Abeta-degrading endopeptidase,neprilysin, in mouse brain: synaptic and axonal localization inversely correlating with Abeta pathology

Metabolism of amyloid-beta peptide (Abeta) is closely associated with the pathology and etiology of Alzheimer's disease (AD). Since neprilysin is the only rate-limiting catabolic peptidase proven by reverse genetics to participate in Abeta metabolism in vivo, we performed detailed immunohistochemical analysis of neprilysin in mouse brain using neprilysin-deficient mice as a negative control. The aim was to assess, at both the cellular and subcellular levels, where Abeta undergoes neprilysin-dependent degradation in the brain and how neprilysin localization relates to Abeta pathology in amyloid precursor protein (APP)-transgenic mice. In hippocampus, neprilysin was present in the stratum pyramidale and stratum lacunosum-moleculare of the CA1-3 fields and the molecular layer of the dentate gyrus. Confocal double immunofluorescence analyses revealed the subcellular localization of neprilysin along axons and at synapses. This observation suggests that after synthesis in the soma, neprilysin, a type II membrane-associated protein, is axonally transported to the terminals, where Abeta degradation is likely to take place. Among various cell types, GABAergic and metabotropic glutamate 2/3 receptor-positive neurons but not catecholaminergic or cholinergic neurons, expressed neprilysin in hippocampus and neocortex, implying the presence of a cell type-specific mechanism that regulates neprilysin gene expression. As expected, Abeta deposition correlated inversely with neprilysin expression in TgCRND8 APP-transgenic mice. These observations not only support the notion that neprilysin functions as a major Abeta-degrading enzyme in the brain but also suggest that down-regulation of neprilysin activity, which may be caused by aging, is likely to elevate local concentrations of Abeta at and around neuronal synapses.     

Contribution of HLA class II genes to susceptibility in achalasia

Abstract: Achalasia is a motor disorder of the esophagus resulting in functional obstruction. The cause of the lesion is unknown although genetic and immunologic factors have been suggested. An association with serological HLA epitopes has been previously reported. In this study, we have further examined this HLA class II association with susceptibility to achalasia by DNA based methods. Achalasia patients ( n =40) and healthy controls ( n = 275), all Caucasians and unrelated, were included in the analysis. The strongest associations were with HLA-DQAl*0101 and two HLA-DQαβ heterodimers having their a chain encoded by this allele. Moreover, relative risk was significantly higher in DQAl*0101 homozygotes as compared to heterozygotes and results suggested that DQB1*02 may have a protective role.     

Heterogeneity of structural abnormalities in the 7q31.3 approximately q34 region in myeloid malignancies

Abnormalities in the long arm of chromosome 7 are a frequent chromosomal aberration in myeloid disorders. Most studies have focused on the analysis of del(7q), demonstrating the presence of several minimal deleted regions in 7q22 approximately q31. By contrast, few studies in myeloid disorders have been devoted to the analysis of translocations, either balanced or unbalanced, involving 7q. In this study, we used fluorescence in situ hybridization (FISH) to characterize the 7q31.3 approximately q34 region (markers D7S480-D7S2227) in patients with deletion or translocation of 7q. A total of 910 cases of myeloid disorders were studied by conventional cytogenetics. Fifty-eight (6%) patients had structural aberrations of 7q. FISH studies were carried out in the 27 patients with involvement of 7q31 approximately q34: 14 cases had an acute myelogenous leukemia and 13 cases had a myelodysplastic syndrome. FISH analysis revealed the existence of high complexity in the 7q31.3 approximately q34 region in patients with unbalanced translocations. No breakpoints in 7q31.3 approximately q34 were found in the cases with deletion or balanced translocation. Nevertheless, studies of unbalanced translocations showed several breakpoints in markers D7S480-D7S2227, which delineate a commonly altered region. The complexity of 7q rearrangements suggests that a synergy of different genetic factors, rather than the alteration of a single tumor suppressor gene, could be involved in the pathogenesis of del(7q) in myeloid disorders.     

Ca2+ permeability of the plasma membrane induced by rotavirus infection in cultured cells is inhibited by tunicamycin and brefeldin A

Rotavirus infection of cultured cells induces a progressive increase in plasma membrane permeability to Ca2+. The viral product responsible for this effect is not known. We have used tunicamycin and brefeldin A to prevent glycosylation and membrane traffic and study the involvement of viral glycoproteins, NSP4 and/or VP7, in rotavirus-infected HT29 and MA104 cells. In infected cells, we observed an increase of plasma membrane Ca2+ permeability and a progressive depletion of agonist-releasable ER pools measured with fura 2 and an enhancement of total Ca2+ content measured as 45Ca2+ uptake. Tunicamycin inhibited the increase in membrane Ca2+ permeability, induced a depletion of agonist-releasable and 45Ca2+-sequestered pools. Brefeldin A inhibited the increase of Ca2+ permeability and the increase in 45Ca2+ uptake induced by infection. We propose that the glycosylated viral product NSP4 (and/or VP7) travels to the plasma membrane to form a Ca2+ channel and hence elevate Ca2+ permeability.     

Hostility and sex differences in the magnitude, duration, and determinants of heart rate response to forehead cold pressor: Parasympathetic aspects of risk

Recent models hypothesize that hostility confers increased risk of CHD through weaker parasympathetic dampening of cardiovascular reactivity (CVR). We tested this possibility using the forehead cold pressor task, a common maneuver which elicits the “dive reflex” characterized by a reflexive decrease in HR presumably through cardiac-parasympathetic stimulation. Participants were initially chosen from the outer quartiles of a sample of 670 undergraduates screened using the hostility subscale of the Aggression Questionnaire ([Buss, A.H., Perry, M., 1992. The Aggression Questionnaire. Journal of Personality and Social Psychology, 63, 452-459.]). The final sample of 80 participants was evenly divided between men and women and high and low hostility. Following a 10-min baseline, participants underwent a 3-min forehead cold pressor task. The task evoked a significant HR deceleration that was mediated by PNS activation, as assessed by respiratory sinus arrhythmia (RSA). Replicating prior research, men displayed greater decrease in HR. More important, low hostiles maintained larger HR deceleration over time compared to high hostiles although the autonomic basis for this effect was unclear. The findings broaden understanding of hostility and sex-related cardiovascular functioning and support the task as a method for evoking PNS-cardiac stimulation.     

Systemic infections in three infants due to a lactose-fermenting strain ofSalmonella virchow

Three previously healthy children developed gastroenteritis which led within a few days to systemic infections, two cases of bacteremia and one of meningitis. A lactose-fermentingSalmonella virchow strain was isolated from cerebrospinal fluid and blood cultures. In one case, this strain was also isolated from stool cultures. All the children had been fed the same milk formula. There was no other relationship between them. The batch of dried-milk formula was confirmed as the source of the infection by isolation of an identical lactose-fermentingSalmonella virchow strain by the Centro Nacional de Alimentación.     

Anti-HBs antibody. A new immunoenzymatic detection. Preliminary results (author's transl)]

The detection of anti-HBs antibody has been carried out using a new technique where a system for the detection of HBs antigen in the solid phase is adopted to demonstrate the presence of anti HBs antibody by inhibition of the immunoenzymatic reaction. The sensitivity of the method has been compared: with that of passive haemagglutination and with that of radioimmunoassay in liquid phase, for human serum obtained from 10 chronic antibody carriers and also serum from 1 rabbit, with that of solid phase radioimmunoassay for 91 positives serums. The method appears: in the range of sensitivity of passive haemagglutination and radioimmunoassay in the liquid phase, less sensitive than that of solid phase radioimmunoassay, far superior to that of electrosyneresis.