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Biomedical subjects

B Weber

Publications and source records attributed to B Weber.

At least 37 records · Page 2Linked to original sources

Optical imaging of the spatiotemporal dynamics of cerebral blood flow and oxidative metabolism in the rat barrel cortex.

Oxidative metabolism and cerebral blood flow (CBF) are two of the most important measures in neuroimaging. However, results from concurrent imaging of the two with high spatial and temporal resolution have never been published. We used flavoprotein autofluorescence (AF) and laser speckle imaging (LSI) in the anaesthetized rat to map oxidative metabolism and CBF in response to single vibrissa stimulation. Autofluorescence responses reflecting oxidative metabolism demonstrated a fast increase with a delay of 0.1 s. The sign-reversed speckle contrast reflecting CBF started to rise with a delay of 0.6 s and reached its maximum 1.4 s after the stimulation offset. The fractional signal changes were 2.0% in AF and 9.7% in LSI. Pixelwise modelling revealed that CBF maps spread over an area up to 2.5-times larger than metabolic maps. The results provide evidence that the increase in cerebral oxidative metabolism in response to sensory stimulation is considerably faster and more localized than the CBF response. This suggests that future developments in functional imaging concentrating on the metabolic response promise an increased spatial resolution.

Animals↗

Salt reduction in municipal sewage allocated for reuse: the outcome of a new policy in Israel.

Israel is presently reusing about 70% of its sewage and the buildup of salts in soils has been detected in some areas of the country. Prevention of sewage salt enrichment is one of the most immediately available solutions. The discharge of brines to municipal sewers is now prohibited and they are discharged to the sea. Discharge of brines to the sea has increased from 580,000 m3 in 1999 to more than 700,000 m3 in 2002. These salts are thus definitively taken out of the reuse cycle, year after year. The concentration of salts in industrial effluents is presently limited by the national water law and a unique regulation limiting the Boron and Sodium content in detergents has been implemented. Israeli industry has undergone a radical change in recent years. Many factories and all the hospitals have adopted K or Ca for softening and neutralization, while others have shifted to reverse osmosis. The concentration of Cl and Na in the sewage reaching treatment plants is declining. The addition of Cl to sewage has dropped from 120 mg/l to 70 mg/l. The average Boron concentration in sewage has dropped from 0.6 mg/l to 0.3 mg/l and should reach 0.2 mg/l by 2008.

Conservation of Natural Resources↗

Water reuse for irrigation from waste water treatment plants with seasonal varied operation modes.

Irrigation periods are usually limited to vegetation periods. The quality requirements for treated wastewater for disposal and for reuse are different. The reuse of water for irrigation allows partly the reuse of the wastewater's nutrients (N and P). Outside the irrigation period the water must be treated for disposal, thus nutrient removal is often required in order to avoid detrimental effects on the receiving surface water body. Only wastewater treatment plants with different operation modes for different seasons can realise these requirements. The nitrification is the most sensitive biological process in the aerobic wastewater treatment process. At low water temperatures the nitrifying bacteria need several weeks to re-start full nitrification after periods without NH4-removal. Therefore it is necessary to develop options for waste water treatment plants which allow a fast re-start of the nitrification process. Based on theoretical considerations and computer simulations of the activated sludge treatment process, one possibility for implementing a wastewater treatment plant with different seasonal operation modes is evaluated.

Agriculture↗

Reduction of the diagnostic window in three cases of human immunodeficiency-1 subtype E primary infection with fourth-generation HIV screening assays.

BACKGROUND AND OBJECTIVES: The influence of genetic variability on the sensitivity of serological diagnosis of human immunodeficiency virus (HIV) infection has, to date, been poorly investigated. The aim of the present study was to assess whether fourth-generation assays for the combined detection of HIV antigen and antibodies to HIV (anti-HIV) permit a reduction of the diagnostic window in comparison to third-generation antibody enzyme immunoassays (EIAs), which so far have shown a poor sensitivity for detection of HIV-1 non-subtype B primary infections. MATERIALS AND METHODS: Three patients with primary HIV-1 subtype E (CRF01-AE) infection were tested with different third- and fourth-generation assays, stand-alone HIV antigen (Ag) EIAs and reverse transcription-polymerase chain reaction (RT-PCR). Additionally, virus lysates from HIV-1 Group M and O and HIV-2, at concentrations of p24 Ag close to the detection limit of licensed HIV Ag EIAs, were investigated with fourth-generation EIAs and HIV Ag EIAs. RESULTS: In the first blood donor, the most sensitive fourth-generation assay detected HIV-1 infection 11 days earlier than five of the eight third-generation antibody assays. Fourth-generation EIAs, with a high sensitivity for HIV antigen, detected HIV-1 subtype E infection simultaneously or 4 days later than HIV-1 RT-PCR on pooled samples. Low concentrations of virus lysates of different HIV-1 subtypes A-H and group O, tested positive with fourth-generation EIAs, with a high sensitivity of the antigen-detection module. CONCLUSIONS: Fourth-generation EIAs, especially those with a high sensitivity for HIV-1 p24 antigen, reduce the diagnostic window for primary HIV-1 subtype E infection in comparison with third-generation antibody-screening assays. These preliminary data from seroconversions and virus lysates indicate that the genetic diversity of HIV-1 does not represent a major challenge for the most sensitive EIAs of this new assay generation.

Antigenic Variation↗

Double stranded RNA virus in South African Trichomonas vaginalis isolates.

AIMS: To screen Trichomonas vaginalis isolates from South Africa for the presence of a small double stranded RNA virus designated T vaginalis virus (TVV). METHODS: TVV was detected by simultaneous extraction of DNA and RNA, and its presence confirmed by electron microscopy and nuclease digestions. RESULTS: TVV was detected in 59 of 72 (81.9%) isolates. CONCLUSIONS: These results indicate a possible higher infection rate of South African T vaginalis isolates by the double stranded RNA virus than has been reported for isolates elsewhere.

Animals↗

White matter glucose metabolism during intracortical electrostimulation: a quantitative [(18)F]Fluorodeoxyglucose autoradiography study in the rat.

[(18)F]Fluorodeoxyglucose (FDG) autoradiography was used to analyze the effects of intracortical electrostimulation on local cerebral metabolic rate for glucose (LCMRglu). The hindleg area in rat brains was electrically stimulated with different frequencies (0, 0.1, 0.25, 0.5, 0.75, 1 stimulus trains per second, two animals per condition). The major result was a strong positive correlation between stimulation frequency and LCMRglu in the callosal fibers originating in the stimulated cortical area. At the highest stimulation frequency callosal LCMRglu was 50.01 micromol/min/100 g compared to 27.87 micromol/min/100 g at baseline. LCMRglu in gray and white matter control areas was stable across conditions. Direct injection of FDG in the stimulated cortex failed to produce increased callosal uptake, excluding the possibility that FDG uptake in the corpus callosum is related to axonal diffusion. Although several previous autoradiographic studies have demonstrated alterations in LCMRglu in white matter, correlations between neural activity and LCMRglu have never been explicitly addressed. Changes in white matter metabolism most likely reflect changes of electrical fiber activity and thus the presented results bear important implications for brain imaging studies.

Animals↗

Seroprevalence of herpes simplex virus types 1 and type 2 in the Frankfurt am Main area, Germany.

Herpes simplex virus (HSV) types 1 and 2 are widespread human infectious agents that are responsible for persistent and latent infections. HSV type 2 (HSV-2) infection is usually transmitted sexually, while HSV type I (HSV-1) is commonly acquired by saliva contact during childhood. In a retrospective study, sera from more than 4,800 patients were analyzed for HSV type-specific IgG antibodies. In people older than 15 years, the seroprevalence of HSV-1 showed no statistically significant discrepancy between the control group (76.3% in females and 75.2% in males), HIV-infected patients (82.8% in females and 84.3% in males), and organ transplant (OTX) recipients (90.3% in females and 86.3% in males) (P>0.05). Age-related analysis of the control group showed that there is an age-dependent increase of HSV-1 seroprevalence in both sexes, reaching its peak in those aged 40 years and older (women 85.4%, men 82.8%). The only age group in which there is a significantly higher seropositivity rate in women than in men is in those aged between 15 and 39 years, with 70.8% versus 63.7% (P<0.05). As with HSV-1, there is an age-related increase of the HSV-2 seroprevalence; however, this increase starts later in life, with the onset of sexual activity. The HSV-2 prevalence across all age groups was highest in female prostitutes (78.0%) and among HIV-infected patients (women 64.1%, men 54.3%); this contrasts with the control group (overall women 17%, men 12.5%; those above 15 years of age, women 18%, men 13.8%) and the OTX patients (women 22.6%, men 9.8%). In the control group the rate of positivity increases with age and peaks in the group older than 40 years (24.2% in women and 16.2% in men). In females the seroprevalence is always elevated compared with males. The data presented show that female sex and older age are independent predictors of HSV-2 seropositivity, while immunosuppression is not. Our additional data show no evidence of a statistically significant humoral HSV-1/HSV-2 cross-immunity. People with HSV-1 serum antibodies have no lower risk of HSV-2 seropositivity than those lacking antibodies to HSV-1. The same is true when investigating HSV-1 seroprevalence rates in HSV-2-seropositive or -negative individuals, retrospectively.

Adolescent↗

[Benzodiazepines and benzodiazepine-like anxiolytics and hypnotics. The implausible contraindication of closed angle glaucoma].

Official drug information materials mention angle-closure glaucoma as an absolute or relative contraindication for benzodiazepines, albeit with some unexplained variability. A systematic review of the literature reveals that this contraindication seems to be based on only one published case while 22 other investigations, some of them controlled, tend to favor that benzodiazepines reduce intraocular pressure. The common 1990-2001 database of the Drug Commission of the German Medical Profession and the Federal Institute for Drugs and Medical Devices contained one spontaneous report of glaucoma related to a benzodiazepine-like hypnotic.

Anti-Anxiety Agents↗

Rapid method for arrayed investigation of IgE-reactivity profiles using natural and recombinant allergens.

BACKGROUND: The availability of increasing numbers of purified natural and recombinant allergens offer the possibility for component-resolved characterization of IgE binding. To make use of this potential, fast and simple methods with high capacity have to be developed. METHODS: A laboratory multiscreen device was used in an innovative two-dimensional approach. In the first step, natural and recombinant allergens were immobilized onto the membrane using the sample chambers as application mask and, after blocking and rotating the membrane through 90 degrees, the same device was used to apply and incubate sera of allergic patients. Enzyme-linked immunoassay (ELISA) quantification of specific IgE was performed for purposes of comparison. RESULTS: Proteins were most efficiently bound onto nitrocellulose in 20 mM sodium hydroxide (NaOH). Up to 45 proteins or extracts could be investigated with a maximum of 45 sera in a single application, resulting in a resolution of 2,025 spots on one membrane with a size comparable to a standard Western blot. A high correlation for IgE-binding between natural and recombinant allergens was observed. Development of the membrane resulted in very evenly distributed square patterns. The results corresponded with the conventional ELISA measurements of specific IgE. CONCLUSIONS: The innovative usage of a standard incubation device for both application of proteins as well as screening of sera provides a simple high throughput method for the characterization of IgE binding to allergens. The results are important for component resolved diagnosis of allergy by means of fast monitoring of IgE- and IgG-reactivity spectra. Recombinant allergens may be used as targets for these purposes.

Allergens↗

Bias in computerized neuropsychological assessment of depressive disorders caused by computer attitude.

OBJECTIVE: Psychiatric patients are increasingly confronted to computerized psychological and psychopathological assessment. Patients' attitude to computers was reported to affect acceptance of computerized assessment. METHOD: In 78 psychiatric in-patients neuropsychological impairment was examined following admission on an open ward by conventional as well as computerized memory and attention tasks. Besides psychopathological assessment, self ratings of computer attitude and acceptance of the computerized assessment were completed. RESULTS: A more negative attitude to computers was found to be significantly correlated to higher nervousness in patients' self report (R=0.38, P=0.0005) as well as to poorer results of computerized attention tasks (R=0.39, P=0.0007). Particularly in patients with depressive disorders computer attitude could be shown to explain 39% of the variance of attention performance. CONCLUSION: Results indicate a significant effect of negative computer attitude on acceptance and thus reliability of computerized examination, resulting in a bias in computerized attention-related assessment in patients with depressive disorders.

Adolescent↗

Food-grade delivery system for controlled gene expression in Lactococcus lactis.

A food-grade system for the delivery of desired genes to Lactococcus lactis, their inducible expression, and their transfer to related strains was established. Based on the thermosensitive pG(+)host replicon, two types of plasmid vectors were constructed which contained sections of either the chromosomal leu operon of L. lactis or the tel operon from the lactococcal sex factor. Genes cloned into the leu or tel sequences of these vectors were delivered to the homologous regions of the chromosome or the sex factor through two single crossovers, leading to integration of the recombinant plasmids and subsequent excision of the vector portions. Inducible transcription of integrated genes was achieved by using the nisin-controlled expression (NICE) system. To establish the signal transduction genes nisRK in L. lactis, the vectors pLNG1363 (targeted to the chromosome) and pUK500 (targeted to the sex factor) were constructed. Fusions of six different peptidase genes (pep) from Lactobacillus delbrueckii with the nisin-inducible promoter P(nisA) were delivered to the sex factor with derivatives of the vector pUK300. Food-grade recombinants of L. lactis were constructed which had the nisRK genes and individual P(nisA)::pep fusions integrated either separately into the chromosome and the sex factor or simultaneously into the sex factor. With both types of recombinants, expression of P(nisA)::pep fusions after induction with nisin was demonstrated. Depending on the loci used for integration of nisRK, variable induction rates were observed. Furthermore, an engineered sex factor carrying a P(nisA)::pepI fusion was transfered by conjugation between two strains of L. lactis at a frequency of 4 x 10(-4).

Bacterial Proteins↗

Hsp90 phosphorylation is linked to its chaperoning function. Assembly of the reovirus cell attachment protein.

Studies on Hsp90 have mainly focused on its involvement in the activation of several families of protein kinases and of steroid hormone receptors. Little is known regarding the role of Hsp90 in the folding of nascent proteins. We previously reported that Hsp90 plays an active role in the posttranslational assembly of the C-terminal globular head of the reovirus attachment protein final sigma1. We show here that Hsp90 becomes phosphorylated in this process. However, only the unphosphorylated form of Hsp90 is complexed with final sigma1, suggesting that Hsp90 phosphorylation is coupled to the release of the chaperone from the target protein. Geldanamycin, which blocks final sigma1 maturation by preventing the release of Hsp90 from final sigma1, also inhibits Hsp90 phosphorylation. Taken together, these results demonstrate that Hsp90 phosphorylation is linked to its chaperoning function.

Animals↗

Constitutional genomic instability with inversions, duplications, and amplifications in 9p23-24 in BRCA2 mutation carriers.

Germ-line mutations of the BRCA2 gene (13q12-13) account for a large proportion of familial breast cancer cases in females and the majority of familial breast cancers in males. Recent studies provide evidence for a role of the BRCA2 protein in the maintenance of genomic integrity by involvement in DNA repair and recombination. In pursuit of identifying in humans genetic damage resulting from mutated BRCA2, we have analyzed constitutional karyotypes of BRCA2 mutation carriers. The present study establishes that constitutional distal 9p rearrangements without obvious additional gross chromosomal alterations are a recurrent feature of independently ascertained families. From our cytogenetic analyses we have no indication of additional gross rearrangements, but we cannot exclude more subtle recombinations in other genomic regions. We also show that the topography of the 9p rearrangements can differ among family members, even within an individual that can have cell populations with different 9p rearrangements. Collectively these results raise point to an association of mutant BRCA2 with genomic instability and gene alteration in 9p23-24 in at least a subset of BRCA2 mutation carriers.

Adult↗

Tubal ligation and risk of ovarian cancer in carriers of BRCA1 or BRCA2 mutations: a case-control study.

BACKGROUND: In several case-control and prospective studies, tubal ligation has been associated with a decreased risk of invasive epithelial ovarian cancer. We aimed to assess the potential of tubal ligation in reducing the risk of ovarian cancer in women who carry predisposing mutations in the BRCA1 or BRCA2 genes. METHODS: We did a matched case-control study among women from Canada, the USA, and the UK who had undergone genetic testing and who carried a pathogenic mutation in BRCA1 or BRCA2. Cases were 232 women with a history of invasive ovarian cancer, and controls were 232 women without ovarian cancer, and who had both ovaries intact. Cases and controls were matched for year of birth, country of residence, and mutation (BRCA1 or BRCA2). The odds ratio for developing ovarian cancer was estimated for tubal ligation, adjusting for oral contraceptive use, parity, history of breast cancer, and ethnic group. FINDINGS: In an unadjusted analysis among BRCA1 carriers, significantly fewer cases than controls had ever had tubal ligation (30 of 173 [18%] vs 60 of 173 [35%], odds ratio 0.37 [95% CI 0.21-0.63]; p=0.0003). After adjustment for oral contraceptive use, parity, history of breast cancer and ethnic group, the odds ratio was 0.39 (p=0.002). Combination of tubal ligation and past use of an oral contraceptive was associated with an odds ratio of 0.28 (0.15-0.52). No protective effect of tubal ligation was seen among carriers of the BRCA2 mutation. INTERPRETATION: Tubal ligation is a feasible option to reduce the risk of ovarian cancer in women with BRCA1 mutations who have completed childbearing.

Adult↗

Hepatitis B virus markers in anti-HBc only positive individuals.

Isolated reactivity to hepatitis B virus (HBV) core antigen (anti-HBc) is observed relatively frequently in immunocompromised individuals, intravenous drug abusers (IVDA), and in the presence of HCV infection. The reason for the lack of HBsAg is not clear. The aim of the present study was to investigate which factors (genetic variability of S gene, low-level HBsAg, and immune complexes may be responsible for the failure of HBsAg detection with commercial HBsAg screening assays. Dilution series of two recombinant HBsAg escape mutants and dilutions of serum samples from chronic HBV carriers with multiple insertions in the a determinant and different HBsAg subtypes were tested with a highly sensitive assay that detects wild-type HBsAg (Elecsys HBsAg, Roche Diagnostics, Penzberg, Germany) and two assays that detect HBV wild-type and escape mutants (Murex HBsAg Version 3, Murex and Enzygnost HBsAg 5.0, Dade Behring, Marburg, Germany). Elecsys HBsAg showed in comparison to Murex HBsAg Version 3 and Enzygnost HBsAg 5.0 a reduced sensitivity for escape mutant detection. On the other hand, the best performance for HBsAg subtype detection was obtained with Elecsys HBsAg. In the second part of the study, a selected panel of isolated anti-HBc reactive (n = 104) serum samples (AxSYM Core) was submitted to testing by Elecsys HBsAg, Murex HBsAg Version 3, Enzygnost HBsAg 5.0, and HBsAg detection after immune complex dissociation (ICD) and anti-HBs determination with two different assays (AxSYM Ausab and Elecsys Anti-HBs). To assess the specificity of anti-HBc test results, all the samples were tested by a second anti-HBc assay (Elecsys Anti-HBc). Quantitative HBV DNA detection was undertaken with a commercially available HBV PCR assay (Amplicor HBV Monitor). HCV infection was present in 65.4% of anti-HBc only reactive individuals. Five AxSYM Core positive samples were negative by Elecsys Anti-HBc. Overall, 15 (14.4%) AxSYM Ausab negative samples gave positive results with Elecsys Anti-HBs (median value: 21 IU/ml). No low-level HBsAg carrier was detected among the isolated anti-HBc reactive individuals with Elecsys HBsAg. There was no evidence for the presence of immune complexes. Only one sample was repeatedly reactive by the Murex HBsAg, suggesting that the a mutant form of HBsAg was responsible for the isolated anti-HBc reactivity, however neutralisation assay was not interpretable and HBV DNA PCR was negative. Fifteen (14.4%) anti-HBc only positive individuals were HBV DNA carriers with concentrations ranging from 800 to more than >4,000,000 copies of viral DNA/ml. In conclusion, the most probable explanations for isolated anti-HBc reactivity in our study group are a possible interference of HBsAg synthesis by HCV infection (65.4%) and divergence of results of anti-HBs assays (14.4%). There is no evidence for the presence of low-level HBsAg carriers and immune complexes. HBsAg mutants cannot be excluded definitively by the test strategy used in the present evaluation.

Amino Acid Sequence↗

Prediction of Sanfilippo phenotype severity from immunoquantification of heparan-N-sulfamidase in cultured fibroblasts from mucopolysaccharidosis type IIIA patients.

Mucopolysaccharidosis type IIIA (MPS-IIIA) is an autosomal recessive lysosomal storage disorder caused by the deficiency of heparan-N-sulfamidase (NS; EC 3.10.1.1), resulting in defective degradation and subsequent storage of heparan sulfate and leading to a clinical phenotype known as Sanfilippo syndrome. A sensitive and specific monoclonal/polyclonal-based immunoquantification assay has enabled the determination of NS protein, down to approximately 3 pg NS protein, in cultured fibroblasts from control and MPS-IIIA patients. Cultured skin fibroblasts from 15 normal controls contained 11.9 to 105 ng of NS protein/mg extracted cell protein, whereas NS protein ranged from "none detected" to 11 ng/mg in fibroblasts from 35 MPS-IIIA patients. A relationship between genotype/phenotype and amount of NS protein present in these MPS-IIIA fibroblasts was established. Immunoquantification, in combination with a specific and highly sensitive tetrasaccharide-based assay of NS activity, enabled the determination of residual specific NS activity in these fibroblasts. Specific NS activity ranged from 28 to 1289 nmol/min/mg NS protein for MPS-IIIA patients, compared to 870 nmol/min/mg of recombinant human NS. It is proposed that this immunoquantification method, in conjunction with the specific NS activity assay, may be used to predict clinical severity in MPS-IIIA patients, allowing for the selection of individuals best suited for gene- and enzyme-replacement therapy when these methods become available. Also proposed is that an enzyme-replacement therapy achieving a correction of approximately 10% of normal NS activity is required to avoid the onset of a Sanfilippo clinical phenotype.

Adolescent↗

Expression and characterization of human recombinant and alpha-N-acetylglucosaminidase.

Mucopolysaccharidosis type IIIB (MPS-IIIB, Sanfilippo type B Syndrome) is a heterosomal, recessive lysosomal storage disorder resulting from a deficiency of [alpha]-N-acetylglucosaminidase (NAGLU). To characterize this enzyme further and evaluate its potential for enzyme replacement studies we expressed the NAGLU-encoding cDNA in Chinese hamster ovary cells (CHO-K1 cells) and purified the recombinant enzyme from the medium of stably transfected cells by a two-step affinity chromatography. Two isoforms of recombinant NAGLU with apparent molecular weights of 89 and 79 kDa were purified and shown to differ in their glycosylation pattern. The catalytic parameters of both forms of the recombinant enzyme were indistinguishable from each other and similar to those of NAGLU purified from various tissues. However, compared to other recombinant lysosomal enzymes expressed from CHO-K1 cells, the mannose-6-phosphate receptor mediated uptake of the secreted form of recombinant NAGLU into cultured skin fibroblasts was considerably reduced. A small amount of phosphorylated NAGLU present in purified enzyme preparations was shown to be endocytosed by MPS-IIIB fibroblasts via the mannose-6-phosphate receptor-mediated pathway and transported to the lysosomes, where they corrected the storage phenotype. Direct metabolic labeling experiments with Na(2) (32)PO(4) confirmed that the specific phosphorylation of recombinant NAGLU secreted from transfected CHO cells is significantly lower when compared with a control lysosomal enzyme. These results suggest that the use of secreted NAGLU in future enzyme and gene replacement therapy protocols will be severely limited due to its small degree of mannose-6-phosphorylation.

Acetylglucosaminidase↗