EVIDENCE FOR CHANGES IN PROTEIN POLYSACCHARIDE ASSOCIATED WITH THE ONSET OF CALCIFICATION IN CARTILAGE

Past work has suggested that protein polysaccharide may play a role in the calcification of cartilage. Recent electron microscopic studies on noncalcified cartilage have indicated that protein polysaccharide in cartilage matrix is represented by granules associated with collagen fibers. The present work has been designed for comparison of the matrix of noncalcified cartilage to that of calcified cartilage, with particular reference to these granules. Small blocks of tibia from 16-day embryos were fixed in cacodylate-buffered glutaraldehyde and postfixed in either phosphate- or Veronal-buffered osmium tetroxide. Special care was taken to maintain the pH above 7.0 at all times. For electron microscopy the tissues were dehydrated, embedded in Epon 812, sectioned, and stained with uranyl acetate or lead citrate. A marked decrease in the size of granules in the matrix of calcified cartilage compared to noncalcified cartilage was noted. Associated with the decrease in the size of granules was a condensation of matrix components and the presence of an amorphous electron-opaque material that was not seen in noncalcified areas. These results are interpreted to represent either a drop in concentration or a change in state of protein polysaccharide with the onset of calcification in cartilage.     

Homology modeling, simulation and molecular docking studies of catechol-2, 3-Dioxygenase from Burkholderia cepacia: Involved in degradation of Petroleum hydrocarbons

Catechol 2, 3-dioxygenase is present in several types of bacteria and undergoes degradation of environmental pollutants through an important key biochemical pathways. Specifically, this enzyme cleaves aromatic rings of several environmental pollutants such as toluene, xylene, naphthalene and biphenyl derivatives. Hence, the importance of Catechol 2, 3-dioxygenase and its role in the degradation of environmental pollutants made us to predict the three-dimensional structure of Catechol 2, 3-dioxygenase from Burkholderia cepacia. The 10ns molecular dynamics simulation was carried out to check the stability of the modeled Catechol 2, 3- dioxygenase. The results show that the model was energetically stable, and it attains their equilibrium within 2000 ps of production MD run. The docking of various petroleum hydrocarbons into the Catechol 2,3-dioxygenase reveals that the benzene, O-xylene, Toluene, Fluorene, Naphthalene, Carbazol, Pyrene, Dibenzothiophene, Anthracene, Phenanthrene, Biphenyl makes strong hydrogen bond and Van der waals interaction with the active site residues of H150, L152, W198, H206, H220, H252, I254, T255, Y261, E271, L276 and F309. Free energy of binding and estimated inhibition constant of these compounds demonstrates that they are energetically stable in their binding cavity. Chrysene shows positive energy of binding in the active site atom of Fe. Except Pyrene all the substrates made close contact with Fe atom by the distance ranges from 1.67 to 2.43 Å. In addition to that, the above mentioned substrate except pyrene all other made π-π stacking interaction with H252 by the distance ranges from 3.40 to 3.90 Å. All these docking results reveal that, except Chrysene all other substrate has good free energy of binding to hold enough in the active site and makes strong VdW interaction with Catechol-2,3-dioxygenase. These results suggest that, the enzyme is capable of catalyzing the above-mentioned substrate.     

IGF-1 modulates gene expression of proteins involved in inflammation,cytoskeleton, and liver architecture

Even though the liver synthesizes most of circulating IGF-1, it lacks its receptor under physiological conditions. However, according to previous studies, a damaged liver expresses the receptor. For this reason, herein, we examine hepatic histology and expression of genes encoding proteins of the cytoskeleton, extracellular matrix, and cell-cell molecules and inflammation-related proteins. A partial IGF-1 deficiency murine model was used to investigate IGF-1’s effects on liver by comparing wild-type controls, heterozygous igf1^+/?, and heterozygous mice treated with IGF-1 for 10 days. Histology, microarray for mRNA gene expression, RT-qPCR, and lipid peroxidation were assessed. Microarray analyses revealed significant underexpression of igf1 in heterozygous mice compared to control mice, restoring normal liver expression after treatment, which then normalized its circulating levels. IGF-1 receptor mRNA was overexpressed in Hz mice liver, while treated mice displayed a similar expression to that of the controls. Heterozygous mice showed overexpression of several genes encoding proteins related to inflammatory and acute-phase proteins and underexpression or overexpression of genes which coded for extracellular matrix, cytoskeleton, and cell junction components. Histology revealed an altered hepatic architecture. In addition, liver oxidative damage was found increased in the heterozygous group. The mere IGF-1 partial deficiency is associated with relevant alterations of the hepatic architecture and expression of genes involved in cytoskeleton, hepatocyte polarity, cell junctions, and extracellular matrix proteins. Moreover, it induces hepatic expression of the IGF-1 receptor and elevated acute-phase and inflammation mediators, which all resulted in liver oxidative damage.     

An analysis of Bat Hawk Macheiramphus alcinus diet in the Melaky Region of lowland western Madagascar

We present information on the prey taken by the Bat Hawk Macheiramphus alcinus in two different areas of lowland western central Madagascar. These are the first dietary data from Madagascar for this widespread Old World species. The recovered remains were almost exclusively of bats and birds, with a few examples of reptiles and insects. In total, 178 pellets were analysed. On the basis of minimum number of individuals and biomass, bats accounted for 58.3% and 30.3%, respectively, and birds 36.1% and 69.7%, respectively. Amongst the nine species of bats recovered from the pellets, four were represented by multiple individuals, particularly taxa belonging to the families Molossidae and Vespertilionidae that fly in open areas, and for the 11 species of identified birds, all were represented by a single individual. These patterns are interpreted as a specialisation of feeding on bats during a narrow window of time at dusk, as they leave day roost sites, and then using birds in a more general manner to fill in nutritional needs.     

Studies on ultrastructural identification and distribution of protein-polysaccharide in cartilage matrix

Previous reports on the ultrastructure of cartilage matrix have described fibers, amorphous ground substance and, in some instances, dense matrix granules. The fibers are presumably collagen, but the nature of the granules is unknown. The primary purpose of this study has been to investigate the ultrastructure of cartilage matrix ih chick embryos with particular emphasis on the distribution and composition of these granules. In matrix of the zone of articular cartilage, mature collagen fibers can be seen but granules are not present. In matrix of all other zones of cartilage, fibers are smaller and granules are present. When the matrix of epiphyseal cartilage is compared to that of the zone of hypertrophic cells, fibers are similar but the granules in the latter zone are larger and more numerous. The granules in both zones were digested by hyaluronidase and positive to colloidal iron staining. Chemical analyses of cartilage from these zones indicate the hexosamine and radiosulfate content of the zone of hypertrophic cells to be higher than that of the zone of epiphyseal cartilage. The increased hexosamine was shown by column chromatography to be principally sulfated mucopolysaccharide, thereby indicating a direct correlation between size and number of granules and sulfated mucopolysaccharide content in the two zones. These data and the results of the electron microscopic histochemical studies are consistent with the concept that the granules in cartilage matrix contain acidic mucopolysaccharide.