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Biomedical subjects

G Wolf

Publications and source records attributed to G Wolf.

At least 397 records · Page 22Linked to original sources

Comparison of double fluorescence staining and LDH-test for monitoring cell viability in vitro.

To examine glutamate-mediated toxic effects in dispersed cultures of the rat cerebral cortex we compared the utility of parameters of cell viability in parallel. A 1 h exposure to glutamate and glutamate analogues (kainate, quinolinate, NMDA) was found to produce typical morphological changes in matured cell cultures. The double-labelling fluorescence technique (fluorescein diacetate/propidium iodide) reflected the degeneration process vividly. A significant increase in LDH-activity released into the medium was noted only when the cells were stressed but still living. When the phase of progressive cell death was running, LDH activity in the medium decreased markedly. Obviously, increasing LDH release in the culture medium must be considered rather to be an indicator for a slowly evolving degenerative process than for cell death.

Animals↗

Glutamate agonist-induced hippocampal lesion and nitric oxide synthase/NADPH-diaphorase: a light and electron microscopical study in the rat.

The 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT)-tetrazolium salt technique for the electron microscopic demonstration of reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) was used to localize nitric oxide synthase in the normal and excitotoxically lesioned rat hippocampus. The reaction product BSPT-formazan was shown to stain membranes predominantly of the endoplasmic reticulum. Apart from singular heavily labeled interneurons, the majority of neurons including pyramidal and granular cells and a few astroglial cells, light microscopically 'unstained', showed labeled membrane portions, but to a by far lesser extent. In lesioned areas some prominantly stained neurons rich in labeled membranes and surrounded by cell debris seemed to be largely preserved. An increased number of ultrahistochemically NADPH-d-stained glial cells, in particular astrocytes, was seen.

Amino Acid Oxidoreductases↗

Mg2+ antagonizes alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid-induced brain damage and convulsions.

alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), an agonist of the non-N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor, was used to imitate glutamate-induced brain injury. A single intracerebroventricular injection of AMPA (9 nmol; 1.7 micrograms) induced convulsive reactions and heavy neurodegeneration in the hippocampal formation. MgSO4 (600 mg/kg), administered 20 min prior to or simultaneously with AMPA exposure, was able to protect completely against this non-NMDA-induced neurotoxicity. Magnesium is suggested to be a hopeful therapeutic principle for glutamate-mediated brain disorders.

Animals↗

Molecular structure of a cross-reactive idiotype on autoantibodies recognizing parenchymal self.

The autoimmune B cell repertoire in anti-tubular basement membrane (alpha TBM) disease is focused on the nephritogenic P1 domain of the 3M-1 target Ag in kidney and normally expresses a disease-modifying cross-reactive Id (IdX). The molecular structure of this Id was determined from a library of rat mAb alpha TBM/alpha 3M-1 by using anchor polymerase chain reactions. Our findings provide the first alignment of V region sequences for rat IgG and reveal that specificity for the P1 domain among alpha 3M-1 antibodies is derived from several recurring germ-line VH genes which have not undergone somatic mutation. The IdX in this repertoire localizes to the H chain on Western blot, and to the CDR3 region as deduced from the cDNA encoding several informative clones. Computer modeling of the Id reveals a conformational structure largely dependent on hydroxyl groups in or near turn position 4 of the H chain CDR3 region. These findings demonstrate that regulatory elements protective of autoimmunity are not encoded in the germ line as IdX, but rather emerge from a recombinatorial diversity engaged by the recognition of parenchymal self.

Amino Acid Sequence↗

Serological and antigenical findings indicating pestivirus in man.

An epidemiological survey for pestivirus was undertaken in Zambia and Europe, in view of the recent serological findings obtained by previous studies in Europe with humans. Collected sera were tested for anti-bovine viral diarrhea virus (BVDV) specific antibodies by IIF and Western Blotting. Of those individuals tested (n = 1272), 15.3% showed a seropositive reaction to the BVDV. Anti-BVDV antibody prevalence in immuno-depressed patients (e.g. HIV positive) was investigated. A higher prevalence was revealed in HIV patients suffering from chronic diarrhoea and in those having developed AIDS Related Complex (ARC). Our of 212 persons tested for pestivirus isolation, a non cytopathic virus strain was detected in 2 buffy coat samples using IIF with a specific anti-BVDV serum. The isolation could be repeated three times during 31 days in one person. The virus was identified as a pestivirus with radioimmuno-precipitation assays and IIF-flow cytometry. A doublet of 120 kD was identified only in cell lysates, indicating a non-structural protein. In order to rule out cross reactivity 30 sera from Hepatitis C seropositive patients were tested against the isolate by IIF-flow cytometry. No antigen-specific binding could be observed. These findings indicated the occurrence of a pestivirus in man and might suggest a relationship with a pestivirus of animal origin.

Animals↗

Regulating factors of renal tubular hypertrophy.

Understanding the mechanisms of tubular hypertrophy is important because these cells simply represent the bulk of the nephron, and there is a convincing link between early tubular enlargement and the progression of renal disease. It seems reasonable to assume that cytokines and polypeptide growth factors including inhibitory factors such as TGF-beta induce in concert, rather than as single factors, tubular hypertrophy. The important observations that the activation of the intrarenal renin-angiotensin axis is altered in situations associated with renal growth, and that ACE inhibitors abolish compensatory hypertrophy in many models provided for us a basis for investigating the growth effects of ANG-II on cultured proximal tubular cells. ANG-II induces, as a single factor, tubular hypertrophy in vitro, and this growth effect has been studied in detail on a molecular levels. Endogenous induction of TGF-beta by ANG-II is important in the peptide-mediated hypertrophy. While some genes induced by ANG-II, such as immediate early genes, are engaged as part of a generalized activation of the nucleus, the hypertrophic effects may be mediated by a set of novel genes which may be part of an identifiable genetic program causing tubular enlargement. The identification of hypertrophy genes may offer new insight into the modulation of cytoplasmic enlargement and its interface with elements that control the cell cycle and may provide a tool for further therapeutic interventions.

Angiotensin II↗

Adenosine influences the high-affinity uptake of transmitter glutamate and aspartate under conditions of hepatic encephalopathy.

The high-affinity uptake of transmitter glutamate and aspartate in hippocampal slices incubated in sera from patients with hepatic encephalopathy was dramatically reduced in neuropil areas by more than 50% compared with the control level. Adenosine compensates for these reduction in reuptake capacity in a concentration-dependent fashion reaching normal values at 500 microM adenosine. The renormalization of glutamate and aspartate uptake caused by adenosine might reasonably be expected to have potential therapeutic implications for the treatment of hepatic encephalopathy.

Adenosine↗

Shedding and repair of renal cell membranes following drug-induced nephrotoxicity in humans.

Nephrotoxic drugs may account for approximately at least 20% of clinically observed cases of acute renal failure in whom tubular lethal or sublethal damage is a predominant finding. Acute toxic tubular cell injury is characterized by loss of cellular polarization, intrinsic energy deficiency, calcium overload, release of toxic proteases and free oxygen radicals, derangement of the cytoskeleton, and vacuolar transformation of brush border microvilli. These events may finally lead to irreversible cell death. Shedding of membrane enzymes and cytoskeletal components in urine (kidney tissue proteinuria) may serve as a noninvasive early marker for assessing tubular cell injury. Successful recovery of renal function depends on early repair of lethally or sublethally damaged nephrons, in which intrinsic nephrogenic adaptive and proliferative responses cooperate in concert with auto/para/-juxtacrine growth promoting factors and cytokines. Exogenously administered growth factors may enhance renal cell recovery, as shown in animal models. Increased expression of immediate early genes in tubular cells after renal injury reflects the ongoing mitogenic activity necessary for reepithelialization and remodeling (new, polarized, differentiated cells). Further progress in understanding the molecular mechanisms of renal tubular injury will probably influence the diagnostic modalities and therapeutic approaches to acute drug induced renal failure.

Animals↗

The impact of geographical, clinical, dietary and radiation-induced features in epidemiology of thyroid cancer.

Cancer of the thyroid accounts for less than 1% of all cancers recognised each year, but the incidence is rising. Much of the early work of the epidemiology and aetiology of thyroid cancer was based on the assumption that thyroid cancer can be treated as an entity. The recognition that two distinct types of endocrine cell occur within the thyroid has made it clear that any discussion of the aetiology and epidemiology of thyroid malignancies must take into account the histological classification of these tumours. Moreover, there are difficult problems to be considered when comparing thyroid cancer incidence across tumour registries, because of a lack of standardisation or morbidity data collection, difficulties in histological diagnosis, varying rates of diagnosis of occult papillary carcinoma, and prevalence and techniques of autopsies. So far only a relatively small proportion of thyroid cancer cases can be explained with adequate certainty as regards epidemiology and aetiology. As in cancer in general, the aetiology and epidemiology of thyroid cancer in detail remains unknown in the majority of cases.

Adult↗

TNF alpha induces expression of the chemoattractant cytokine RANTES in cultured mouse mesangial cells.

We investigated the effect of several immune-relevant cytokines on expression of the chemoattractant intercrine/chemokine RANTES in a mouse mesangial cell line (MMC). Fifty ng/ml recombinant tumor necrosis factor alpha (TNF alpha) induced a marked increase in RANTES transcripts after two hours. RANTES mRNA remained elevated for 24 to 48 hours after stimulation, and could be abolished by co-incubation with 30 micrograms/ml of a neutralizing rabbit anti-TNF alpha antibody. Protein expression of RANTES, as assessed by indirect immunofluorescence and Western blotting, increased in MMCs 24 hours after TNF alpha stimulation. Interleukin-1 beta, tumor necrosis factor beta (TNF beta), and lipopolysaccharide (LPS) also increased expression of RANTES mRNA. In addition, RANTES mRNA expression was stimulated in glomeruli harvested from rats following renal in vivo perfusion with TNF alpha. Our results indicate that mesangial cells produce the small cytokine RANTES. This factor, in concert with other chemoattractants, may play a role in the glomerular recruitment of inflammatory cells like macrophages/monocytes.

Animals↗

Monoclonal antibodies to human chorionic gonadotropin (HCG) and their use in two-site binding enzyme immunoassays.

A panel of mouse monoclonal antibodies (MABs) was produced against human chorionic gonadotropin (HCG) and its isolated beta-subunit (beta-HCG). According to their binding specificities the antibodies could be divided into HCG-specific and cross-reactive MABs. The HCG-specific antibodies reacted with antigenic sites on holo-HCG or holo-HCG and beta-HCG, or exclusively with the non-associated beta-HCG chain. The cross-reactive antibodies reacted with either HCG and luteinizing hormone (LH) or with HCG, LH, follicle-stimulating hormone (FSH) and thyroid-stimulating hormone (TSH). According to the binding specificities of the MABs and their reciprocal inhibition detected in two-site binding enzyme immunoassays (EIA), altogether 13 epitopes (including the 3 hidden epitopes detectable only on free non-associated beta-HCG) were distinguished by the antibodies described here. Antibody combinations resulting in most effective and specific HCG- or beta-HCG-determination were used as clinical assays and proved their reliability and correctness for monitoring patients with HCG- and/or beta-HCG-producing tumors before and after therapy.

Animals↗

Quinolinate and kainate facilitate magnesium penetration into brain tissue.

To study the penetration of magnesium ions from blood into brain tissue, magnesium content in serum and hippocampus of normal and of excitotoxically affected rats was estimated after a single subcutaneous injection of magnesium sulphate (600 mg kg-1). In normal rats Mg2+ levels in serum rose from 1 to 6 mM, while that of the hippocampus remained constant, provided the brains were perfused before magnesium measurement. Following unilateral intracerebroventricular injection of the excitotoxic glutamate analogues, quinolinate or kainate acid, Mg2+ levels increased up to 38% on the (unaffected) contralateral side. Since magnesium is known to prevent glutamate-mediated neurodegeneration, our findings on the accessibility of exogenously applied magnesium may justify further investigations on the utility of magnesium for a therapeutic approach to limiting excitotoxic brain injury in human patients.

Animals↗

The newly discovered retinoic acid-X receptors (RXRs).

All-trans-retinoic acid functions by binding to nuclear receptor proteins (RAR alpha,beta,gamma) that interact with response elements in various genes involved in development and differentiation. A new family of nuclear receptors (RXR alpha,beta,gamma) has been discovered that uses 9-cis-retinoic acid as ligand and may function particularly in regulation of genes involved in intermediary metabolism.

Carrier Proteins↗

Lipofuscin, the age pigment.

Lipofuscin is the fluorescent mixture of compounds that accumulates increasingly with age in human tissues, including the pigment epithelium of the retina. A substantial component of lipofuscin has been identified in the retina as N-retinylidene-N-retinylethanolamine. This compound presumably is derived from the reaction of retinaldehyde with the membrane lipid phosphatidylethanolamine.

Adult↗

Uptake of ascorbic acid by human neutrophils.

Human neutrophils produce bacterial oxidants that could have a damaging effect on the neutrophils themselves. For protection, neutrophils acquire a high (10-20 mmol/L) level of intracellular ascorbic acid by oxidizing extracellular ascorbic acid to dehydroascorbic acid, rapidly taking up the dehydroascorbic acid and re-reducing it to ascorbic acid.

Ascorbic Acid↗