Dmitry Goldgaber

Mutations in familial Creutzfeldt-Jakob disease and Gerstmann-Sträussler-Scheinker's syndrome

A host protein encoded by the gene specifying the scrapie amyloid precursor affects pathogenesis of the transmissible spongiform encephalopathies: Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinkers syndrome (GSS), and kuru in man, and scrapie in animals. We found a mutation in this gene of two patients with CJD from one family and a second mutation in the same gene in three patients with GSS from another family. The mutation in two related familial CJD patients changed glutamine in position 200 tolysine. This mutation was absent in other individuals including unrelated patients with familial CJD, sporadic CJD, and GSS. The other mutation in three GSS patients changed proline in position 102 to leucine, the same mutation described recently in some GSS families. We did not find it in six unaffected relatives of the GSS patients or in other individuals including sporadic and familial CJD patients. A rare insertion described earlier in one CJD family was also absent in all tested individuals.

Mutant ubiquitin expressed in Alzheimer’s disease causes neuronal death1

Ubiquitin‐B+1 (UBB+1) is a mutant ubiquitin that accumulates in the neurones of patients with Alzheimers disease (AD). Here we report on the biochemical and functional differences between ubiquitin and UBB+1 and the effect of the mutant protein on neuronal cells. UBB+1 lacks the capacity to ubiquiti‐nate, and although it is ubiquitinated itself, UBB+1 is not degraded by the ubiquitin‐proteasomal system and is quite stable in neuronal cells. Overexpression of UBB+1 in neuroblastoma cells significantly induces nuclear fragmentation and cell death. Our results demonstrate that accumulation of UBB+1 in neurones is detrimental and may contribute to neuronal dysfunction in AD patients.—de Vrij, F. M. S., Sluijs, J. A., Gregori, L., Fischer, D. F., Hermens, W. T. J. M. C., Goldgaber, D., Verhaagen, J., van Leeuwen, F. W., Hol, E. M. Mutant ubiquitin expressed in Alzheimers disease causes neuronal death. FASEB J. 15, 2680–2688 (2001)

Cdc2 phosphorylation of nucleolin demarcates mitotic stages and Alzheimer’s disease pathology ☆

Nucleolin is a major multifunctional nuclear phosphoprotein that is phosphorylated by Cdc2 kinase in mitosis and that participates in a number of cellular processes. The monoclonal antibody TG-3 generated against neurofibrillary tangles (NFT) found in Alzheimers disease (AD) is highly specific for mitotic cells in culture. We here demonstrate that phosphorylation of nucleolin by Cdc2 kinase generates the TG-3 epitope. The unique pool of TG-3 immunoreactive nucleolin appears abruptly during the prophase. It is associated with chromosomes through the metaphase and it gradually disappears during separation of chromosomes and exit from mitosis. In the brain, nucleolin was localized not only to nuclei but also to neuronal cytoplasm, and it is a marker for early NFT. In patients with AD, Cdc2 phosphorylated nucleolin was present in NFT. These findings suggest that phosphorylation of nucleolin by Cdc2 kinase is a critical event and the point of convergence of two distinct pathways, mitosis and neurodegeneration.

Clinical and molecular genetic study of a large German kindred with Gerstmann‐Straussler‐Scheinker syndrome

We have verified, by full open reading frame sequencing, the presence of an amino-acid-altering mutation in codon 102 of the scrapie amyloid protein gene in three affected members of a large and well-documented German family with experimentally transmitted Gerstmann-Straussler-Scheinker syndrome. In addition, we identified the mutation by partial sequencing or DNA restriction enzyme analysis in three of 12 presently healthy family members with an affected parent, and none of 12 members without an affected parent. Thus, a total of six of 15 family members at risk for the disease (including the three established cases) had the same codon 102 mutation, a proportion consistent with the autosomal dominant inheritance pattern of disease expression. It is undetermined whether the mutation influences susceptibility to infection by an exogenous agent or is itself a proximate cause of disease.

Alzheimer β/A4-Amyloid Precursor Protein: Evidence for Putative Amyloidogenic Fragment

Abstract: Recombinant baculovirus was used to overexpress human Alzheimer β/A4‐amyloid precursor protein (APP) in Spodoptera frugiperda (Sf9) cells. Lysates of these cells were then analyzed for the presence of carboxyl‐terminal fragments of APP by an immuno‐blotting assay using either an antibody against the APP cytoplasmic domain (rabbit anti‐human 695APP645–694 or an antibody against the amino terminus of β/A4‐amyloid (rabbit anti‐human 695APP586–606). Anti‐human 695APP645–694 identified APP holoprotein, a 25‐kDa species, and a prominent group of carboxyl‐terminal fragments of 17, 16, and 14 kDa, whereas anti‐human 695APP586–606 identified APP holoprotein and a single prominent low‐molecular‐mass protein species comigrating with the 17‐kDa carboxyl‐terminal fragment identified by anti‐human 695APP645–694. No immunoreactive species was detected at these molecular mass positions when either antibody was used for analysis of lysates of either uninfected Sf 9 cells or Sf 9 cells infected with wild‐type Autographa californica baculovirus. For each antibody, specific immunoreactivity was abolished by preabsorption with the corresponding peptide immunogen. The incorporation of a β/A4‐amyloid amino‐terminal epitope into a 17‐kDa fragment of APP suggests that, in the baculoviral overexpression system, the electrophoretic microheterogeneity of APP carboxyl‐terminal fragments is due, at least in part, to alternative proteolysis of APP. If such carboxyl‐terminal fragments of APP containing an intact β/A4‐amyloid domain are produced in human brain, then they may represent intermediates in the conversion of APP to deposited β/A4‐amyloid. The identification of potentially amyloidogenic fragments in recom‐binantly engineered Sf 9 cells may provide a useful experimental system for determination of alternative sites of APP proteolysis and investigation of the processing mechanisms involved.

Stress and well-being in mothers of young children 11 years after the Chornobyl nuclear power plant accident"

BACKGROUND This paper examines the association between exposure to the Chornobyl nuclear power plant explosion and the psychological and physical well-being of mothers with young children. The study also examines whether exposure to Chornobyl increased the vulnerability of mothers to subsequent economic and social stress, and thus represents a unique test of the stress-vulnerability model in a non-Western setting. METHOD The sample consisted of mothers evacuated from the contamination zone surrounding the plant (evacuees) and mothers who had never lived in a radiation-contaminated area (controls). In addition to exposure status, the interview obtained data on perceived economic stress, social stress and stress moderators. The dependent variables were measured by the SCL-90 global severity index (GSI), perceived physical health and number of days unable to work due to illness. RESULTS Overall, evacuees reported fewer stressors and greater personal and social resources than control mothers. Nevertheless, evacuees scored higher on the GSI, reported lower perceived physical health and took more sick days relative to control mothers, even after controlling for demographic factors, stressors and stress moderators. Tests of interaction effects were not statistically significant. CONCLUSIONS The findings confirmed that married women with young children evacuated to Kyiv following the Chornobyl nuclear power plant explosion reported significantly poorer psychological and perceived physical health than controls 11 years later. Although perceived social and economic adversities also affected these outcomes, there was no evidence that exposure to the Chornobyl accident increased the vulnerability of mothers to these stressors, giving support to the additive burden model of stress.

Amyloidogenic and anti-amyloidogenic properties of recombinant transthyretin variants

Most transthyretin (TTR) mutations lead to TTR amyloid depositions in patients with familial amyloidotic polyneuropathy and familial amyloidotic cardiomyopathy. However, though an amyloidogenic protein itself, TTR inhibits aggregation of Alzheimers amyloid beta protein (Aβ) in vitro and in vivo. The pathogenic relationship between two amyloidogenic processes remains unclear. To understand how TTR mutations influence the ability of TTR to inhibit Aβ amyloidosis, forty-seven recombinant TTR variants were produced and analyzed. We showed that all recombinant proteins formed tetramers and were functional in thyroxine binding. Acid denaturation at pH 3.8 resulted in aggregation and fibril formation of all TTR variants. However, only TTR G42 and TTR P55 formed fibrils at pH 6.8. Most TTR variants bound to Aβ and inhibited Aβ aggregation in vitro. TTR variants S64, A71, Q89, V107, H114 and I122 revealed decreased binding to Aβ and decreased inhibition of Aβ aggregation. Only TTR G42 and TTR P55 completely failed to bind Aβ and to inhibit Aβ aggregation. We suggest that TTR variants characterized by decreased binding to Aβ or by decreased inhibition of Aβ aggregation in vitro may contribute to Aβ amyloid formation in vivo. These TTR variants might be important targets for epidemiological studies in Alzheimers disease.

Creutzfeldt-Jakob disease and kuru patients lack a mutation consistently found in the Gerstmann-Sträussler-Scheinker syndrome

We and others have recently reported that patients with the Gerstmann-Sträussler-Scheinker syndrome have a mutation at codon 102 of the gene coding for amyloid protein that accumulates in this disease. We report here that this mutation was not found in 5 familial and 27 sporadic cases of Creutzfeldt-Jakob disease or in 3 patients with kuru, so that although this mutation may be responsible for amyloidogenesis and transmissibility in Gerstmann-Sträussler-Scheinker syndrome, it cannot be the only cause of human spongiform encephalopathy.

Isolation and propagation of nephropathia epidemica virus in bank voles.

Three strains of nephropathia epidemica (NE) virus were isolated from lung tissues of bank voles (Clethrionomys glareolus) and a grey-sided vole (C. rufocanus) trapped in Västerbotten county, Sweden. Two of these isolates were serially passaged in seronegative laboratory-bred bank voles. Experimentally infected animals developed a subclinical infection characterized by virus persistence, particularly in lung tissue. Attempts to infect other species of colonized rodents with NE virus and to isolate NE virus from acute phase patient blood were unsuccessful. The serial propagation of NE virus in colonized bank voles provides opportunities to study experimental infection in its reservoir rodent host.

Ukrainian application of the Children's Somatization Inventory: psychometric properties and associations with internalizing symptoms.

This paper examines the psychometric properties of the Childrens Somatization Inventory (CSI) in 600 10–12-year old children in Kyiv, Ukraine, replicating and extending the original findings from a sample in Nashville, Tennessee (J. Garber et al. 1991). The Kyiv children had significantly lower CSI total scores and reported significantly fewer symptoms than the American children. The Kyiv mothers, however, reported significantly more somatization symptoms in their children than did the American mothers. A factor analysis of the childrens data yielded four similar factors encompassing pseudoneurologic, cardiovascular, gastrointestinal, and pain/weakness symptoms. Consistent with the findings from the Nashville study, the CSI was significantly related to the childrens self-reports of health and depressive and anxiety symptoms and to maternal reports of child depression and anxiety symptoms. In addition, although more children with the highest CSI scores (25+) reported various illness experiences than those with 0–1 symptoms, no differences were found in the school absentee records. Thus, the results were congruent with the findings of the Nashville study, indicating that the CSI reliably measured somatization in this Ukrainian sample.