Mechanism of endotoxin-induced reduction in the number of beta-adrenergic receptors in dog livers: role of phospholipase A

The role of phospholipase A on the endotoxin-induced reduction in the number of beta-adrenergic receptors in dog liver plasma membranes was investigated. The results show that digestion of control liver plasma membranes with exogenous phospholipase A2 (0.2 unit/200 micrograms protein) decreased the specific binding of (-)-[3H]dihydroalprenolol by 37.3% (P less than 0.01) and reduced the number of receptor sites by 31.7% (P less than 0.05). These decreases in the specific binding and the number of beta-adrenergic receptors were completely reversible by the addition of phosphatidylcholine (0.2 mM). Endotoxin administration (2 hr postendotoxin) decreased the specific binding by 36% (P less than 0.05) and reduced the number of beta-adrenergic receptors by 33% (P less than 0.05), and these decreases were completely reversible by the addition of 0.2 mM phosphatidylcholine. Digestion of control liver membranes with exogenous phospholipase A2 decreased phosphatidylcholine and phosphatidylethanolamine levels by 50.6 and 51.2%, respectively, but increased lysophosphatidylcholine and lysophosphatidylethanolamine levels by 12- and 8.4-fold, respectively. Endotoxin administration decreased phosphatidylcholine and phosphatidylethanolamine contents by 21.4 and 23.8%, respectively, but increased lysophosphatidylcholine and lysophosphatidylethanolamine contents by 2.1- and 1.4-fold, respectively. In addition, endotoxin administration increased endogenous phospholipase A activity by 73.5%. Based on these results, it is suggested that the decreases in the specific binding and the number of beta-adrenergic receptors in dog livers during endotoxic shock are a result of phospholipase A activation.     

The role of Asn14 in the stability and conformation of the reactive-site loop of winged bean chymotrypsin inhibitor: crystal structures of two point mutants Asn14-->Lys and Asn14-->Asp.

A double-headed chymotrypsin inhibitor, WCI, from winged bean seeds was cloned for structural and biochemical studies. The inhibitor was subjected to two point mutations at a conserved position, Asn14. This residue, known to have a pivotal role in stabilizing the first reactive-site loop (Gln63-Phe68) of the inhibitor, is highly conserved in the sequences of the other members of Kunitz (STI) family as well as in the sequences of Kazal family of serine protease inhibitors. The mutants, N14K and N14D, were subjected to biochemical assay and their characteristics were compared with those of the recombinant inhibitor (rWCI). Crystallographic studies of the recombinant and the mutant proteins are discussed. These studies were primarily aimed at understanding the importance of the protein scaffolding towards the conformational rigidity of the reactive-site loop. Our analysis reveals that, as the Lys14 side chain takes an unusual fold in N14K and the Asp14 side chain in N14D interacts with the loop residues by water-mediated hydrogen bonds, the canonical conformation of the loop has remained effectively intact in both the mutant structures. However, minor alterations such as a 2-fold increase in the inhibitory affinity towards the cognate enzyme were observed.     

Seasonal Abundance of Acanthamoeba rhysodes (Singh, 1952) (Protozoa: Gymnamoebia) in a Mangrove Litter-Soil Ecosystem of Gangetic-Estuary,India1,2

Acanthamoeba rhysodes has been found to be a predominant intertidal benthic gymnamoeba in the mangrove ecosystem of Sundarbans of lower deltaic Bengal, facing the Bay. The sampling zones under study were the highest high tide regions, with characteristic mangrove litter-soil, inundated twice per month during the highest ebb of spring tide. Population abundance of this species, both in its trophic and cystic forms in the three distinct seasonal periods of pre-monsoon (March to June), monsoon (July to October), and post-monsoon (November to February) has been surveyed for over two years. These seasonal periods affect the physico-chemical parameters of the habitat substrata, including temperature, pH, and salinity. It has been found that the overall number of organisms per gram of soil attains peak value during the monsoon period. This value comes down in post-monsoon samples and is the least in pre-monsoon ones.     

Vasculature of the head and respiratory organs in an obligate air-breathing fish,the swamp eel Monopterus (=Amphipnous) cuchia

Methyl methacrylate vascular corrosion replicas were used to examine the macrocirculation in the head region and the microcirculation of respiratory vessels in the air-breathing swamp eel Monopterus cuchia. Fixed respiratory tissue was also examined by SEM to verify capillary orientation. The respiratory and systemic circulations are only partially separated, presumably resulting in supply of mixed oxygenated and venous blood to the tissues. A long ventral aorta gives rise directly to the coronary and hypobranchial arteries. Two large shunt vessels connect the ventral aorta to the dorsal aorta, whereas the remaining ventral aortic flow goes to the respiratory islets and gills. Only two pairs of vestigial gill arches remain, equivalent to the second and third arches, yet five pairs of aortic arches were identified. Most aortic arches supply the respiratory islets. Respiratory islet capillaries are tightly coiled spirals with only a fraction of their total length in contact with the respiratory epithelium. Valve-like endothelial cells delimit the capillary spirals and are unlike endothelial cells in other vertebrates. The gills are highly modified in that the lamellae are reduced to a single-channel capillary with a characteristic three-dimensional zig-zag pathway. There are no arterio-arterial lamellar shunts, although the afferent branchial artery supplying the gill arches also supplies respiratory islets distally. A modified interlamellar filamental vasculature is present in gill tissue but absent or greatly reduced in the respiratory islets. The macro- and micro-circulatory systems of M. cuchia have been considerably modified presumably to accommodate aerial respiration. Some of these modifications involve retention of primitive vessel types, whereas others, especially in the microcirculation, incorporate new architectural designs some of whose functions are not readily apparent.     

Keratinophilic fungi from Orissa, India, II: Isolations from feathers of wild birds and domestic fowls

Feather samples from 117 birds, representing 11 species, were examined for keratinophilic fungi. Of these, 63 (53.8%) were found to be positive. Nine species of fungi classified in 6 genera were isolated: Aphanoascus terreus (1.7%), Chrysosporium indicum (26.4%), Chrys. tropicum (11.1%) the Chrysosporium state of Aphanoascus spp. (2.5%), the Chrysosporium state of Arthroderma tuberculatum (3.4%), Ctenomyces serratus (0.8%), Gymnoascoideus petalosporus (7.6%), Malbranchea spp. (6.8%), Rollandina hyalinospora (6.8%). Passer domesticus (the house sparrow) harboured the largest varieties of keratinophilic fungi: the Chrysosporium state of Arthroderma tuberculatum was found to be associated with it selectively. The associations of these keratinophilic fungi with birds were non-clinical in nature.     

Anemia in experimental visceral leishmaniasis in hamsters.

Experimental infection of hamsters with Leishmania donovani caused visceral leishmaniasis in which hematological changes occurred. The infected hamsters were anemic and reticulocyte counts were high. No significant change in the serum erythropoietin level was noted. Red cell membrane Na(+)-K(+)-ATPase and acetylcholinesterase activities increased. Osmotic fragility of the erythrocytes from infected animals increased. The level of 2,3-diphosphoglycerate of the red cells increased with the degree of anemia.     

Phenazine ethosulfate as a preferred electron acceptor to phenazine methosulfate in dye-linked enzyme assays.

Rapid nonenzymatic reduction of 2,6-dichlorophenolindophenol by N-methyl phenazonium methosulfate has been observed in aqueous solution and has been found to increase with increasing pH and ionic strength. The instability of N-methyl phenazonium methosulfate in aqueous solution has been explored in terms of change of absorption spectrum and formation of free radicals as evidenced by EPR spectroscopy. N-Ethyl phenazonium ethosulfate has been found to be much more stable than the methyl analog and did not reduce dichlorophenolindophenol nonenzymatically. The implications of these findings with respect to use of these dyes as artificial electron acceptors are discussed and the recommendation made that, wherever possible, use of N-methyl phenazonium methosulfate be discontinued in favor of use of the N-ethyl analog.     

A study of the relative efficiency of mustard oil cake and urea and the effect of salinity in plankton production in indian fishponds

Augmentation of plankton production in Indian fishponds by application of manure and inorganic fertilisers was studied in laboratory experiments. The effects of the salinity of the medium were also examined.Laboratory trials were made with mustard oil cake and urea on equivalent nutrient bases in media with varying degrees of salinity (0–30 ppt at 2 ppt. intervals). It was observed that Closterium, Fragilaria, Pinnularia and Gyrosigma grew well in the higher salinity range between 24 to 30 ppt. whereas lower salinities ranging from a trace to 8 ppt. are suitable for improved production of Anabaena, Synedra, Navicula, Amphipleura, Amphora and Nitzschia. Comparatively better production of plankton was recorded with urea than with mustard oil cake.