Differences in collagen metabolism between normal and osteoarthritic human articular cartilage.

Normal human articular cartilage synthesizes only one type of a chain, which exhibits the chromatographic behavior of the alpha(l)(II) chains described for chick and bovine cartilage. Osteoarthritic cartilage, on the other hand, synthesizes in addition a collagen containing alpha(2) chains and beta components. The different structural features of the two types of collagen may account for some of the functional defects of osteoarthritic cartilage.

Effects of calcitonin and parathyroid hormone on the metabolism of chondrocytes in culture.

Rabbit articular chondrocytes in suspension culture synthesize Type II collagen [3alpha1(II)] in the absence of extracellular Ca2+ and Type I collagen [2alpha1(I) - alpha2] in the complete medium. As a result of pre-treatment in monolayer culture with calcitonin or parathyroid hormone in the complete medium, an influx of Ca2+ into the cells occurs. These cells produce mainly Type I collagen when transferred to suspension cultures in the medium devoid of CaCl2. If added directly to the suspension culture medium containing no CaCl2, calcitonin stimulates an active efflux of Ca2+ from the cells into the medium and leads the cells to synthesize Type I collagen. Under similar conditions, parathyroid hormone does not change the collagen-phenotype.

Hepatoprotective potential of ether insoluble phenolic components of n-butanol fraction (EPC-BF) of flaxseed against CCl(4) -induced liver damage in rats.

OBJECTIVE: to investigate the hepatoprotective potential of ether insoluble phenolic components of n-butanol fraction (EPC-BF) of flaxseed against CCl(4) -induced liver damage in rats. MATERIALS AND METHODS: Hepatotoxicity was induced to Wistar rats by administration of 0.2% CCl(4) in olive oil (8 mL/kg, i.p.) on the seventh day of treatment. Hepatoprotective potential of EPC-BF at doses, 250 and 500 mg/kg, p.o. was assessed through biochemical and histological parameters. RESULTS: EPC-BF and silymarin pretreated animal groups showed significantly decreased activities of Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and level of total bilirubin, elevated by CCl(4) intoxication. Hepatic lipid peroxidation elevated by CCl(4) intoxication were also found to be alleviated at almost normal level in the EPC-BF and silymarin pretreated groups. Histological studies supported the biochemical findings and treatment of EPC-BF at doses 250 and 500 mg/kg, p.o. was found to be effective in restoring CCl(4) -induced hepatic damage. However, EPC-BF did not show dose-dependent hepatoprotective potential. EPC-BF depicted maximum protection against CCl(4) -induced hepatic damage at lower dose 250 mg/kg than higher dose (500 mg/ kg). CONCLUSION: EPC-BF possesses the significant hepatoprotective activity against CCl(4) induced liver damage, which could be mediated through increase in antioxidant defenses.

Classification of nonenzymatic homologues of protein kinases.

Protein Kinase-Like Non-kinases (PKLNKs), which are closely related to protein kinases, lack the crucial catalytic aspartate in the catalytic loop, and hence cannot function as protein kinase, have been analysed. Using various sensitive sequence analysis methods, we have recognized 82 PKLNKs from four higher eukaryotic organisms, namely, Homo sapiens, Mus musculus, Rattus norvegicus, and Drosophila melanogaster. On the basis of their domain combination and function, PKLNKs have been classified mainly into four categories: (1) Ligand binding PKLNKs, (2) PKLNKs with extracellular protein-protein interaction domain, (3) PKLNKs involved in dimerization, and (4) PKLNKs with cytoplasmic protein-protein interaction module. While members of the first two classes of PKLNKs have transmembrane domain tethered to the PKLNK domain, members of the other two classes of PKLNKs are cytoplasmic in nature. The current classification scheme hopes to provide a convenient framework to classify the PKLNKs from other eukaryotes which would be helpful in deciphering their roles in cellular processes.

Metabolic changes in rabbit articular cartilage due to inflammation.

The effect of inflammation on the articular cartilage of rabbit knee joints were studied. The inflammation was induced by intraarticular injections of corton oil or rabbit peritoneal leukocyte lysates. An increase in the activities of various lysosomal enzymes was observed in the synovial fluid as well as in the cartilage of the inflamed joints. Loss of proteoglycans, increased rate of degradation of collagen and proteoglycans, and increased rate of their synthesis were evident in the treated cartilage. The rate of uptake of 3H-thymidine was also increased. A significant change was observed in the type of collagen synthesized by these explants in vitro. In addition to synthesizing their characteristics Type II collagen, the cartilage explants from the treated joints synthesized Type I collagen.

Characterization of collagen and its precursors synthesized by rabbit-articular-cartilage cells in various culture systems.

Rabbit articular cartilage synthesizes type II collagen, comprised of alpha1(II) chains, in vivo or in vitro. Chondrocytes from the same tissue have the ability to produce type I chollagen with the chain composition 2alpha(I)-alpha2 or type II collagen, depending upon the culture systems in which they are maintained. Type I collagen and its precursors have been identified in the medium of monolayer cultures. Upon transfer from monolayer to suspension cultures, the cells synthesize type II collagen in the medium devoid of CaCl2 and type I collagen and its precursors in the complete medium.

Chemical changes associated with aging of collagen in vivo and in vitro.

Collagen extracted from rat skin by neutral-salt solutions contains less aldehydes than the more insoluble collagen fractions. The concentration of aldehydes in collagen is directly related to its capacity to form stable cross-linked gels, which do not redissolve on cooling and become more insoluble in a variety of reagents. Whereas the absorption spectrum of neutral-salt-soluble collagen treated with N-methylbenzothiazolone hydrazone resembles that of acetaldehyde, the more insoluble collagen fractions show increasing amounts of a component that behaves like an alphabeta-unsaturated aldehyde. The ratio between alpha- and beta-sub-units present in a particular fraction of soluble collagen seems to be constant and independent of the age of the animal. Neutral-salt-soluble collagen, which has a low concentration of beta-components, will generate intramolecular bonds if gelled at 37 degrees . These intramolecular bonds seem to precede the formation of stable intermolecular cross-links, since these gels can redissolve when cooled to yield a soluble collagen with a higher content of beta-components of intramolecular origin.

Synthesis of collagen by chondrocytes in suspension culture: modulation by calcium, 3':5'-cyclic AMP, and prostaglandins.

Rabbit articular chondrocytes synthesize type II collagen [3alpha(1)(II)] in vivo and type I collagen [2alpha(1)(I).alpha(2)] in monolayer cultures. In suspension culture the nature of phenotype depends on extracellular Ca(2+). The relationship of Ca(2+) and 3':5'-cyclic AMP (cAMP) in regulation of collagen synthesis has been investigated. In suspension culture, cAMP levels of chondrocytes increase by 2- to 3-fold and then reach basal values regardless of the presence or absence of extracellular Ca(2+). The cells, however, synthesize primarily type II collagen in the absence of CaCl(2) in the medium and type I collagen in medium containing 1.8 mM CaCl(2). If CaCl(2) is added when intracellular cAMP levels are low, the phenotype is type I collagen. These observations minimize the role of cAMP as a second messenger in the chondrocyte culture system. Increasing endogenous cAMP with a phosphodiesterase inhibitor or adding exogenous dibutyryl-cAMP leads the cells to synthesize type I collagen, although this effect is significantly less pronounced if the medium contains ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA). Increased concentrations of cAMP may mobilize the intracellular calcium pools and activate the cells to switch their phenotypic expression. Prostaglandins E(2) and F(2)alpha, thought to be involved in rheumatoid arthritis and bone resorption, have no significant effect on cAMP content of chondrocytes and alter their collagen phenotype to a small extent.

Isolation and characterization of cyanogen bromide peptides from the collagen of bovine articular cartilage.

Significant amounts of native collagen can be extracted from bovine articular cartilage after removal of the acid mucopolysaccharides by controlled proteolysis. The fraction thus solubilized upon denaturation gives rise to three identical alpha chains. Cleavage of these chains with CNBr generated nine peptides, all of which contain glycine as one-third of their total amino acid residues. Two of the smaller peptides CB-1 and CB-2 contain partially hydroxylated proline. A similar CNBr digest of intact cartilage also gives a series of peptides identical with those obtained from the soluble cartilage collagen. The absence of cross-linking peptides, the fact that only few beta components are seen in articular cartilage collagen and the similarity in peptide pattern between the two collagen fractions investigated, suggests that this collagen is stabilized by a different cross-linking mechanism, possibly involving an association with the tissue proteoglycans.

Mortality associated with induced abortion.

PIP: 5 deaths occurred out of the 3752 induced abortions carried out at J.J. Hospital attached to Grant Medical College, Bombay, between April 1972 and March 1977. 4 of these deaths occurred during the second trimester, indicating that delayed abortion brings increased risks. Most of the cases in the second trimester (total 1152) were terminated by intra-amniotic hypertonic saline (IAS), including the 4 deaths. The fifth death occurred after a suCtion curettage at 12 weeks. There was a relatively high incidence of complications mainly involving infection after administration of IAS. To minimize the mortality due to intra-amniotic hypertonic saline injection it is essential that 1) the patient is carefully evaluated by the specialist, who will do the procedure in a well equipped hospital; and 2) the patient should be kept under constant observation throughout the procedure and afterwards. Although induced abortion will never be without some risk, the risk may be minimized if the procedure is carried out in the first trimester under suitable conditions by an experienced person using well-established techniques.^ieng