• Volume 12, Issue 3

      September 1987,   pages  165-320

    • General relationships of mammalian orders and evolutionary development of primates inferred from best-fit α-globin phylogenies

      Shama Barnabas R Usha T N Guru Row John Barnabas

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      A best-fit α-globin phylogeny was identified by means of a global minimization approach from among the topologies generated by a parsimony strategy. Zip parsimony method was used to derive a set of near-parsimonious trees. For each of these topologies, a difference matrix was computed; and the topology with the best goodness of fit with the original matrix was retained as the best tree. Based on this phylogenetic scheme inter-relationships among eutherian orders and the evolutionary development of primates has been discussed.

    • Molecular mechanism of facilitated transport by carrier ionophores:a study of energetics

      N Sreerama Saraswathi Vishveshwara

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      The mechanism of ion transport by carrier ionophores is investigated. The electrostatic potential is used as index of the binding energy of a cation with valinomycin and enniatin B. The ion binding capacities of these ionophores are studied as functions of conformation and of distance of an approaching ion-complex. The energetics of dirnerisation and the binding energy profile of an ion in dimers of valinomycin and enniatin B are examined. The binding energy profiles and the electrostatic potential surfaces of valinomycin and enniatin B are compared in relation to their biological activities.

    • Domain II + III of bovine serum albumin: Isolation and its characterization

      Sudhir K Agarwal A Salahuddin

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      Some properties of a fragment of bovine serum albumin containing residues 184–582 of the protein sequence, produced by cyanogen bromide cleavage, have been reported. Urea-induced difference spectra of the fragment showed considerable exposure of aromatic chromophores by 8 M urea. Reversible unfolding of the fragment by urea, as followed by difference spectral measurements at 30°C, pH 7.0, occurred in two distinct steps involving at least 3 major conformational states, namely the native (N), intermediate (X) and completely denatured (D) states. The co-operativity values for the two transitions, N⇌X and X⇌Dwere found to be 4.0 and 16.4, respectively. Analysis of the data on bilirubin binding to bovine serum albumin and its fragment suggested that the fragment retains significant amount of its native structure. However, hydrodynamic parameters such as Stokes radius (3.f14 nm), diffusion coefficient (6.98 × 10−7cm2/s) and frictional ratio (1.32) obtained by analytical gel chromatography as well as intrinsic viscosity (4.31 ml/g) indicates some asymmetry in the fragment molecule.

    • Effect of chemical modification on the binding of gossypol by gossypin (11S protein) and congossypin (7S protein) of cottonseed

      I Mohan Reddy M S Narasinga Rao

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      Binding of gossypol by gossypin and congossypin and their succinylated and sulfhydryl group-blocked derivatives has been measured. The binding by gossypin and congossypin is characterized by weak interaction. Succinylation of gossypin decreases the binding affinity whereas that of congossypin increases it. Blocking of sulfhydryl groups of both the proteins does not significantly affect gossypol binding, Succinylation dissociates gossypin and causes conformational changes whereas it does not dissociate congossypin but causes conformational changes. Sulfhydryl group blocking does not dissociate gossypin or congossypin, nor does it cause any conformational changes.

    • Immediate effects of pyridazinone herbicides on photosynthetic electron transport in algal systems

      Karunyal Samuel Salil Bose

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      Pyridazinone herbicides, SANDOZ 9785 (4-chloro-5-dimethylamino2-phenyl-3-(2H) pyridazinone), SANDOZ 9789 (4-chloro-5 (methylamino)-2-(α, α, α-trifluoro-m-tolyl-3-(2H) pyridazinone) and SANDOZ 6706 (4-chloro-5-(methylamino)-2-(α, α, α-trifluoro-m-tolyl-3-(2H) pyridazinone) inhibited photosystem II electron transport inChlorella protothecoides, when the herbicides were added to the assay medium. The inhibitory eficiency varied with the algal species and the nature of substitution of pyridazinones. Using 3 algal systemsviz., Chlorella, Scenedesmus andAnacystis, the I50 value of for the inhibition of photosynthesis of 3 substituted pyridazinones (SANDOZ 9785, SANDOZ 6706 and SANDOZ 9789) were determined. SANDOZ 9789 was found to be the weakest inhibitor of photosystem II electron transport (H2O→ benzoquinone) as compared to SANDOZ 9785 and SANDOZ 6706. In general, the order of inhibition could be given as SANDOZ 6706 >- SANDOZ 9785 > SANDOZ 9789. The I50 value of photosynthetic particles obtained fromChlorella cells was similar to that of whole cells, suggesting that the cell wall ofChlorella did not act as a barrier for the herbicide action. Studies on the light intensity dependence of SANDOZ 9785 inhibition of electron transport (H2O→ benzoquinone) showed that the light-dependent portion of the curve was more sensitive than the light independent portion of the curve. It is suggested that the site of action was on the reducing side of photosystem II.

    • Heat shock response ofChlorella protothecoides during greening

      T Valliammai A Gnanam R Mannar mannan

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      Heterotrophically grown cells ofChlorella protothecoides were transferred to autotrophic medium and allowed to green at 25°C. The protein synthetic activity of the greening cells measured in terms of incorporation of [35S]-methionine showed a maximum around 20 h of greening and thereafter started declining. Similarly, an analysis of densitometric tracings of the fluorographic profile of the polypeptides associated with both total cellular fraction and membrane fractions during different hours of greening revealed that maximum number of polypeptides were getting labelled around 20 h of greening. At 20 h of greening, the cells were shifted to 40°C and the effect of heat shock on protein synthesis was studied. The heat shock treatment caused a definite decrease in the incorporation of [35S]-methionine into proteins. Due to heat shock, the synthesis of total soluble proteins was affected much more than that of the thylakoid membrane bound proteins. When the cells were transferred back to 25°C after a brief period of heat shock at 40°C, there was a considerable recovery in the protein synthesis and this recovery was found to be significant in the case of soluble proteins, while there was no such definite recovery in the synthesis of thylakoid membrane bound proteins.

    • Modulation of gene expression by the product offitA gene inEscherichia coli

      S Balachandra Dass R Jayaraman

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      Physiological parameters such as viability, gross RNA synthesis,β-galactosidase induction, development of phages T4, T7 andλ have been studied in temperature-sensitiveEscherichia coli strains harbouring fit A76,fit A24 andfit A76fit A24 mutations in rpoB+ andrpoB240 genetic backgrounds. The efficiently of expression of these functions is influenced by thefit A alleles depending upon the medium of growth and/or temperature. Strains harbouring therpoB240 mutation and thefit A76 mutation, either alone or together with thefit A24 mutation, are rifampicin-sensitive even at the perfssive temperature. The results suggest possible interaction between thefit A gene product and RNA polymerase invivo.

    • Enzymes of carbohydrate metabolism ofCotugnia digonopora and their activity in the presence of anthelmintics,in vitro

      N A Pampori V M L Srivastava

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      Cotugnia digonopora, a fowl cycllophyllidean cestode, was found to possess most of the enzymes, associated with the glycolytic sequence and phosphoenolpyruvate branch point, in the cytosol fraction. Enzymes of malate metabolism were predominantly mitrochondrial. Anthelmintic agents inhibited hexokinase, phosphofructokinase, glucose-6-phosphate dehydrogenase, malate dehydrogenase, fumarate reductase, and malic enzyme. In intact worms this effect was significantly reduced. However, the activities of glycogen Phosphorylase and pyruvate kinase were significantly enhanced.

    • Alteration of lipid composition, Na+, K+-ATPase and Δ5-β-hydroxy steroid dehydrogenase activities in microsomal membranes of mature toad ovary in different seasons

      Shyamali Mukherjee Parul Chakrabarti Parimal C Sen

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      Microsomal membranes isolated by sucrose density gradient centrifugation from mature toad ovary has been found to vary significantly in lipid composition and various enzyme activities in different seasons. Na+, K+—ATPase activity is the highest in breeding season (rainy season). Significantly the optimum temperature for enzyme activity is 30°C. The other enzyme Δ5-3β-hydroxysteroid dehydrogenase activity is also lower in hibernation period than other seasons. The total phospholipid, sterol and fatty acid contents differ significantly between seasons. The poly-unsaturated fatty acid, except arachidonic acid content in hibernation period is much lower than that during other seasons. The sterol content is also the lowest in this season. The present findings indicate that during hibernation period the membrane is more rigid and the metabolic activity of the animal is slow because of a lower level of various functionally important enzyme activities.

    • Effect of steroid hormones on vaginal epithelial cells: Anin vitro model for steroid hormone action

      S Vijayasaradhi A Khar P D Gupta

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      Conditions have been standardized to maintain rat vaginal epithelial cellsin vitro with more than 95% viability. Cultured epithelial cells were used to study the effects of normal fetal calf scrum, estradiol and progesterone on the incorporation of [3H]-uridine in RNA and incorporation of [14C]-aminoacids in proteins. While fetal calf serum and estradiol stimulate the incorporation of both uridine and afno acids, progesterone did not show any effect. Estradiol treated vaginal cells show typical fcroridges (indicative of keratinization of cells) in contrast to estradiol deprived cells, which show microvilli on cell surface when examined in scanning electron microscope.

    • Lipid peroxidation in plumbagin administered rats

      R Sankar P S Devamanoharan G Raghupathi M Krishnasamy C S Shyamala Devi

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      Plumbagin was administered to rats at a concentration of 1,2,4,8 and 16 mg per kg body weight. After 24 h lipid peroxide levels were found to decrease in subcellular fractions of liver. Plumbagin inhibited ascorbate and nicotinafde adenine dinucleotide phosphate (reduced) dependent lipid peroxidation but was without any effect on cumene hydroperoxide dependent lipid peroxidation. Injection of 16 mg of plumbagin per kg body weight was found to decrease liver total reduced glutathione and also fcrosomal glucose-6-phosphatase. The results are discussed with reference to the anti- and prooxidant properties of plumbagin.

    • Superoxide production from macrophages of leprosy patients after stimulation withMycobacterium leprae

      Jolly Marolia P R Mahadevan

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      The macrophages from peripheral blood of normal healthy individuals respond to live or killedMycobacterium leprae by producing superoxide. On the other hand, the macrophages from bacteriologically positive (B +LL) or long term treated bacteriologically negative (B -LL) and tuberculoid leprosy patients are unable to produce superoxide when stimulated with liveMycobacterium leprae. While killedMycobacterium leprae induce superoxide with the cells from tuberculoid andB(-)LL patients, cells fromB(+)LL patients fail to respond. The deficiency inB(-)LL patients to produce superoxide appears to be specific withMycobacterium leprae and the defect can be counteracted by the addition of colchicine. These observations indicate a preexisting membrane disposition which does not favour superoxide production. A similar situation is seen in the cells from tuberculoid leprosy patients. Thus it appears that both cured and active lepromatous leprosy patients have defective macrophages, unable to respond to liveMycobacterium leprae to produce superoxide anion, in contrast to the situation with the cells from normal healthy individuals.

    • Secreted proteins of quiescent, serum-stimulated and over-confluent mouse embryo fibroblasts

      M Subramaniam G Shanmugam

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      Quiescent and proliferating cultures of Swiss mouse embryo fibroblasts were pulse labelled with [14C]-amino acids and the newly synthesized proteins that were secreted into the medium were resolved by electrophoresis on Polyacrylafde gradient gels. Conditioned media obtained from quiescent cultures that were stimulated to grow by the addition of 20% fetal calf serum showed the presence of two unique polypeptides of molecular weights 48000 and 26000. A polypeptide of molecular weight 45000 was present in increased amounts in serum-stimulated cells than in quiescent cells. This protein was also superinduced in quiescent cells by cycloheximide treatment. Mouse embryo fibroblasts grown under over-crowded conditions secreted two proteins of molecular weights 35000 and 11000. The 35 K polypeptide was shown to be related to the major excreted protein of transformed cells, since it was immunoprecipitated by an antiserum to major excreted protein. These results indicate that the 48 K and 26 K proteins may be proliferation specific proteins, while the 35 K protein present in the conditioned media of over-confluent cells may be a marker of morphological transformation.

    • Peanut agglutinin induced alterations in capsular and extracellular polysaccharide synthesis andex -planta nitrogenase activity of cowpea rhizobia

      Bina Mody Vinod Modi

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      The root exudate ofArachis hypogea (groundnut) and its seed lectin peanut agglutinin were found to stimulate the synthesis of exopolysaccharide and capsular polysaccharide of the microsymbiont cowpeaRhizobium strain JLn (c). The synthesis of capsular polysaccharide was enhanced 1.5-fold and 2-fold in the presence of peanut agglutinin and root exudate, respectively. The synthesis of capsular polysaccharide was suppressed in the presence of different forms of combined nitrogen. Quantitative differences were also detected between the exopolysaccharide of cells grown in the presence and absence of root exudate. Electron microscopic examination of negatively stained lectin-treated JLn (c) cells showed an increased deposition of capsular polysaccharide surrounding the cells. Hurthermore,ex planta nitrogenase activity of JLn(c) cells in the presence of lectin was found to be enhanced by 63% in correlation with the increased synthesis of polysaccharides.

    • Effect of environmental factors on the limpid composition membrane structure and permeability ofMicrosporum gypseum

      Dipak Kapur Rakesh Pandey Seema Vaidya G K Khuller

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      Supplementation with unsaturated fatty acids, substitution of glucose by glycerol as carbon source and lowered growth temperature (20°C) increased the total phospholipid content ofMicrosporum gypseum spheroplasts. Levels of sterols increased with glycerol substitution and decreased in other growth conditions. Substantial changes were seen in the ratios of unsaturated to saturated fatty acids and phosphatidylcholine to phosphatidylethanolafne under all the experimental conditions. Changed lipid composition resulted in altered uptake of amino acids (L-lysine, L-aspartic acid and L-glycine) and increased number of binding sites for a fluorescent probe, 1-anilinonaphthalene-8-sulfonate.

    • Nitrogen assimilation in opaque and normal sorghum during grain development

      Chitra Narasimhan R P Johari S L Mehta

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      Activity of key nitrogen assimilating enzymes was studied in developing grains of high-lysine opaque sorghum P-721 and normal sorghum CSV-5. The higher percentage of protein in opaque sorghum was mainly due to lower starch content since protein per grain was less than in CSV-5. During grain development, albufn and globulin decreased while prolafne and glutelin increased. Prolafne content in CSV-5 was higher than in opaque sorghum. Average nitrate reductase activity in flag and long leaf were similar in both the varieties. The nitrate reductase activity decreased during grain development. Glutamate dehydrogenase activity was higher during early development and lower at later stages in opaque sorghum than in CSV-5. Glutamate oxaloacetate transaminase activity was higher and glutamine synthetase lower in opaque sorghum than in CSV-5 grains during development. Glutamate synthase activity was higher in opaque sorghum up to day 20 and lower thereafter than in CSV-5. It is suggested that reduced activities of glutamine synthetase as well as glutamate synthase in opaque sorghum as compared to CSV-5 during later stages of development may restrict protein accumulation in the former.

    • Interaction energy studies on pyrazolopyrimidine nucleoside antibiotics — A theoretical study: Oxoformycin B

      Rajendra Prasad Ojha M Roychoudhury Nitish K Sanyal

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      The biological activity of oxoformycin B has been exafned on the basis of the model developed for the incorporation of nucleoside analogues during transcription. Claverie’s simplified formula has been employed for intermolecular interaction energy calculation. The pairing energy of oxoformycin B base with complementary bases as well as the association energy with nucleic acid base pairs have been calculated. The results are compared with those of similar computation with normal bases. In addition to the in-plane interaction the vertical interaction energy between the analogue and the normal bases has been computed to specify the particular position of the analogue in the chain. On the basis of the model an attempt has been made to explain the mechanism of the biological action of oxoformycin B and to compare the biological activity of pyrazolopyrimidine nucleoside analogues.

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