• Volume 20, Issue 2

      June 1995,   pages  141-287

    • Relationship between glycerol-3-phosphate dehydrogenase, fatty acid synthase and fatty acid binding proteins in developing human placenta

      Anup K Bandyopadhyay Tanya Das Gouri Sankar Sa Manju Mukherjea

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      The activities of the enzymes glycerol-3-phosphate dehydrogenase and fatty acid synthase are inhibited by palmitoyl-coenzyme A and oleate. The two isoforms of fatty acid binding proteins (PI 6.9 and PI 5.4) enhance the activities of glycerol-3-phosphate dehydrogenase and fatty acid synthase in the absence of palmitoyl-coenzyme A or oleate and also protect them against palmitoyl-coenzyme A or oleate inhibition. Levels of fatty acid binding proteins, the activities of the enzymes fatty acid synthase and glycerol-3-phosphate dehydrogenase increase with gestation showing a peak at term. However, the activity of fatty acid synthase showed the same trend up to the 30th week of gestation and then declined slightly at term. With the advancement of pregnancy when more lipids are required for the developing placenta, fatty acid binding proteins supply more fatty acids and glycerol-3-phosphate for the synthesis of lipids. Thus a correlation exists between glycerol-3-phosphate dehydrogenase, fatty acid synthase and fatty acid binding proteins in developing human placenta.

    • Effects of actinomycin d and cycloheximide on gonadotropin- or 17α, 20β-dihydroxy-4-pregnen-3-one-inducedin vitro maturation in oocytes of the catfish,Clarias batrachus

      K Balamurugan S Haider

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      The effect of inhibitors in the oocyte maturation ofClarias batrachus, was investigatedin vitro using actinomycin D and cycloheximide. Full-grown immature oocytes incubated with salmon gonadotropin (SG-G100) and 17α, 20ß-dihydroxy-4-pregnen-3-one at the concentration of 1 μg/ml induced 86.0 ± 1.2% and 91.3 ± 2.4% of germinal vesicle breakdown, respectively. When the oocytes were incubated with SG-G100 (1 μg/ml) + different concentrations of actinomycin D or cycloheximide, a significant drop in the frequency of germinal vesicle breakdown was observed. Thus, gonadotropin-induced maturation was inhibited by both transcriptional and translational inhibitors. When the oocytes were incubated with 17α, 20ß-dihydroxy-4-pregnen-3-one (1μg/ml) + different concentrations of cycloheximide, a significant inhibition of germinal vesicle breakdown was recorded. However, the maturation was not inhibited when the oocytes were incubated in the presence of 17α, 20ß-dihydroxy-4-pregnen-3-one (1 μg/ml) and different concentrations of actinomycin D. This suggests that mRNA synthesis is not obligatory for 17α, 20β-dihydroxy-4-pregnen-3-one induced oocyte maturation. Based on the time course experiment, it was observed that the inhibition of maturation in cycloheximide treated oocytes extends up to 12 h after which the effect becomes slowly subdued.

    • Antigenicity of filarial superoxide dismutase in human bancroftian filariasis

      Madhusmita Bal Manoj K Das

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      Superoxide dismutase activity was measured in different stages of growth of filarial parasites (human and cattle). The activity was almost undetected or very low in microfilarial stage but in adult worms, the enzyme activity was high. The enzyme was characterized to be a Cu/Zn superoxide dismutase. Most of the enzyme activity was associated with a detergent extractable fraction of adult (Setaria) parasite. The enzyme was also detected in thein vitro released products of adult worms. The superoxide dismutase activity was completely inhibited with IgG antibody from chronic filarial patients in contrast to IgG from normal people. Filarial patients particularly have high IgG and IgM antibody levels to purified enzyme. However, individuals from non-filarial regions of Orissa are sero-negative for superoxide dismutase antibodies. Antibody response to superoxide dismutase could thus be used for filarial diagnosis.

    • Age-related prevalence of antibodies to infective larvae ofWuchereria bancrofti in normal individuals from a filaria-endemic region

      M K Beuria M Bal A P Dash Manoj K Das

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      Antibody isotypic levels (IgM, IgE and IgG subclasses) to infective larvae (L3) ofWuchereria bancrofti were measured in 104 normal individuals from a filaria-endemic region in Orissa. The titres of antibodies were considerably higher in adults (n = 25, 25.1± 3.8 year) than in children (n = 52, 7.1 ± 2.1 year). Young children (n = 14) less than four years were seronegative to all isotypes other than IgM, the sero-conversion to which was achieved in the children (n=15) by the age of 7.5±1.2 years. The prevalence of other isotypes increased with age and reached a maximum in early adulthood (18.6 ±1.6 years), which persisted in older adults (> 30 years). However, the increase in IgG3 prevalence with age was less marked. IgG2 was detected only after 10 years of age. Compared to the high prevalence (100%) of IgM, IgE, IgG1, and IgG2, in adults. IgG3 and IgG4 prevalences were low, 35% and 28% respectively. IgA level to L3 antigen was found to be extremely low even in adults. These data indicate that the prevalence of L3 antibodies was different for different isotypes and the acquisition of antibody response essentially followed an age dependent pattern.

    • Cell cycle and DNA content of mitotic cells in brain ganglia ofdrosophila larvae

      S C Lakhotia J K Roy Sujata kar Chowdhuri (neeRoy)

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      The programmes of replication of hetero- and euchromatin regions, mitotic cell cycle and the DNA content in metaphases in brain ganglia from late third instar larvae ofDrosophila melanogaster (wild type and a tumour bearing mutant, 1(2)gl, strain) and ofDrosophila nasuta were examined by autoradiography of [3H]thymidine labelled (continuous or pulse) cells and by cytophotometry, respectively. Brain ganglia labelled continuously with [3H]thymidine for 24 hin vitro showed a significantly high proportion of cells with incorporation of radioactivity restricted to heterochromatin only. Pulse labelling of brain ganglia from larvae ofDrosophila melanogaster andDrosophila nasuta followed by chase for different time intervals showed that (i) the frequency of labelled metaphases was more than 50% within 15 to 30 min of chase and remained higher than 50% in nearly all the chase samples till 24 h, (ii) euchromatin labelled metaphases appeared with a low frequency within 1 to 4 h chase period but the heterochromatin labelled metaphases continued to be more common in the later chase samples also, (iii) single chromatid labelled second cycle metaphases were seen within 1 to 4 h after the pulse, but their frequency did not increase in the later samples. Cytophotometry of feulgen-DNA and Hoechst 33258 stained metaphases in late third instar larval brain ganglia revealed a greater variation in the DNA content of individual metaphases, although the means were close to the expected 4 C content. It appears that in relation to the known asymmetric cell divisions of neuroblast and other neural cells, the mitotically active cells in brain ganglia comprise a heterogenous population with widely varying lengths of the different phases of cell cycle; some of them may not cycle regularly and may possibly have a discontinuous S-phase.

    • Comparative analysis of prosomes and multicatalytic proteinases from rabbit erythrocytes

      Jayanta K Pal Koko Murakami

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      Prosomes and multicatalytic proteinases were purified from rabbit erythrocyte lysates and were analysed to determine their relationship. During purification by sucrose density gradient centrifugation using low salt buffer, they sedimented at 20–26S. Upon further purification, using high salt buffer, prosomes were recovered as 20S complexes as determined by their characteristic polypeptide pattern. Interestingly, both the 26S and 20S components had protease activity. Therefore, in order to determine their relationship with the multicatalytic proteinases, which are reported to contain a similar set of polypeptides, highly pure prosomes and the multicatalytic proteinases were analysed. Both 20S prosomes and multicatalytic proteinases showed protease activity and also had identical protein subunits of molecular weight ranging from 21 kDa to 35 kDa. Among these, at least two were immunologically identical as determined by Western blot using two monoclonal antibodies prepared against duck prosomes. Furthermore, protease activities of both components were inhibited almost to the same extent by an endogenous inhibitor specific for high molecular weight proteases and calpain. These results thus establish that the 20S prosomes and multicatalytic proteinases are identical, and suggest further that proteolytic activity could be the principal function of prosomes.

    • Developmental regulation of silk protein P25 in the silkwormBombyx mori

      K Muthumani S Mathavan S Mayilvahanan

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      P25 protein was extracted from cocoons of the silkwormBombyx mori by alkali solubilization and purified by gel elution. The purity and authenticity of the protein were confirmed by SDS-PAGE, 2-dimensional gel electrophoresis and peptide mapping. Polyclonal anti-P25 sera were raised in rabbit and mice. The relative abundance of P25 protein present in the larva during different developmental stages was analysed by SDS-PAGE, and quantified by sandwich ELISA. The minimum level (0.2 μg/animal) of this protein was recorded at the beginning of the first instar and maximum (16.7mg/pair of silkgland) on the final day of the V instar. During each moult period, P25 protein level was suppressed; the level increased with the initiation of feeding and reached maximum on the 3rd day of each instar except the final instar where the maximum was recorded prior to pupal moult. Western blot analysis also confirmed the developmental stage-specific accumulation of P25 protein in the silkwormBombyx mori.

    • Variability in ionization state, stoichiometry and aggregation in histidine complexes with formic acid

      Stephen Suresh M Vijayan

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      The crystal structures of the complexes of L and DL histidine with formic acid have been determined as part of an effort to define biologically and evolutionarily important interactions and aggregation patterns. In terms of ionization state and stoichiometry they may be described as L-histidine formate formic acid and DL-histidine formate monohydrate respectively. In the L-histidine complex, amino acid molecules arranged in head-to-tail sequences centred around 21 screw axes are interconnected by formic acid molecules and formate ions. Histidine-formate interactions in the structure gives rise to a characteristic interaction pattern involving a linear array of alternating imidazole groups and formate ions. In DL-histidine formale monohydrate, head-to-tail sequences involving glide related molecules are interconnected through main chain-side chain interactions leading to amino acid layers. The layers are held together by formate ions and water molecules arranged in strings along which the ion and the molecule alternate. The patterns of amino acid aggregation in histidine complexes exhibit considerably higher variability than those in complexes involving arginine and lysine do.

    • Prediction of B-cell epitopes forSalmonella typhi OmpC

      A Arockiasamy S Krishnaswamy

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      The possible B-cell epitopes of the outer membrane porin OmpC ofSalmonella typhi have been identified, using the primary structure of the protein, by means of multiple sequence alignment and the known molecular structure of two other porins. From the analysis, 8 regions were identified as immunodominant and these were ranked based on antigenic index and the ratio of the number of nonconserved residues to the fragment length. Model building of the top two ranked regions show the tendency to form loop structures supporting the possibility of these being candidate epitopes.

    • Reconsideration of some results for linear and circular DNA

      G A Rodley

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      A reassessment is made of some results for linear and circular DNA. The alternative side-by-side configuration is used in an analysis of situations where some doubt about current double helix based interpretations may exist. The following specific areas are considered: (i) certain linear DNA examples involving duplex interactions, (ii) nucleosome sequence data and (iii) the banding effect observed in gel electrophoresis studies ofin vitro circular DNA.

    • The branching pattern of major groups of land plants inferred from parsimony analysis of ribosomal RNA sequences

      S Barnabas S Krishnan J Barnabas

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      The parsimony and bootstrap branching pattern of major groups of land plants derived from relevant 5S rRNA sequence trees have been discussed in the light of paleobotanical and morphological evidences. Although 5S rRNA sequence information is not useful for dileneating angiosperm relationships, it does capture the earlier phase of land plant evolution. The consensus branching pattern indicates an ancient split of bryophytes and vascular plants from the charophycean algal stem. Among the bryophytes,Marchantia andLophocolea appear to be phylogenetically close and together withPlagiomnium form a monophyletic group.Lycopodium andPsilotum arose early in vascular land plant evolution, independent of fem-sphenopsid branch. Gymnosperms are polyphyletic; conifers, Gnetales and cycads emerge in that order with ginkgo joiningCycas. Among the conifers,Metasequoia,Juniperus andTaxus emerge as a branch independent ofPinus which joins Gnetales.

      The phylogeny derived from the available ss-RNA sequences shows that angiosperms are monophyletic with monocots and dicots diverging from a common stem. The nucleotide replacements during angiosperm descent from the gymnosperm ancestor which presumably arose around 370 my ago indicates that monocots and dicots diverged around 180 my ago, which is compatible with the reported divergence estimate of around 200 my ago deduced from chloroplast DNA sequences.

    • The line transect method for estimating densities of large mammals in a tropical deciduous forest: An evaluation of models and field experiments

      K Surendra Varman R Sukumar

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      We have evaluated techniques of estimating animal density through direct counts using line transects during 1988–92 in the tropical deciduous forests of Mudumalui Sanctuary in southern India for four species of large herbivorous mammals, namely, chital (Axis axis). sambar (Cervus unicolor). Asian elephant (Elephas maximus) and gaur (Bos gaurus)

      Density estimates derived from the Fourier Series and the Half-Normal models consistently had the lowest coefficient of variation. These two models also generated similar mean density estimates. For the Fourier Series estimator, appropriate cut-off widths for analyzing line transect data for the four species are suggested. Grouping data into various distance classes did not produce any appreciable differences in estimates of mean density or their variances, although model fit is generally better when data arc placed in fewer groups. The sampling effort needed to achieve a desired precision (coefficient of variation) in the density estimate is derived. A sampling effort of 800 km of transects returned a 10% coefficient of variation on estimate for ehital; for the other species a higher effort was needed to achieve this level of precision. There was no statistically significant relationship between detectability of a group and the size of the group for any species. Density estimates along roads were generally significantly different from those in the interior of the forest, indicating that road-side counts many not be appropriate for most species.

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