Biomedical subjects

G Wolf

Publications and source records attributed to G Wolf.

At least 253 records · Page 14Linked to original sources

PubMed1997

Patterns of chromosomal alterations in metastasizing and nonmetastasizing primary head and neck carcinomas.

In an attempt to define chromosomal alterations that are associated with the metastatic phenotype, we investigated a total of 29 metastasizing (pN+) and 19 non-metastasizing (pN0) head and neck squamous cell carcinomas by comparative genomic hybridization (CGH). The analysis indicated that the pN0 tumors carried preferentially overrepresentations of chromosomes 5p, 6p, and 7p and that the pN+ tumors were frequently characterized by deletions on chromosomes 7q, 10q, 11p, 11q, 15q, and 20p and overrepresentations of the chromosomes 19q and 20q. In particular, the use of difference histograms and statistical analysis indicated that the deletions on chromosomes 10q25-q26 and 11p13-p14 were highly significant for metastasizing carcinomas. The findings on chromosome 10q were supported by loss of heterozygosity analysis in the primary tumors and eight synchronous lymph node metastases using four microsatellite polymorphisms. The data suggest that distinct patterns of genetic lesions are responsible for the metastatic phenotype of head and neck squamous cell carcinomas.

Carcinoma, Squamous Cell↗

PubMed1997

Cytotoxic T-lymphocyte responses in cattle infected with bovine viral diarrhea virus.

A system for a reproducible in vitro restimulation of bovine viral diarrhea virus (BVDV)-specific cytotoxic T-cells (CTL) was developed. Lymphocyte cultures of BVDV-immunised cattle were stimulated with infectious BVDV isolate PT810 and recombinant bovine interleukin-2 for 12 to 25 days. A specific lysis of Concanavalin A-stimulated BVDV-infected autologous target cells was observed, whereas allogeneic BVDV-infected target cells were only marginally lysed as detected by flow cytometry. BVDV-specific lymphocyte transformation was further characterised by the expression of bovine lymphocyte activation antigens and bovine MHC class-II molecules. Secondary stimulation of CTL was influenced by in vitro production of BVDV-specific neutralising antibodies, which were secreted exclusively in BVDV-inoculated lymphocyte cultures of immunised cattle. These results demonstrate the presence of CTL in peripheral blood mononuclear cells (PBMC) of immunised cattle which can kill autologous BVDV-infected antigen-presenting cells after in vitro restimulation.

Animals↗

PubMed1997

2,7-Dihydrodichlorofluorescein diacetate as a fluorescent marker for peroxynitrite formation.

Reactive oxygen species (ROS) have been implicated as an important causative factor in cell damage, including apoptosis and necrosis. Their proposed actions comprise lipid peroxidation, DNA damage, destruction of the mitochondrial respiratory chain and protein modifications. Recent experiments underline the importance of peroxynitrite, the reaction product of the two potent reactive species nitric oxide and superoxide. Several fluorogenic compounds have been used in order to determine ROS formation in living cells. Besides dihydrorhodamine-123 (DHR-123), at present mostly applied to monitor peroxynitrite, 2,7-dihydrodichlorofluorescein (DCF-H) is used for detection of hydrogen peroxide and nitric oxide. We employed a cell free approach to evaluate the specificity and sensitivity of DCF-H to various oxidizing compounds. Our studies imply that DCF-H is much more sensitive to peroxynitrite oxidation than any other compound tested. In order to study peroxynitrite generation within individual cells, primary glial cultures loaded with DCF-H were monitored with a laser scanning microscope. Microglia, stimulated to simultaneously produce the peroxynitrite precursors nitric oxide and superoxide, displayed the greatest increase in DCF fluorescence, whereas microglia producing either nitric oxide or superoxide alone showed a relatively small increase in DCF fluorescence. In conclusion, DCF-H was demonstrated to be an excellent peroxynitrite marker with the potential to detect peroxynitrite formation in living cells.

Animals↗

PubMed1997

ISCOM vaccine against contagious bovine pleuropneumonia (CBPP). 1. Biochemical and immunological characterization.

A better vaccine than the existing ones against contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) would improve the chances for eradication of CBPP. In such an effort, immunostimulating complexes (ISCOMS) have been prepared from the whole detergent-solubilized cells of MmmSC and characterized biochemically and immunologically. The most efficient detergent for solubilization of the mycoplasma was MEGA-10 which yielded a high recovery of proteins in the ISCOMS. The ISCOMS showed the typical cage-like structure by EM and sedimented as 19S by sucrose gradient centrifugation. The protein pattern of the ISCOMS, analyzed in SDS-PAGE, revealed a great number of bands distributed along the gel as high and low molecular weight polypeptides. The Western blot developed with a serum from a CBPP infected animal detected a reduced number of polypeptides. In samples from whole mycoplasma cells and in ISCOMS, lectin blots revealed more than 20 carbohydrate structures. The ISCOMS induced a strong primary antibody response in mice measured by ELISA and the boost resulted in a 6-fold increase of the serum antibody response. The IgG response was distributed into various IgG subclasses with high IgG1, IgG2a and IgG2b titres while the IgG3 response was low. In cattle the ISCOM vaccine induced strong primary and long lasting secondary antibody responses of similar magnitudes as those of naturally infected animals as recorded by ELISA which persisted more than a year. IgG response was equally distributed in IgG1 and IgG2 subclasses. Also a cell-mediated immune response measured by proliferation assay was induced by low dose of ISCOMS. In the growth inhibition test, sera from vaccinated cattle readily inhibited colony growth already after the first immunization.

Animals↗

PubMed1997

Angiotensin II stimulates expression of the chemokine RANTES in rat glomerular endothelial cells. Role of the angiotensin type 2 receptor.

Glomerular influx of monocytes/macrophages (M/M) occurs in many immune- and non-immune-mediated renal diseases. The mechanisms targeting M/M into the glomerulus are incompletely understood, but may involve stimulated expression of chemokines. We investigated whether angiotensin II (ANG II) induces the chemokine RANTES in cultured glomerular endothelial cells of the rat and in vivo. ANG II stimulated mRNA and protein expression of RANTES in cultured glomerular endothelial cells. The ANG II-induced RANTES protein was chemotactic for human monocytes. Surprisingly, the ANG II-stimulated RANTES expression was transduced by AT2 receptors because the AT2 receptor antagonists PD 123177 and CGP-42112A, but not an AT1 receptor blocker, abolished the induced RANTES synthesis. Intraperitoneal infusion of ANG II (500 ng/h) into naive rats for 4 d significantly stimulated glomerular RANTES mRNA and protein expression compared with solvent-infused controls. Immunohistochemistry revealed induction of RANTES protein mainly in glomerular endothelial cells and small capillaries. Moreover, ANG II- infused animals exhibited an increase in glomerular ED-1- positive cells compared with controls. Oral treatment with PD 123177 (50 mg/liter drinking water) attenuated the glomerular M/M influx without normalizing the slightly elevated systolic blood pressure caused by ANG II infusion, suggesting that the effects on blood pressure and RANTES induction can be separated. We conclude that the vasoactive peptide ANG II may play an important role in glomerular chemotaxis of M/M through local induction of the chemokine RANTES. The observation that the ANG II- mediated induction of RANTES is transduced by AT2 receptors may influence the decision as to which substances might be used for the therapeutic interference with the activity of the renin-angiotensin system.

Angiotensin II↗

PubMed1997

Patterns of chromosomal imbalances in adenocarcinoma and squamous cell carcinoma of the lung.

Comparative genomic hybridization was used to screen 25 adenocarcinomas and 25 squamous cell carcinomas of the lung for chromosomal imbalances. DNA copy number decreases common to both entities were observed on chromosomes 1p, 3p, 4q, 5q, 6q, 8p, 9p, 13q, 18q, and 21q. Similarly, DNA gains were observed for chromosomes 5p, 8q, 11q13, 16p, 17q, and 19q. Adenocarcinomas showed more frequently DNA overrepresentations of chromosome 1q and DNA losses on chromosomes 3q, 9q, 10p, and 19, whereas squamous cell carcinomas were characterized by increased overrepresentations of chromosome 3q and 12p as well as deletions of 2q. For the first time, we used a histogram representation and statistical analysis to evaluate the differences between both tumor groups. In particular, the overrepresentation of the chromosomal band 1q23 and the deletion at 9q22 were significantly associated with adenoid differentiation, whereas the DNA loss of chromosomal band 2q36-37 and the overrepresentations at 3q21-22 and 3q24-qter were statistically significant markers for the squamous cell type. The study strengthens the notion that different tumor subgroups of the respiratory tract are characterized by distinct patterns of chromosomal alterations.

Adenocarcinoma↗

PubMed1997

Prognostic significance of polo-like kinase (PLK) expression in non-small cell lung cancer.

Our previous data indicate that the expression of the PLK gene which codes for a serine/threonine kinase is restricted to proliferating cells. In Northern blot experiments PLK mRNA expression was at the limit of detection in normal lung tissue but elevated in most samples of non-small cell lung cancer (NSCLC). A very low frequency of PLK transcripts was only found in bronchiolo-alveolar carcinomas. NSCLC patients whose tumors showed moderate PLK expression survived significantly longer (5 year survival rate=51.8%) than those with high levels of PLK transcripts (24.2%, P=0.001). No statistically significant correlation was found between PLK mRNA expression and age, sex, TNM status, histological type or degree of differentiation. Interestingly, the prognosis of patients in post-surgical stages I and II was correlated with PLK expression (5 year survival rates in stage I: 69.1% (moderate PLK) - 43.5% (high PLK), P=0.03 or in stage II: 51.9% (moderate PLK) - 9.9% (high PLK), P=0.006). These results suggest that PLK mRNA expression provides a new independent prognostic indicator for patients with NSCLC.

Adenocarcinoma, Bronchiolo-Alveolar↗

PubMed1997

Preimplantation diagnosis of single gene disorders by two-step oocyte genetic analysis using first and second polar body.

Previous work on preimplantation genetic diagnosis (PGD) of single gene disorders by the first polar body (IPB) analysis has demonstrated that the genotype of a considerable number of embryos resulting from heterozygous oocytes cannot be predicted without testing their second PB (IIPB). To overcome this limitation we introduce a two-step DNA analysis of oocytes using both IPB and IIPB to identify hemizygous mutation-free oocytes following the second meiotic division. In the application of the approach to PGD of cystic fibrosis (CF) Delta F-508 mutation, sickle cell disease, and hemophilia B, 80 oocytes were studied by both PBs, resulting in the identification and transfer of 32 homozygous normal embryos. A follow-up genotyping of 52 embryos, resulting from oocytes tested by both IPB and IIPB demonstrated the accuracy of the predicted genotypes. In addition to a nested PCR analysis of the mutant genes in PBs and resulting embryos, simultaneous amplification of different polymorphic markers was performed, demonstrating the reliability of the two-step polar body analysis of oocytes.

Alleles↗

PubMed1997

Oxidized low density lipoprotein stimulates protein kinase C (PKC) activity and expression of PKC-isotypes via prostaglandin-H-synthase in P388D1 cells.

Treatment of P388D1 macrophage-like cells with oxLDL enhanced protein kinase C (PKC) activity in cell extracts. Similar effects were induced by acetylated LDL (acLDL) and maleylated albumin (malBSA). Treatment with oxLDL, acLDL and malBSA was also accompanied by increased production of prostaglandins as well as by an enhanced level of prostaglandin H synthase 2 (cyclooxygenase 2, COX 2). Modified (lipo)proteins differentially affected the levels of individual cytosolic PKC-isoenzymes. Effects of oxLDL on PKC activity/expression were abrogated by indometacin, by pre-exposure to the dual lipoxygenase/cyclooxygenase inhibitor ML 3000 and by treatment with N-(2-cyclohexyloxy-4-nitrophenyl)methane sulfonamide (NS-398). These results suggest a predominantly COX 2-dependent and isotype-specific effect of modified (lipo)proteins on PKC.

Acetates↗

PubMed1997

Olfactory neuroblastoma: detection of genomic imbalances by comparative genomic hybridization.

Olfactory neuroblastoma (esthesioneuroblastoma) is a very rare tumour of the olfactory mucosa. Morphological features and cytogenetic studies strongly suggest a neuro-ectodermal origin. Up to now, cytogenetic studies are inconsistent. Some of them have proposed that the tumour belongs to the pPNET family. In the present study we describe genomic imbalances in olfactory neuroblastoma in a 46-year-old woman by using the molecular cytogenetic technique--comparative genomic hybridization (CGH)--in order to define the spectrum of genetic abnormalities in the tumour. The anatomical location and morphological findings were the basis for the diagnosis of esthesionearoblastoma. Immunohistochemical reactions for NSE, synaptophysin, chromogranin A, HNK-1/Leu-7 and S-100 revealed a characteristic immunophenotype. The CGH analysis showed multiple changes including DNA overrepresentations of chromosomes 4, 8, 11 and 14, partial DNA gains of the long arms of chromosomes 1 and 17, deletions of the entire chromosomes 16, 18, 19 and X, and partial losses of chromosomes 5q and 17p. This study represents an early utilisation of the CGH technique in olfactory neuroblastoma and demonstrates that the tumour carries complex chromosomal aberrations.

Biomarkers, Tumor↗

PubMed1997

Parathyroid scintigraphy: first experiences with technetium (III)-99m-Q12.

We report the preliminary results of a prospective study demonstrating technetium(III)-99m furifosmin (Q12) uptake in histologically proven parathyroid adenomas. Scintigraphy was performed in 12 patients with hyperparathyroidism. Q12 correctly identified the parathyroid adenomas by focal prolonged tracer retention in ten of the 12 patients. In the two patients without localized tracer retention, no parathyroid adenoma could be identified surgically, either. The retention half-times ranged from 0.5 to 1.8 h (mean 1.27) in the parathyroid adenomas and from 0.3 to 1.2 h (mean 1.05) in the thyroid gland. Our preliminary results show that Q12 is a feasible, sensitive tracer for parathyroid scintigraphy. In comparison to sestamibi (MIBI) and tetrofosmin, Q12 displays short retention in the thyroid gland, which would seem rather advantageous. Further evaluation is needed to determine which among Q12, tetrofosmin and MIBI is most sensitive for the detection especially of small parathyroid adenomas, and which tracer properties will best reflect the degree of endocrine activity.

Adenoma↗

PubMed1997

[Sitting or supine stereotaxic core biopsy of the breast? A comparison based on a randomized, prospective study].

UNLABELLED: OBJECTIVE/MATERIAL AND METHODS: In a prospective randomized study, the techniques of stereotactic breast biopsies in prone and sitting position were compared. Part of the data has already been published. A total of 103 women underwent stereotactic breast biopsies, either prone (n = 51; using TRC-Mammotest, Sweden) or in the sitting position (n = 52; using Stereotix 2, General Electric Medical Systems, Milwaukee, Wisconsin, USA). With the help of pre- and post-biopsy questionnaires, anxiety, pain, and subjective experience were recorded in all patients. Vasovagal reactions were scored from 0 to 2 according to their severity. All biopsy results were verified by surgery. The specificities and sensitivities for the two positions were calculated and statistically compared. RESULTS: With regard to overall tolerance no statistically significant difference between biopsies performed in the sitting or the prone position was noted. Significantly more patients (p = 0.04) in the prone position stated they would prefer premedication prior to a repeat biopsy. Three patients (prone; n = 1; sitting; n = 2) fainted during the procedure. There was no statistically significant difference between the two biopsy positions regarding sensitivity (95%) and specificity (100%). CONCLUSIONS: More attention should be paid to patient care and, especially, preintervention information. Biopsies in the prone or sitting position are equally well tolerated. Somatic reactions are not a major problem during breast biopsy. Success and validity are independent of the biopsy position.

Adult↗

PubMed1997

Genomic (5'UTR) and serological differences among German BVDV field isolates.

Isolates of bovine viral diarrhoea virus (BVDV) collected in Germany were examined for their genomic heterogeneity in sequences from the 5'untranslated region (UTR) of the viral genome. Polymerase chain reaction (PCR) tests based on the 5'UTR and the region coding for the NS2-3/4A polypeptide were used to differentiate between BVDV I and BVDV II genotypes. Eleven out of 96 BVDV-isolates were identified as BVDV II. Virus neutralization tests with BVDV I- or II-specific antisera raised in cattle were done. The mean titers were reduced by 7.2-fold (BVDV I-antiserum versus type II-isolates) or 35-fold (BVDV II-antiserum versus type I-isolates) when using the respective heterologous virus.

Animals↗

PubMed1997

Reduction of adverse events in MRI of the breast by personal patient care.

PURPOSE: To determine the difference in anxiety reactions in patients undergoing standard (non-breast) magnetic resonance imaging (MRI) compared to breast magnetic resonance imaging (MRM) and to evaluate the influence of patient information before the breast imaging examination on the rate of premature termination of the procedure. MATERIALS AND METHODS: Over 2 years, 5837 non-breast and 336 breast magnetic resonance examinations were performed at our institution. One group of breast MRM patients (n = 144) received detailed information and a second group (n = 189) received only routine information before MRI. The rates of premature termination were recorded for all groups. RESULTS: In 0.5% (27/5837) of patients undergoing standard MRI examinations the study had to be stopped prematurely. Of the breast MRM patients, those who had received only routine information had a significantly higher rate of premature termination when compared to the better-informed patients and those undergoing standard MRI (5.5%, 10/189, P= 0.01). A significantly lower rate of premature termination occurred in the better-informed breast group (0%, 0/144). CONCLUSION: MRM is associated with an increase in patient anxiety and higher rates of incomplete examination than other MR procedures. We recommend careful patient preparation including detailed verbal information before MRM and support during the procedure to obtain optimal patient compliance.

Anxiety↗

PubMed1997

Immunocytochemistry of endothelial nitric oxide synthase in the rat brain: a light and electron microscopical study using the tyramide signal amplification technique.

There are many inconsistencies in the literature about the cellular and subcellular distribution of the endothelial isoform of nitric oxide synthase (eNOS) in the brain. We have re-investigated its localization by light and electron microscopical (LM, EM) immunocytochemistry and the NADPH-diaphorase reaction. Using bovine aortic tissue as a positive control the protocols for the fixation and staining procedure were optimized. Only cryosections immersion-fixed with aceton and a mixture of aldehydes exhibited a clear-cut immunostaining. In rat brain tissue the endothelium of the entire vasculature showed immunoreactivity and, in addition to that, the epithelial cells of the choroid plexuses, whereas neurons never displayed any signs of immunostaining. EM immunoprecipitates were seen irregularly distributed in the cytosol or attached to endocellular membranes. EM NADPH-diaphorase histochemistry using the tetrazolium salt BSPT provided incoherent pictures in so far as the reaction product was exclusively bound to membranes. The restriction of eNOS within brain tissue to the vasculature may have implications for the differential significance of NOS isoforms in brain function.

Animals↗

PubMed1997

The role of angiotensin II in diabetic nephropathy: emphasis on nonhemodynamic mechanisms.

Several systemic or intrarenal networks of cytokines and growth factors can be modulated by the diabetic state. We summarize the status of the renin-angiotensin system in diabetes mellitus and review the evidence of its involvement in the pathogenesis of diabetic nephropathy. Particular emphasis is placed on the nonhemodynamic properties of this vasoactive agent as both a renal growth factor and a profibrogenic peptide. Antagonizing the effects of angiotensin II with converting enzyme inhibitors is an established protective strategy in the management of diabetic nephropathy even in the absence of systemic hypertension. This and other indirect evidence from experimental animal studies suggest that the intrarenal concentration of angiotensin II may be increased as a result of increased synthesis and despite enhanced breakdown, that this peptide participates in the progression of diabetic nephropathy. However, down-regulation of angiotensin type 1 (AT1)-receptors is one of the abnormalities of both tubules and glomeruli in diabetic renal disease. A heightened bioactivation of the intrarenal angiotensin II system is therefore likely but not certain. Studies in cultured proximal tubular and glomerular mesangial cells have disclosed striking similarities between the effects of high glucose-containing medium and of treatment with angiotensin II on the growth properties and the induction of cytokines in these cells. There may also exist additive effects of angiotensin II and high glucose on signal-transduction pathways, such as activation of protein kinase C, although the contractile response to angiotensin II may be blunted by high glucose in mesangial cells. An important downstream mediator of the effects of both angiotensin II and high glucose is the activation of transforming growth factor-beta that can mediate at least some of the hypertrophic and profibrotic effects of either angiotensin II or high glucose in the diabetic kidney.

Angiotensin II↗

PubMed1997

Eduard Schwarz, a neglected pioneer in the history of nutrition.

An account of the journey around the world by the Austrian ship's doctor Eduard Schwarz on a sailing ship from 1857 to 1859, his successful cure of nightblindness among the sailors, and how he was maligned by some of the Viennese medical press for his view that nightblindness is a nutritional disorder.

Ascorbic Acid↗

PubMed1997

Small-cell lung cancer is characterized by a high incidence of deletions on chromosomes 3p, 4q, 5q, 10q, 13q and 17p.

The genetic mechanisms that define the malignant behaviour of small-cell lung cancer (SCLC) are poorly understood. We performed comparative genomic hybridization (CGH) on 22 autoptic SCLCs to screen the tumour genome for genomic imbalances. DNA loss of chromosome 3p was a basic alteration that occurred in all tumours. Additionally, deletions were observed on chromosome 10q in 94% of tumours and on chromosomes 4q, 5q, 13q and 17p in 86% of tumours. DNA loss was confirmed by loss of heterozygosity (LOH) analysis for chromosomes 3p, 5q and 10q. Simultaneous mutations of these six most abundant genetic changes were found in 12 cases. One single tumour carried at least five deletions. DNA under-representations were observed less frequently on chromosome 15q (55%) and chromosome 16q (45%). The prevalent imbalances were clearly indicated by the superposition of the 22 tumours to a CGH superkaryogram. In our view, the high incidence of chromosomal loss is an indication that SCLC is defined by a pattern of deletions and that the inactivation of multiple growth-inhibitory pathways contributes in particular to the aggressive phenotype of that type of tumour.

Aged↗