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Biomedical subjects

M Rojkind

Publications and source records attributed to M Rojkind.

At least 73 records · Page 4Linked to original sources
PubMed1984

Entamoeba histolytica: collagenolytic activity and virulence.

Several axenic strains of pathogenic and nonpathogenic Entamoeba histolytica were tested for their capacity to digest native radioactive type I collagen gels and to produce liver abscesses when injected into the liver of newborn hamsters. The results demonstrate that the pathogenic strains of amebas (HM1:IMSS, HM3:IMSS, HM38:IMSS and HK9) have a collagenolytic activity that closely correlates with their in vivo capacity to produce liver lesions. The nonpathogenic isolate (Laredo) did not show collagenolytic activity and failed to produce lesions in the liver of newborn hamsters. The results also demonstrate that type I collagen obtained from rodents and cats is degraded less by amebic collagenase than is bovine collagen, which is similar to human collagen. These findings suggest that species susceptibility to invasive infection may depend, among other factors, on the characteristics of the extracellular components of host tissues.

Animals↗
PubMed1983

Liver fibrosis.

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Amino Acids↗
PubMed1983

In vivo collagen synthesis and deposition in fibrotic and regenerating rat livers.

The content of liver collagen increases during regeneration posthepatectomy as well as during scar formation after chronic administration of CCl4. However, while in the former system restitution at integrum of the liver is obtained and a normal ratio of connective tissue to cellular elements is established, in the later condition homeostasis is lost and an absolute increase in connective tissue takes place. Therefore, it was important to determine if this difference was associated with differences in the amount and types of collagen produced and deposited in vivo. In this communication we describe experiments performed to determine the amount of collagen types I, III and V present in the connective tissues prepared from control and experimental animals, and to measure the extent of radioactivity in each collagen type after an in vivo pulse with tritiated proline. Our results showed that 5 days post-hepatectomy the net collagen content per g of fresh liver decreased to 50% of control values, the predominant collagen was type I, and the ratio of type I/III collagen was increased as compared to normal liver; in addition, a relative increase in type V collagen (23.6%) as compared to control animals (6.6%) was also seen. At 11 days post-hepatectomy, the amount of collagen per g of tissue increased to normal values, the ratio of type I/III collagen decreased due to an increase in type III collagen, and the amount of type IV collagen decreased to 12.8%. In the cirrhotic liver the net collagen content showed a five-fold increase, and the predominant collagen present in the extracellular matrix was type I collagen. The amount of radioactivity in collagens deposited into the extracellular matrix of normal, regenerating and fibrotic livers closely followed the percent concentration of liver collagen types. Our results suggest that collagens produced during tissue growth are those needed for remodeling and therefore they may be synthesized by the regenerating cells, mainly parenchymal and endothelial cells. In fibrosis, the predominant collagen is type I, and accordingly, it may be produced preferentially by the fibroblastic cells that are increased in number in the injured liver.

Animals↗
PubMed1983

Liver collagen-type characterization in human schistosomiasis. A histological, ultrastructural, and immunocytochemical correlation.

Liver biopsies of four patients with hepatosplenic schistosomiasis, two patients with schistosomiasis and chronic active hepatitis, two patients with chronic active hepatitis and four control patients with no clinical evidence of either disease, were examined by standard light microscopic techniques, electron microscopy and immunocytochemical staining for collagen type I, III and B. Pure schistosomiasis showed the classical "clay-pipe stem fibrosis" and granulomata composed of eosinophils, macrophages and lymphocytes. In that group, the hepatocellular damage was less conspicuous than in the groups with chronic hepatitis and was usually confined to the granulomatous or fibrotic areas. Destruction of the normal architecture and infiltration by macrophages and lymphocytes with severe damage of hepatocytes was found only in the cases of chronic active hepatitis, with or without associated schistosomiasis. Increased collagen deposits were demonstrated in all three groups. Types I, III and B were found in the enlarged portal triads and fibrotic septa. The intranodular or intralobular collagen stained negatively for type I and strongly positive for types III and B.

Collagen↗
PubMed1982

The collagenase of Entamoeba histolytica.

The present work was designed to investigate the capacity of trophozoites of Entamoeba histolytica to adhere to and digest human collagen types I and III in vitro. The time-course of binding of ameba to both human collagen types I and III was similar. However, the kinetics of detachment were different for each collagen type. Trophozoites of E. histolytica cultured on heat-reconstituted type I collagen gels produced a well-defined area of lysis. Quantitative studies using 14C-labeled collagen revealed that after 24 h of incubation, Entamoeba digested three and a half times more type I than type III collagen, thus suggesting the presence of a collagenase with higher specificity for type I collagen. This activity was optimum with trophozoites harvested after 42 h in culture (1.5 X 10(5) trophozoites/ml). The digestion of type I collagen was a function of the number of trophozoites, and was inhibited by EDTA, L-cysteine, and serum, but not by soybean trypsin inhibitor, phenylmethanesulfonyl fluoride, or N-ethylmaleimide (NEM). Electrophoretic analysis of the type I collagen fragments revealed three main classes of polypeptides of 75,000, 50,000, and 25,000 daltons. Subsequent proteolysis of these collagen fragments was probably carried out by other proteases derived from trophozoites. This activity was inhibited with 10 mM NEM. Collagenase activity appeared to be located at the plasma membrane and direct contact of the ameba with the substrate is required for collagen digestion. The results suggest that collagenase activity of E. histolytica may play an important role in tissue invasion.

Adhesiveness↗
PubMed1981

Colchicine improves the alterations in the liver adenylate cyclase system of cirrhotic rats.

The adenylate cyclase system and the number and affinities of receptors for insulin and glucagon were studied in rats treated with CCl4 and in rats that received colchicine in addition to CCl4. Liver glycogen, cAMP and total collagen content were also measured in those animals. Rats received only mineral oil or only colchicine were used as controls. In this latter group, all parameters measured were normal. Differences in liver collagen content between groups treated with CCl4 and CCl4 + colchicine were not statistically significant. Basal adenylate cyclase activity was 2-fold increased in the CCl4 group. In the animals receiving CCl4 and colchicine, adenylate cyclase activity was normal. Adenylate cyclase activity stimulated by fluoride or by glucagon was also increased in the CCl4 group. However, this increase was due to to the enhanced basal activity. The number of receptors and the affinities of the receptors of glucagon and insulin were normal in all groups. cAMP levels were found increased in the CCl4 treated animals and this was accompanied by a 90% reduction in liver glycogen. In the group treated with CCl4 + colchicine, cAMP was normal and liver glycogen was only reduced 25%. These results suggest that part of the clinical and biochemical improvements observed in the colchicine treated animals were due in part to a reversal of the alterations of the adenylate cyclase system induced by CCl4.

Adenylyl Cyclases↗
PubMed1980

Connective tissue biomatrix: its isolation and utilization for long-term cultures of normal rat hepatocytes.

A new procedure is introduced for the isolation of connective tissue fibers, called biomatrix, containing a significant portion of the extracellular matrix (basement membrane components and components of the ground substance). Biomatrix isolated from normal rat liver contains >90% of the tissue's collagens and all of the known collagen types, including types I and III and basement membrane collagens. The purified collagenous fibers are associated with noncollagenous acidic proteins (including fibronectins and possibly small amounts of glycosaminoglycans). Procedures are also described for preparing tissue culture substrates with these fibers by either smearing tissue culture dishes with frozen sections or by shredding the biomatrix into small fibrils with a homogenizer. The biomatrix as a substrate has a remarkable ability to sustain normal rat hepatocytes long-term in culture. The hepatocytes, which on tissue culture plastic or on type I collagen gels do not survive more than a few weeks, have been maintained for more than 5 mo in vitro when cultured on biomatrix. These cells cultured on rat liver biomatrix show increased attachment and survival efficiencies, long-term survival (months) and retention of some hepatocyte-specific functions.

Animals↗
PubMed1980

Immunocytochemical localization of type B collagen: a component of basement membrane in human liver.

An indirect immunofluorescent method for localizing specific types of collagen was applied to unfixed cryostat sections of autopsy and biopsy specimens of normal and fibrotic human livers. Monospecific antibodies to Types I, III, and B collagens were raised in goats by injecting collagens extracted from normal and fibrotic human livers. The antibody against B collagen stained the delicate sinusoidal meshwork within the lobule that closely paralleled in distribution the staining of reticulin fibers as seen in classical silver preparations. Antibody to Type III collagen stained at the sinusoidal aspect of the hepatocytes in a pattern similar to anti-B antibody. All three antibodies tested strongly stained the portal tracts and the fibrotic bands in specimens with cirrhosis. These observations indicate that reticulin fibers within the liver lobule contain both Type III and Type B collagen.

Antibodies↗