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Biomedical subjects

A G Nerlich

Publications and source records attributed to A G Nerlich.

At least 55 records · Page 3Linked to original sources
PubMed1993

Immunohistochemical detection of interstitial collagens in bone and cartilage tissue remnants in an infant Peruvian mummy.

We investigated the immunohistochemical presence of various collagen types in bone and cartilage tissue from an infant Peruvian mummy dating between 500 and 1000 A.D. which had been excavated at the necropolis of Las Trancas in the Nazca region in Peru. Following careful rehydration and decalcification of the tissue, the mummy tissue showed morphologically good preservation of the matrix, which could be shown to be composed of various collagen types in a typical pattern. Bone consisted of a collagen I matrix with a small rim of collagen III and V at the endosteal lining and a pericellular collagen V staining around osteocytic holes. In the hypertrophic cartilage of the epiphyseal growth plate, a typical pattern of collagen types II and X could be found. These observations provide evidence that in well-preserved mummy tissue the antigenic determinants of major matrix components are still adequately preserved for an immunohistochemical analysis. This technique may thus be a very helpful tool for the analysis of pathologic processes of historic bone tissue. It may also allow in certain circumstances a distinction between pseudopathologic tissue destruction and pathologic tissue alteration.

Bone and Bones↗
PubMed1993

Immunohistochemical analysis of interstitial collagens in cartilage of different stages of osteoarthrosis.

The distribution of the interstitial collagens I, II and III was analyzed immunohistochemically in cartilage and bone samples from 32 patients with degenerative osteoarthrosis at various morphological stages. The alterations observed showed a very patchy, focal distribution demonstrating significant heterogeneity in the tissue reaction. In minor osteoarthrotic lesions a focal pericellular deposition of collagens III and I was seen, while the majority of the interterritorial matrix reacted exclusively with collagen II antibodies. These changes were first seen in the superficial cartilage layer. At the more advanced stages of osteoarthrosis, particularly when osteophytic bone spur formation was present, extensive changes in the expression of collagen types in the pericellular matrix was revealed with extensive and overlapping localization of collagens I, II and III in the whole cartilage. These observations support the suggestion that degenerative cartilage shows a collagen type "switch". In addition, it was demonstrated that the interterritorial cartilage matrix was still mainly composed of collagen II even in advanced lesions. These observations may explain some of the previous discrepancies reported.

Adult↗
PubMed1992

Localization of collagen X in human fetal and juvenile articular cartilage and bone.

The tissue localization was analysed of collagen X during human fetal and juvenile articular cartilage-bone metamorphosis. This unique collagen type was found in the hypertrophic cartilage zone peri- and extracellularly and in cartilage residues within bone trabeculae. In addition, occasionally a slight intracellular staining reaction was found in prehypertrophic proliferating chondrocytes and in chondrocytes surrounding vascular channels. A slight staining was also seen in the zone of periosteal ossification and occasionally at the transition zone of the perichondrium to resting cartilage. Our data provide evidence that the appearance of collagen X is mainly associated with cartilage hypertrophy, analogous to the reported tissue distribution of this collagen type in animals. In addition, we observed an increased and often "spotty" distribution of collagen X with increasing cartilage "degeneration" associated with the closure of the growth plate. In basal hypertrophic cartilage areas, a co-distribution of collagens II and X was found with very little and "spotty" collagen III. In juvenile cartilage areas around single hypertrophic chondrocytes, co-localization of collagens X and I was also detected.

Adolescent↗
PubMed1992

Comparison of various basement membrane components in benign and malignant peripheral nerve tumours.

Immunohistochemical methods were used to analyse benign and malignant tumours of peripheral nerve tissue. We tested for the distribution of basement membrane (BM) components collagen IV, laminin, heparan sulphate proteoglycan, fibronectin, for S100 protein and for the presence of interstitial collagens III and V. Laminin was generally noted in association with Schwann cells, but collagen IV occurred with perineural cells. When tested for BM components, fibroblasts were notably non-reactive except for fibronectin. Three specific area-dependent BM patterns were observed in the benign tumours: (a) Schwann cell-like, in fascicular areas (Antoni A areas of schwannoma, central fibrous bundles of plexiform neurofibromas and central areas of cutaneous neurofibroma), (b) perineural cell-like (capsular structures of schwannoma) and (c) fibroblast-like (myxoid and fibrously transformed areas). Most malignant tissues showed a variably fragmentary focal deposition of laminin. Other BM components were present only in well-differentiated areas. Poorly differentiated tumours demonstrated fibronectin reactivity alone. Our results provide evidence that the specific staining pattern for BM components helps to differentiate the various cellular proliferations in neurogenic tumours. Schwann cells are not only distinguishable from perineural cells by S100 protein staining, but also by their specific BM staining. In addition, perineural cells can be separated from fibroblasts, which do not express BM material. The "tropism" of laminin in normal nerves and benign neural tumours--which persists in neurogenic sarcomas--indicates preferential Schwann cell differentiation in these cells.

Basement Membrane↗
PubMed1992

Excessive collagen formation in fibrolamellar carcinoma of the liver: a morphological and biochemical study.

We have studied the excessive deposition of extracellular matrix in a patient with fibrolamellar carcinoma of the liver. The collagen matrix was predominantly composed of collagens I, III, and V. Since specific mRNAs for collagens I and III were detected by in situ hybridization, we also provide evidence that the fibroblastoid stromal cells were the major source of this collagen. Occasionally, also tumor cells could be shown to express collagen III-mRNA. Furthermore, some tumor cells showed positive signals for TGF-beta 1, while isolated stromal cells expressed interleukin-6. This cytokine expression may probably be related to the altered control of collagen gene expression.

Adolescent↗
PubMed1991

Identification of lymph and blood capillaries by immunohistochemical staining for various basement membrane components.

In the present report we analyzed the presence and distribution of various basement membrane (BM) proteins in normal blood and lymph vessels with special emphasis on BM-associated heparan sulfate proteoglycan (HSPG) when compared to the BM-components collagen IV, laminin and fibronectin. We found that normal lymph capillaries have a BM that contains only collagen IV and small amounts of laminin, but almost no BM-associated HSPG and fibronectin, while blood capillaries showed a BM comprising of all components tested for. Larger lymphatics, however, were indistinguishable from blood vessels on the basis of BM staining. Lymphangiomas showed a BM pattern similar to that of lymph capillaries. Our findings provide evidence that the differential staining of BM-components may represent a reliable method for morphological distinction between blood and lymph capillaries. A comparison of these results with the BM-pattern in other functionally specialized blood vessels (glomerulus, sinusoids) provides evidence that the BM-composition may have some major impact on the functional properties. Thus, it is conceivable that the lack of HSPG in lymph capillaries may be essential for a free influx of fluid and proteins into these capillaries, which may have been extravasated into the interstitium.

Basement Membrane↗
PubMed1990

Collagen cross-linking in adult patients with acute and chronic fibrotic lung disease. Molecular markers for fibrotic collagen.

Lung tissue from patients with interstitial lung disease (ILD), adult respiratory distress syndrome (ARDS), and control subjects with no obvious fibrotic lung disease was analyzed for its content of the collagen cross-links hydroxylysinonorleucine (HLNL), dihydroxylysinonorleucine (DHLNL), and hydroxypyridinium (OHP). We observed significant elevations of the DHLNL:HLNL ratio in patients with ARDS, and significant increases in the content of OHP in lungs of patients with ILD. These results are consistent with data from animal models of fibrotic lung disease, suggesting that increases in the DHLNL:HLNL ratio of lung collagen may serve as a marker of an acute fibrotic episode, whereas increased lung collagen OHP content serves as a marker of chronic lung fibrosis. We suggest that the underlying mechanism for the changes in DHLNL content in (pre)fibrotic acutely injured lung tissue and in OHP content in long-term fibrosis may be an increase in the activity of lysyl hydroxylase, a key intracellular enzyme responsible for a specific post-translational modification of collagen.

Acute Disease↗
PubMed1989

Characterization and localization of basement membrane-associated heparan sulfate proteoglycan in human tissues.

A polyclonal antiserum was raised in rabbits against basement membrane heparan sulfate proteoglycan (HSPG) purified from extracts of isolated porcine glomeruli. The antiserum was characterized by enzyme immunoassay, immunoprecipitation, and immunohistological methods. It was shown to recognize specifically the core protein of HSPG from porcine, rat, bovine, and human glomerular basement membrane, but it did not bind to HSPG from guinea pig or rabbit kidney. The affinity-purified antiserum did not cross-react with other basement membrane proteins like laminin, fibronectin, or collagen type IV. Immunohistochemical studies on tissue sections from several human organs revealed specific basement membrane staining, although the intensity of the reaction differed among the organs tested. Whereas glomerular and other capillary basement membranes showed prominent staining with antibody to the core protein of the proteoglycan, those from nerve, skeletal, cardiac, and smooth muscle reacted only weakly. Adipocytes and liver sinusoid walls were not stained. Independent of the extent of HSPG staining the basement membranes of all different tissues tested reacted strongly with an antiserum against type IV collagen.

Antibody Specificity↗
PubMed1987

Studies on collagen metabolism in the Marfan syndrome.

The pattern of collagen metabolism was studied in nine fibroblast cultures from Marfan patients. The cellular synthesis of collagen and non-collagenous proteins was significantly increased, whereas secretion and degradation remained unchanged. Other steps of post-translational processing such as hydroxylation of prolyl or lysyl residues, affecting triple helix stability, were found to be normal. Furthermore, peptide mapping of isolated a 1(I), a 2(I) and a 1(III) gave no evidence for structural defects. Hence, our study would support the notion that defects other than those affecting collagen type I or III metabolism must represent the molecular basis of the Marfan syndrome.

Adolescent↗
PubMed1987

Pattern of collagen types and molecular structure of collagen in acute post-traumatic pulmonary fibrosis.

Analyses of collagen types and their amino acid structures have been made with lungs obtained from eight patients who died from the adult respiratory distress syndrome after non-pulmonary trauma. Collagen in lungs from patients with the adult respiratory distress syndrome was twice as soluble as that in control lungs (p less than 0.01). The proportion of type III collagen in the whole organ as well as in the pepsin solubilised fraction was slightly but significantly raised (net increase of about 5-10% type III collagen (p less than 0.05]. Both type I and type III collagen from the patients contained less hydroxylysine than collagen from control lungs. The alterations in tissue composition described here and observed in normal wound healing support the notion that acute post-traumatic pulmonary fibrosis resembles a wound healing process in the lungs.

Acute Disease↗
PubMed1984

Release of amino-terminal procollagen peptides in paraquat-induced acute pulmonary fibrosis.

Acute pulmonary fibrosis following intoxication with paraquat is characterized by increased collagen synthesis and deposition in the lungs. In the present study, a chronic lung lymph fistula preparation in sheep was used to investigate the changes in procollagen type III peptide level in the lung lymph in acute pulmonary distress associated with paraquat application. The procollagen type III peptide is supposed to be an indicator of the changing biosynthetic pattern of pulmonary collagen in lung fibrosis. All animals tested showed a progressive pulmonary distress following the application of paraquat characterized by increasing lymph flows and lymph protein flows which monitored the microvascular membrane damage. In parallel to these findings, procollagen peptide levels in the lymph fluid increased, even before manifest biochemically or histologically detectable pulmonary fibrosis occurred. The present results suggest that the release of procollagen type III peptides into lymph fluid is an early indicator of beginning fibrotic tissue disarrangement, even before completed pulmonary fibrosis can be detected by common tissue analysis.

Acute Disease↗
PubMed1984

Biochemical changes in parenchymal lung tissue associated with posttraumatic respiratory failure.

We analysed the composition of parenchymal lung tissue from eleven multiply injured patients who died of protracted respiratory failure after an accident and six normal lungs. Lung wet and dry weights, protein, DNA, RNA and collagen amounts were determined in the lungs. Concentrations per dry weight of all parameters revealed only minor changes. Since there was a marked, time-dependent increase in both wet and dry weight, total lung contents of all components showed highly significant increases with survival time. Protein and RNA contents were elevated linearly in all patients, whereas DNA and collagen increased only in patients surviving more than 2 weeks. No correlation existed between biochemical data and a morphological fibrosis score, while morphometrically estimated cellularity and DNA concentration agreed well. Our results provide evidence that the reparative process for lung injury associated with traumatic shock is a tissue remodelling involving all tissue components in the long run.

Adolescent↗
PubMedDate not supplied

Human keratinocyte cell lines differ in the expression of the collagenolytic matrix metalloproteinases-1,-8, and -13 and of TIMP-1.

We investigated cells and conditioned media of the three human keratinocyte cell lines HaCaT (non-tumorigenic), A5 (benign, tumorigenic) and II-4RT (malignant, tumorigenic) with regard to production and secretion of the collagenases-1 to -3 (MMP-1, MMP-8 and MMP-13) and TIMP-1 using semi-nested RT-PCR, Western blots, ELISA, immunocytochemistry and casein zymography. Transcripts of MMP-1, -8, -13 and TIMP-1 were detected in all cell lines by RT-PCR and the corresponding proteins were found in the cytoplasm of all three cell lines by Western blot analysis and/or immunocytochemistry. The conditioned media of the malignant II-4RT cells contain significantly more MMP-1 and MMP-8 than those of HaCaT or A5 as evidenced by immunoblotting and ELISA. In addition to the presence of latent MMP-1, zymography also detected the active form of this enzyme. TIMP-1 was found only in extracts of all three cell lines, predominantly in A5. This study clearly indicates that the epithelial tumor cells synthesize different collagenases and TIMP-1. The malignant clone secretes increased amounts of distinct collagenases compared to the non-tumorigenic cell line, thereby verifying a correlation between biological behaviour and the amount of collagenases. In addition, we provide clear evidence that MMP-8 is not exclusively found in polymorphonuclear granulocytes, but also in keratinocyte cell lines.

Base Sequence↗
PubMedDate not supplied

Tissue distribution of major matrix metalloproteinases and their transcripts in human breast carcinomas.

The key event of invasive growth of malignant epithelial tumors is the dissolution of the peritumoral basement, membrane (BM). Accordingly, numerous immunohistochemical studies have shown that particularly in breast carcinomas there is an almost complete loss of the BM, even in well-differentiated carcinomas. In order to find out the significance of tumor-associated BM-degradation we localized major matrix metalloproteinases (MMPs) as the most important proteolytic enzymes for connective tissue dissolution. As a prerequisite, we had to identify antibodies reacting specifically on paraffin-embedded tissue material. Extensive pretesting of MMP-2, -3 and -9 antibodies of various sources provided evidence that only a small proportion of the antibodies analyzed showed a specific, positive staining result, as most of the commercial antibodies did not react on the paraffin material or revealed non-specific staining results. Using the specifically reacting antibodies, we analyzed material from 65 cases of invasive ductal breast cancer by immunohistochemistry for the localization of MMP-2, -3 and -9 and by the non-radioactive in-situ hybridization technique for the localization of the MMP-3-mRNA. The specificity of the in-situ hybridization was analyzed using the sense control. We observed a distinct positive immunoreaction for MMP-2, -3 and -9 over both invasive, as well as non-invasive tumor cells, without apparent differences in the staining intensity. Remarkably, there was a significant staining of tumor cell complexes undergoing lymphangiotic dissemination. In addition to this tumor cell staining pattern, a positive immunoreaction, although to reduced proportion, was observed over peritumoral fibroblastic and endothelial stroma cells. Normal breast tissue also revealed a positive immunostaining of epithelial and stromal cells. Using in-situ hybridization, we observed mRNA expression for MMP-3 both in tumor and stroma cells, comparable to the protein data. Normal breast epithelia reacted weakly positive for MMP-3-mRNA. Our data indicate that there is a major active expression of important MMPs in invasive breast carcinomas as the possible cause for the matrix dissolution. These MMPs are synthesized both by tumor and peritumoral stroma cells which may interact with each other. However, the de-novo synthesis and the amount of immunoreactive enzyme protein does not seem to be significantly enhanced in invasive versus noninvasive tumor areas or normal breast epithelia, indicating that other mechanisms, such as enzyme activation and/or differences in the levels of proteinase inhibitors may be biologically essential factors.

Breast↗