodies specific to the ^ABri peptide labeled Congo red positive vascular lesions in several peripheral organs; in addition, parenchymal immunoreactivity was also seen in many of the tissues tested. Biochemical analysis of the deposited material isolated from pancreas, uterus and skeletal muscle identified ^ABri species similar to those found in amyloid lesions in the CNS (Nature, 399:776,1999). The main component was full-length ABri featuring N-terminus pyroglutamate. The data indicate that amyloid formation in FBD occurs not only in the brain but also peripherally. This is the first case of cerebral amyloidosis associated with neurodegeneration in demented patients where the amyloid deposition is also systemic
— id: 101621, year: 2000, vol: 26, page: Abstract No., stat: Journal Article,

Familial British dementia: Immunohistochemical and immunoelectron microscopic study
Holton, JL; Lashley, T; Vidal, R; Rostagno, A; Guerin, CJ; Houlden, H; Plant, G; Frangione, B; Ghiso, J; Revesz, T
2000 SEP ;10(4):713-714, Brain pathology
— id: 73961, year: 2000, vol: 10, page: 713, stat: Journal Article,

Familial British dementia with amyloid angiopathy: early clinical, neuropsychological and imaging findings
Mead S; James-Galton M; Revesz T; Doshi RB; Harwood G; Pan EL; Ghiso J; Frangione B; Plant G
2000 May;123(Pt 5):975-991, Brain
Familial British dementia with amyloid angiopathy (FBD) is an autosomal dominant condition characterized by a dementia, progressive spastic tetraparesis and cerebellar ataxia with onset in the sixth decade. A point mutation in the BRI gene has been shown to be the genetic abnormality. Genealogical work with the large family originally reported by Worster-Drought and updated by Plant has identified nine generations dating back to the late eighteenth century. The pedigree now includes six living affected patients, 35 historical cases, and 52 descendants at risk of having inherited the disease. A common ancestor has been identified between the large pedigree and a case report of 'familial cerebellar ataxia with amyloid angiopathy'. An autopsy case from a separate family with an identical condition is described but no common ancestor with the large pedigree has been found. Case histories have been researched and updated in each pedigree. Eleven individuals at risk of FBD, aged between 44 and 56 years, agreed to undergo a clinical and neuropsychological assessment along with MRI brain imaging in order to clarify early diagnostic features. Five of the eleven were thought to show early clinical signs of the disease. Neurological examination was abnormal in three, with limb and gait ataxia and mild spastic paraparesis. Three had impaired recognition and recall memory and another had mild impairment of delayed visual recall. All affected individuals had an abnormal MRI of the brain, consisting of deep white-matter hyperintensity (T(2)-weighted scans) and lacunar infarcts, but no intracerebral haemorrhage. The corpus callosum was affected particularly, and in one patient it was severely atrophic
— id: 9380, year: 2000, vol: 123, page: 975, stat: Journal Article,

Substitutions at codon 22 of Alzheimer's abeta peptide induce diverse conformational changes and apoptotic effects in human cerebral endothelial cells [In Process Citation]
Miravalle L; Tokuda T; Chiarle R; Giaccone G; Bugiani O; Tagliavini F; Frangione B; Ghiso J
2000 Sep 1;275(35):27110-27116, Journal of biological chemistry
Cerebral amyloid angiopathy is commonly associated with normal aging and Alzheimer's disease and it is also the principal feature of hereditary cerebral hemorrhage with amyloidosis Dutch type, a familial condition associated to a point mutation G to C at codon 693 of the amyloid beta (Abeta) precursor protein gene resulting in a Glu to Gln substitution at position 22 of the Abeta (E22Q). The patients carrying the AbetaE22Q variant usually present with lobar cerebral hemorrhages before 50 years of age. A different mutation described in several members of three Italian kindred who presented with recurrent hemorrhagic strokes late in life, between 60 and 70 years of age, also associated with extensive cerebrovascular amyloid deposition has been found at the same position 22, this time resulting in a Glu to Lys substitution (E22K). We have compared the secondary structure, aggregation, and fibrillization properties of the two Abeta40 variants and the wild type peptide. Using flow cytometry analysis after staining with propidium iodide and annexin V, we also evaluated the cytotoxic effects of the peptides on human cerebral endothelial cells in culture. Under the conditions tested, the E22Q peptide exhibited the highest content of beta-sheet conformation and the fastest aggregation/fibrillization properties. The Dutch variant also induced apoptosis of cerebral endothelial cells at a concentration of 25 &mgr;m, whereas the wild type Abeta and the E22K mutant had no effect. The data suggest that different amino acids at position 22 confer distinct structural properties to the peptides that appear to influence the onset and aggressiveness of the disease rather than the phenotype
— id: 9376, year: 2000, vol: 275, page: 27110, stat: Journal Article,

An assessment of the antioxidant and the antiamyloidogenic properties of melatonin: implications for Alzheimer's disease [In Process Citation]
Pappolla MA; Chyan YJ; Poeggeler B; Frangione B; Wilson G; Ghiso J; Reiter RJ
2000 ;107(2):203-231, Journal of neural transmission
This review summarizes recent advancements in our understanding of the potential role of the amyloid beta protein in Alzheimer's disease. It also discusses the significance of amyloid beta in initiating the generation of partially reduced oxygen species and points out their role in damaging essential macromolecules in the CNS which leads to neuronal dysfunction and loss. Recently acquired experimental data links these destructive oxidative processes with some neurodegenerative aspects of Alzheimer's disease. The experimental findings related to the free radical scavenging and antioxidative properties of melatonin are tabulated and its efficacy and the likely mechanisms involved in its ability to reduce neuronal damage mediated by oxygen-based reactive species in experimental models of Alzheimer's disease are summarized. Besides the direct scavenging properties and indirect antioxidant actions of melatonin, its ability to protect neurons probably also stems from its antiamyloidogenic properties. Melatonin is also unique because of the ease with which it passes through the blood-brain barrier
— id: 9375, year: 2000, vol: 107, page: 203, stat: Journal Article,

Dementia associated with amyloid beta angiopathy, tau perivascular pathology and APOE epsilon 4 genotype
Piccardo, P; Vidal, R; Calero, M; Farlow, MR; Unverzagt, FW; Gomez-Tortosa, E; Ghiso, J; Hyman, B; Frangione, B; Ghetti, B
2000 SEP ;10(4):714-714, Brain pathology
— id: 54519, year: 2000, vol: 10, page: 714, stat: Journal Article,

Cerebral deposition of ABRI amyloid in Familial British Dementia
Revesz T; Holton J; Vidal R; Rostagno A; Lashley T; Plant G; Frangione B; Ghiso J
2000 ;21:S191-S191, Neurobiology of aging
— id: 73972, year: 2000, vol: 21, page: S191, stat: Journal Article,

Clearance of Alzheimer's amyloid-ss(1-40) peptide from brain by LDL receptor-related protein-1 at the blood-brain barrier
Shibata M; Yamada S; Kumar SR; Calero M; Bading J; Frangione B; Holtzman DM; Miller CA; Strickland DK; Ghiso J; Zlokovic BV
2000 Dec;106(12):1489-1499, Journal of clinical investigation
Elimination of amyloid-ss peptide (Ass) from the brain is poorly understood. After intracerebral microinjections in young mice, (125)I-Ass(1-40) was rapidly removed from the brain (t(1/2) </= 25 minutes), mainly by vascular transport across the blood-brain barrier (BBB). The efflux transport system for Ass(1-40) at the BBB was half saturated at 15.3 nM, and the maximal transport capacity was reached between 70 nM and 100 nM. Ass(1-40) clearance was substantially inhibited by the receptor-associated protein, and by antibodies against LDL receptor-related protein-1 (LRP-1) and alpha(2)-macroglobulin (alpha(2)M). As compared to adult wild-type mice, clearance was significantly reduced in young and old apolipoprotein E (apoE) knockout mice, and in old wild-type mice. There was no evidence that Ass was metabolized in brain interstitial fluid and degraded to smaller peptide fragments and amino acids before its transport across the BBB into the circulation. LRP-1, although abundant in brain microvessels in young mice, was downregulated in older animals, and this downregulation correlated with regional Ass accumulation in brains of Alzheimer's disease (AD) patients. We conclude that the BBB removes Ass from the brain largely via age-dependent, LRP-1-mediated transport that is influenced by alpha(2)M and/or apoE, and may be impaired in AD
— id: 42015, year: 2000, vol: 106, page: 1489, stat: Journal Article,

In vitro evidence that beta-amyloid peptide 1-40 diffuses across the blood-brain barrier and affects its permeability
Strazielle N; Ghersi-Egea JF; Ghiso J; Dehouck MP; Frangione B; Patlak C; Fenstermacher J; Gorevic P
2000 Jan;59(1):29-38, Journal of neuropathology & experimental neurology
Beta amyloid peptides are major insoluble constituents of amyloid fibrils in senile plaques and cerebrovascular deposits, both characteristic of Alzheimer disease (AD). Low concentrations of soluble forms of amyloid peptides are also present in normal CSF. We previously demonstrated that the 40 amino acid form of soluble beta-amyloid peptide (sAbeta) is rapidly cleared from rat CSF into blood. Herein we hypothesized that a saturable, outwardly directed flux of this peptide occurs at the blood-brain barrier (BBB) and tested whether supraphysiological (possibly pathological) concentrations of sAbeta could alter the permeability of this barrier to a paracellular tracer, polyethylene glycol (PEG). Using an in vitro model of BBB, we showed that influx and efflux of sAbeta were equal, modest (60%-160% greater than that of PEG), and not saturable. These observations suggest that sAbeta moved across the monolayer by a diffusional process, and not via a transporter. PEG flux was doubled immediately after the luminal concentration of cold sAbeta was raised to 5 microM, and was doubled 150 min after the abluminal concentration of sAbeta was increased to 5 microM. Pathological elevations of sAbeta concentration in plasma or brain interstitial fluid may, therefore, alter the permeability of brain capillaries in vivo
— id: 57562, year: 2000, vol: 59, page: 29, stat: Journal Article,

Lipidation of apolipoprotein E influences its isoform-specific interaction with Alzheimer's amyloid beta peptides
Tokuda T; Calero M; Matsubara E; Vidal R; Kumar A; Permanne B; Zlokovic B; Smith JD; Ladu MJ; Rostagno A; Frangione B; Ghiso J
2000 Jun 1;348 Pt 2:359-365, Biochemical journal
The inheritance of the apolipoprotein E (apoE) epsilon4 allele is a prevailing risk factor for sporadic and familial Alzheimer's disease (AD). ApoE isoforms bind directly to Alzheimer's amyloid beta (Abeta) peptides both in vitro and in vivo. Recent studies suggest that association of apoE with lipids may modulate its interaction with Abeta. We examined the binding of lipid-associated and delipidated apoE3 and apoE4 isoforms to Abeta utilizing a solid-phase binding assay and estimated the dissociation constants for the interaction of various apoE and Abeta species. Using native apoE isoforms from stably transfected RAW 264 and human embryonic kidney 293 cells, apoE3 had greater affinity than apoE4 for both Abeta1-40 and Abeta1-42. Delipidation of apoE decreased its affinity for Abeta peptides by 5-10-fold and abolished the isoform-specificity. Conversely, incorporation of apoE isoforms produced by baculovirus-infected Sf9 cells into reconstituted human high-density-lipoprotein lipoparticles restored the affinity values for Abeta peptides and resulted in preferential binding of apoE3. The data demonstrate that native lipid-associated apoE3 binds to Abeta peptides with 2-3-fold higher affinity than lipid-associated apoE4. Since the isoforms' binding efficiency correlate inversely with the risk of developing late-onset AD, the results suggest a possible involvement of apoE3 in the clearance or routing out of Abeta from the central nervous system as one of the mechanisms underlying the pathology of the disease
— id: 9378, year: 2000, vol: 348 Pt 2, page: 359, stat: Journal Article,

Senile dementia associated with amyloid beta protein angiopathy and tau perivascular pathology but not neuritic plaques in patients homozygous for the APOE-epsilon4 allele [In Process Citation]
Vidal R; Calero M; Piccardo P; Farlow MR; Unverzagt FW; Mendez E; Jimenez-Huete A; Beavis R; Gallo G; Gomez-Tortosa E; Ghiso J; Hyman BT; Frangione B; Ghetti B
2000 Jul;100(1):1-12, Acta neuropathologica
Amyloid beta protein deposition in cortical and leptomeningeal vessels, causing the most common type of cerebral amyloid angiopathy, is found in sporadic and familial Alzheimer's disease (AD) and is the principal feature in the hereditary cerebral hemorrhage with amyloidosis, Dutch type. The presence of the Apolipopriotein E (APOE)-epsilon4 allele has been implicated as a risk factor for AD and the development of cerebral amyloid angiopathy in AD. We report clinical, pathological and biochemical studies on two APOE-epsilon4 homozygous subjects, who had senile dementia and whose main neuropathological feature was a severe and diffuse amyloid angiopathy associated with perivascular tau neurofibrillary pathology. Amyloid beta protein and ApoE immunoreactivity were observed in leptomeningeal vessels as well as in medium-sized and small vessels and capillaries in the parenchyma of the neocortex, hippocampus, thalamus, cerebellum, midbrain, pons, and medulla. The predominant peptide form of amyloid beta protein was that terminating at residue Val40, as determined by immunohistochemistry, amino acid sequence and mass spectrometry analysis. A crown of tau-immunopositive cell processes was consistently present around blood vessels. DNA sequence analysis of the Amyloid Precursor Protein gene and Presenilin-1 (PS-1) gene revealed no mutations. In these APOE-epsilon4 homozygous patients, the pathological process differed from that typically seen in AD in that they showed a heavy burden of perivascular tau-immunopositive cell processes associated with severe amyloid beta protein angiopathy, neurofibrillary tangles, some cortical Lewy bodies and an absence of neuritic plaques. These cases emphasize the concept that tau deposits may be pathogenetically related to amyloid beta protein deposition
— id: 9373, year: 2000, vol: 100, page: 1, stat: Journal Article,

New familial forms of cerebral amyloid and dementia
Vidal R; Ghiso J; Frangione B
2000 Nov;5(6):575-576, Molecular psychiatry
— id: 42014, year: 2000, vol: 5, page: 575, stat: Journal Article,

A decamer duplication in the BRI gene originates a de novo amyloid peptide that causes dementia in a Danish kindred
Vidal R; Revesz T; Rostagno A; Bek T; Braengaard H; Plant G; Ghiso J; Frangione B
2000 ;21:S58-S58, Neurobiology of aging
— id: 101630, year: 2000, vol: 21, page: S58, stat: Journal Article,

A decamer duplication in the 3' region of the BRI gene originates an amyloid peptide that is associated with dementia in a Danish kindred
Vidal R; Revesz T; Rostagno A; Kim E; Holton JL; Bek T; Bojsen-Moller M; Braendgaard H; Plant G; Ghiso J; Frangione B
2000 Apr 25;97(9):4920-4925, Proceedings of the National Academy of Sciences of the United States of America
Familial Danish dementia (FDD), also known as heredopathia ophthalmo-oto-encephalica, is an autosomal dominant disorder characterized by cataracts, deafness, progressive ataxia, and dementia. Neuropathological findings include severe widespread cerebral amyloid angiopathy, hippocampal plaques, and neurofibrillary tangles, similar to Alzheimer's disease. N-terminal sequence analysis of isolated leptomeningeal amyloid fibrils revealed homology to ABri, the peptide originated by a point mutation at the stop codon of gene BRI in familial British dementia. Molecular genetic analysis of the BRI gene in the Danish kindred showed a different defect, namely the presence of a 10-nt duplication (795-796insTTTAATTTGT) between codons 265 and 266, one codon before the normal stop codon 267. The decamer duplication mutation produces a frame-shift in the BRI sequence generating a larger-than-normal precursor protein, of which the amyloid subunit (designated ADan) comprises the last 34 C-terminal amino acids. This de novo-created amyloidogenic peptide, associated with a genetic defect in the Danish kindred, stresses the importance of amyloid formation as a causative factor in neurodegeneration and dementia
— id: 9379, year: 2000, vol: 97, page: 4920, stat: Journal Article,

Vascular transport of Alzheimer's amyloid ? peptides and apolipoproteins
Zlokovic B; Ghiso J; Frangione B
Cerebral amyloid angiopathy in Alzheimer's disease and related disorders Boston : Kluwer, 2000,
— id: 5151, year: 2000, vol: , page: 325, stat: Chapter,

Clearance of amyloid beta-peptide from brain: transport or metabolism?
Zlokovic, B V; Yamada, S; Holtzman, D; Ghiso, J; Frangione, B
2000 Jul;6(7):718-719, Nature medicine
— id: 101674, year: 2000, vol: 6, page: 718, stat: Journal Article,

Functional and structural properties of lipid-associated apolipoprotein J (clusterin)
Calero M; Tokuda T; Rostagno A; Kumar A; Zlokovic B; Frangione B; Ghiso J
1999 Dec 1;344 Pt 2:375-383, Biochemical journal
Apolipoprotein J (apoJ, clusterin) is a multifunctional protein normally associated with lipids in plasma and cerebrospinal fluid, and secreted as lipoparticles by hepatocytes and astrocytes. To investigate whether the structural and functional properties of apoJ are modulated upon binding to lipids, we prepared apoJ high-density lipoprotein (HDL)-like particles employing either synthetic or plasma HDL-derived lipids. The majority of the resulting lipoparticles contained one molecule of apoJ per particle and exhibited the same alpha2 electrophoretic mobility characteristic of apoJ-containing plasma HDL. Particle size seemed to be dependent on the presence of cholesterol in the lipid mixture and ranged from diameters of 10 nm in the presence of cholesterol to 20 nm in the absence of cholesterol. CD analysis and Fourier-transform infrared spectroscopy revealed similar changes in the apoJ secondary structure induced by its incorporation into lipoparticles, namely a decrease in alpha-helix content and an increase in beta-turn structures. Two functional assays, the binding interaction with Alzheimer's amyloid beta peptides and the inhibitory activity of the complement membrane-attack complex, did not detect any changes in apoJ activity following its lipidation (P>0.05). On the contrary, the binding affinity to the cellular receptor megalin was enhanced significantly (P<0.01) after the association with lipids; the K(d) value decreased from 78.8+/-10.7 nM for the delipidated form to 37. 0+/-7.3 nM for apoJ-HDL. Although it is not known whether the structural changes observed are directly responsible for the higher receptor-binding affinity, the data suggest that the complement inhibition and amyloid beta-binding motifs are located in areas of the molecule different from those involved in the apoJ-megalin interaction
— id: 9384, year: 1999, vol: 344 Pt 2, page: 375, stat: Journal Article,

Potent neuroprotective properties against the Alzheimer beta-amyloid by an endogenous melatonin-related indole structure, indole-3-propionic acid
Chyan YJ; Poeggeler B; Omar RA; Chain DG; Frangione B; Ghiso J; Pappolla MA
1999 Jul 30;274(31):21937-21942, Journal of biological chemistry
Widespread cerebral deposition of a 40-43-amino acid peptide called the amyloid beta-protein (Abeta) in the form of amyloid fibrils is one of the most prominent neuropathologic features of Alzheimer's disease. Numerous studies suggest that Abeta is toxic to neurons by free radical-mediated mechanisms. We have previously reported that melatonin prevents oxidative stress and death of neurons exposed to Abeta. In the process of screening indole compounds for neuroprotection against Abeta, potent neuroprotective properties were uncovered for an endogenous related species, indole-3-propionic acid (IPA). This compound has previously been identified in the plasma and cerebrospinal fluid of humans, but its functions are not known. IPA completely protected primary neurons and neuroblastoma cells against oxidative damage and death caused by exposure to Abeta, by inhibition of superoxide dismutase, or by treatment with hydrogen peroxide. In kinetic competition experiments using free radical-trapping agents, the capacity of IPA to scavenge hydroxyl radicals exceeded that of melatonin, an indoleamine considered to be the most potent naturally occurring scavenger of free radicals. In contrast with other antioxidants, IPA was not converted to reactive intermediates with pro-oxidant activity. These findings may have therapeutic applications in a broad range of clinical situations
— id: 9387, year: 1999, vol: 274, page: 21937, stat: Journal Article,

Distinct secretases, a cysteine protease and a serine protease, generate the C termini of amyloid beta-proteins Abeta1-40 and Abeta1-42, respectively
Figueiredo-Pereira ME; Efthimiopoulos S; Tezapsidis N; Buku A; Ghiso J; Mehta P; Robakis NK
1999 Apr;72(4):1417-1422, Journal of neurochemistry
The carboxy-terminal ends of the 40- and 42-amino acids amyloid beta-protein (Abeta) may be generated by the action of at least two different proteases termed gamma(40)- and gamma(42)-secretase, respectively. To examine the cleavage specificity of the two proteases, we treated amyloid precursor protein (APP)-transfected cell cultures with several dipeptidyl aldehydes including N-benzyloxycarbonyl-Leu-leucinal (Z-LL-CHO) and the newly synthesized N-benzyloxycarbonyl-Val-leucinal (Z-VL-CHO). All dipeptidyl aldehydes tested inhibited production of both Abeta1-40 and Abeta1-42. Changes in the P1 and P2 residues of these aldehydes, however, indicated that the amino acids occupying these positions are important for the efficient inhibition of gamma-secretases. Peptidyl aldehydes inhibit both cysteine and serine proteases, suggesting that the two gamma-secretases belong to one of these mechanistic classes. To differentiate between the two classes of proteases, we treated our cultures with the specific cysteine protease inhibitor E-64d. This agent inhibited production of secreted Abeta1-40, with a concomitant accumulation of its cellular precursor indicating that gamma(40)-secretase is a cysteine protease. In contrast, this treatment increased production of secreted Abeta1-42. No inhibition of Abeta production was observed with the potent calpain inhibitor I (acetyl-Leu-Leu-norleucinal), suggesting that calpain is not involved. Together, these results indicate that gamma(40)-secretase is a cysteine protease distinct from calpain, whereas gamma(42)-secretase may be a serine protease. In addition, the two secretases may compete for the same substrate. Dipeptidyl aldehyde treatment of cultures transfected with APP carrying the Swedish mutation resulted in the accumulation of the beta-secretase C-terminal APP fragment and a decrease of the alpha-secretase C-terminal APP fragment, indicating that this mutation shifts APP cleavage from the alpha-secretase site to the beta-secretase site
— id: 9389, year: 1999, vol: 72, page: 1417, stat: Journal Article,

Familial cerebrovascular amyloidosis with neurofibillary tangles causing dementia in British patients is due to a stop codon mutation of a novel gene BRI mapped to chromosome 13
Ghiso, J; Vidal, R; Rostagno, A; Mead, S; Revesz, T; Plant, G; Frangione, B
1999 Oct 23-28;25(1-2):297-297, Abstracts (Society for Neuroscience)
— id: 15871, year: 1999, vol: 25, page: 297, stat: Journal Article,

Immunochemical microanalysis of amyloid proteins in fine-needle aspirates of abdominal fat
Kaplan B; Vidal R; Kumar A; Ghiso J; Gallo G
1999 Sep;112(3):403-407, American journal of clinical pathology
Diagnosis of systemic amyloidosis by fine-needle aspiration biopsy of abdominal fat is well established. A complete diagnosis now must include determination of the chemical type of amyloid. Microextracts of amyloid proteins from 11 Congo red-positive aspirate samples were analyzed with immunochemical methods. There was correspondence of the results obtained by immunohistologic and Western blotting analyses in 3 of 4 specimens with kappa light chain amyloid, 5 of 6 with lambda amyloid, and 1 with amyloid A. The method provides rapid and reliable diagnostic information necessary for classification of the chemical type of amyloid required for initiation of specific modes of therapy, with little discomfort to the patient
— id: 9386, year: 1999, vol: 112, page: 403, stat: Journal Article,

Lipoprotein-free amyloidogenic peptides in plasma are elevated in patients with sporadic Alzheimer's disease and Down's syndrome
Matsubara E; Ghiso J; Frangione B; Amari M; Tomidokoro Y; Ikeda Y; Harigaya Y; Okamoto K; Shoji M
1999 Apr;45(4):537-541, Annals of neurology
About 90% of the soluble amyloid beta (sA beta) that circulates in normal human plasma is associated with lipoprotein particles. In sporadic Alzheimer's disease patients, free sA beta42 but not sA beta40 is increased approximately 2.3-fold compared with age-matched controls, although a more marked elevation (approximately 8-fold for free sA beta40 and about 20-fold for sA beta42) is found in Down's syndrome patients. The data suggest that lipoprotein-sA beta dissociation may contribute to the influx of sA beta into the brain as a result of decreased plasma clearance
— id: 9388, year: 1999, vol: 45, page: 537, stat: Journal Article,

Abeta1-40 peptide bearing the APP693 Dutch mutation induces apoptosis in human cerebral endothelial cells
Miravalle, L; Tokuda, T; Chiarle, R; Kramer, H; Frangione, B; Ghiso, J
1999 Oct 23-28;25(1-2):2124-2124, Abstracts (Society for Neuroscience)
— id: 15838, year: 1999, vol: 25, page: 2124, stat: Journal Article,

Dual anti-amyloidogenic and anti-oxidant properties of Melatonin : a new therapy for Alzheimer's disease?
Papolla M; Chyan X; Bozner P; Soto C; Shao H; Reiter R; Brewer G; Robakis N; Zagorski M; Frangione B; Ghiso J
Alzheimer's disease and related disorders New York : Wiley, 1999,
— id: 5148, year: 1999, vol: , page: 661, stat: Chapter,

Alzheimer beta protein mediated oxidative damage of mitochondrial DNA: prevention by melatonin
Pappolla MA; Chyan YJ; Poeggeler B; Bozner P; Ghiso J; LeDoux SP; Wilson GL
1999 Nov;27(4):226-229, Journal of pineal research
Most contemporary progress in Alzheimer's disease (AD) stems from the study of a 42 43 amino acid peptide. called the amyloid beta protein (Abeta), as the main neuropathologic marker of the disorder. It has been demonstrated that Abeta has neurotoxic properties and that such effects are mediated by free-radicals. Exposure of neuronal cells to Abeta results in a spectrum of oxidative lesions that are profoundly harmful to neuronal homeostasis. We had previously shown that Abeta25-35 induces oxidative damage to mitochondrial DNA (mtDNA) and that this modality of injury is prevented by melatonin. Because Abeta25 35 does not occur in AD and because the mode of toxicity by Abeta25-35 may be different from that of Abeta1-42 (the physiologically relevant form of Abeta), we extended our initial observations to determine whether oxidative damage to mtDNA could also be induced by Abeta1-42 and whether this type of injury is prevented by melatonin. Exposure of human neuroblastoma cells to Abeta1-42 resulted in marked oxidative damage to mtDNA as determined by a quantitative polymerase chain reaction method. Addition of melatonin to cell cultures along with Abeta completely prevented the damage. This study supports previous findings with Abeta25-35, including a causative role for Abeta in the mitochondrial oxidative lesions present in AD brains. Most important, the data confirms the neuroprotective role of melatonin in Abeta-mediated oxidative injury. Because melatonin also inhibits amyloid aggregation, lacks toxicity, and efficiently crosses the blood-brain barrier, this hormone appears superior to other available antioxidants as a candidate for pharmacologic intervention in AD
— id: 9385, year: 1999, vol: 27, page: 226, stat: Journal Article,

Melatonin abolishes the pro-aggregatory effects of apoE4 on the Alzheimer beta-protein
Poeggeler, B.; Chyan, Y.-J.; Bryant, T.; Wisniewski, T.; Frangione, B.; Ghiso, J.; Pappolla, M.
1999 ;25(1-2):1805-78, Abstracts (Society for Neuroscience)
— id: 97638, year: 1999, vol: 25, page: 1805, stat: Journal Article,

Characterization of a novel trypanosome lytic factor from human serum
Raper J; Fung R; Ghiso J; Nussenzweig V; Tomlinson S
1999 Apr;67(4):1910-1916, Infection & immunity
Natural resistance of humans to the cattle pathogen Trypanosoma brucei brucei has been attributed to the presence in human serum of nonimmune factors that lyse the parasite. Normal human serum contains two trypanosome lytic factors (TLFs). TLF1 is a 500-kDa lipoprotein, which is reported to contain apolipoprotein A-I (apoA-I), haptoglobin-related protein (Hpr), hemoglobin, paraoxonase, and apoA-II, whereas TLF2 is a larger, poorly characterized particle. We report here a new immunoaffinity-based purification procedure for TLF2 and TLF1, as well as further characterization of the components of each purified TLF. Immunoaffinity-purified TLF1 has a specific activity 10-fold higher than that of TLF1 purified by previously described methods. Moreover, we find that TLF1 is a lipoprotein particle that contains mainly apoA-I and Hpr, trace amounts of paraoxonase, apoA-II, and haptoglobin, but no detectable hemoglobin. Characterization of TLF2 reveals that it is a 1,000-kDa protein complex containing mainly immunoglobulin M, apoA-I, and Hpr but less than 1% detectable lipid
— id: 6065, year: 1999, vol: 67, page: 1910, stat: Journal Article,

pH-dependent fibrillogenesis of a VkappaIII Bence Jones protein
Rostagno A; Vidal R; Kaplan B; Chuba J; Kumar A; Elliott JI; Frangione B; Gallo G; Ghiso J
1999 Dec;107(4):835-843, British journal of haematology
Disorders of immunoglobulin (Ig) synthesis that occur in malignant plasma-cell proliferation may result in either granular (LCDD) or fibrillar (AL) tissue deposition of light-chain monoclonal components. The structural features that govern the transition from soluble polypeptides to either fibrillar or granular conformational states remain undefined. Among the many factors presumed to play a role in these transitions the net charge of the molecule has been associated with folding conformation changes. The majority of the proteins involved in AL amyloidosis show acidic isoelectric points (pI 3.8-5.2), whereas most L chains with basic pIs deposit in granular patterns. In our studies a 12 kD VkappaIII fragment was purified as the main component of the fibrils isolated from myocardium and adipose tissue of the pericardium obtained post-mortem from an individual with systemic AL amyloidosis. An apparently identical 12 kD VL fragment with the same N-terminal sequence constituted the BJ protein present in the urine. This urinary protein exhibited strikingly cathodic electrophoretic mobility on agarose gels and lacked retention by anionic exchange chromatography matrices, indicative of a highly basic pI (>10). When it was subjected to in vitro fibril-formation experiments, the BJ protein adopted a fibrillar conformation only at acidic pHs, remaining aggregated but not fibrillar at physiological pH. The data indicate that a specific tissue deposition pattern involves not only structural properties of the protein but rather more complex mechanisms in which acidic micro-environments may contribute to the stabilization of amyloidogenic conformations
— id: 9382, year: 1999, vol: 107, page: 835, stat: Journal Article,

Lipidation of apoE influences isoform specific interaction with Alzheimer's Abeta peptides
Tokuda, T; Calero, M; Matsubara, E; Vidal, R; Ferris, S; Smith, J; Ladu, M; Rostagno, A; Frangione, B; Ghiso, J
1999 Oct 23-28;25(1-2):1348-1348, Abstracts (Society for Neuroscience)
— id: 15846, year: 1999, vol: 25, page: 1348, stat: Journal Article,

A stop-codon mutation in the BRI gene associated with familial British dementia
Vidal R; Frangione B; Rostagno A; Mead S; Revesz T; Plant G; Ghiso J
1999 Jun 24;399(6738):776-781, Nature
Familial British dementia (FBD), previously designated familial cerebral amyloid angiopathy-British type, is an autosomal dominant disorder of undetermined origin characterized by progressive dementia, spasticity, and cerebellar ataxia, with onset at around the fifth decade of life. Cerebral amyloid angiopathy, non-neuritic and perivascular plaques and neurofibrillary tangles are the predominant pathological lesions. Here we report the identification of a unique 4K protein subunit named ABri from isolated amyloid fibrils. This highly insoluble peptide is a fragment of a putative type-II single-spanning transmembrane precursor that is encoded by a novel gene, BRI, located on chromosome 13. A single base substitution at the stop codon of this gene generates a longer open reading frame, resulting in a larger, 277-residue precursor. Release of the 34 carboxy-terminal amino acids from the mutated precursor generates the ABri amyloid subunit. The mutation creates a cutting site for the restriction enzyme XbaI, which is useful for detecting asymptomatic carriers. Antibodies against the amyloid or homologous synthetic peptides recognize both parenchymal and vascular lesions in FBD patients. A point mutation at the stop codon of BRI therefore results in the generation of the ABri peptide, which is deposited as amyloid fibrils causing neuronal disfunction and dementia
— id: 56965, year: 1999, vol: 399, page: 776, stat: Journal Article,

Somatic mutations of the L12a gene in V-kappa(1) light chain deposition disease: potential effects on aberrant protein conformation and deposition
Vidal R; Goni F; Stevens F; Aucouturier P; Kumar A; Frangione B; Ghiso J; Gallo G
1999 Dec;155(6):2009-2017, American journal of pathology
Light chain deposition disease (LCDD) and light chain amyloidosis (AL) are disorders of monoclonal immunoglobulin deposition in which normally soluble serum precursors form insoluble deposits in tissues. A common feature in both is the clonal proliferation of B-cells that produce pathogenic light chains. However, the deposits in LCDD differ from those in AL in that they are ultrastructurally granular rather than fibrillar and do not bind Congo red or colocalize with amyloid P component or apolipoprotein E. The reason(s) for their differences are unknown but are likely multifactorial and related to their protein conformation and their interaction with other molecules and tissue factors in the microenvironment. Knowledge of the primary structure of the light chains in LCDD is very limited. In the present study two new kappa(1) light chains from patients with LCDD are described and compared to seven other reported kappa-LCDD proteins. The N-terminal amino acid sequences of light chain GLA extracted from the renal biopsy and light chain CHO from myocardial tissue were each identical to the respective light chains isolated from the urines and to the V-region amino acid sequences translated from the cloned cDNAs obtained from bone marrow cells. The germline V-region sequences, determined from the genomic DNA in both and in MCM, a previously reported kappa(1) LCDD light chain, were identical and related to the L12a germline gene. The expressed light chains in all three exhibit amino acid substitutions that arise from somatic mutation and result in increased hydrophobicity with the potential for protein destabilization and disordered conformation
— id: 9383, year: 1999, vol: 155, page: 2009, stat: Journal Article,

Biotechnological production of A? peptides
Vidal R; Shao X; Ghiso J; Gorevic P; Frangione B
Alzheimer's disease and related disorders New York : Wiley, 1999,
— id: 5147, year: 1999, vol: , page: 723, stat: Chapter,

Neurovascular interactions of Alzheimer's amyloid ? peptide with apolipoproteins J and E
Zlokovic B; Frangione B; Ghiso J
Clusterin in normal brain functions and during neurodegeneration Austin TX : RG Landes, 1999,
— id: 5149, year: 1999, vol: , page: 71, stat: Chapter,

Antibodies directed to the carboxyl terminus of amyloid beta peptides recognize sequence epitopes and distinct immunoreactive deposits in Alzheimer's disease brain
Jimenez-Huete A; Alfonso P; Soto C; Albar P; Rabano A; Ghiso J; Frangione B; Mendez E
1998 ;1:41-48, Alzheimer's reports
— id: 102367, year: 1998, vol: 1, page: 41, stat: Journal Article,

Human blood-brain barrier receptors for Alzheimer's amyloid-beta 1- 40. Asymmetrical binding, endocytosis, and transcytosis at the apical side of brain microvascular endothelial cell monolayer
Mackic JB; Stins M; McComb JG; Calero M; Ghiso J; Kim KS; Yan SD; Stern D; Schmidt AM; Frangione B; Zlokovic BV
1998 Aug 15;102(4):734-743, Journal of clinical investigation
A soluble monomeric form of Alzheimer's amyloid-beta (1-40) peptide (sAbeta1-40) is present in the circulation and could contribute to neurotoxicity if it crosses the brain capillary endothelium, which comprises the blood-brain barrier (BBB) in vivo. This study characterizes endothelial binding and transcytosis of a synthetic peptide homologous to human sAbeta1-40 using an in vitro model of human BBB. 125I-sAbeta1-40 binding to the brain microvascular endothelial cell monolayer was time dependent, polarized to the apical side, and saturable with high- and low-affinity dissociation constants of 7.8+/-1.2 and 52.8+/-6.2 nM, respectively. Binding of 125I-sAbeta1-40 was inhibited by anti-RAGE (receptor for advanced glycation end products) antibody (63%) and by acetylated low density lipoproteins (33%). Consistent with these data, transfected cultured cells overexpressing RAGE or macrophage scavenger receptor (SR), type A, displayed binding and internalization of 125I-sAbeta1-40. The internalized peptide remains intact > 94%. Transcytosis of 125I-sAbeta1-40 was time and temperature dependent, asymmetrical from the apical to basolateral side, saturable with a Michaelis constant of 45+/-9 nM, and partially sensitive to RAGE blockade (36%) but not to SR blockade. We conclude that RAGE and SR mediate binding of sAbeta1-40 at the apical side of human BBB, and that RAGE is also involved in sAbeta1-40 transcytosis
— id: 7669, year: 1998, vol: 102, page: 734, stat: Journal Article,

Cerebrovascular accumulation and increased blood-brain barrier permeability to circulating Alzheimer's amyloid beta peptide in aged squirrel monkey with cerebral amyloid angiopathy
Mackic JB; Weiss MH; Miao W; Kirkman E; Ghiso J; Calero M; Bading J; Frangione B; Zlokovic BV
1998 Jan;70(1):210-215, Journal of neurochemistry
Senescent squirrel monkey is a valuable model to study pathogenesis of cerebrovascular amyloid angiopathy (CAA). Cerebrovascular sequestration and blood-brain barrier (BBB) permeability to 121I-amyloid beta(1-40) synthetic peptide (sA beta(1-40)) were studied in adult versus aged squirrel monkey 1 h after a single intravenous injection. In aged monkey, the half-time of elimination of sA beta(1-40), t(1/2)e, was prolonged by 0.6 h, the systemic clearance, ClSS, was reduced from 1.8 to 1.1 ml/min/kg, and the mean residence time of intact peptide in the circulation was increased by 1 h (45%). In adult monkey, cerebrovascular sequestration of intact sA beta(1-40) was significant, and the BBB permeability was 18.6-fold higher than for inulin. In aged monkey, the sequestration of intact sA beta(1-40) by cortical and leptomeningeal microvessels and the BBB permeability were increased by 5.9, 1.8-, and 2.1-fold, respectively, in the presence of an unchanged barrier to inulin. In brain parenchyma of aged animals, 76.1% of circulating sA beta(1-40) remained intact versus 45.7% in adult. We conclude that multiple age-related systemic effects, i.e., reduced body elimination and systemic clearance of sA beta(1-40), and reduced peripheral metabolism, may act in concert with BBB mechanisms, i.e., increased transendothelial transport and microvascular accumulation of blood-borne sA beta(1-40), and reduced brain metabolism to enhance the development of CAA
— id: 57200, year: 1998, vol: 70, page: 210, stat: Journal Article,

Inhibition of Alzheimer beta-fibrillogenesis by melatonin
Pappolla M; Bozner P; Soto C; Shao H; Robakis NK; Zagorski M; Frangione B; Ghiso J
1998 Mar 27;273(13):7185-7188, Journal of biological chemistry
It is generally postulated that the amyloid beta protein (Abeta) plays a central role in the progressive neurodegeneration observed in Alzheimer's disease. Important pathologic properties of this protein, such as neurotoxicity and resistance to proteolytic degradation, depend on the ability of Abeta to form beta-sheet structures or amyloid fibrils. We report that melatonin, a hormone recently found to protect neurons against Abeta toxicity, interacts with Abeta1-40 and Abeta1-42 and inhibits the progressive formation of beta-sheets and amyloid fibrils. These interactions between melatonin and the amyloid peptides were demonstrated by circular dichroism and electron microscopy for Abeta1-40 and Abeta1-42 and by nuclear magnetic resonance spectroscopy for Abeta1-40. Inhibition of beta-sheets and fibrils could not be accomplished in control experiments when a free radical scavenger or a melatonin analog were substituted for melatonin under otherwise identical conditions. In sharp contrast with conventional anti-oxidants and available anti-amyloidogenic compounds, melatonin crosses the blood-brain barrier, is relatively devoid of toxicity, and constitutes a potential new therapeutic agent in Alzheimer's disease
— id: 9390, year: 1998, vol: 273, page: 7185, stat: Journal Article,

IgG1-kappa biclonal gammopathy associated with multiple myeloma suggests a regulatory mechanism
Pizzolato M; Bragantini G; Bresciani P; Pavlovsky S; Chuba J; Vidal R; Rostagno A; Ghiso J
1998 Jul;102(2):503-508, British journal of haematology
Multiclonal gammopathies associated with multiple myeloma may result either from a neoplastic transformation of a cell clone undergoing immunoglobulin class switching or from independent transforming events yielding proliferation of unrelated plasma cell clones. The simultaneous presence of more than one neoplastic clone may possess regulatory implications in terms of cell proliferation, clonal expansion, secretion of M-components or response to chemotherapy. We report a patient, diagnosed with multiple myeloma stage IIIa, who presented with two well-defined homogeneous IgG1-kappa components in the serum (designated WER-1 and WER-2) with striking differences in their plasma concentration and response to the classic melphalan/prednisone treatment. Immunochemical characterization and amino terminal sequence analysis of both the heavy and light chains of each M-component undoubtedly determined their biclonal origin. WER-1 was identified as IgG1(VHII)-kappaI while WER-2 was classified as IgG1(VHIII)-kappaIII. The plateau phase was characterized by very low or undetectable levels of WER-2, a high, almost constant, concentration of WER-1 and the absence of Bence Jones proteinuria, whereas these parameters were completely reversed during the escape phase with levels resembling those observed at the time of diagnosis. The statistically significant negative correlation between the biclonal components and the different susceptibility to the treatment clearly suggests regulatory interactions between the clones WER-1 and WER-2
— id: 7746, year: 1998, vol: 102, page: 503, stat: Journal Article,

A transgenic model of human normal sequence transthyretin (TTR) cardiac amyloidosis showing age and gender dependent tissue deposition
Teng, M; Yin, J; Vidal, R; Ghiso, J; Gallo, G; Buxbaum, J
1998 MAR ;46(3):192A-192A, Journal of investigative medicine
— id: 53491, year: 1998, vol: 46, page: 192A, stat: Journal Article,

Gender-dependent amyloid deposition in aging male mice transgenic for human transthyretin (TTR)
Teng, M; Yin, J; Vidal, R; Ghiso, J; Gallo, G; Buxbaum, J
1998 MAR 17 ;12(4):A325-A325, FASEB journal
— id: 53726, year: 1998, vol: 12, page: A325, stat: Journal Article,

Biology of A beta amyloid in Alzheimer's disease (vol 4, pg 313, 1997)
Wisniewski, T; Ghiso, J; Frangione, B
1998 ;5(1):65-65, Neurobiology of disease
— id: 97597, year: 1998, vol: 5, page: 65, stat: Journal Article,

In vitro and in vivo evidence for a rapid clearance of cerebrospinal fluid-borne amyloid beta-peptide into blood and appreciable accumulation by cerebral arteries
Ghersi-Egea, JF; Strazielle, N; Dehouck, MP; Ghiso, J; Gorevic, P; Fenstermacher, JD
1997 DEC ;505P(2):53P-54P, Journal of physiology
— id: 53595, year: 1997, vol: 505P, page: 53P, stat: Journal Article,

Alzheimer's soluble amyloid beta is a normal component of human urine
Ghiso J; Calero M; Matsubara E; Governale S; Chuba J; Beavis R; Wisniewski T; Frangione B
1997 May 12;408(1):105-108, FEBS letters
Soluble A beta (Sa beta) is normally present at a low concentration in human plasma and cerebrospinal fluid. Although the factors involved in the regulation of Sa beta plasma levels are still unknown, we have explored its excretion in the urine as one of the possible homeostatic mechanisms. The presence of Sa beta in the urine was investigated via immunoprecipitation experiments with anti-A beta antibodies followed by detection and identification by immunoblot, MALDI mass spectrometry and sequence analysis. Soluble A beta (4.3 kDa) immunoreactivity was present in the urine of normal donors, Down's syndrome individuals as well as in patients with renal disorders exhibiting glomerular or mixed proteinuria. Edman degradation of the immunoprecipitated material yielded the intact A beta N-terminus and mass spectra analysis indicated the existence of a major component at mlz 4327, corresponding to the molecular mass of A beta1-40. Semiquantitative data obtained from the immunoprecipitation experiments indicate that under normal conditions the daily excretion of intact Sa beta in the urine represents less than 1% of the circulating pool
— id: 7153, year: 1997, vol: 408, page: 105, stat: Journal Article,

In vivo blood-brain barrier uptake of Alzheimer's A? peptides in guinea pigs : evidence for vascular sequestration of A?1-40 (but not A?1-42)
Ghiso J; Martel C; Mackic J; McComb JG; Frangione B; Zlokovic B
Alzheimer's disease : biology, diagnosis, and therapeutics New York : Wiley, 1997,
— id: 5146, year: 1997, vol: , page: 375, stat: Chapter,

Amino-terminal identity of co-existent amyloid and non-amyloid immunoglobulin kappa light chain deposits. A human disease to study alterations of protein conformation
Kaplan B; Vidal R; Kumar A; Ghiso J; Frangione B; Gallo G
1997 Dec;110(3):472-478, Clinical & experimental immunology
Tissue deposition of monoclonal immunoglobulin light chains is a serious complication in some patients with B cell proliferative disorders. The deposits are typically fibrillar and Congophilic in amyloid (AL) and non-fibrillar and Congophobic in light chain deposition disease (LCDD), and rarely coexist in the same patient. From post-mortem tissue of an individual with fibrillar and non-fibrillar kappa light chain deposits in different sites, we separately extracted and analysed biochemically and immunochemically the non-amyloid deposits from isolated glomeruli, the amyloid from isolated renal arteries and the amyloid from myocardium in which the only deposits were amyloid restricted to mural arteries. Western blotting analysis of both the extracted amyloid and the non-amyloid deposits demonstrated 25-kD bands immunoreactive with anti-kappa antibody, and the identity of the N-terminal amino acid sequences that belong to the variable region kappaIV light chain subgroup. This is the first human disease in which antigenically similar but morphologically different deposits have been separately biochemically analysed. We propose that combined LCDD and AL is an ideal human disease to study the relationships and the factors that influence the conversion of non-amyloidogenic to amyloidogenic conformations
— id: 7942, year: 1997, vol: 110, page: 472, stat: Journal Article,

Isoform-specific effects of apolipoproteins E2, E3, and E4 on cerebral capillary sequestration and blood-brain barrier transport of circulating Alzheimer's amyloid beta
Martel CL; Mackic JB; Matsubara E; Governale S; Miguel C; Miao W; McComb JG; Frangione B; Ghiso J; Zlokovic BV
1997 Nov;69(5):1995-2004, Journal of neurochemistry
Cerebral capillary sequestration and blood-brain barrier (BBB) permeability to apolipoproteins E2 (apoE2), E3 (apoE3), and E4 (apoE4) and to their complexes with sA beta(1-40), a peptide homologous to the major form of soluble Alzheimer's amyloid beta, were studied in perfused guinea pig brain. Cerebrovascular uptake of three apoE isoforms was low, their blood-to-brain transport undetectable, but uptake by the choroid plexus significant. Binding of all three isoforms to sA beta(1-40) in vitro was similar with a K(D) between 11.8 and 12.9 nM. Transport into brain parenchyma and sequestration by BBB and choroid plexus were negligible for sA beta(1-40)-apoE2 and sA beta(1-40)-apoE3, but significant for sA beta(1-40)-apoE4. After 10 min, 85% of sA beta(1-40)-apoE4 taken up at the BBB remained as intact complex, whereas free sA beta(1-40) was 51% degraded. Circulating apoE isoforms have contrasting effects on cerebral capillary uptake of and BBB permeability of sA beta. ApoE2 and apoE3 completely prevent cerebral capillary sequestration and blood-to-brain transport of sA beta(1-40). Conversely, apoE4, by entering brain microvessels and parenchyma as a stable complex with sA beta, reduces peptide degradation and may predispose to cerebrovascular and possibly enhance parenchymal amyloid formation under pathological conditions
— id: 9391, year: 1997, vol: 69, page: 1995, stat: Journal Article,

Transport of apolipoproteins E and J at the blood-brain barrier - Relevance to Alzheimer's disease
Martel, CL; Ghiso, J; Frangione, B; Zlokovic, BV
1997 JAN-FEB ;7(1):28-36, S.T.P. pharma pratiques : techniques reglementations
It has been shown that the blood-brain barrier regulates uptake of apolipoprotein E and apolipoprotein J, and of their complexes with soluble amyloid beta(1-40), a synthetic peptide homologous to the major form of soluble Alzheimer's amyloid beta peptide. Apolipoprotein E isoforms E3 and E4 do not enter. brain parenchyma, and exhibit minimal sequestration by cerebral capillaries. There is negligible brain uptake and sequestration of the soluble amyloid beta(1-40)-apolipoprotein E3 complex, but there is moderate uptake of intact soluble amyloid beta(1-40)-apolipoprotein E4 which is mediated by an as yet unknown mechanism. Both apolipoprotein J and soluble amyloid beta(1-40)-apolipoprotein J exhibit a remarkable cerebrovascular permeability which appears to be mediated by the glycoprotein 330 (megalin) receptor. It is therefore suggested that blood-brain barrier transport of apolipoproteins complexed with amyloid beta may play a role in either preventing (in the case of apolipoproteins J and E3) or enhancing (in the case of apolipoprotein E4) the formation of amyloid deposits in cerebral microvessels and brain parenchyma under certain pathological conditions
— id: 53196, year: 1997, vol: 7, page: 28, stat: Journal Article,

Inhibition of Alzheimer's ?-amyloid fibril formation
Pappolla M; Bozner P; Soto C; Zagorski M; Shao H; Frangione B; Ghiso J
1997 ;1:?-?, Emerging therapeutic targets
— id: 102368, year: 1997, vol: 1, page: ?, stat: Journal Article,

Biology of A beta amyloid in Alzheimer's disease
Wisniewski T; Ghiso J; Frangione B
1997 ;4(5):313-328, Neurobiology of disease
The genetic associations with the pathological features of AD are diverse: A rapidly growing number of mutations in presenilin 1 and 2 on chromosomes 14 and 1, respectively, are found in many early-onset FAD patients (Lendon et al., 1997). In addition, beta PP mutations are found in a small percentage of early-onset FAD kindreds. The apoE4 allele on chromosome 19 is associated with the presence of the most common form of AD, sporadic AD (Wisniewski & Frangione, 1992; Namba et al., 1991). However, it is clear that other proteins are also involved in the pathogenesis of AD, since some early-onset FAD kindreds do not have linkage to PS1, PS2, apoE, or beta PP, while at least 50% of late-onset AD is unrelated to apoE. Other proteins which have been implicated in the formation of senile plaques, but so far are not known to have any genetic linkage to AD, include proteoglycans (Snow et al., 1987), apoA1 (Wisniewski et al., 1995a), alpha 1-antichymotrypsin (Abraham et al., 1988), HB-GAM (Wisniewski et al., 1996a), complement components (McGeer & Rogers, 1992), acetylcholinesterase (Friede, 1965), and NAC (Ueda et al., 1993). Which of these proteins will be the most important for the etiology of the most common form of AD, late-onset sporadic AD, remains an open question. Three of the genes which are now known to be linked to AD, including PS1, beta PP, and apoE, have been established immunohistochemically and biochemically to be components of senile plaques (see Fig. 1). This raises at least two possibilities: either each of these proteins is part of one pathway with A beta-related amyloid formation as a final causative pathogenic event or amyloid deposition in AD is a reactive process related to dysfunction of a number of different CNS proteins. Whether or not amyloid formation is directly causative in the pathogenesis of AD, current data suggest that new therapeutic approaches which may inhibit the aggregation and/or the conformational change of sA beta to A beta fibrils (Soto et al., 1996) have the greatest likelihood to make a significant impact on controlling amyloid accumulation in AD
— id: 7854, year: 1997, vol: 4, page: 313, stat: Journal Article,

C-terminal fragments of alpha- and beta-tubulin form amyloid fibrils in vitro and associate with amyloid deposits of familial cerebral amyloid angiopathy, British type
Baumann MH; Wisniewski T; Levy E; Plant GT; Ghiso J
1996 Feb 6;219(1):238-242, Biochemical & biophysical research communications
Familial amyloidosis, British type, is an autosomal dominant disease characterized by progressive dementia, spastic paralysis and ataxia. The identity of the accumulating amyloid is not known, thus preventing the definitive classification of the disease. Biochemical methods were used to characterize the nature of the amyloid deposits from the brain tissue of one individual who died with this disease. The purified tissue material was subjected to trypsin digestion and subsequent N-terminal sequence analysis. Major tryptic fragments yielded the sequences VGINYQPPTVVPGGDLAK, FDLMYAK, GLTVPEL and GYLTVAAVFR, which are all tryptic fragments of the C-termini of human tubulin subunits alpha and beta. Synthetic peptides based on the sequences of these fragments formed amyloid fibrils in vitro fitting the characteristic definition of amyloid. These findings suggest that the C-terminal fragments of both alpha- and beta-tubulin are closely associated to the amyloid deposits of familial amyloidosis, British type
— id: 6886, year: 1996, vol: 219, page: 238, stat: Journal Article,

Clearance of Alzheimer's soluble beta-peptide from ventricular cerebrospinal fluid into blood
Fenstermacher, J; Strazielle, N; GhersiEgea, JF; Ghiso, J; Dehouck, MP; Patlak, C; Frangione, B; Gorevic, P
1996 MAR 8 ;10(3):3912-3912, FASEB journal
— id: 98397, year: 1996, vol: 10, page: 3912, stat: Journal Article,

Chaperoning amyloid in Alzheimer's disease: the art of avoiding sticky situations?
Frangione B; Castano EM; Prelli F; Soto C; Ghiso J; Wisniewski T
Apolipoprotein E and Alzheimer's disease Berlin : Springer, 1996,
— id: 4976, year: 1996, vol: , page: 151, stat: Chapter,

Apolipoprotein E and amyloidogenesis
Frangione B; Castano EM; Wisniewski T; Ghiso J; Prelli F; Vidal R
1996 ;199:132-141, CIBA Foundation symposium
Alzheimer's amyloid beta-protein (A beta) is a modified, pathogenic form of a constitutive host protein, soluble amyloid beta-protein (sA beta). Both are conformational isomers encoded by the gene for the beta-amyloid precursor protein (APP), located on chromosome 21. sA beta and A beta have identical sequence but are thought to differ in their secondary structure and physicochemical properties, hence they are conformational isomers. sA beta is easily degraded, while A beta is particularly resistant. A beta has a high beta-pleated sheet content, while sA beta is thought to be more random-coil and/or alpha-helical. A beta, unlike sA beta, adopts an amyloidogenic conformation, forms aggregates and gives rise to fibrils. Most early-onset forms of Alzheimer's disease (AD) have been linked to mutations of the presenilin 1, presenilin 2 or APP genes, located on chromosomes 14, 1 and 21, respectively. Their relationship to amyloidogenesis is being investigated. On the other hand, the major risk factor for the most common form, sporadic and familial late-onset AD, is the presence of the apoE epsilon 4 allele. Recent studies have shown that a 10 kDa C-terminal fragment of apoE is complexed to A beta in neuritic plaques and that apoE isoforms can modulate amyloid formation in vitro. Moreover, thrombin cleavage of apoE generates a similar C-terminal fragment that can form amyloid-like fibrils. Thus neuritic plaques may contain both A beta and apoE amyloid fibrils. AD can be neuropathologically defined by the presence of several interacting proteins that can adopt an amyloidogenic conformation. This has led us to hypothesize that in AD, amyloidosis may be reactive rather than causative
— id: 9396, year: 1996, vol: 199, page: 132, stat: Journal Article,

Light chain cardiomyopathy. Structural analysis of the light chain tissue deposits [see comments]
Gallo G; Goni F; Boctor F; Vidal R; Kumar A; Stevens FJ; Frangione B; Ghiso J
1996 May;148(5):1397-1406, American journal of pathology
Cardiomyopathy due to monoclonal light chain deposits is a complication of plasma cell disorders. The deposits may be either fibrillar as in light chain amyloid or nonfibrillar as in light chain deposition disease. The reasons for these structural differences are still unknown. We characterized the myocardial deposits by immunohistochemical examination of sections and extraction and biochemical analysis of the tissue deposits in a patient (MCM) who died of myeloma and systemic light chain deposition disease. Amino acid sequence analysis of the extracted nonfibrillar MCM kappa-light chain reveals that it belongs to the L12a germline subset of the kappa(I) protein and contains five distinctive amino acid substitutions (three in the framework region III and two in the complementarity-determining region III) that have not been reported previously in the same positions in other kappa(I) light chains. The theoretically determined isoelectric point (pI 8.21) of the MCM light chain is high compared with the low isoelectric point of other Bence Jones proteins from subjects without light chain deposition disease. The diffuse binding to basement membranes and the high isoelectric point of the MCM kappa-light chain suggest electrostatic interaction as a possible mechanism of tissue deposition. The spatial locations of the five distinctive residues and a sixth rare substitution of the MCM protein modeled on the backbone structure of REI, a kappa(I)-soluble Bence Jones light chain of known three-dimensional structure, may be responsible for protein destabilization, partial unfolding, and aggregation leading to tissue deposition
— id: 9393, year: 1996, vol: 148, page: 1397, stat: Journal Article,

Fate of cerebrospinal fluid-borne amyloid beta-peptide: rapid clearance into blood and appreciable accumulation by cerebral arteries
Ghersi-Egea JF; Gorevic PD; Ghiso J; Frangione B; Patlak CS; Fenstermacher JD
1996 Aug;67(2):880-883, Journal of neurochemistry
In Alzheimer's disease, the neuritic or senile amyloid plaques in hippocampus and association cortex, the diffuse plaques in brain areas such as the cerebellum and sensorimotor cortex, and the amyloid deposits in the walls of pial and parenchymal blood vessels are mainly composed of amyloid beta-peptides. In the present study, either soluble 40-residue amyloid beta-peptide radiolabeled with 125I (I-sAbeta) or [14C]polyethylene glycol ([14C]PEG, a reference material) was briefly infused into one lateral ventricle of normal rats. By 3.5 min, 30% of the I-sAbeta was cleared from ventricular CSF into blood; another 30% was removed over the next 6.5 min. No [14C]PEG was lost from the CSF-brain system during the first 5 min, and only 20% was cleared by 10 min. Much of the I-sAbeta that reached the subarachnoid space was retained by pial arteries and arterioles. Virtually no I-sAbeta was found in brain. The clearance of amyloid beta-peptides from the CSF-brain system, reported herein for normal rats, may be reduced in Alzheimer's disease, thus contributing to amyloid deposition in cerebral tissue and blood vessels
— id: 9392, year: 1996, vol: 67, page: 880, stat: Journal Article,

beta PP and Tau interaction. A possible link between amyloid and neurofibrillary tangles in Alzheimer's disease
Giaccone G; Pedrotti B; Migheli A; Verga L; Perez J; Racagni G; Smith MA; Perry G; De Gioia L; Selvaggini C; Salmona M; Ghiso J; Frangione B; Islam K; Bugiani O; Tagliavini F
1996 Jan;148(1):79-87, American journal of pathology
Extracellular deposition of amyloid fibrils and intraneuronal accumulation of paired helical filaments (PHFs) are the neuropathological hallmarks of Alzheimer's disease. The major constituent of amyloid fibrils is a 39- to 43-residue peptide (termed A beta), which is derived from a 695- to 770-amino-acid precursor protein (termed beta PP). The main component of PHFs identified so far is the microtubule-associated protein tau. Yet, there is no direct evidence of interconnection between these two pathological states. We report here that antibodies to an epitope located between residues 713 and 723 of beta PP770 (ie, the transmembrane region of beta PP distal to A beta) consistently labeled PHFs in the brain of Alzheimer patients. Solid phase immunoassay showed that a peptide homologous to residues 713 to 730 of beta PP770 bound tau proteins. This beta PP peptide spontaneously formed fibrils in vitro and, in the presence of tau, generated dense fibrillary assemblies containing both molecules. These data suggest that beta PP or beta PP fragments containing the tau binding site are involved in the pathogenesis of PHFs in Alzheimer's disease
— id: 9397, year: 1996, vol: 148, page: 79, stat: Journal Article,

Proteasome activity is required for the stage-specific transformation of a protozoan parasite
Gonzalez J; Ramalho-Pinto FJ; Frevert U; Ghiso J; Tomlinson S; Scharfstein J; Corey EJ; Nussenzweig V
1996 Nov 1;184(5):1909-1918, Journal of experimental medicine
A prominent feature of the life cycle of intracellular parasites is the profound morphological changes they undergo during development in the vertebrate and invertebrate hosts. In eukaryotic cells, most cytoplasmic proteins are degraded in proteasomes. Here, we show that the transformation in axenic medium of trypomastigotes of Trypanosoma cruzi into amastigote-like organisms, and the intracellular development of the parasite from amastigotes into trypomastigotes, are prevented by lactacystin, or by a peptide aldehyde that inhibits proteasome function. Clasto-lactacystin, an inactive analogue of lactacystin, and cell-permeant peptide aldehyde inhibitors of T. cruzi cysteine proteinases have no effect. We have also identified the 20S proteasomes from T. cruzi as a target of lactacystin in vivo. Our results document the essential role of proteasomes in the stage-specific transformation of a protozoan
— id: 8375, year: 1996, vol: 184, page: 1909, stat: Journal Article,

Biochemical characterization of Alzheimer's soluble amyloid beta protein in human cerebrospinal fluid: association with high density lipoproteins
Koudinov AR; Koudinova NV; Kumar A; Beavis RC; Ghiso J
1996 Jun 25;223(3):592-597, Biochemical & biophysical research communications
The soluble form of Alzheimer's amyloid beta protein (sA beta) is associated with high density lipoproteins (HDL) in normal human plasma (BBRC, 1994, 205, 1164-1171). Since sA beta is also present in cerebrospinal fluid (CSF) and the lipoprotein pattern of CSF is different from that of plasma, it was of interest to ascertain whether the interaction of sA beta with HDL also occurs in CSF. Normal human CSF lipoproteins were obtained by sequential flotation ultracentrifugation and analyzed for the presence of sA beta via immunoblot, size-exclusion chromatography, immunoelectron microscopy, N-terminal sequence and mass-spectrometry analyses. Soluble A beta was associated with CSF-HDL particles of 16.8 +/- 3.2 nm in diameter and approximately 200 kDa of relative molecular mass. A approximately 4.3 kDa component purified by HPLC was immunoreactive with anti-A beta antibodies and exhibited an N-terminal sequence identical to the A beta peptide with a mass of 4325.1 Da, indicating that the main sA beta specie associated with CSF-HDL is A beta 1-40
— id: 7011, year: 1996, vol: 223, page: 592, stat: Journal Article,

Blood-brain barrier uptake of the 40 and 42 amino acid sequences of circulating Alzheimer's amyloid beta in guinea pigs
Martel CL; Mackic JB; McComb JG; Ghiso J; Zlokovic BV
1996 Mar 15;206(2-3):157-160, Neuroscience letters
An intracarotid brain infusion/capillary depletion technique was used in guinea pigs to examine cerebral capillary sequestration and transport into brain parenchyma of sA beta 1-40 and sA beta 1-42, synthetic peptides identical to two forms of the amyloid beta peptide found in Alzheimer's disease lesions: the 40 residue form, found primarily in vascular deposits, and the 42 residue form, found primarily in senile plaques. The peptides crossed well into the brain parenchyma via a specific transport mechanism for which sA beta 1-40 had an approximately two-fold greater affinity than sA beta 1-42. There was significant capillary sequestration of sA beta 1-40, but retention by the microvasculature of sA beta 1-42 was negligible. These data suggest that the level of the 40 residue peptide in cerebral vasculature and of the 42 residue peptide in parenchyma could be regulated by blood-brain barrier sequestration and transport of their respective circulating precursors
— id: 9395, year: 1996, vol: 206, page: 157, stat: Journal Article,

Apolipoprotein J and Alzheimer's amyloid beta solubility
Matsubara E; Soto C; Governale S; Frangione B; Ghiso J
1996 Jun 1;316 ( Pt 2):671-679, Biochemical journal
Apolipoprotein J (apoJ) has been found associated with soluble amyloid beta (sA beta) in plasma and cerebrospinal fluid in normal individuals and co-deposited with fibrillar A beta in Alzheimer's cerebrovascular and parenchymal lesions. Although studies in vitro and in vivo indicate that apoJ is a major carrier protein for sA beta, its role in the fibrillogenesis process is not known. We report herein that apoJ in its native high-density lipoprotein lipidic environment is fully active to interact with A beta peptides. Furthermore, apoJ prevents aggregation and polymerization of synthetic A beta in vitro. The interaction was stable for at least 14 days at 37 degrees C in physiologic buffers, and the peptide retrieved after complex dissociation at low pH retained its inherent aggregation properties. In addition, the binding to apoJ protects synthetic A beta from proteolytic degradation; both A beta 1-42 and A beta 1-40 were more resistant to proteolysis by trypsin and chymotrypsin when complexed to apoJ. The data suggest that the interaction may preclude sA beta aggregation in biological fluids and point to a protecting role of apoJ for complexed A beta species
— id: 57410, year: 1996, vol: 316 ( Pt 2), page: 671, stat: Journal Article,

Alzheimer's soluble amyloid beta is a normal component of urine
Matsubara, E.; Governale, S.; Calero, M.; Wisniewski, T.; Frangione, B.; Ghiso, J.
1996 ;22(1-3):1170-33631, Abstracts (Society for Neuroscience)
— id: 97642, year: 1996, vol: 22, page: 1170, stat: Journal Article,

Fibrillary glomerulonephritis related to serum fibrillar immunoglobulin-fibronectin complexes
Rostagno A; Vidal R; Kumar A; Chuba J; Niederman G; Gold L; Frangione B; Ghiso J; Gallo G
1996 Nov;28(5):676-684, American journal of kidney diseases
Fibrillary glomerulonephritis is a disease of uncertain origin and pathogenesis characterized by nonamyloidotic fibrils in glomeruli. We report immunohistological, immunochemical, and biochemical studies of a serum fibrillar cryoprecipitate obtained from a patient with fibrillary glomerulonephritis, that formed on prolonged storage at 4 degrees C. By Western blot and amino acid sequence analysis, the cryoprecipitated fibril components consisted of immunoglobulins, heavy chains gamma and mu, light chains kappa and lambda, and fibronectin, similar to the proteins identified by immunofluorescence and immunoelectron microscopy in the glomerular fibrils. These findings support the hypothesis that serum precursors may be the source of the fibrillar deposits and suggest a role for immunoglobulin-fibronectin complexes in the pathogenesis of fibrillary glomerulonephritis
— id: 7254, year: 1996, vol: 28, page: 676, stat: Journal Article,

Alzheimer's presenilin 1 gene expression in platelets and megakaryocytes. Identification of a novel splice variant
Vidal R; Ghiso J; Wisniewski T; Frangione B
1996 Sep 9;393(1):19-23, FEBS letters
The presenilin 1 (PS1) gene located on chromosome 14 has been linked with the majority of early-onset FAD. The normal biological role of PS1 as well as the mechanism by which mutations in PS1 cause FAD remains unknown. PS1 expression in platelets and the Dami megakaryocytic cell line was examined by Western blot analysis and RT-PCR. Using an anti-N-terminus PS1 antibody we detected PS1 immunoreactive bands of 44, 32 and 27 kDa in both cell types. After RT-PCR we observed that platelets and megakaryocytes carry at least four different PS1 transcripts. One of them is a novel PS1 splice variant that lacks the coding sequence for exon 10 resulting in a shorter 409 amino acid protein
— id: 7919, year: 1996, vol: 393, page: 19, stat: Journal Article,

Glycoprotein 330/megalin: probable role in receptor-mediated transport of apolipoprotein J alone and in a complex with Alzheimer disease amyloid beta at the blood-brain and blood-cerebrospinal fluid barriers
Zlokovic BV; Martel CL; Matsubara E; McComb JG; Zheng G; McCluskey RT; Frangione B; Ghiso J
1996 Apr 30;93(9):4229-4234, Proceedings of the National Academy of Sciences of the United States of America
A soluble form of Alzheimer disease amyloid beta-protein (sA beta) is transported in the blood and cerebrospinal fluid mainly complexed with apolipoprotein J (apoJ). Using a well-characterized in situ perfused guinea pig brain model, we recently obtained preliminary evidence that apoJ facilitates transport of sA beta (1-40)-apoJ complexes across the blood-brain barrier and the blood-cerebrospinal fluid barrier, but the mechanisms remain poorly understood. In the present study, we examined the transport process in greater detail and investigated the possible role of glycoprotein 330 (gp330)/megalin, a receptor for multiple ligands, including apoJ. High-affinity transport systems with a Km of 0.2 and 0.5 nM were demonstrated for apoJ at the blood-brain barrier and the choroid epithelium in vivo, suggesting a specific receptor-mediated mechanism. The sA beta (1-40)-apoJ complex shared the same transport mechanism and exhibited 2.4- to 10.2-fold higher affinity than apoJ itself. Binding to microvessels, transport into brain parenchyma, and choroidal uptake of both apoJ and sA beta (1-40)-apoJ complexes were markedly inhibited (74-99%) in the presence of a monoclonal antibody to gp330/megalin and were virtually abolished by perfusion with the receptor-associated protein, which blocks binding of all known ligands to gp330. Western blot analysis of cerebral microvessels with the monoclonal antibody to gp330 revealed a protein with a mass identical to that in extracts of kidney membranes enriched with gp330/megalin, but in much lower concentration. The findings suggest that gp330/megalin mediates cellular uptake and transport of apoJ and sA beta (1-40)-apoJ complex at the cerebral vascular endothelium and choroid epithelium
— id: 9394, year: 1996, vol: 93, page: 4229, stat: Journal Article,

Amyloids, genes and chaperones
Frangione B; Wisniewski T; Castano E; Ghiso J
Research advances in Alzheimer's disease and related disorders New York : Wiley, 1995,
— id: 4974, year: 1995, vol: , page: 563, stat: Chapter,

Familial cerebral amyloid angiopathy (British type) with nonneuritic amyloid plaque formation may be due to a novel amyloid protein
Ghiso J; Plant GT; Revesz T; Wisniewski T; Frangione B
1995 Mar;129(1):74-75, Journal of the neurological sciences
— id: 7904, year: 1995, vol: 129, page: 74, stat: Journal Article,

Amyloidosis
Ghiso, Jorge; Vidal, Ruben; Gallo, Gloria; Fragione, Blas
1995 ;35(2):93-102, Revista brasileira de reumatologia
Amiloidose e um termo generico que descreve amplo espectro de doencas caracterizadas pela deposicao de proteinas fibrilares insoluveis em diversos orgaos. As fibrilas depositadas compoem-se predominantemente de proteinas de baixo peso molecular que sao normalmente soluveis sob condicoes fisiologicas. Todos os tipos de amiloide compartilham propriedades fisicas, estruturais e tintoriais comuns: conformacao em placa beta, alto grau de insolubilidade e aparencia fibrilar a microscopia eletronica...
— id: 90048, year: 1995, vol: 35, page: 93, stat: Journal Article,

Characterization of apolipoprotein J-Alzheimer's A beta interaction
Matsubara E; Frangione B; Ghiso J
1995 Mar 31;270(13):7563-7567, Journal of biological chemistry
The main component of Alzheimer's amyloid deposits, A beta, has been found also as a soluble (sA beta) normal constituent of biological fluids and cell culture supernatants. Whether or not sA beta is the immediate precursor of A beta, it is clear that peptides with the same amino acid sequence can have both fibrillar and non-fibrillar conformations. The interconversion mechanism from one form to another is presently under intensive investigation. We have previously described that (i) a synthetic peptide A beta 1-40 immobilized on affinity matrices was able to retrieve apolipoprotein J (apoJ) from plasma and cerebrospinal fluid; and (ii) the interaction of sA beta with apoJ occurs in vivo, as demonstrated by the ability of anti-apoJ to co-precipitate sA beta from normal cerebrospinal fluid. We have characterized the binding between A beta 1-40 and apoJ and found that the interaction is saturable, specific, and reversible. The dissociation constant of 2 x 10(-9) M is indicative of high affinity binding. The stoichiometry of the reaction is 1:1; apoJ has five times more affinity for fresh A beta 1-40 than for the aggregated peptide. Competitive inhibition studies carried out with apolipoprotein E (isoforms E2, E3, and E4), transthyretin, vitronectin, and alpha 1-antichymotrypsin indicate that the complex apoJ.A beta 1-40 cannot be dissociated by any of these competitors at physiologic concentrations. The data strongly suggest that apoJ plays an important role as a carrier protein for sA beta
— id: 8065, year: 1995, vol: 270, page: 7563, stat: Journal Article,

Tau protein directly interacts with the amyloid beta-protein precursor: implications for Alzheimer's disease
Smith MA; Siedlak SL; Richey PL; Mulvihill P; Ghiso J; Frangione B; Tagliavini F; Giaccone G; Bugiani O; Praprotnik D; et al
1995 Apr;1(4):365-369, Nature medicine
The simultaneous presence of intracellular neurofibrillary tangles (NFT) and extracellular senile plaques in Alzheimer's disease (AD) suggests that the two lesions could be synergistically interrelated. However, although the main protein components of NFT and senile plaques, tau (tau) and amyloid beta-protein, respectively, are well characterized, the molecular mechanisms responsible for their deposition in lesions are unknown. We demonstrate, using four independent techniques, that tau directly interacts with a conformation-dependent domain of the amyloid beta-protein precursor (beta PP) encompassing residues beta PP714-723. The putative tau-binding domain includes beta PP717 mutation sites that are associated with familial forms of AD. Our findings strongly suggest that NFT and senile plaques, often thought of as independent structures, may play a role in each other's formation during the pathogenesis of AD
— id: 9398, year: 1995, vol: 1, page: 365, stat: Journal Article,

High-density-lipoprotein-independent killing of Trypanosoma brucei by human serum
Tomlinson S; Jansen AM; Koudinov A; Ghiso JA; Choi-Miura NH; Rifkin MR; Ohtaki S; Nussenzweig V
1995 Mar;70(1-2):131-138, Molecular & biochemical parasitology
The cattle pathogen Trypanosoma brucei brucei is morphologically indistinguishable from the human pathogens T.b. rhodesiense and T.b. gambiense. However, unlike the human pathogens, T.b. brucei is lysed by normal human serum (NHS). The trypanolytic factor in NHS co-purifies with high-density lipoproteins (HDL), but its precise nature is unknown. Using a new fluorescence-based viability assay to assess T.b. brucei killing, we find that the HDL-deficient sera from two patients with Tangier disease are as trypanolytic as NHS. Fractionation of the Tangier sera by density ultracentrifugation revealed that the activity resides only in lipoprotein-depleted fractions. Tangier and NHS were also subjected to molecular sieving chromatography, and the activity profiles were identical. Lytic fractions to T. brucei (but not to T. rhodesiense) appeared under two distinct peaks of 100-600 kDa and > 1000 kDa. Neither peak coincided with the position of the major serum lipoproteins, as determined by cholesterol titrations. The high-molecular-mass peak did not contain the HDL-associated apolipoprotein-A1. Further, we did not find that purified apolipoproteins A1 or J are lytic for the trypanosomes. We conclude that the killing of T. brucei by human serum can be independent of HDL
— id: 56721, year: 1995, vol: 70, page: 131, stat: Journal Article,

S182 protein in Alzheimer's disease neuritic plaques
Wisniewski T; Palha JA; Ghiso J; Frangione B
1995 Nov 18;346(8986):1366-1366, Lancet
— id: 7909, year: 1995, vol: 346, page: 1366, stat: Journal Article,

The blood brain barrier regulates transport of Alzheimer's amyloid ? and apolipoproteins E and J
Zlokovic B; Mackic J; Martel C; Wisniewski T; Frangione B; Ghiso J
Research advances in Alzheimer's disease and related disorders New York : Wiley, 1995,
— id: 4975, year: 1995, vol: , page: 585, stat: Chapter,

CHARACTERIZATION BY RADIOSEQUENCING OF THE CARBOXY-TERMINAL DERIVATIVES PRODUCED FROM NORMAL AND MUTANT AMYLOID-BETA PROTEIN PRECURSORS
CHEUNG, TT; GHISO, J; SHOJI, M; CAI, XD; GOLDE, T; GANDY, S; FRANGIONE, B; YOUNKIN, S
1994 JUL ;15(7):S54-S54, Neurobiology of aging
— id: 52404, year: 1994, vol: 15, page: S54, stat: Journal Article,

CHARACTERIZATION BY RADIOSEQUENCING OF THE CARBOXYL-TERMINAL DERIVATIVES PRODUCED FROM NORMAL AND MUTANT AMYLOID-BETA PROTEIN PRECURSORS
CHEUNG, TT; GHISO, J; SHOJI, M; CAI, XD; GOLDE, T; GANDY, S; FRANGIONE, B; YOUNKIN, S
1994 MAR ;1(1):30-38, Amyloid
The 39-43 amino acid (similar to 4 kD) amyloid beta-protein (A beta) deposited as amyloid Alzheimer's disease is an infernal peptide beginning 99 residues from the COOH end of a much larger amyloid beta-protein precursor (beta APP). In cultured cells, normal processing of the beta APP produces similar to 8.7, similar to 9.6, similar to 10.9, and similar to 11.4 kD COOH-terminal derivatives that appear to contain all or part of the A beta domain. In this study, we metabolically labeled transfected human neuroblastoma (M17) cells with [H-3]phenylalanine plus [S-35]methionine and then radiosequenced the immunoprecipitated COOH-terminal beta APP derivatives faking advantage of the fact that the A beta has phenylalanines at positions 4, 19, and 20, and a single methionine at position 35. Our analysis of the derivatives produced by transfected M17 cells expressing beta APP(695) confirms that the similar to 8.7 kD COOH-terminal derivative begins at A beta(17) and indicates that the similar to 9.6 and similar to 10.9 kD derivatives begin at A beta(10) and A beta(4) respectively. Significantly we find that the 11.4 kD derivative begins at A beta(1). Thus normal beta APP processing produces a potentially amyloidogenic COOH-terminal derivative that has the AP domain intact at its amino terminus. We have previously shown that cells expressing beta APP(Delta NL) a mutant linked to familial Alzheimer's disease, produce an increased amount of the 11.4 kD COOH-terminal derivative and secrete more A beta. Radiosequencing of these derivatives showed that the Delta NL mutant is cleaved at the same location as wild type beta APP producing an 11.4 kD COOH-terminal derivative and A beta that both begin at A beta(1). Thus the Delta NL mutation appears to accelerate a cleavage that releases an 11.4 kD COOH-terminal derivative identical to that normally produced from wild type beta APP, and it appears that this 11.4 kD derivative is further processed to release excess A beta
— id: 87453, year: 1994, vol: 1, page: 30, stat: Journal Article,

Chaperoning Alzheimer's amyloids
Frangione B; Wisniewski T; Ghiso J
1994 ;15 Suppl 2:S97-S99, Neurobiology of aging
— id: 6619, year: 1994, vol: 15 Suppl 2, page: S97, stat: Journal Article,

Potential role of apolipoprotein-E in fibrillogenesis
Gallo G; Wisniewski T; Choi-Miura NH; Ghiso J; Frangione B
1994 Sep;145(3):526-530, American journal of pathology
Immunohistochemical and biochemical studies have demonstrated several different proteins in amyloid deposits that are not intrinsic components of the fibril itself but may play a role in their deposition and fibril formation. We compared the distribution of several amyloid-associated proteins, ie, amyloid P component, apolipoprotein-E, apolipoprotein-J, and vitronectin, in the deposits of several different amyloids, in particular light chain amyloid, with those in the deposits of nonamyloid monoclonal immunoglobulin, which may be considered a form of preamyloid disease. Although 100% of amyloid specimens (7 amyloid A, 15 immunoglobulin light chain, and 1 transthyretin) had amyloid P component and 100% had apolipoprotein-E (2 amyloid A, 10 immunoglobulin light chain, and 1 transthyretin) co-localized with the primary amyloid protein, none of the monoclonal nonamyloid cases (14 light chain deposition disease and 6 light and heavy chain deposition disease) had amyloid P component and only 1 of 11 had apolipoprotein-E. On the other hand, staining for apolipoprotein-J and vitronectin was positive in 100% of cases of amyloid and nonamyloid monoclonal deposits. The association between the presence of apolipoprotein-E and amyloid P component in the fibrillar form of monoclonal light chain deposits and their absence in the nonfibrillar form of deposits suggest a role for these proteins in the process of fibrillogenesis. This lends support for the previously proposed concept that apolipoprotein-E functions as a pathological chaperone by altering the conformation of amyloidogenic proteins
— id: 9401, year: 1994, vol: 145, page: 526, stat: Journal Article,

Characterization of a novel processing pathway for Alzheimer's amyloid beta precursor protein
Ghiso J; Gardella JE; Liem L; Gorevic PD; Frangione B
1994 Apr 25;171(1-2):213-216, Neuroscience letters
Amyloid beta (A beta) is a normal proteolytic fragment of a large precursor protein (beta PP) which undergoes altered conformation, leading to fibril formation. Two main beta PP processing pathways have been described, and we are now reporting the characterization of a third beta PP pathway. A membrane-associated 16 kDa component identified in human platelets isolated from normal donors. Based on size, immunoreactivity and amino acid sequence analysis, the fragment is a C-terminal beta PP component which starts at position 642 (APP770 numbering) and contains the intact A beta sequence. The presence of this novel pathway of beta PP processing in resting platelets suggest that it occurs as a normal event
— id: 9402, year: 1994, vol: 171, page: 213, stat: Journal Article,

Unifying features of systemic and cerebral amyloidosis
Ghiso J; Wisniewski T; Frangione B
1994 Feb;8(1):49-64, Molecular neurobiology
Amyloidosis is a generic term for a group of clinically and biochemically diverse diseases that are characterized by the deposition of an insoluble fibrillar protein in the extracellular space. Over 16 biochemically distinct amyloids are known. Despite this diversity, all amyloids have a particular ultrastructural and tinctorial appearance, a beta-pleated sheet structure, and are codeposited with a group of amyloid-associated proteins. The most common amyloidosis is Alzheimer's disease (AD), where A beta is the main component of the amyloid. Recently it has been found that A beta exists as a normal soluble protein (sA beta) in biological fluids. This links AD more closely to some of the systemic amyloidoses, where the amyloid precursor is found in the circulation normally. Numerous mutations have been found in the A beta precursor (beta PP) gene, associated with familial AD. Many mutations are also found in some of the hereditary systemic amyloidoses. For example, over 40 mutations in the transthyretin (TTR) gene are associated with amyloid. However, both A beta and TTR related amyloid deposition can occur with no mutation. The pathogenesis of amyloid is complex, and appears to be associated with genetic and environmental risk factors that can be similar in the systemic and cerebral amyloidoses
— id: 9403, year: 1994, vol: 8, page: 49, stat: Journal Article,

Binding of soluble beta-amyloid in vitro and in vivo
Golabek, A.; Marques, M.; Koji, I.; Ghiso, J.; Herbert, J.; Frangione, B.; Wisniewski, T.
1994 ;20(1-2):1643-64, Abstracts (Society for Neuroscience)
— id: 97648, year: 1994, vol: 20, page: 1643, stat: Journal Article,

Ocular amyloid deposition in familial amyloidosis, Finnish: an analysis of native and variant gelsolin in Meretoja's syndrome
Kivela T; Tarkkanen A; Frangione B; Ghiso J; Haltia M
1994 Sep;35(10):3759-3769, Investigative ophthalmology & visual science. IOVS
PURPOSE: To analyze the deposition of amyloid and its precursors in eyes of patients with familial amyloidosis, Finnish (FAF; Meretoja's syndrome), a hereditary systemic amyloidosis. METHODS. Autopsy eyes from three patients with FAF and ten control eyes were studied by Congo red staining and with antibodies to the nonmutated part of gelsolin (GS-2C4), the mutated gelsolin Asn-187 fragment (AGel), and amyloid-P component (AP). RESULTS. Congo red and antisera to AP and AGel bound to amyloid deposits in the cornea and conjunctiva, the sclera, the perineurium of ciliary nerves, the walls of ciliary vessels, the optic nerve sheaths, the stroma of the ciliary body, and along the choriocapillaris. mAb GS-2C4 bound weakly and focally to most deposits and strongly around the choriocapillaris. It labeled the corneal epithelium and endothelium, keratocytes, scleral fibroblasts, trabecular and lens epithelial cells, the ciliary muscle and epithelium, the iris sphincter and dilator, and stromal cells of the conjunctiva and uveal tract. CONCLUSIONS. Local production, especially in the cornea, conjunctiva, sclera, and ciliary muscle, and systemic deposition, particularly in blood vessles and in the sclera, may contribute to amyloid deposits in FAF. To explain the complex pattern of deposition, microenvironmental factors such as lamellar architecture of the cornea and sclera, altered processing of gelsolin, or blood-tissue barriers must be invoked. In addition to corneal lattice dystrophy type II, the observed deposits help to explain glaucoma in patients with FAF
— id: 9400, year: 1994, vol: 35, page: 3759, stat: Journal Article,

The soluble form of Alzheimer's amyloid beta protein is complexed to high density lipoprotein 3 and very high density lipoprotein in normal human plasma
Koudinov A; Matsubara E; Frangione B; Ghiso J
1994 Dec 15;205(2):1164-1171, Biochemical & biophysical research communications
The amyloid fibrils of Alzheimer's neuritic plaques and cerebral blood vessels are mainly composed of aggregated forms of a 39 to 44 amino acids peptide, named amyloid beta (A beta). A similar although soluble form of A beta (sA beta) has been identified in plasma, cerebrospinal fluid and cell culture supernatants, indicating that it is produced under physiologic conditions. We report here that sA beta in normal human plasma is associated with lipoprotein particles, in particular to the HDL3 and VHDL fractions where it is complexed to ApoJ and, to a lesser extent, to ApoAI. This was assessed by immunoprecipitation experiments of purified plasma lipoproteins and lipoprotein-depleted plasma and confirmed by means of amino acid sequence analysis. Moreover, biotinylated synthetic peptide A beta 1-40 was traced in normal human plasma in in vitro experiments. As in the case of sA beta, biotinylated A beta 1-40 was specifically recovered in the HDL3 and VHDL fractions. This data together with the previous demonstration that A beta 1-40 is taken up into the brain via a specific mechanism and possibly as an A beta 1-40-ApoJ complex indicate a role for HDL3- and VHDL-containing ApoJ in the transport of the peptide in circulation and suggest their involvement in the delivery of sA beta across the blood-brain barrier
— id: 6664, year: 1994, vol: 205, page: 1164, stat: Journal Article,

BETA-PROTEIN PRECURSOR INTERACTION WITH TAU
PERRY, G; SIEDLAK, S; MULVIHILL, P; RICHEY, PL; PRAPROTNIK, D; GHISO, J; FRANGIONE, B; KALARIA, R; SMITH, MA
1994 JUL ;15(7):S78-S78, Neurobiology of aging
— id: 52406, year: 1994, vol: 15, page: S78, stat: Journal Article,

Alzheimer's disease and soluble A beta
Wisniewski T; Ghiso J; Frangione B
1994 Mar-Apr;15(2):143-152, Neurobiology of aging
The discovery of soluble amyloid beta (sA beta) suggests that the role of amyloid in Alzheimer's disease (AD) is similar to the previously studied systemic amyloidoses and alters the notion that membrane damage is the initial event in AD. The disease state is characterized by the abnormal accumulation of a normal degradative peptide, which becomes resistant to further proteolysis due to a conformational change. Mutations in the beta PP gene have been found in a very small percentage of AD cases; hence other factors, both genetic and environmental, need to be identified. Priority needs to be given to detailed studies of the structural differences between sA beta and the A beta in amyloid deposits. This will help uncover the determining factors governing the aggregation of sA beta. These structural alterations may be critical for the possible toxic effects A beta and/or associated proteins (molecular chaperones, e.g., apolipoprotein E) have on brain cell function
— id: 6778, year: 1994, vol: 15, page: 143, stat: Journal Article,

Brain uptake of circulating apolipoproteins J and E complexed to Alzheimer's amyloid beta
Zlokovic BV; Martel CL; Mackic JB; Matsubara E; Wisniewski T; McComb JG; Frangione B; Ghiso J
1994 Dec 15;205(2):1431-1437, Biochemical & biophysical research communications
Amyloid beta (A beta) is a fibrillar component in Alzheimers' disease amyloid deposits and a soluble peptide (sA beta) normally present in body fluids. We have recently reported that the blood-brain barrier (BBB) has a capability to control cerebrovascular sequestration and transport of circulating sA beta. In this study, we examined whether two circulating amyloid-associated proteins shown to bind sA beta, apolipoproteins J (apo J) and E (apo E), can cross the BBB alone and/or complexed to a synthetic peptide homologous to a major form of sA beta, sA beta 1-40. Brain perfusion experiments in guinea pigs showed significant uptake of both apo J and sA beta 1-40-apo J complexes. In contrast, blood-brain transport of sA beta 1-40-apo E was negligible, while apo E had a limited access across the BBB, indicating that the apo E found within the brain is produced locally. It is concluded that sA beta 1-40 binding to apo J and apo E results in significant (> 100-fold) difference in brain uptake of their respective complexes. We hypothesize that in normal brain apo J facilitates sA beta transport
— id: 9399, year: 1994, vol: 205, page: 1431, stat: Journal Article,

Transport of Alzheimer's amyloid beta and apolipoproteins E and J at the blood-brain barrier
Zlokovic, B. V.; Mackic, J. B.; Martell, C. J.; Wisniewski, T.; Frangione, B.; Ghiso, J.
1994 ;20(1-2):1642-1437, Abstracts (Society for Neuroscience)
— id: 97649, year: 1994, vol: 20, page: 1642, stat: Journal Article,

THE BLOOD-BRAIN-BARRIER REGULATES TRANSPORT OF ALZHEIMERS AMYLOID-BETA AND APOLIPOPROTEIN-E AND APOLIPOPROTEIN-J
ZLOKOVIC, BV; MACKIC, JB; MARTELL, CL; WISNIEWSKI, T; FRANGIONE, B; GHISO, J
1994 JUL ;15(7):S80-S80, Neurobiology of aging
— id: 52407, year: 1994, vol: 15, page: S80, stat: Journal Article,

Alzheimer's disease and amyloid
Frangione B; Wisniewski T; Ghiso J
Amyloid and amyloidosis 1993 New York : Parthenon, 1993,
— id: 5144, year: 1993, vol: , page: 310, stat: Chapter,

Alzheimer's disease and Dutch variant: Opposing faces of a single coin
Frangione, Blas; Wisniewski, Thomas; Tagliavini, Fabrizio; Bugiani, Orso; Ghiso, Jorge
Alzheimer's disease : advances in clinical and basic research New York : Wiley, 1993,
— id: 4969, year: 1993, vol: , page: 387, stat: Chapter,

Light Chain Deposition disease : biochemical characterization of tissue deposits
Gallo G; Boctor F; Frangione B; Ghiso J
Amyloid and amyloidosis 1993 New York : Parthenon, 1993,
— id: 5145, year: 1993, vol: , page: 280, stat: Chapter,

Characterization and fibrillogenesis of a construct (C109) homologus to the carboxyl end of the amyloid precusor protein of Alzheimer's disease
Gardella JE; Gorgone G; Ghiso J; Castano E; Frangione B; Gorevic PD
Alzheimer's disease : advances in clinical and basic research New York : Wiley, 1993,
— id: 5143, year: 1993, vol: , page: 411, stat: Chapter,

High-level expression and in vitro mutagenesis of a fibrillogenic 109-amino-acid C-terminal fragment of Alzheimer's-disease amyloid precursor protein
Gardella JE; Gorgone GA; Candela L; Ghiso J; Castano EM; Frangione B; Gorevic PD
1993 Sep 15;294(Pt 3):667-674, Biochemical journal
We amplified DNA encoding the 3' 109 codons of Alzheimer's-disease amyloid precursor protein (APP) inclusive of the beta protein (A beta) and cytoplasmic domains from cDNA using oligonucleotide primers designed to facilitate cloning into the T7 expression vector pT7Ad23K13. We also modified this construct to generate recombinant molecules incorporating two recently described APP mutants by site-directed mutagenesis. Both native C109 (deletion construct inclusive of the C-terminal 109 residues of APP) and constructs with a single mutation at codon 642 (T-->G, resulting in a substitution of glycine for valine) or a double mutation at codons 595 (G-->T, substituting asparagine for lysine) and 596 (A-->C, substituting leucine for methionine) were expressed in Escherichia coli to levels of 5-20% of total bacterial protein after induction. The major constituent of expressed C109 protein had an apparent molecular mass of 16-18 kDa by SDS/PAGE and appeared to be the full-length construct by size and N-terminal microsequencing. Also present was a 4-5 kDa species that co-purified with C109, constituting only approximately 1% of expressed protein, which was revealed by Western-blot analysis with antibodies specific for A beta epitopes and after biotinylation of purified recombinant C109. This fragment shared N-terminal sequence with, and appeared to arise by proteolysis of, full-length C109 in biosynthetic labelling experiments. C109 spontaneously precipitated after dialysis against NaCl or water, and with prolonged (> 20 weeks) standing was found by electron microscopy to contain a minor (< 5%) fibrillar component that was reactive with antibodies to a C-terminal epitope of APP. Recombinant C109 appears to duplicate some of the biochemical and physicochemical properties of C-terminal A beta-inclusive fragments of APP that have been found in transfected cells, brain cortex and cerebral microvessels
— id: 9406, year: 1993, vol: 294, page: 667, stat: Journal Article,

The cerebrospinal-fluid soluble form of Alzheimer's amyloid beta is complexed to SP-40,40 (apolipoprotein J), an inhibitor of the complement membrane-attack complex
Ghiso J; Matsubara E; Koudinov A; Choi-Miura NH; Tomita M; Wisniewski T; Frangione B
1993 Jul 1;293(Pt 1):27-30, Biochemical journal
The amyloid fibrils deposited in Alzheimer's neuritic plaque cores and cerebral blood vessels are mainly composed of aggregated forms of a unique peptide, 39-42 amino acids long, named amyloid beta (A beta). A similar, although soluble, A beta ('sA beta') has been identified in cerebrospinal fluid, plasma and cell supernatants, indicating that it is normally produced by proteolytic processing of its precursor protein, amyloid precursor protein (APP). Using direct binding experiments we have isolated and characterized an 80 kDa circulating protein that specifically interacts with a synthetic peptide identical with A beta. The protein was unmistakably identified as SP-40,40 or ApoJ, a cytolytic inhibitor and lipid carrier, by means of amino acid sequence and immunoreactivity with specific antibodies. Immunoprecipitation with anti-SP-40,40 retrieved soluble A beta from cerebrospinal fluid, indicating that the interaction occurs in vivo
— id: 8397, year: 1993, vol: 293, page: 27, stat: Journal Article,

Epitope map of two polyclonal antibodies that recognize amyloid lesions in patients with Alzheimer's disease
Ghiso J; Wisniewski T; Vidal R; Rostagno A; Frangione B
1993 ;1:19-20, Parkinson/Alzheimer digest
— id: 101632, year: 1993, vol: 1, page: 19, stat: Journal Article,

The cerebrospinal fluid soluble form of Alzheimer's amyloid beta is complexed to SP-40,40 (APO J), an inhibitor of the complement membrane attack complex
Ghiso, Jorge; Matsubara, Etsuro; Koudinov, Alexei; Wisniewski, Thomas; Frangione, Blas
1993 ;19(1-3):19-S80, Abstracts (Society for Neuroscience)
— id: 97650, year: 1993, vol: 19, page: 19, stat: Journal Article,

Synthetic peptides corresponding to different mutated regions of the amyloid gene in familial Creutzfeldt-Jakob disease show enhanced in vitro formation of morphologically different amyloid fibrils
Goldfarb LG; Brown P; Haltia M; Ghiso J; Frangione B; Gajdusek DC
1993 May 15;90(10):4451-4454, Proceedings of the National Academy of Sciences of the United States of America
We synthesized polypeptides corresponding to sequences encoded by normal and mutant alleles in the regions of codon 178 (Asp-->Asn) and codon 200 (Glu-->Lys) of the chromosome 20 amyloid gene that have been linked to familial Creutzfeldt-Jakob disease. Peptide suspensions from both regions spontaneously formed amyloid fibrils with different morphological characteristics and aggregation tendencies. Fibrillar arrays were denser and more profuse in mutant than in normal peptide suspensions and were even more marked when the homologous mutant and normal peptides were mixed together. Preparations from the region of codon 200 were in all cases more fibrillogenic than corresponding peptides from the region of codon 178. These in vitro observations support the hypothesis that amino acid changes from pathogenic single-allele point mutations in Creutzfeldt-Jakob disease may nucleate the in vivo folding behavior of the normal host protein to favor formation of insoluble amyloid fibrils
— id: 9407, year: 1993, vol: 90, page: 4451, stat: Journal Article,

Immunohistochemical analysis of lattice corneal dystrophies types I and II
Kivela T; Tarkkanen A; McLean I; Ghiso J; Frangione B; Haltia M
1993 Dec;77(12):799-804, British journal of ophthalmology
Corneal buttons from four patients with lattice corneal dystrophy (LD) type I, thought to be an isolated corneal amyloidosis, and from six patients with LD type II, part of systemic familial amyloidosis, Finnish type (FAF; Meretoja's syndrome), were studied by immunohistochemistry to determine the differential distribution in the amyloid deposits of amyloid P component (AP), mutated gelsolin specific for FAF, and native gelsolin. In both types of LD, antibodies to AP labelled lattice lines and a discontinuous layer of amyloid deposits under Bowman's layer. In LD type II, particularly, they also reacted with streak-like amyloid deposits between corneal almellae, especially in the limbal region. While the anti-FAF antiserum strongly labelled all amyloid deposits in LD type II, it failed to react unequivocally with them in LD type I. Both in LD type I and in two control specimens representing granular dystrophy, the monoclonal antibody (MAb) GS-2C4 to gelsolin faintly labelled some deposits, while in LD type II it reacted non-homogeneously with most amyloid deposits. In all specimens, MAb GS-2C4 labelled corneal epithelial cells and occasional stromal keratocytes and endothelial cells. The results suggest that Meretoja's syndrome, a systemic disease, can be diagnosed even retrospectively from corneal buttons subjected to histopathological study
— id: 9405, year: 1993, vol: 77, page: 799, stat: Journal Article,

Proteolytic processing of human amyloid beta protein precursor in insect cells. Major carboxyl-terminal fragment is identical to its human counterpart
Ramabhadran TV; Gandy SE; Ghiso J; Czernik AJ; Ferris D; Bhasin R; Goldgaber D; Frangione B; Greengard P
1993 Jan 25;268(3):2009-2012, Journal of biological chemistry
The predominant component of amyloid plaques of Alzheimer's disease is the amyloid beta protein (A beta), a 39-42-amino-acid peptide derived by proteolysis of a family of precursors known as amyloid precursor proteins (APP). In mammalian brain and in cultured mammalian cells, the release of APP amino-terminal fragments into the extracellular medium occurs by a proteolytic cleavage within the A beta domain, thereby precluding amyloidogenesis. Infection of Sf9 insect cells with baculovirus vectors containing APP cDNAs results in high levels of APP expression. The concomitant release of amino-terminal fragments of APP and the production of carboxyl-terminal, cell-associated cleavage products are observed. Here we demonstrate by direct protein microsequencing that the proteolytic processing of APP in the Sf9 cells generates a prominent carboxyl-terminal species that is identical to that produced in human cells, suggesting that the major pathway for proteolytic processing of APP is conserved among metazoans
— id: 9409, year: 1993, vol: 268, page: 2009, stat: Journal Article,

Cerebrospinal fluid inhibits Alzheimer beta-amyloid fibril formation in vitro
Wisniewski T; Castano E; Ghiso J; Frangione B
1993 Oct;34(4):631-633, Annals of neurology
Alzheimer's disease is characterized by the deposition of beta-protein (A beta) as amyloid. Recently, it was found that A beta is a normal component of serum and cerebrospinal fluid. Synthetic peptides homologous to A beta form amyloid-like fibrils spontaneously in water or physiological solutions. Using a peptide homologous to A beta 1-40, we find that fibril formation is inhibited by the presence of cerebrospinal fluid
— id: 6554, year: 1993, vol: 34, page: 631, stat: Journal Article,

Apolipoprotein E: binding to soluble Alzheimer's beta-amyloid
Wisniewski T; Golabek A; Matsubara E; Ghiso J; Frangione B
1993 Apr 30;192(2):359-365, Biochemical & biophysical research communications
Apolipoprotein E (apo E) is associated with Alzheimer's beta-amyloid (A beta) in senile plaques. A beta is now known to be a normal soluble peptide (sA beta) found in the cerebrospinal fluid (CSF) and other biological fluids. We have used synthetic A beta peptides bound to affinity membranes in order to determine whether apo E or any other amyloid associated protein will bind to these membranes, when they are bathed in CSF. Under these conditions apo E, as well as another apolipoprotein, apolipoprotein J (Apo J), bound to the membranes. Using ELISA and ligand binding studies, we found a high avidity binding of A beta peptides to apo E. This suggests that apo E, as well as other related proteins may bind not only amyloid A beta but also sA beta. This interaction may be critical in amyloid formation
— id: 9408, year: 1993, vol: 192, page: 359, stat: Journal Article,

Apolipoprotein E and Alzheimer's beta-amyloid fibril formation
Wisniewski, Thomas; Golabek, Adam; Ghiso, Jorge; Frangione, Blas
1993 ;19(1-3):1034-633, Abstracts (Society for Neuroscience)
— id: 97651, year: 1993, vol: 19, page: 1034, stat: Journal Article,

Blood-brain barrier transport of circulating Alzheimer's amyloid beta
Zlokovic BV; Ghiso J; Mackic JB; McComb JG; Weiss MH; Frangione B
1993 Dec 30;197(3):1034-1040, Biochemical & biophysical research communications
The origin of amyloid beta (A beta) deposited in brain and cerebral blood vessels of patients with Alzheimer's Disease (AD) is not known. In this study, we tested whether soluble A beta (sA beta) can cross the blood-brain barrier (BBB). An in vivo vascular brain perfusion model and capillary depletion technique in guinea-pigs were used to determine cerebral capillary sequestration and blood-brain transport of a synthetic peptide identical with residues 1-40 (SP-40) of A beta. A saturable, specific binding of SP-40 has been demonstrated at the luminal side of the BBB, with the Kd of 25 +/- 2 nM, and Bmax of 188 +/- 11 fmol/min/g of isolated microvessels. A specific transcellular BBB transport of SP-40 into brain parenchyma exhibited the Km of 49 +/- 10 nM, and Vmax of 111 +/- 19 fmol/min/g of capillary depleted brain. We concluded that the BBB has the capability to control the cerebrovascular sequestration and blood-to-brain transport of circulating sA beta. Hence, sA beta can contribute to both cerebrovascular and parenchymal amyloid formation
— id: 9404, year: 1993, vol: 197, page: 1034, stat: Journal Article,

Distribution of Alzheimer's disease amyloid protein precursor in normal human and rat nervous system
Coria F; Moreno A; Torres A; Ahmad I; Ghiso J
1992 Feb;18(1):27-35, Neuropathology & applied neurobiology
Alzheimer's disease and cerebral amyloid angiopathies (CAA) are clinically heterogeneous diseases, but pathogenically related by the deposition of beta A4-amyloid in the brain in the form of neuritic plaques and/or vascular infiltrates. Antibodies directed against the N-terminal region of the predicted sequence of the beta A4 amyloid protein precursor (APP) were used to investigate the cellular distribution of this protein in the brain of normal humans and rats. We found a widespread presence of APP throughout the nervous tissue, including neurons, blood vessels, meningeal membranes, choroid plexus and ependymal cells. The highest APP immunoreactivity in both species was found in neuronal cell bodies and their processes, and around blood vessels. These findings may account for the clinical, pathological and aetiological differences found among the beta A4-amyloidosis
— id: 9415, year: 1992, vol: 18, page: 27, stat: Journal Article,

ACCELERATED INSTRUCTIVE FIBRILLOGENESIS IN THE DUTCH VARIANT OF ALZHEIMER'S DISEASE
FRANGIONE B; WISNIEWSKI T; GHISO J
1992 ;43(16 PART E):200-A422, Journal of cellular biochemistry. Supplement
— id: 97652, year: 1992, vol: 43, page: 200, stat: Journal Article,

ACCELERATED INSTRUCTIVE FIBRILLOGENESIS IN THE DUTCH VARIANT OF ALZHEIMER'S DISEASE AND THE ROLE OF PATHOLOGICAL CHAPERONES IN AMYLOID FORMATION
FRANGIONE B; WISNIEWSKI T; GHISO J
1992 ;13(SUPPL. 1):S73-S80, Neurobiology of aging
— id: 97600, year: 1992, vol: 13, page: S73, stat: Journal Article,

Beta protein precursor expression in human platelets and a megakaryocyte cell line. Possible implications for the origin of cerebral amyloidosis in Alzheimer's disease
Gardella JE; Gorgone GA; Munoz PC; Ghiso J; Frangione B; Gorevic PD
1992 Sep;67(3):303-313, Laboratory investigation
BACKGROUND: The origin of the amyloid beta protein (A beta) that is the main constituent of amyloid fibrils occurring in the senile plaques and cerebrovasculature of individuals afflicted with Alzheimer's Disease, Down's Syndrome, Hereditary Cerebral Hemorrhage with Amyloidosis--Dutch Type, and Sporadic Cerebral Amyloid Angiopathy, is central to the pathogenesis of these disorders. Evidence exists to support a neuronal and/or a vascular origin. We have reported that platelets may serve as one possible source of the A beta sequence via an intact, membrane-associated Beta Protein Precursor (beta PP) which is encoded by a platelet transcript (BBRC 173:1292-1298, 1990). EXPERIMENTAL DESIGN: Immunoaffinity chromatography and western blotting of extracted cellular proteins, polymerase chain reaction amplification of beta PP mRNA, fluorescence activated flow cytometric analysis, and confocal scanning laser microscopy have been employed to characterize the presence and distribution of thrombocytic beta PP. RESULTS: Immunoblot analysis with antibodies specific for the carboxyl-terminal end of beta PP indicates that platelets and the Dami megakaryocyte cell line express membrane-associated species of intact beta PP ranging in molecular weight from 110 to 140 kilodaltons (kd), as well as carboxyl-terminal reactive forms ranging from 16 to 22 kd. Thrombin stimulation of platelets induces the release of five soluble beta PP species, which possess apparent isofocusing points in the range of 4.1-5.5. By contrast, extracts of peripheral blood mononuclear cells enriched by ficoll centrifugation, endothelial cells and a B cell line were not immunoreactive by western blot, even though beta PP transcripts could be amplified by polymerase chain reaction. The distribution of platelet beta PP was localized by flow cytometric analysis and scanning laser microscopy, using fluorescein-labeled antibodies. Study of the subcellular distribution of platelet beta PP indicates that these translation products are accumulated in discrete foci throughout the thrombocyte, possibly corresponding to secretory granules. CONCLUSIONS: The size of the carboxyl-terminal forms of beta PP indicate that the A beta sequence is present as a membrane associated constituent in unstimulated platelets, and may represent alternative pathways of beta PP processing. Cleavage or other abnormal processing of platelet-associated beta PP in Alzheimer's disease provides one mechanism whereby cerebral amyloid might derive from the circulation
— id: 9413, year: 1992, vol: 67, page: 303, stat: Journal Article,

A 109-amino-acid C-terminal fragment of Alzheimer's-disease amyloid precursor protein contains a sequence, -RHDS-, that promotes cell adhesion
Ghiso J; Rostagno A; Gardella JE; Liem L; Gorevic PD; Frangione B
1992 Dec 15;288(Pt 3):1053-1059, Biochemical journal
Amyloid beta (A beta), the major constituent of the fibrils composing senile plaques and vascular amyloid deposits in Alzheimer's disease (AD) and related disorders, is a 39-42-residue self-aggregating degradation peptide of a larger multidomain membrane glycoprotein designated amyloid precursor protein (APP). An array of biological functions has been assigned to different APP domains, including growth regulation, neurotoxicity, inhibitory activity of serine proteinases and promotion of cell-cell and cell-matrix interactions. A beta is generated through an as-yet-unknown catabolic pathway that by-passes or inhibits the cleavage of APP within the A beta sequence. We have identified a 16 kDa intermediate APP C-terminal fragment containing A beta in leptomeningeal vessels of aged normal individuals and AD patients by means of its immunoreactivity with a panel of four different anti-(APP C-terminal) antibodies, indicating a different pathway of APP processing. Previous studies have indicated that the APP C-terminal domain is the most likely to be involved in cell-matrix interactions. A 109-amino-acid construct C109 with a sequence analogous to the C-terminal of APP (positions 587-695 of APP695), similar in length and immunoreactivity to the 16 kDa fragment, was found to promote cell adhesion. By use of synthetic peptides, this activity was initially located to the extracellular 28 residues of A beta. Inhibition studies demonstrated that the sequence RHDS (amino acids 5-8 of A beta, corresponding to residues 601-604 of APP695 was responsible for the adhesion-promoting activity. The interaction is dependent on bivalent cations and can be blocked either by the tetrapeptides RHDS and RGDS or by an anti-(beta 1 integrin) antibody. Thus, through integrin-like surface receptors, APP or its derivative proteolytic fragments containing the sequence RHDS may modulate cell-cell or cell-matrix interactions
— id: 9410, year: 1992, vol: 288, page: 1053, stat: Journal Article,

Epitope map of two polyclonal antibodies that recognize amyloid lesions in patients with Alzheimer's disease
Ghiso J; Wisniewski T; Vidal R; Rostagno A; Frangione B
1992 Mar 1;282(Pt 2):517-522, Biochemical journal
Two synthetic peptides with sequences identical with those of fragments of the extracellular domain of the Alzheimer's-disease amyloid precursor protein (APP) were used to raise antibodies. SP28 comprises positions 597-624 of the APP695 isoform, whereas SP41 extends towards the N-terminus (amino acids 584-624) and contains the entire SP28 peptide. Using e.l.i.s.a. and inhibition experiments we identified the two beta-turn-containing segments 602-607 and 617-624 as the epitopes recognized by anti-SP41 and anti-SP28 respectively. Both antibodies immunolabelled amyloid lesions in brains from Alzheimer's-disease patients and patients with related disorders, whereas they were unreactive in control brains. However, when probed on immunoblots, anti-SP28 failed to detect full-length APP from baculovirus-infected Sf9 cells, and anti-SP41 reacted weakly compared with other anti-APP antisera. The data suggest that these antibodies are directed to conformational epitopes not existent in the native molecules but present after alternative APP processing
— id: 9414, year: 1992, vol: 282, page: 517, stat: Journal Article,

Production of the Alzheimer amyloid beta protein by normal proteolytic processing
Shoji M; Golde TE; Ghiso J; Cheung TT; Estus S; Shaffer LM; Cai XD; McKay DM; Tintner R; Frangione B; et al
1992 Oct 2;258(5079):126-129, Science
The 4-kilodalton (39 to 43 amino acids) amyloid beta protein (beta AP), which is deposited as amyloid in the brains of patients with Alzheimer's diseases, is derived from a large protein, the amyloid beta protein precursor (beta APP). Human mononuclear leukemic (K562) cells expressing a beta AP-bearing, carboxyl-terminal beta APP derivative released significant amounts of a soluble 4-kilodalton beta APP derivative essentially identical to the beta AP deposited in Alzheimer's disease. Human neuroblastoma (M17) cells transfected with constructs expressing full-length beta APP and M17 cells expressing only endogenous beta APP also released soluble 4-kilodalton beta AP, and a similar, if not identical, fragment was readily detected in cerebrospinal fluid from individuals with Alzheimer's disease and normal individuals. Thus cells normally produce and release soluble 4-kilodalton beta AP that is essentially identical to the 4-kilodalton beta AP deposited as insoluble amyloid fibrils in Alzheimer's disease
— id: 9411, year: 1992, vol: 258, page: 126, stat: Journal Article,

Amyloidoma of the CNS. II. Immunohistochemical and biochemical study
Vidal RG; Ghiso J; Gallo G; Cohen M; Gambetti PL; Frangione B
1992 Oct;42(10):2024-2028, Neurology
We present the immunohistochemical and biochemical identification of an amyloidoma localized to the cerebral white matter in a patient who shows no evidence of systemic or extracranial localized amyloid deposits. Immunohistologic and immunochemical studies, using antibodies against biochemically different amyloid fibrils and amyloid-associated proteins, showed reactivity with antibodies only to lambda light chain and serum amyloid P-component. Amino acid sequence analysis of the purified amyloid fibrils extracted from the brain tumor indicates that the amyloid protein is an unusual immunoglobulin lambda light chain, starting at residue five of the variable domain. These fibrils consist of lambda chain fragments of different lengths (10 to 30 kd) very likely arising by polymerization of the amyloid subunit or sequential proteolytic cleavage of the light chain, or both. After exposure to denaturing agents, the 10-kd subunit retains the characteristics of native amyloid fibrils by electron microscopy
— id: 9412, year: 1992, vol: 42, page: 2024, stat: Journal Article,

ACCELERATED FIBRILLOGENESIS IN THE DUTCH VARIANT OF ALZHEIMER'S DISEASE
WISNIEWSKI T; GHISO J; FRANGIONE B
1992 ;42(4 SUPPL. 3):304-238, Neurology
— id: 97654, year: 1992, vol: 42, page: 304, stat: Journal Article,

I corpi di Lewy immunoreagiscono con gli anticorpi dell'amiloide di tipo finnico omologo alla gelsolina
Wisniewski T; Haltia M; Ghiso J; Frangione B
1992 ;2:59-60, Update on Parkinson
— id: 102366, year: 1992, vol: 2, page: 59, stat: Journal Article,

Lewy bodies and gelsolin
Wisniewski T; Haltia M; Ghiso J; Frangione B
1992 ;1:6-8, Parkinson/Alzheimer digest
— id: 97677, year: 1992, vol: 1, page: 6, stat: Journal Article,

Familial amyloidosis - Finish type - and its relationship to Lewy bodies in Parkinson's and Diffuse Lewy Body disease
Frangione B; Haltia M; Levy E; Ghiso J; Kiuru S; Prelli F; Wisniewski T
Proceedings of the XIth International Congress of Neuropathology, September 2-8, 1990, Kyoto, Japan [Tokyo] : Japanese Society of Neuropathology, 1991,
— id: 4980, year: 1991, vol: , page: 150, stat: Chapter,

Immunoreactivity of Alzheimer amyloid precusor protein (APP) specific antisera with platelet granule constituents
Gardella JE; Ghiso J; Gorgone GA; Marratta D; Kaplan AP; Frangione B; Gorevic PD
Amyloid and amyloidosis 1990 Boston : Kluwer, 1991,
— id: 5141, year: 1991, vol: , page: 723, stat: Chapter,

ALZHEIMER'S AMYLOID PRECURSOR PROTEIN CONTAINS A TETRAPEPTIDE SEQUENCE-RHIDS THAT PROMOTES CELL ADHESION
GHISO J; ROSTAGNO A; FRANGIONE B
1991 ;17(1-2):913-913, Abstracts (Society for Neuroscience)
— id: 101623, year: 1991, vol: 17, page: 913, stat: Journal Article,

Amyloidosis due to a mutation of the gelsolin gene in an American family with lattice corneal dystrophy type II [see comments]
Gorevic PD; Munoz PC; Gorgone G; Purcell JJ Jr; Rodrigues M; Ghiso J; Levy E; Haltia M; Frangione B
1991 Dec 19;325(25):1780-1785, New England journal of medicine
— id: 9416, year: 1991, vol: 325, page: 1780, stat: Journal Article,

Shared gelsolin antigenicity between familial amyloidosis, Finnish type (FAF) and one form of familial lattice corneal dystrophy (LCD) with polyneuropathy from the United States
Gorevic PD; Munoz PC; Rodrigues M; Haltia M; Ghiso J; Frangione B
Amyloid and amyloidosis 1990 Boston : Kluwer, 1991,
— id: 5140, year: 1991, vol: , page: 423, stat: Chapter,

Polymerization of gelsolin variant fragment in tissue causes familial amyloidosis, Finnish type (FAF)
Haltia M; Ghiso J; Prelli F; Levy E; Gallo G; Kiuru S; Somer H; Palo J; Frangione B
Amyloid and amyloidosis 1990 Boston : Kluwer, 1991,
— id: 5142, year: 1991, vol: , page: 409, stat: Chapter,

Gelsolin variant and beta-amyloid co-occur in a case of Alzheimer's with Lewy bodies
Haltia M; Ghiso J; Wisniewski T; Kiuru S; Miller D; Frangione B
1991 Jul-Aug;12(4):313-316, Neurobiology of aging
Familial amyloidosis, Finnish type (FAF), is an autosomal dominant form of amyloidosis which is related to a point mutation in the gelsolin gene localized on chromosome 9. The mutation corresponds to codon 187 of the secreted form of gelsolin, and is expressed in the amyloid fibril at residue 15. Our original FAF patient was demented, and neuropathological analysis showed Alzheimer type brain lesions associated with both classical and cortical Lewy bodies. Furthermore, antiserum against the gelsolin-derived FAF amyloid reacted strongly with both classical and cortical Lewy bodies of this FAF patient. In preliminary experiments similar results were obtained in cases of Parkinson's disease and diffuse Lewy body disease. These observations may indicate a role for gelsolin in the pathogenesis of Parkinson's disease and related conditions
— id: 9421, year: 1991, vol: 12, page: 313, stat: Journal Article,

Isolation and amino terminal sequence of beta-trace, a novel protein from human cerebrospinal fluid
Kuruvilla AP; Hochwald GM; Ghiso J; Castano EM; Pizzolato M; Frangione B
1991 Nov 29;565(2):337-340, Brain research
beta-Trace, a 23.5 kDa glycoprotein of unknown biological functions, is present in all body fluids tested. It is found in higher concentration in human seminal fluid and cerebrospinal fluid (CSF) than in serum. A one-step procedure for the isolation of beta-trace from pooled CSF is described, by affinity chromatography using a specific antibody made against beta-trace. Amino terminal sequence analysis yields the sequence A P E A Q V S V Q P N F Q Q D K F L G with no homology to known proteins, indicating that beta-trace is a novel CSF protein
— id: 9417, year: 1991, vol: 565, page: 337, stat: Journal Article,

The primary structure of the Fab fragment of protein KAU, a monoclonal immunoglobulin M cold agglutinin
Leoni J; Ghiso J; Goni F; Frangione B
1991 Feb 15;266(5):2836-2842, Journal of biological chemistry
The complete amino acid sequence of the Fab fragment of protein KAU, a human monoclonal cold agglutinin (IgMk) with anti-I activity, was determined. The light chain (L-chain) consists of 215 residues; the variable (V)L region belongs to the Hum/Kv325/kIIIb sub-subgroup that is preferentially selected in human IgM autoimmune response. The joining (J) region is encoded by the Jk4 gene, and the constant region (C)L domain expresses the km3 allotypic marker. The Fd fragment contains 232 amino acids, and 120 of them comprise the variable domain. The VH region corresponds to the VHIV subgroup and is closely related to the VHIV 2.1 gene isolated from genomic DNA expressed in peripheral blood of a healthy Caucasian. The complementary-determining region 1 has a unique amino acid (Asp) at position 31, and the complementary-determining region 3 codified by the diversity segment (D) gene, shows poor homology with other known D sequences. The joining segment with two unusual substitutions at the D-J junction is encoded by the JH4 gene. Thus, cold agglutinin KAU is an IgM, VkIIIb-Jk4-km3; VHIV-JH4-C mu
— id: 9424, year: 1991, vol: 266, page: 2836, stat: Journal Article,

Alzheimer patients: preamyloid deposits are immunoreactive with antibodies to extracellular domains of the amyloid precursor protein
Tagliavini F; Giaccone G; Verga L; Ghiso J; Frangione B; Bugiani O
1991 Jul 8;128(1):117-120, Neuroscience letters
In patients with Alzheimer's disease, in patients with Down's syndrome and in aged non-demented individuals, anti-beta-protein antibodies label not only the fibrillary amyloid, but also preamyloid deposits. The latter are made up of amorphous material lacking the tinctorial, optical and ultrastructural properties of amyloid fibrils. To investigate the antigenic profile of preamyloid deposits, we have carried out an immunohistochemical study on specimens of cerebral cortex from 4 Alzheimer patients and two non-demented individuals, using antibodies to the beta-protein (anti-SP28), the C-terminal region of the amyloid precursor protein (APP) (anti-SP20) and an APP extracellular epitope between residues 50 and 100 (anti-preA4). Anti-preA4 and anti-SP28 immunoreactivity was found to be present in preamyloid deposits, whereas anti-SP20 immunoreactivity was not. These findings suggest that an extracellular portion of APP, close to the N-terminus of the molecule, participates with beta-protein in the composition of preamyloid deposits
— id: 9420, year: 1991, vol: 128, page: 117, stat: Journal Article,

Amyloid protein of Gerstmann-Straussler-Scheinker disease (Indiana kindred) is an 11 kd fragment of prion protein with an N-terminal glycine at codon 58
Tagliavini F; Prelli F; Ghiso J; Bugiani O; Serban D; Prusiner SB; Farlow MR; Ghetti B; Frangione B
1991 Mar;10(3):513-519, EMBO journal
Gerstmann-Straussler-Scheinker (GSS) disease is a familial neurological disorder pathologically characterized by amyloid deposition in the cerebrum and cerebellum. The GSS amyloid is immunoreactive to antisera raised against the hamster prion protein (PrP) 27-30. This is a proteinase K-resistant glycoprotein of 27-30 kd that is derived from an abnormal isoform of a neuronal glycoprotein of 33-35 kd designated PrPSc and is a molecular marker of amyloid fibrils isolated from animals with scrapie and humans with related disorders. We have purified and characterized proteins extracted from amyloid plaque cores isolated from two patients of the Indiana kindred of GSS disease. We found that the major component of GSS amyloid is an 11 kd degradation product of PrP, whose N-terminus corresponds to the glycine residue at position 58 of the amino acid sequence deduced from the human PrP cDNA. In addition, amyloid fractions contained larger PrP fragments with apparently intact N-termini and amyloid P component. These findings suggest that the disease process leads to proteolytic cleavage of PrP, generating an amyloidogenic peptide that polymerizes into insoluble fibrils. The N-terminal cleavage of PrP in GSS disease occurs at a tryptophan-glycine peptide bond identical to that cleaved by proteinase K in vitro to generate PrP 27-30 from hamster PrPSc at codon 90. Since no mutations of the structural PrP gene have been found in the Indiana family of GSS disease, it is conceivable that factors other than the primary structure of PrP play a crucial role in the process of amyloid formation and the development of clinical neurologic dysfunction
— id: 9423, year: 1991, vol: 10, page: 513, stat: Journal Article,

Peptides homologous to the amyloid protein of Alzheimer's disease containing a glutamine for glutamic acid substitution have accelerated amyloid fibril formation
Wisniewski T; Ghiso J; Frangione B
1991 Nov 14;180(3):1528-1528, Biochemical & biophysical research communications
— id: 9418, year: 1991, vol: 180, page: 1528, stat: Journal Article,

Peptides homologous to the amyloid protein of Alzheimer's disease containing a glutamine for glutamic acid substitution have accelerated amyloid fibril formation [published erratum appears in Biochem Biophys Res Commun 1991 Nov 14;180(3):1528]
Wisniewski T; Ghiso J; Frangione B
1991 Sep 30;179(3):1247-1254, Biochemical & biophysical research communications
beta-Amyloid (A beta) deposition in fibril form is the central event in a number of diseases, including Alzheimer's disease (AD) and hereditary cerebral hemorrhage with amyloidosis - Dutch type (HCHWA-D). A beta is produced by degradation of a larger amyloid precursor protein (APP). Recently a mutation in the APP gene has been found in HCHWA-D causing a glutamine for glutamic acid substitution at residue 22 of A beta. The influence of this mutation on fibrillogenesis is not known, although it is clear that affected patients have accelerated cerebrovascular amyloid deposition, with disease symptoms early in life. We report the in vitro demonstration of accelerated fibril formation in a 28 residue synthetic peptide homologous to the Dutch variant A beta. Furthermore, in eight residue peptides homologous to A beta the presence of the mutation is necessary for fibril formation. These findings provide a mechanism for accelerated amyloid formation in the Dutch variant of APP
— id: 9419, year: 1991, vol: 179, page: 1247, stat: Journal Article,

LEWY BODIES ARE IMMUNOREACTIVE WITH ANTIBODIES RAISED TO GELSOLIN AMYLOID
WISNIEWSKI T; HALTIA M; GHISO J; FRANGIONE B
1991 ;41(3 SUPPL. 1):177-316, Neurology
— id: 97655, year: 1991, vol: 41, page: 177, stat: Journal Article,

Lewy bodies are immunoreactive with antibodies raised to gelsolin related amyloid-Finnish type
Wisniewski T; Haltia M; Ghiso J; Frangione B
1991 May;138(5):1077-1083, American journal of pathology
Lewy bodies (LB) are intraneuronal, cytoplasmic inclusion bodies. They are invariably present in Parkinson's and diffuse LB diseases. Their composition by direct biochemical methods is unknown, although LBs are immunoreactive with a number of antibodies, including anti-ubiquitin and anti-neurofilament antibodies. Familial amyloidosis, Finnish type (FAF), is an autosomal-dominant form of systemic amyloidosis. The authors have isolated and partially sequenced the amyloid. The protein has significant sequence identity with gelsolin, an actin-modulating protein. Rabbit polyclonal antibodies raised to the FAF amyloid not only immunostain the amyloid but also LBs in the cortex and substantia nigra of Parkinson's and diffuse LB disease brains. Immunoreactivity is absorbed by the purified amyloid but is unaffected by ubiquitin. This provides a link between the LB and one of the human amyloidoses, FAF
— id: 9422, year: 1991, vol: 138, page: 1077, stat: Journal Article,

COOCCURRENCE OF 2 AMYLOID PROTEINS, GELSOLIN AND BETA-PROTEIN, IN A PATIENT WITH FAMILIAL AMYLOIDOSIS, FINNISH TYPE, AND ALZHEIMERS-DISEASE
Frangione, B; Haltia, M; Ghiso, J; Kiuru, S; Prelli, F
1990 May-Jun;11(3):300-300, Neurobiology of aging
— id: 31940, year: 1990, vol: 11, page: 300, stat: Journal Article,

Intact Alzheimer amyloid precursor protein (APP) is present in platelet membranes and is encoded by platelet mRNA
Gardella JE; Ghiso J; Gorgone GA; Marratta D; Kaplan AP; Frangione B; Gorevic PD
1990 Dec 31;173(3):1292-1298, Biochemical & biophysical research communications
Using antibodies directed against N-terminal and C-terminal epitopes we have immunologically detected APP species in the membrane and saline-soluble fractions of unstimulated platelets, and in the conditioned medium of thrombin-stimulated platelets. These studies demonstrate an intact 140 kD membrane-associated form of APP that is released on degranulation. Evidence that platelets synthesize at least one form of APP (APP751) was obtained by enzymatic amplification of specific mRNA using Polymerase Chain Reaction (PCR) and direct sequence analysis of PCR product. Processing of APP for release may occur via successive C-terminal truncations, and/or by the release and proteolysis of an intact membrane associated form. An intact form of APP in platelets provides a circulating substrate upon which proteases from many tissues may act to produce beta protein (AB) during pathologic conditions
— id: 9425, year: 1990, vol: 173, page: 1292, stat: Journal Article,

Gelsolin variant (Asn-187) in familial amyloidosis, Finnish type
Ghiso J; Haltia M; Prelli F; Novello J; Frangione B
1990 Dec 15;272(3):827-830, Biochemical journal
Familial amyloidosis, Finnish type (FAF), is an inherited form of systemic amyloidosis clinically characterized by cranial neuropathy and lattice corneal dystrophy. We have demonstrated that the protein subunit isolated from amyloid fibrils shows considerable sequence identity with gelsolin, an actin-binding protein. We have purified the amyloid subunit from a second case and further analysed different fractions from the previous one. Sequence analysis shows that, in both cases, the amyloid subunit starts at position 173 of the mature molecule; it has a heterogeneous N-terminus and contains one amino acid substitution, namely asparagine for aspartic acid, at position 15 (gelsolin residue 187), that is due to a guanine-to-adenine transversion corresponding to nucleotide-654 of human plasma gelsolin cDNA. The substitution maps in a fragment with actin-binding activity and is located in a repetitive motif highly conserved among species. Thus FAF is the first human disease known to be caused by an internal abnormal degradation of a gelsolin variant. We designate this variant of gelsolin-associated amyloidosis 'Agel Asn-187'
— id: 9426, year: 1990, vol: 272, page: 827, stat: Journal Article,

Binding of cystatin C to C4: the importance of sense-antisense peptides in their interaction
Ghiso J; Saball E; Leoni J; Rostagno A; Frangione B
1990 Feb;87(4):1288-1291, Proceedings of the National Academy of Sciences of the United States of America
Hydropathic anticomplementarity of amino acids indicates that peptides derived from complementary DNA strands may form amphiphilic structures and bind one another. By using this concept, we have found that the antisense peptide Ser-Tyr-Asp-Leu complementary to the segment Gln-Ile-Val-Ala-Gly (residues 55-59) in cystatin C (an inhibitor of cysteine proteases) is located at positions 611-614 of the beta chain of human C4, the fourth component of complement. Here we describe and characterize the specific interaction between cystatin C and C4 by ligand affinity chromatography and ELISA. Interaction between the two native proteins was mimicked on replacement of one of them with the corresponding sense-antisense peptide coupled to a carrier protein, and the binding was inhibited by these synthetic peptides in solution. Through the interaction with C4, cystatin C may play a regulatory role in complement activation that might be of particular importance at tissue sites where both proteins are produced by macrophages
— id: 9432, year: 1990, vol: 87, page: 1288, stat: Journal Article,

Amyloid in familial amyloidosis, Finnish type, is antigenically and structurally related to gelsolin
Haltia M; Ghiso J; Prelli F; Gallo G; Kiuru S; Somer H; Palo J; Frangione B
1990 Jun;136(6):1223-1228, American journal of pathology
Immunohistochemical studies of six patients with familial amyloidosis, Finnish type, showed that their amyloid deposits did not react with polyclonal antibodies against the amyloid proteins of other, established forms of systemic or cerebral amyloidosis. However, strong immunoreactivity was observed with rabbit antiserum raised against a low molecular weight purified amyloid subunit isolated from one of the patients. This immunoreactivity was abolished by absorption with the low molecular weight amyloid fraction. The amino terminal sequence of the amyloid protein subunit was homologous to gelsolin, an actin-modulating protein, and the amyloid deposits in tissues reacted with a monoclonal antibody against gelsolin. These studies show that the amyloid protein in familial amyloidosis, Finnish type, is not related to previously identified forms of amyloid, including prealbumin (transthyretin) variants, but represents a novel amyloidogenic protein related to gelsolin, a plasma and cytoplasmic protein
— id: 9429, year: 1990, vol: 136, page: 1223, stat: Journal Article,

Amyloid protein in familial amyloidosis (Finnish type) is homologous to gelsolin, an actin-binding protein
Haltia M; Prelli F; Ghiso J; Kiuru S; Somer H; Palo J; Frangione B
1990 Mar 30;167(3):927-932, Biochemical & biophysical research communications
Familial amyloidosis, Finnish type, is clinically characterized by cranial neuropathy and lattice corneal dystrophy. It is an autosomal dominant form of systemic amyloidosis with small deposits of congophilic material occurring in most tissues, particularly in association with blood vessel walls and basement membranes. Amyloid fibrils were extracted from the kidney of patient VUO, and rabbit antiserum raised against the 12 kDa purified amyloid subunit displayed strong immunohistochemical reactivity with the amyloid deposits. The amino terminal sequence of this 12 kDa amyloid protein (ATEVPVSWESFNNGD) showed homology with gelsolin (or actin depolymerizing factor), a 93 kDa plasma protein. The amyloid peptide is a degradation product, starting at position 173, of the gelsolin molecule
— id: 9431, year: 1990, vol: 167, page: 927, stat: Journal Article,

Mutation in gelsolin gene in Finnish hereditary amyloidosis
Levy E; Haltia M; Fernandez-Madrid I; Koivunen O; Ghiso J; Prelli F; Frangione B
1990 Dec 1;172(6):1865-1867, Journal of experimental medicine
Familial amyloidosis, Finnish type (FAF), is an autosomal dominant form of familial amyloid polyneuropathy. The novel amyloid fibril protein found in these patients is a degradation fragment of gelsolin, an actin-binding protein. We found a mutation (adenine for guanine) at nucleotide 654 of the gelsolin gene in genomic DNA isolated from five FAF patients. This site is polymorphic since the normal allele was also present in all the patients tested. This mutation was not found in two unaffected family members and 11 normal controls. The A for G transition causes an amino acid substitution (asparagine for aspartic acid) that was found at position 15 of the amyloid protein. The mutation and consequent amino acid substitution may lead to the development of FAF
— id: 9427, year: 1990, vol: 172, page: 1865, stat: Journal Article,

The complete amino acid sequence of toxin TsTX-VI isolated from the venom of the scorpion Tityus serrulatus
Marangoni S; Ghiso J; Sampaio SV; Arantes EC; Giglio JR; Oliveira B; Frangione B
1990 Oct;9(5):595-601, Journal of protein chemistry
The complete sequence of the toxin TsTX-VI from the venom of the scorpion Tityus serrulatus Lutz and Mello is presented. The sequence has been determined by automated Edman analysis of the reduced and carboxymethylated protein as well as of the resulting peptides, obtained from S. aureus protease and tryptic digestions. TsTX-VI is composed of 62 residues and has a calculated molecular weight of 6717. Homology studies with other scorpion toxins show that TsTX-VI is more similar to the Old World than to the North American scorpion toxins. The hydropathic index indicates that TsTX-VI is more hydrophobic than Ts-gamma. Toxicity studies carried out in mice demonstrate that i.v. injection of TsTX-VI is unable to evoke the usual symptoms induced by the typical neurotoxins of this venom, but only a generalized allergic reaction. These properties are important in clarifying the relationship between primary structure and biological function of scorpion toxins
— id: 9428, year: 1990, vol: 9, page: 595, stat: Journal Article,

Coexistence of Alzheimer's amyloid precursor protein and amyloid protein in cerebral vessel walls
Tagliavini F; Ghiso J; Timmers WF; Giaccone G; Bugiani O; Frangione B
1990 Jun;62(6):761-767, Laboratory investigation
Polyclonal antibodies to synthetic peptides homologous to amino acid residues 45-62, 597-624, and 676-695 of the predicted sequence of Alzheimer's amyloid precursor protein (APP) were used to investigate the site of origin of APP, and the relationship between APP and amyloid protein in Alzheimer's disease (AD) and hereditary cerebral hemorrhage with amyloidosis, Dutch type (HCHWA-D). Cortical sections as well as homogenates of isolated leptomeningeal and cortical microvessels from three patients with AD, two patients with HCHWA-D, and two nondemented controls were probed. In vessel extracts of both groups of patients and the controls, APP was detected as a set of proteins with electrophoretic mobility of 105 to 135 kilodaltons. In cortical sections of all subjects, APP immunoreactivity was found in leptomeningeal and cortical vessel walls. In patients with AD and HCHWA-D, APP and amyloid fibrils coexisted in the same vessels. Moreover, APP immunoreactivity was found in association with 50% of senile plaques in AD brains, but was not evidenced in parenchymal amyloid deposits in patients with HCHWA-D. These data suggest that the vascular system is a source of APP and that the processing of APP into insoluble fibrils in AD and HCHWA-D may take place in situ
— id: 9430, year: 1990, vol: 62, page: 761, stat: Journal Article,

ALZHEIMERS-DISEASE AND HEREDITARY (DUTCH-TYPE) CEREBRAL- HEMORRHAGE - COEXISTENCE OF AMYLOID PRECURSOR PROTEIN AND AMYLOID PROTEIN IN CEREBRAL VESSEL WALLS
Tagliavini, F; Ghiso, J; Timmers, WF; Giaccone, G; Bugiani, O; Frangione, B
1990 May;49(3):332-332, Journal of neuropathology & experimental neurology
— id: 31946, year: 1990, vol: 49, page: 332, stat: Journal Article,

INDIANA KINDRED OF GERSTMANN-STRAUSSLER-SCHEINKER DISEASE - ISOLATION OF LOW-MOLECULAR-WEIGHT PROTEIN FROM AMYLOID PLAQUE CORES
Tagliavini, F; Prelli, F; Ghiso, J; Bugiani, O; Farlow, MR; Ghetti, B; Frangione, B
1990 May-Jun;11(3):304-304, Neurobiology of aging
— id: 31942, year: 1990, vol: 11, page: 304, stat: Journal Article,

Alzheimer's disease amyloid precursor protein is present in senile plaques and cerebrospinal fluid: immunohistochemical and biochemical characterization
Ghiso J; Tagliavini F; Timmers WF; Frangione B
1989 Aug 30;163(1):430-437, Biochemical & biophysical research communications
The amyloid fibrils deposited in cerebral vessel walls and senile plaques in Alzheimer's disease are polymeric forms of a 4 kDa fragment produced by proteolysis of a putative precursor protein (APP). Using antibodies to several fragments of the deduced precursor, we were able to demonstrate the presence of APP in senile plaques, brain extracts and cerebrospinal fluid. Membrane-associated APP is detected as a group of 105-135 kDa proteins while soluble APP is predominantly 105 kDa, does not react with an anti C-terminal antibody, and is 10 kDa shorter than the membrane-bound APP. Amino terminal sequence of the tissue 105 kDa protein indicates that APP begins at residue 18 of the cDNA sequence. These findings imply that i) two forms of APP are detected: membrane-bound and secreted, and ii) APP can be processed in situ
— id: 9433, year: 1989, vol: 163, page: 430, stat: Journal Article,

Relation of the amyloid beta protein precursor to heparan sulfate proteoglycans
Gowda DC; Margolis RK; Frangione B; Ghiso J; Larrondo-Lillo M; Margolis RU
1989 May 19;244(4906):826-828, Science
— id: 9434, year: 1989, vol: 244, page: 826, stat: Journal Article,

Stroke in Icelandic patients with hereditary amyloid angiopathy is related to a mutation in the cystatin C gene, an inhibitor of cysteine proteases
Levy E; Lopez-Otin C; Ghiso J; Geltner D; Frangione B
1989 May 1;169(5):1771-1778, Journal of experimental medicine
Cystatin C is an inhibitor of lysosomal cysteine proteases and consists of 120 amino acids. A variant of cystatin C lacking the first NH2-terminal residues and having one amino acid substitution at position 68 forms amyloid deposits mainly in the walls of brain arteries, causing fatal strokes in Icelandic patients with familial cerebral hemorrhage secondary to a form of an autosomal dominant amyloidosis. To understand the molecular basis of the genetic defect, the gene encoding cystatin C was isolated from genomic DNA libraries made from normal tissue and the brain of an Icelandic patient with hereditary cerebral hemorrhage with amyloidosis (HCHWA-I). The data indicate that the cystatin C gene encodes a polypeptide of 146 amino acids, of which the first 26 correspond to a secretory peptide signal sequence. The gene contains two intervening sequences that interrupt the coding region at amino acids 55 and 93. Comparison with genes encoding salivary cystatins and kininogen proteins show sequence homology and conservation of exon-intron structure. Except for a mutation in the second exon (CAG instead of CTG in the normal gene, resulting in the substitution of glutamine for a leucine residue), the gene cloned from the brain of the Icelandic patient is identical to the normal cystatin C gene. Thus, HCHWA-I is the first familial type of amyloidosis related to a point mutation in a gene encoding for an inhibitor. The mutation in the structural gene encoding cystatin C appears to be the primary defect in this inherited disorder causing amyloid fibril formation and accumulation followed by cerebral hemorrhage
— id: 9435, year: 1989, vol: 169, page: 1771, stat: Journal Article,

HEREDITARY STROKE IN ICELANDIC PATIENTS WITH AMYLOID ANGIOPATHY IS RELATED TO A MUTATION IN THE CYSTATIN-C GENE, AN INHIBITOR OF CYSTEINE PROTEASES
Levy, E; Lopezotin, C; Ghiso, J; Geltner, D; Frangione, B
1989 Apr;37(2):A541-A541, Clinical research
— id: 31712, year: 1989, vol: 37, page: A541, stat: Journal Article,

Isolation of a sequence encoding human cystatin C. Conservation of exon-intron structure between members of the cysteine proteinase inhibitors superfamily
Ghiso J; Cowan N; Frangione B
1988 May;369 Suppl:205-208, Biological chemistry
The human cystatin C gene was cloned using a synthetic oligonucleotide predicted from a portion of its amino-acid sequence. The nucleotide sequence of the restriction fragment hybridizing with the oligonucleotide confirms the existence of one exon encoding amino acids 56-93 of human cystatin C and its relationship to kininogens. However the deduced amino-acid sequence differs in one position from the sequence of the cystatin C fragment deposited as amyloid fibrils in hereditary cerebral hemorrhage with amyloidosis of icelandic origin
— id: 9436, year: 1988, vol: 369 Suppl, page: 205, stat: Journal Article,

[Amyloidosis of the central nervous system]
Castano EM; Ghiso JA; Frangione B
1987 ;47(5):534-542, Medicina: organo de la Socieda Argentina de Investigation Clinica
— id: 57573, year: 1987, vol: 47, page: 534, stat: Journal Article,

Constitutive secretion of cystatin C (gamma-trace) by monocytes and macrophages and its downregulation after stimulation
Warfel AH; Zucker-Franklin D; Frangione B; Ghiso J
1987 Dec 1;166(6):1912-1917, Journal of experimental medicine
Cystatin C (gamma-trace) was found to be a constitutively secreted protein of isolated human monocytes and mouse peritoneal macrophages, as well as the histiocytic lymphoma cell lines U937, P388D.1, and J774. This proteinase inhibitor is not uniquely secreted by monocytes/macrophages, but was also identified in the conditioned media from several primary cells, including brain cells, and diverse established cell lines. In vitro treatment of resident mouse peritoneal macrophages with either LPS or IFN-gamma caused a downregulation in cystatin C secretion. Elaboration of this protein was also diminished by macrophages that had been stimulated by thioglycollate in vivo, and treatment of these cells with LPS led to further decline. It is suggested that, under some inflammatory conditions, downregulation of cystatin C may contribute to tissue pathology
— id: 9437, year: 1987, vol: 166, page: 1912, stat: Journal Article,

MONOCYTE MACROPHAGE SECRETION OF PROTEINASE-INHIBITOR CYSTATIN- C (GAMMA-TRACE) - ITS DOWN-REGULATION BY INFLAMMATORY STIMULI
Warfel, AH; Zuckerfranklin, D; Frangione, B; Ghiso, J
1987 Apr;35(3):A640-A640, Clinical research
— id: 31202, year: 1987, vol: 35, page: A640, stat: Journal Article,

In vitro formation of amyloid fibrils from two synthetic peptides of different lengths homologous to Alzheimer's disease beta-protein
Castano EM; Ghiso J; Prelli F; Gorevic PD; Migheli A; Frangione B
1986 Dec 15;141(2):782-789, Biochemical & biophysical research communications
Two synthetic peptides corresponding to the reported 28-residue sequence of Alzheimer's Disease beta-protein (SP28) and to residues 12-28 (SP17) were used to form fibrils in vitro. Synthetic fibrils bound Congo Red and closely resembled amyloid fibrils isolated from leptomeninges and senile plaques of Alzheimer's brain by electron microscopy. A polyclonal antiserum to SP28 specifically decorated both synthetic and native amyloid by colloidal gold immunoelectron microscopy. Amyloid fibrils isolated from tissue were insoluble on SDS-Polyacrylamide gels, and tended to aggregate while synthetic amyloid fibrils were completely solubilized, releasing only monomers of SP28 and SP17. Anti-SP28 immunostained cerebrovascular and plaque core amyloid, but not neurofibrillary tangles, in tissue section. Western blot analysis showed that anti-SP28 reacted with a 4 kDa band released from amyloid core-enriched preparations and leptomeninges. By contrast, a 16 kDa band corresponding to the tetramer of beta-protein was not recognized. These data suggest that as little as a 17 residue sequence of beta-protein may be required to form fibrils and that the complete sequence of the 4 kDa beta-protein may be important in determining insolubility and the formation of intermediate size polymers.
— id: 9438, year: 1986, vol: 141, page: 782, stat: Journal Article,

Amyloid fibrils in hereditary cerebral hemorrhage with amyloidosis of Icelandic type is a variant of gamma-trace basic protein (cystatin C)
Ghiso J; Jensson O; Frangione B
1986 May;83(9):2974-2978, Proceedings of the National Academy of Sciences of the United States of America
A gamma-trace variant protein is the major constituent of the amyloid fibrils in patients from Iceland with hereditary cerebral hemorrhage with amyloidosis. The protein consists of 110 residues and is similar to human urinary gamma-trace basic protein (or cystatin C) beginning at its 11th amino-terminal residue. It has an amino acid substitution (glutamine for leucine) at position 58 (position 68 in gamma-trace numbering), which is near the proposed active site of related proteins--namely, cysteine protease inhibitors and kininogens. It is postulated that a point mutation has occurred, leading to the production of an unusual protein that is abnormally degraded, bound, and/or precipitated. Alternatively, gamma-trace basic protein may be genetically polymorphic, and the variant described here may represent an as-yet-undiscovered isotype or an allelic form that is linked to, but not responsible for, the deposition disease. Our data on the structure of a gamma-trace variant protein suggests that its gene expresses a polyprotein precursor in which active peptides are flanked by basic amino acid residues that permit cleavage to liberate small internal peptides. It is likely that the nucleotide sequence coding for Arg-Xaa and Lys-Xaa repeated several times in the molecule may function as alternative splicing sites for mRNA processing.
— id: 9439, year: 1986, vol: 83, page: 2974, stat: Journal Article,

Hereditary cerebral amyloid angiopathy: the amyloid fibrils contain a protein which is a variant of cystatin C, an inhibitor of lysosomal cysteine proteases
Ghiso J; Pons-Estel B; Frangione B
1986 Apr 29;136(2):548-554, Biochemical & biophysical research communications
Hereditary Cerebral Hemorrhage With Amyloidosis is an autosomal dominant form of amyloidosis restricted to the cerebral vasculature. We have previously demonstrated that the amyloid protein subunit is similar to Cystatin C (or gamma-trace), an inhibitor of lysosomal cysteine proteinases, and homologous to kininogens. High pressure liquid chromatography tryptic fingerprint analysis was developed to distinguish Cystatin C from the amyloid protein. Moreover, we isolated and sequenced tryptic peptides in which the differences were detected. The data prove that the amyloid protein is 10 residues shorter than Cystatin C and has one amino acid substitution at residue 58.
— id: 9440, year: 1986, vol: 136, page: 548, stat: Journal Article,

Association of human lambda light chain V/J/C segments: serologic analysis and primary structure of the lambda VI Bence Jones protein THO
Ghiso J; Solomon A; Frangione B
1986 Jan;136(2):716-719, Journal of immunology
The complete amino acid sequence of the human monoclonal lambda VI light chain Bence Jones protein THO was determined. We have found it to have remarkable similarities to the previously sequenced lambda VI Bence Jones protein SUT. Immunochemical analyses demonstrated that both lambda VI chains belong to a V lambda VI sub-subgroup. The 98-residue V gene-encoded segments of proteins THO and SUT are closely homologous and are distinguished from other lambda VI chains by a one-residue deletion at the V-J recombination site. Proteins THO and SUT have identical 13-residue J segments and therefore are encoded by the same J lambda gene. Further, both proteins have identical 105-residue C regions that by sequence represent products of the C lambda 3 (Kern-, Oz+) gene. The primary structure and serologic properties of proteins THO and SUT imply at the protein level of association between certain types of V lambda, J lambda, and C lambda segments.
— id: 9441, year: 1986, vol: 136, page: 716, stat: Journal Article,

HEREDITARY CEREBRAL ANGIOPATHY WITH AMYLOIDOSIS - AMYLOID FIBERS CONTAIN A VARIANT OF CYSTATIN-C, AN INHIBITOR OF LYSOSOMAL PROTEASE CYSTEINE
GHISO, J; ESTEL, BP; JENSSON, O; FRANGIONE, B
1986 OCT ;46(5):530-530, Medicina: organo de la Socieda Argentina de Investigation Clinica
— id: 41331, year: 1986, vol: 46, page: 530, stat: Journal Article,

AMYLOID FIBRILS IN HEREDITARY CEREBRAL-HEMORRHAGE WITH AMYLOIDOSIS IS A VARIANT OF GAMMA TRACE (CYSTATIN-C)
GHISO, J; JENSSON, O; FRANGIONE, B
1986 MAR ;73(3):309-309, Acta neurologica Scandinavica
— id: 41458, year: 1986, vol: 73, page: 309, stat: Journal Article,

HEREDITARY CEREBRAL AMYLOID ANGIOPATHY - THE AMYLOID FIBRILS CONTAIN A PROTEIN WHICH IS A VARIANT OF CYSTATIN-C, AN INHIBITOR OF LYSOSOMAL CYSTEINE PROTEINASES
GHISO, J; JENSSON, O; FRANGIONE, B
1986 APR ;34(2):A652-A652, Clinical research
— id: 41409, year: 1986, vol: 34, page: A652, stat: Journal Article,

INTERACTION OF CYSTATIN-C WITH THE COMPLEMENT-SYSTEM
SABALL, E; ROSTAGNO, A; GHISO, J
1986 FEB ;46(5):530-531, Medicina: organo de la Socieda Argentina de Investigation Clinica
— id: 73965, year: 1986, vol: 46, page: 530, stat: Journal Article,