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C Röder

Publications and source records attributed to C Röder.

34 records · Page 2Linked to original sources

Expression of laminin alpha1, alpha2, alpha4, and alpha5 chains, fibronectin, and tenascin-C in skeletal muscle of dystrophic 129ReJ dy/dy mice.

The dy/dy mouse is an animal model for human merosin-negative congenital muscular dystrophy (CMD), which has been reported to have reduced or no expression of the basement membrane protein laminin alpha2. We here investigate various myogenic and nonmyogenic tissues of mature dy/dy and control 129ReJ mice histologically and for laminin alpha2 expression. In addition, expression patterns of laminin alpha1, alpha2, alpha4, and alpha5 chains, the interstitial proteins fibronectin and tenascin-C, and the adhesion molecules VCAM-1, ICAM-1, and alpha4 integrin were characterized in skeletal muscle of 1- and 7-day and mature (>6 weeks old) dy/dy and control 129ReJ mice. The laminin alpha2 chain remained detectable in myogenic tissues of dy/dy mice by immunofluorescence using two different monoclonal antibodies and by Northern blot analysis. However, laminin alpha2 expression was significantly reduced or not detectable in nonmyogenic tissues of dy/dy mice, including skin, lung, kidney, brain, thymus, and eye. Focal lesions were observed in mature skeletal muscle only, characterized by necrotic tissue, isolated VCAM-1- and ICAM-1-positive cells indicative of inflammatory processes, and regenerating muscle fibers surrounded by intense tenascin-C and fibronectin expression. In contrast to studies on human CMD muscle, laminin alpha1 was not detectable in either dy/dy or control skeletal muscle using immunofluorescence or Northern blot analysis. Immunofluorescence localized laminin alpha4 to basement membranes of blood vessels, the endoneurium of the intramuscular nerves, and the neuromuscular junction in skeletal muscle of 1- and 7-day-old dy/dy and control mice. In mature muscle, laminin alpha4 expression shifted to the perineurium of intramuscular nerves in both dy/dy and control mice. Furthermore, strong upregulation of laminin alpha4 in the basement membranes of blood vessels, the perineurium of intramuscular nerves, and of isolated regenerating muscle fibers in the dy/dy mice was apparent. Investigation of 1-day-old animals revealed expression of laminin alpha5 in skeletal muscle fiber basement membranes of dy/dy but not control animals. This difference between dy/dy and control animals was no longer apparent at 7 days after birth, indicating a temporary shift in expression pattern of laminin alpha5 in dy/dy animals. Analysis of the extracellular matrix components of 1- and 7-day-old dy/dy and control skeletal muscle revealed an early onset of the dystrophy, even before histopathological features of the disease were evident. Our data confirm the absence of laminin alpha1 chain in myogenic tissues of both dy/dy and control mice and suggest compensation for reduced laminin alpha2 in dy/dy skeletal muscle by laminin alpha4 and, in early development, also laminin alpha5. These results have significant ramifications in the diagnosis of human merosin-negative CMD.

Age of Onset↗

Photoallergic skin reaction to ribavirin.

A 65-yr-old woman with chronic hepatitis C was treated with three million units interferon-alpha t.i.w. and 1000 mg ribavirin daily. At wk 16 of combination therapy the patient developed an itchy eczematous erythema, partly of urticarial character, which was almost confined to ultraviolet (UV)-exposed sites. Histopathological examination of the skin lesions was consistent with a photoallergic reaction. The minimal erythematous dose for UVA and UVB was assessed on healthy skin. After 24 h, a distinct erythema at the UVB irradiated site was found, whereas no reaction was seen with UVA provocation up to a dose of 10 J/cm2. Correspondingly, determination of the absorption spectrum of ribavirin revealed maximum absorption within UVB at 282.5 nm. Ribavirin was stopped, and the cutaneous lesions and pruritus completely disappeared without subsequent hyperpigmentation. This case indicates that ribavirin is a potential photosensitizer for UVB, which may become increasingly relevant in patients with chronic hepatitis C undergoing combination therapy for 6-12 months with interferon-alpha and ribavirin.

Aged↗

Differential mucin MUC7 gene expression in invasive bladder carcinoma in contrast to uniform MUC1 and MUC2 gene expression in both normal urothelium and bladder carcinoma.

Mucins (MUCs) are high molecular weight membrane glycoproteins. The gene expression of MUCs (MUC1-MUC8) may change characteristically during malignant transformation of epithelial tissues. Total RNA was isolated from the four bladder cancer cell lines RT4, 647V, HT1376, and 486P (pathological gradings between G1 and G4) and 17 samples of transitional cell carcinomas, as well as 16 samples of normal human urothelium of the bladder from surgically removed specimens. The RNA samples were studied with MUC1-, MUC2- and MUC7-specific nested reverse transcription-PCRs. Gene expression of MUC1 and MUC2 was found positive in all normal, as well as in malignant, tissue samples and in the tumor cell lines. In contrast, gene expression of MUC7 was only detected in bladder cancer cell lines and samples of invasive transitional cell carcinomas, but neither in superficial, noninvasive bladder tumors nor normal bladder urothelium. Only one of the samples of normal urothelium obtained from 16 different tumor-bearing bladders was positive for MUC7 gene expression. These results suggest a differential MUC7 gene expression with the onset of malignant transformation of the bladder urothelium.

Carcinoma, Transitional Cell↗

Comparative analysis of bone marrow and venous blood isolates from gastrointestinal cancer patients for the detection of disseminated tumor cells using reverse transcription PCR.

We investigated 141 bone marrow and 104 venous blood isolates from gastrointestinal cancer patients with a cytokeratin (CK) 20-specific nested reverse transcription PCR for the detection of disseminated tumor cells at time of primary tumor resection. In colorectal cancer patients, 20 of 65 (31%) bone marrow and 9 of 52 (17%) venous blood isolates yielded a CK 20 mRNA-positive result in a stage-dependent manner. The detection rates for gastric cancer patients were 11 of 49 (22%) and 5 of 30 (17%) for bone marrow and venous blood, respectively. In pancreatic cancer patients, positive signals were found in advanced tumor stage. A duplex PCR system improved the feasibility of the test. After analyzing 70 sets of bone marrow and venous blood isolates from colorectal, gastric, and pancreatic cancer patients, we observed a higher detection rate in bone marrow isolates. Survival of patients with CK 20 mRNA-positive findings was significantly shorter than that of negatively tested patients.

Biomarkers, Tumor↗

Cloning of the mouse laminin alpha 4 cDNA. Expression in a subset of endothelium.

Endothelial cells express a 400-kDa or 240-kDa laminin alpha chain, depending on their tissue of origin or physiological state [1, 2]. Using differential display and subsequent screening of a mouse endothelial cell cDNA library we here identify the gene coding for the 240-kDa laminin chain as the laminin alpha 4 gene. The complete mouse laminin alpha 4 cDNA sequence is reported and compared with other laminin alpha chains. In situ hybridization of embryonic and new born mouse tissues revealed expression of laminin alpha 4 mRNA in a subset of endothelium, in particular aortic endothelium, endocardium and endothelium of blood vessels in the skin and in the brain. Strong laminin alpha 4 expression by aortic endothelia was confirmed by data obtained from cultured bovine aortic endothelial cells (BAEC). Isolation of laminin from BAEC conditioned medium revealed a Y-shaped molecule in rotary shadowing. Subsequent sequencing of BAEC laminin resulted in laminin alpha 4, beta 1 and gamma 1 amino acid sequences, confirming that laminin alpha 4 is one of the major laminin alpha chains expressed by aortic endothelium not only in the mouse. In addition, strong laminin alpha 4 mRNA expression occurred in peripheral nerves, cardiac muscle, fat, the dermis of the skin and lung stroma of mouse tissues. The data demonstrate a cytokine and progesterone-regulated differential expression of laminin alpha 4 mRNA in mouse endothelium, suggesting a distinct functional role for this laminin chain in endothelium.

Amino Acid Sequence↗

The detection of disseminated tumor cells in bone marrow from colorectal-cancer patients by a cytokeratin-20-specific nested reverse-transcriptase-polymerase-chain reaction is related to the stage of disease.

The dissemination of cancer cells is a prerequisite in the development of micrometastases and solid metastases. Our previous examinations of these cells were based on immunocytological staining of tumor-associated antigens and cytokeratins. We have now developed a highly sensitive and specific detection method based on a nested reverse-transcriptase-polymerase-chain reaction (RT-PCR) of cytokeratin-20 (CK-20) mRNA. Using this method, we examined the bone marrow of 57 patients with colorectal cancer and detected increasing numbers of CK-20-positive samples, depending on the UICC stage. Some 35% of all bone-marrow samples tested positive for CK-20: none were found in colorectal cancer stage 1, 24% were in stage II, 31% in stage III and 71% in stage IV. Investigation of bone-marrow specimens of patients with pancreatic cancer showed that 4 out of 11 patients were positive for CK-20 mRNA. To confirm that sample positivity for CK-20 expression was due to disseminated tumor cells, we examined bone marrow from a control group (n = 16) without apparent carcinoma. In this group, 15 out of 16 donors were CK-20-negative, while one donor with familial adenomatous polyposis showed a CK-20-specific signal.

Base Sequence↗

Autoradiographic analysis of neuropeptide Y receptor binding sites in the rat hippocampus after kainic acid-induced limbic seizures.

Changes in peptide YY receptor binding were investigated at various intervals after limbic seizures induced in rats by an intraperitoneal injection of kainic acid (10-12 mg/kg). Six to 24 h after kainic acid, specific peptide YY binding, representing Y1 and Y2 neuropeptide Y receptor subtypes, was markedly enhanced in the strata radiatum and oriens CA3 (increase by up to 185% and 178% of control values, respectively). Seven and 30 days after kainic acid, a reduction by up to 63% was found. The basal and kainic acid-induced changes in peptide YY binding were mainly represented by Y2 receptor sites. In the hilus of the dentate gyrus, an increase of global peptide YY binding by up to 400% was observed after 24 h which became attenuated to 125% after 30 days. In the molecular layer of the dentate gyrus global peptide YY binding increased by up to 87% between six and 24 h after kainic acid injection and was reduced by 37% after 30 days. Similar changes were observed in the cerebral cortex. Whereas in the hilus of the dentate gyrus peptide YY binding consisted mainly of Y2 sites, it represented predominantly Y1 receptors in the molecular layer and the cortex. The decline in global and Y2 specific peptide YY binding observed at 30 days in the hippocampus proper was prevented in animals protected from seizure-induced brain damage by an anticonvulsant dose of phenobarbital 3 h after injection of kainic acid. In the stratum moleculare of the dentate gyrus, Y2 specific binding was significantly enhanced while global peptide YY binding was slightly decreased compared to controls. These results show lasting changes in neuropeptide Y receptor binding sites after the acute seizures induced by kainic acid. Since neuropeptide Y modulates glutamatergic neurotransmission, these modifications may play an important role in the hippocampal excitability of chronically epileptic rats.

Animals↗

[Kaposi sarcoma in Caucasian women. Clinical, chemical laboratory and endocrinologic studies in 8 women with HIV-associated or classical Kaposi sarcoma].

Classic and HIV-associated Kaposi's sarcoma (KS) are predominantly found in male Caucasians. It is unclear why KS is so rare in women. At the Department of Dermatology, University of Frankfurt, epidemic KS was diagnosed in 299/751 (40%) male and 5/72 (18%) female HIV-infected patients. To detect common parameters regarding immunodeficiency, hormonal disturbances, abnormalities in clinical course, or sexual practices, data of eight women with KS (five HIV+, three HIV-) were compared. In HIV+ women KS is more aggressive than in HIV- women and more aggressive than in HIV+ men. Early and frequent visceral involvement and no preference for the lower extremities are found. Compared to HIV- women with KS, HIV+ women are generally younger (mean age 38 vs 71 years) and significantly immunodeficient (reduced absolute CD4 cell count and CD4/CD8 ratio). All women with KS investigated for sexual hormones (7/8) showed abnormalities: 7/7 had low oestrogens, 4/6 low LH and 3/5 low progesterone. Controls (n = 11, HIV+ without KS) showed a clear trend for higher serum levels of sexual hormones, but the difference was not significant. Although different reasons for low levels of oestrogens (drug abuse, marasmus, menopause) must be considered, the lack of oestrogens seems to be a common finding in HIV+ and HIV- women with KS. There was no correlation between sexual practices (anal or oral-faecal contacts) and the manifestation or aggressiveness of KS.

Acquired Immunodeficiency Syndrome↗

Kainic acid induced seizures cause a marked increase in the expression of neurokinin-3 receptor mRNA in the rat cerebellum.

Marked changes in the expression of the tachykinin peptide neurokinin B (NKB) have been recently observed in animal models of epilepsy. In this study we investigated mRNA levels encoding the receptor for NKB, the neurokinin-3 receptor (NK-3R), after limbic seizures induced by kainic acid (KA) in the rat. NK-3R mRNA levels were determined by nuclease protection assay at various time intervals after i.p. injection of KA in the rat. Increases of more than 200% were observed in NK-3R mRNA in the cerebellum after 7 and 30 days. In the hippocampus a moderate, reversible increase (of 70%, 1 day after KA) was seen. In the frontal cortex a reduction of NK-3R mRNA (2 days after KA) was found. In the amygdala, levels of the transcript were decreased (by 50% and more) at all intervals investigated. The decreases in mRNA levels in the amygdala are consistent with the severe damage observed in this brain area. The increases in NK-3R mRNA in the cerebellum point to the development of receptor supersensitivity and suggest a functional role of NKB in this animal model of epilepsy.

Animals↗

Detection of the anti-p53 antibody response in malignant and benign pancreatic disease.

Genomic alterations in the p53 tumour-suppressor gene and overexpression of p53 protein, resulting from gene mutations, are frequently found in pancreatic cancer. In this study we analysed the sera of 160 patients with malignant and benign pancreatic diseases for the presence of circulating antibodies to the p53 protein. The analysis of the sera was performed using two different enzyme-linked immunosorbent assay (ELISA) systems. To further substantiate the results, all sera were analysed by the Western blot technique using a cell lysate of PancTu-1 cells (p53 mutation at codon 176) as antigen source. Additionally, all positive sera were analysed by the Western blot technique using recombinant p53 as the antigen source. Although the rate of p53 mutations in pancreatic tumours is of the same order as in other adenocarcinomas (> or = 50%), an antibody response was found in only 5/78 (6.4%) sera from patients with pancreatic cancer. Two out of 82 (2.4%) sera of patients with benign pancreatic diseases were clearly positive for p53 antibodies. One additional specimen was weakly positive, i.e. only in one ELISA and Western blot system.

Adult↗

Kainic acid seizures in the rat: differential expression of chromogranin A, carboxypeptidase H and peptidylglycine alpha-amidating monooxigenase in subfields of the hippocampal formation.

Using in situ hybridization histochemistry concentrations of mRNAs encoding chromogranin A (ChA), carboxypeptidase H (CPH) and peptidylglycine alpha-amidating monooxigenase (PAM) have been investigated in the hippocampus after kainic acid (KA)-induced limbic seizures in the rat. Increased concentrations (by 150%) of ChA and CPH mRNAs were found in the granule cell layer 24 h after KA injection. At the same time PAM mRNA levels were only slightly elevated (by 50%). Whereas the increases in CPH and PAM transcripts were only transient, ChA mRNA concentrations in the granule cell layer were elevated up to 2 months after the initial seizures. In contrast, in the pyramidal cell layers of all hippocampal subfields (CA1 to CA3) ChA mRNA concentrations were significantly reduced (by 40-70%) 1-60 days after KA. PAM and CPH messages were slightly reduced in the pyramidal cell layer of CA1 but not in CA2 and CA3. The experiments demonstrate that KA-induced limbic seizures cause sustained changes in the expression of ChA mRNA. At the same time the expression of two enzymes involved in post-translational processing of neuropeptides, PAM and CPH, becomes only transiently altered. Synthesis of ChA may be regulated differently in the strata granulosum and pyramidale during epileptic seizures.

Animals↗

Cell surface proteins binding to recombinant soluble HIV-1 and HIV-2 transmembrane proteins.

OBJECTIVE: To further characterize cell surface proteins binding to recombinant soluble (rs) forms of the transmembrane glycoproteins gp41 of HIV-1 (rsgp41) and gp36 of HIV-2 (rsgp36). METHODS: Various human and murine cell lines of different lineages were surface-labelled with 125I. rsgp41 and rsgp36 were bound to CnBr-Sepharose and used as an affinity matrix for the surface-labelled cell lysates. The bound cell surface proteins were separated on sodium dodecyl sulphate polyacrylamide gel electrophoresis under reducing conditions. A rabbit serum was produced against one of the cell surface proteins and flow cytometry used to compare the results with those obtained from affinity chromatography. RESULTS: We have confirmed and extended the results obtained by Qureshi et al. [1]. In addition to the 44kD protein, we identified cell surface proteins with molecular weights of 98 and 106 kD binding with high affinity to both rsgp41 and rsgp36. We have demonstrated differences between human and murine cell lines in the expression of the cell surface proteins that interact with rsgp41 and rsgp36. Furthermore, a correlation between the level of rsgp41 and rsgp36 binding proteins, detected either by affinity chromatography or by reactivity with an antiserum directed against one of the cell surface binding components was shown. CONCLUSIONS: Three cell surface proteins, with molecular weights of 44, 98 and 106 kD, bind with high affinity to rs forms of gp41 and gp36. Their expression decreases from a T-lymphoid cell line, to a monoblastoid cell line, to a cell line representing mature monocytes. Human T-cell lymphotropic virus-infected cell lines show a predominance of the 44 kD protein. There are species-specific differences, in that murine cell lines lack the 44 kD protein.

Animals↗