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Biomedical subjects

R Schäfer

Publications and source records attributed to R Schäfer.

At least 19 recordsLinked to original sources
PubMed1995

Nuclear matrix condensation and c-myc and c-fos expression are specifically altered in culture rat hepatocytes after exposure to cyproterone acetate and phenobarbital.

Regenerative or hyperplastic growth promotes carcinogenesis and can be induced by many nongenotoxic carcinogens. The mitogenic potential of the rodent liver tumor promoters, cyproterone acetate and phenobarbital was investigated in primary rat hepatocyte cultures. Two premitotic markers were analyzed, the expression of two immediate-early genes (c-fos and c-myc) and the decrease in the nuclear quinacrine dihydrochloride fluorescence indicative for a G0-G1 cell cycle shift. C-fos expression and decrease in nuclear fluorescence could be induced by both chemicals, phenobarbital being the lesser potent, whereas c-myc expression was only inducible by cyproterone acetate. In situ hybridization with c-myc revealed that both chemicals enhanced c-myc mRNA levels in individual cells, however the number of responding hepatocytes was increased by cyproterone acetate only. The chemical-induced premitotic changes in hepatocytes were highly specific in terms of affected genes and ploidy levels of responding hepatocytes.

Animals↗
PubMed1995

Diagnosis of penile hemangioma by use of biopty gun for corpus cavernosum biopsy.

Penile hemangiomas are very uncommon mesenchymal tumors with less than 20 cases described in the literature. We report on a case of sclerosing hemangioma of the corpus cavernosum diagnosed by MRI and needle biopsy. MRI could delineate the exact anatomical relationship of the hemangioma, corpus cavernosum, tunica albuginea and Buck's fascia. However, the tumor revealed an unspecific inhomogeneous high and low signal intensity shown to be typical for carcinoma, inflammation, fibrosis or plaques. MRI cannot distinguish between benign and malignant penile tumors. A needle biopsy was performed with the Biopty gun and a good tissue sample was obtained for pathological analysis. Our case report demonstrates that this technique is rapid, safe, painless and effective in obtaining adequate tissue samples for microscopic studies. Since sclerosing hemangioma is a benign tumor and the patient was asymptomatic, no further treatment had to be performed.

Adolescent↗
PubMed1994

p53 mutations in oropharyngeal carcinomas: a comparison of solitary and multiple primary tumours and lymph node metastases.

Despite the steadily increasing number of patients suffering from squamous cell carcinomas of the oropharyngeal region, little is known about the molecular steps involved in the induction of these neoplasms. The aim of this study was to determine the incidence and type of p53 mutations in oropharyngeal squamous cell carcinomas. Five of 38 patients had lymph node metastases, three patients had multiple primary carcinomas and two patients presented with multiple primary tumours and lymph node metastases. Exons 5 through 8 of the p53 gene were screened by single-strand conformation polymorphism analysis followed by direct DNA sequencing. A total of 16 tumours (42%) contained point mutations scattered throughout exons 5 to 8. Most mutations (56%) were transitions, predominantly G-->A. G-->T mutations prevailed among the transversions that have also been found in smoking-related lung cancer. One carcinoma of the soft palate showed a mutation which was retained in a lymph node metastasis. The status of p53 differed in primary carcinomas of patients with multiple tumours indicating an independent generation of these neoplasms. The frequent occurrence of p53 mutations in oropharyngeal carcinomas supports the view that this gene plays a role in initiation or progression of the malignant phenotype.

Base Sequence↗
PubMed1993

Establishment of a cellular assay system for G protein-linked receptors: coupling of human NK2 and 5-HT2 receptors to phospholipase C activates a luciferase reporter gene.

A functional cellular assay system was developed for the detection of substances modulating the activity of G protein-coupled receptors, linked to the phospholipase C second messenger system. The human adenocarcinoma cell line A549 was transformed with the Photinus pyralis luciferase gene under the control of the ICAM-1 gene 5'regulatory region and, subsequently, stably transfected with the human neurokinin 2 (NK2) receptor gene. The ICAM-1 promoter is known to be inducible via the phospholipase C signal transduction pathway. In this NK2 receptor test cell line, expression of luciferase was inducible by neurokinin A and other NK2-specific agonists. The order of potency of the three neurokinins substance P, neurokinin A and neuromedin K was consistent with published data and results from ligand binding studies performed with the same NK2 test cell line. The agonistic effect of neurokinin A could be inhibited in a dose-dependent manner by simultaneous addition of NK2-specific antagonists or protein kinase C-inhibitors. Similarly, a stable test cell line expressing the human serotonin 2 receptor was established. Agonist-induced luciferase expression in this cell line was abolished in the presence of 5-HT2-specific antagonists. These cellular assay systems can be employed for the identification of competitive, non-competitive and allosteric modulators of the NK2 and the 5-HT2 receptor, and they represent prototypes for analogous test cell lines for other phospholipase C-coupled receptors.

Biological Assay↗
PubMed1992

Fatigue behavior of direct post-and-core-restored premolars.

Evaluation of long-term mechanical behavior of new types of restorations in clinical trials is time-consuming. A partial alternative can be found in experimental fatigue-testing, which simulates accelerated mechanical deterioration. The aim of this study was to determine the feasibility of using fatigue-testing of a complex dental restoration and to evaluate the mechanical fatigue behavior of premolar teeth restored with a titanium alloy post and an amalgam or composite core. Eighty-seven human upper premolar teeth were decoronated, embedded, and restored with a prefabricated post of 1 mm diameter. The teeth were randomly assigned to one of two groups corresponding with a core build-up of amalgam or chemically-cured core composite, respectively. Five to 21 days after restoration, the specimens were subjected to cyclic loading (frequency, 5 Hz), at an angle of 45 degrees to the long axis of the tooth. The boundary technique was used for determination of the mean fatigue strengths of the restorations at 10(4), 10(5), and 10(6) cycles, simulating up to 1-3 years of clinical functioning. Mean fatigue strength was expressed in percentage of initial strength: For 10(4), 10(5), and 10(6) cycles, the results were 66%, 58%, and 52%, respectively, for the amalgam and 62%, 62%, and 53% for the composite group. It is concluded that fatigue-testing of more complex systems is possible, if a suitable testing method is selected. The restorations showed a comparable strength reduction after 10(6) cycles of about 50% of their initial strength. The composite core build-up showed a behavior less predictable than that of the amalgam, which might be attributed to handling parameters.

Alloys↗
PubMed1992

[The synchronous appearance of multiple testicular epidermoid cysts and of a malignant mixed germinal tumor in the contralateral testis].

We describe the simultaneous appearance of multiple epidermoid cysts in the right and an adult teratoma containing embryonal carcinoma and choriocarcinoma in the left testis. No similar case has previously been described. Epidermoid cysts of the testis are rare, accounting for about 1% of all testicular tumors. Epidermoid cysts are now regarded as monoepidermally developed teratomas of germ cell origin. Testicular teratomas in adults, however, are always malignant. Because epidermoid cysts are rare tumors, primary therapy often consists in ablation of the testis. In the case described, excision of the epidermoid cyst protected the patient from complete castration. This case demonstrates the simultaneous appearance of a malignant and a benign testicular germ cell tumor.

Adult↗
PubMed1992

Lack of effect of diacetyl-splenopentin (Berlopentin) on the pharmacokinetics of caffeine.

The influence of Berlopentin on caffeine clearance was measured after a 6 week i.v. treatment or a 4 week s.c. administration within a phase I trial. It was demonstrated that therapy with Berlopentin did not affect significantly caffeine elimination. Thus, clearance of drugs which are biotransformed by hepatic microsomal oxidation are unlikely to be affected by the coadministration of Berlopentin.

Adult↗
PubMed1991

A high-affinity inositol 1,3,4,5-tetrakisphosphate receptor protein from brain is specifically labelled by a newly synthesized photoaffinity analogue, N-(4-azidosalicyl)aminoethanol(1)-1-phospho-D-myo-inositol 3,4,5-trisphosphate.

A photolabile arylazido analogue of Ins(1,3,4,5)P4 selectively substituted at the 1-phosphate group was synthesized by coupling 2-aminoethanol(1)-1-phospho-D-myo-inositol 4,5-bisphosphate with N-hydroxysuccinimidyl-4-azidosalicylic acid [Schäfer, Nehls-Sahabandu, Grabowsky, Dehlinger-Kremer, Schulz & Mayr (1990) Biochem. J. 272, 817-825] and subsequently phosphorylating the product by bovine brain Ins(1,4,5)P3 3-kinase. The product, N-(4-azidosalicyl)-aminoethanol(1)-1-phospho-D-myo-inositol 3,4,5-trisphosphate [AsaIns(1,3,4,5)P4] was radioiodinated and purified by anion-exchange chromatography. AsaIns(1,3,4,5)P4 bound to a high-affinity Ins(1,3,4,5)P4 receptor from pig cerebellum with an affinity only 3-fold lower than that of Ins(1,3,4,5)P4. Photoirradiation of 125I-AsaIns(1,3,4,5)P4 in the presence of the receptor preparation revealed that the radioactive label was specifically associated with a protein band of apparent molecular mass 42 kDa, which Donié & Reiser [(1991) Biochem. J. 275, 453-457] had previously tentatively assigned to the Ins(1,3,4,5)P4 receptor protein. The radioactive label was displaced from the receptor when the binding reaction with 125I-AsaIns(1,3,4,5)P4 was carried out in the presence of 5 microM-Ins(1,3,4,5)P4.

Affinity Labels↗
PubMed1991

[The occurrence of Cryptococcus neoformans in fecal samples from birds kept in human living areas].

With help of Guizotia creatinine agar (syn. bird seed agar) Cryptococcus (Cr.) neoformans var. neoformans was isolated from 71 (7.7%) of 925 investigated droppings of birds kept within human living area. Cr. neoformans was detected in droppings of 1.2% from 164 psittacines, in droppings of 1.7% from 118 budgerigars and in droppings of 18.4% from (carrier) pigeons. This yeast was not isolated from droppings of 13 small parrots, 10 eclectus parrots, 13 rosellas, 21 cockatiels, 6 Polytelidus sp., 32 Cacatua sp., 13 Carduelis sp., 5 waxbills, 5 starlings, 120 chicken, 9 turkeys, 2 geese, 2 Turdus sp., 1 duck and 1 gull.

Animals↗
PubMed1991

[Incidence of antibodies to Borrelia burgdorferi in dogs in small animal practice in North Bavaria].

With help of indirect immunofluorescent assay antibodies were detected to Borrelia burgdorferi, agent of Lyme borreliosis in 46 (35.5%) serum samples of 130 randomly selected dogs of a small animal practice in Northern Bavaria. A positive reaction of the indirect immunofluorescent assay was demonstrated in 15 cases in a serum dilution of 1:64, in 9 cases in a serum dilution of 1:128, in 5 cases in a serum dilution of 1:256, in 2 cases in a serum dilution of 1:512, in 3 cases in a serum dilution of 1:1024, in 2 cases in a serum dilution of 1:2048 and in 10 cases in a serum dilution of 1:4096 and higher. There was significant correlation between the symptoms pain of segments or muscles in dorsal region, between painful arthritis, between paralysis of hind legs or between chronic erythema ("hot spot") and the positive reaction of the immunofluorescent assay in the serum dilution of 1:256 ("borderline titre") and higher.

Animals↗
PubMed1991

Cloning of the human gene for intercellular adhesion molecule 1 and analysis of its 5'-regulatory region. Induction by cytokines and phorbol ester.

Human intercellular adhesion molecule-1 (ICAM-1), a specific ligand for the lymphocyte function-associated Ag-1 (LFA-1), plays an important role in leukocyte-endothelial cell interactions. It is induced by proinflammatory cytokines such as IL-1, TNF-alpha, or IFN-gamma. However, little is known concerning the intracellular regulatory mechanisms which trigger ICAM-1 up-regulation. In order to study potential regulatory elements involved in ICAM-1 induction we have cloned the human ICAM-1 gene and 5 kb of its 5'-regulatory region. The sequence of the cDNA was found to be distributed over seven exons separated by six introns, whereby each of the five extracellular Ig-like domains of ICAM-1 is encoded by its own exon. The upstream sequence harbors a number of sequence motifs implicated in the regulation and expression of eukaryotic genes, including binding sites for the transcription factors SP-1, AP-1, and NF-kB. Primer extension and S1 nuclease analysis revealed two transcription initiation sites 319 bp and 41 bp upstream of the translation start site. Consensus TATA boxes were found at the expected positions about 25 bp upstream of both start sites. Reverse transcriptase polymerase chain reaction showed differential use of the two TATA boxes in A549 and HS913T cells. Both RNA seem to code for the same for of ICAM-1 protein. For regulation studies a 1.3-kb EcoRI/SalI fragment of the 5'-flanking region was used to promote transcription of a linked luciferase reporter gene in transient-transfection assays in A549 and HS913T cells. Treatment of A549 cells with IL-1 or TNF-alpha resulted in a two- or fourfold increase in luciferase activity. Furthermore, a sixfold induction could be achieved after treatment with the phorbol ester PMA. In contrast, agents that increase intracellular cAMP levels did not induce luciferase activity. Northern blot analysis was used to investigate the kinetics of ICAM-1 mRNA synthesis upon induction with TNF-alpha and PMA. These data suggest that the up-regulation of ICAM-1 by cytokines occurs at least partly at the transcriptional level. Deletion analysis of the 1.3-kb fragment of the 5'-flanking region revealed sequences responsible for promotion and inhibition of transcription. In particular, two functionally distinct regions have been characterized: a short fragment containing an NF-kB binding site has been shown to function as an activator, followed immediately downstream by a sequence acting as a silencer element. Therefore, ICAM-1 gene expression seems to be modulated by multiple cis-acting elements.

Base Sequence↗
PubMed1991

[Quality control by autopsy. How often do the postmortem examination findings correct the clinical diagnosis?].

To ascertain whether autopsies still provide important information in the face of modern diagnostic procedures, the findings in 780 autopsies (543 men and 237 women: mean age 58 [1-94] years), performed between 1977 and 1990, were compared with the clinical diagnoses. Autopsy confirmed the clinical diagnosis in 74.9% of cases, a clinically not diagnosed basic disease was demonstrated in 13.6%, and a not recognized final complication in 11.5%. In 9.7% of autopsies previously unrecognized but clinically important additional diseases were found. There was no decrease in the proportion of unrecognized basic diseases in the course of the observation period. It was unrelated to the duration of hospital stay, but increased with increasing age of the patients. These observations indicate that even nowadays, in the age of highly technical medicine, autopsies point to any limitations of diagnosis and are thus of importance for clinical quality control.

Age Factors↗
PubMed1991

Alpha B-crystallin is a small heat shock protein.

Sequence similarity between alpha B-crystallin and small heat shock proteins (HSPs) has prompted us to investigate whether alpha B-crystallin expression is induced by heat shock. Indeed, accumulation of alpha B-crystallin was detected immunologically in NIH 3T3 cells after incubation at elevated temperatures and after addition of Cd2+ or sodium arsenite to these cells. Two-dimensional gel electrophoresis revealed identity between alpha B-crystallin from eye lenses and from heat-treated fibroblasts. The promoter of the alpha B-crystallin gene was fused to the bacterial chloramphenicol acetyltransferase gene and was shown to confer heat inducibility on this reporter gene in transient transfection assays. A perfect heat shock element within the promoter region is likely to mediate this response. Small HSPs and alpha B-crystallin were shown to share the following two physical properties: (i) they form supramolecular structures with sedimentation values around 17 S and (ii) they are associated with the nucleus at high temperatures and are localized in the cytoplasm under normal conditions. We conclude that alpha B-crystallin has to be considered a member of the class of small HSPs.

Animals↗
PubMed1991

Short-term treatment with gamma interferon induces stable reversion of ras-transformed mouse fibroblasts.

Persistent revertants have been generated from NIH 3T3 cells transformed by an activated human Ha-ras gene after short-term gamma interferon treatment in the presence of the cardiac aminoglycoside ouabain. Normal fibroblastlike morphology and anchorage dependence are restored in revertants. Tumorigenicity in nude mice is abolished. The revertants continue to express high steady-state levels of the ras oncogene. Partial retransformation of reverted cells is induced after 5-azacytidine treatment or after infection with retrovirus vectors carrying the v-abl, v-fes, v-myc, or v-src oncogene. The revertants resist the transforming activities of the v-Ha-ras and v-mos oncogenes.

Animals↗
PubMed1991

Alpha B crystallin accumulation is a specific response to Ha-ras and v-mos oncogene expression in mouse NIH 3T3 fibroblasts.

The conditional expression of the v-mos and Ha-ras(EJ) oncogenes in NIH 3T3 cells leads to the accumulation of a 23-kDa protein (p23) (R. Klemenz, S. Hoffmann, R. Jaggi, and A.-K. Werenskiold, Oncogene 4:799-803, 1989). We purified p23 to homogeneity and determined part of the amino acid sequence. The obtained sequence is identical with that of the eye lens protein alpha B crystallin. Northern (RNA) blot and Western immunoblot experiments were performed to demonstrate that alpha B crystallin mRNA and protein do indeed accumulate as a consequence of v-mos and Ha-ras oncogene expression. Comparison of cDNA clones obtained from the mRNA of eye lenses and of oncogene-expressing fibroblasts revealed identity between them. The major transcription initiation site of the alpha B crystallin gene in our experimental system was shown by primer extension experiments to be identical with the one used in eye epithelial cells. In addition, we identified a second minor initiation site 49 nucleotides further upstream. Serum growth factors did not stimulate alpha B crystallin expression in growth-arrested cells.

Amino Acid Sequence↗
PubMed1991

Molecular and functional analysis of tumor-suppressor genes by transfection.

The transformed and tumorigenic phenotype of H-ras transfected rat FE-8 cells can be suppressed by cell fusion with normal rat embryo fibroblasts. Transfection into FE-8 cells of DNA prepared from normal human placenta followed by selective elimination of tumorigenic transfected cell clones resulted in the isolation of phenotypically normal revertants. These cells exhibited a fibroblastlike, normal morphology; were anchorage-dependent; and were unable to proliferate in medium with reduced serum concentrations. Their tumorigenicity was also reduced. The suppressed phenotype has been transferred in a second transfection cycle. Human repetitive DNA sequences were detected in secondary transfectant DNA. A putative human suppressor gene, designated NTS-1, has been molecularly cloned. Reintroduction of cloned NTS-1 sequences into FE-8 cells resulted in suppression of the neoplastic phenotype in spite of a high ras expression.

Animals↗
PubMed1991

Endothelin-evoked contractions in bovine ciliary muscle and trabecular meshwork: interaction with calcium, nifedipine and nickel.

In the present study, we compared contractile responses of isolated bovine ciliary muscle and trabecular meshwork strips to endothelin-1 and carbachol. 1. Endothelin-1 is a potent contracting agent for ciliary muscle and trabecular meshwork. The EC50 was 5 x 10(-9) mol/l for both tissues. The maximal force evoked by endothelin was 73% of the maximal carbachol response in trabecular meshwork and 52% in ciliary muscle. 2. Carbachol contracted both tissues with an EC50 of 2 x 10(-7) mol/l. 3. In ciliary muscle, the tension was completely dependent on extracellular calcium. 4. In trabecular meshwork, 23 +/- 4% of the endothelin- and 42 +/- 10% of the carbachol-induced force response remained after removal of extracellular calcium. 5. Nifedipine (10(-5) mol/l) had only a slight relaxing effect in both tissues. 6. Nickel (10(3) mol/l) inhibited the development of force in both tissues. The relaxation induced by nickel was more pronounced in endothelin- than in carbachol-induced contractions. Different intracellular mechanisms mediating the action of endothelin and carbachol can be described: i) a calcium- and a nickel-sensitive pathway in both tissues and ii) an additional mechanism independent of external calcium in trabecular meshwork. These results indicate functional differences between the contractile region of trabecular meshwork and ciliary muscle. Endothelin may participate in accommodation and regulation of the intraocular pressure.

Animals↗