• Volume 18, Issue 3

      September 1993,   pages  303-422

    • The structure of duplex DNA in the nucleosome core particle: An alternative model

      A T Baker V Mckee G A Rodley

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      Many studies indirectly indicate that the conformation ofin vivo duplex DNA is the double helix. The most direct view, from the X-ray analysis of the nucleosome core particle, has also been interpreted in terms of the double helix structure. However, an alternative possibility exists; that the duplex adopts a metastable side-by-side conformation which readily converts to the double helix on removal of protein. Evidence for the existence of this conformation has been obtained from a reanalysis of the electron density map for the nucleosome particle.

    • Synthesis and release of proteins homologous to excretory-secretory antigens during embryogenesis ofSetaria digitata

      K P Lala Dhas S William Decruse Kaleysa Raj

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      Localization of antigens homologous to excretory-secretory proteins in developing embryos ofSetaria digitata has been carried out by indirect fluorescent antibody test, [14C] labelling studies and Western blotting. Indirect fluorescent antibody test showed binding of excretory-secretory antibodies at perivitelline space. The fluorescent antibody binding was almost absent at small morulae stage and increasing in intensity in the successive developmental stages with maximum at coiled microfilaria stage. Hatched microfilaria did not show the presence of antigens by immunofluorescence. Immuno-complex of excretory-secretory antiserum against “amniotic fluid” collected from developing embryos ofSetaria digitata labelled with [14C] amino acids showed highest radioactivity at coiled and tadpole stages and differed significantly from small morulae, big morulae and hatched microfilaria. Immunoblot analysis of amniotic fluid showed two proteins, 16.5 and 11 kDa, to be highly antigenic. The antigenic protein (11 kDa) content as seen by immuno blotting increased during embryogenesis and decreased at the stage of hatching.

    • Diagnostic utility of fractionated urinary filarial antigen

      V Chenthamarakshan U M Padigel P Ramaprasad M V R Reddy B C Harinath

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      Urinary filarial antigen isolated from urine samples of microfilaraemic patients was analysed for its antigenic activity by immunoblotting and enzyme linked immunosorbent assay techniques. SDS-PAGE fractionation of urinary filarial antigen showed 11 protein bands, of which two showed reactivity with immunoglobulin-G fraction of filarial serum immunoglobulin in immunoblotting. Antigenic analysis of SDS-PAGE fractions of urinary filarial antigen by inhibition enzyme linked immunosorbent assay using filarial serum immunoglobulin-G andWuchereria bancrofti microfilarial excretory-secretory antigen revealed 3 fractions, numbers 5, 6 and 9 with significant activity. In indirect enzyme linked immunosorbent assay using fractions 5 and 6, filarial immunoglobulin-G antibody was detected in about 90% of microfilaraemics, 80% clinical filariasis and 20% of endemic normal individuals. Further, there was no phosphorylcholine epitope in these fractions. Fractions 5 and 6 can be a candidate antigens for the immunodiagnosis of filariasis.

    • Binding of thyroid hormone to mouse granulosa cell nuclei and its biological relevance

      R Biswas A Bandyopadhyay S Guin S Bhattacharya

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      Thyroid hormone showed specific binding ability to mouse granulosa cells from immature mice, primed with post menopausal gonadotropin. Saturation of specific binding sites was reached by 2 nM concentration of the hormone. A Scatchard analysis of thyroid hormone binding exhibited a Kd of 42 x l0-9M/mg nuclear DNA and a maximum binding capacity of 1 pmol/mg nuclear DNA. Competitive inhibition studies showed thyroid hormone binding to be analogue specific. Addition of 100 ng of thyroid hormone to granulosa cell incubations (1 x 106 cells/well) resulted in a three-fold increase in cellular protein synthesis. Thyroid hormone resulted in a dose dependant increase in progesterone release from granulosa cell. It also stimulated the formation of pregnenolone (83%) and progesterone (81%) from radiolabeled cholesterol as compared to control. This stimulation by thyroid hormone was completely inhibited by cycloheximide. Results indicate a direct effect of thyroid hormone on granulosa cells, its binding to nuclei causing an increase in steroidogenesis through the mediation of protein(s).

    • Changes in insulin receptor, hexokinase and NADPH producing enzymes in Choroid plexus during experimental diabetes

      Mansoor A Ansari Biharil Gupta Najma Z Baquer

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      The activities of insulin receptor and the enzymes hexokinase (EC and NADP-dependent malic enzyme (EC1.1.1.40), glucose 6-phosphate dehydrogenase (EC and isocitrate dehydrogenase (EC were measured in rat choroid plexus in alloxan induced diabetes. A significant decrease was observed in the activities of all the enzymes except isocitrate dehydrogenase and also the choroid plexus insulin receptor activity was decreased. A reversal of the efect was observed with insulin administration to diabetic rats. It may be concluded that the enzymes of choroid plexus together with insulin receptor are directly controlled by-the concentration of insulin.

    • Thermophilic fungi: An assessment of their potential for growth in soil

      A K Rajasekaran R Maheshwari

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      An attempt has been made to forecast the potential of thermophilic fungi to grow in soil in the laboratory and in the field in the presence of a predominantly mesophilic fungal flora at usual temperature. The respiratory rate of thermophilic fungi was markedly responsive to changes in temperature, but that of mesophilic fungi was relatively independent of such changes. This suggested that in a thermally fluctuating environment, thermophilic fungi may be at a physiological disadvantage compared to mesophilic fungi. In mixed cultures in soil plates, thermophilic fungi outgrew mesophilic fungi under a fluctuating temperature regime only when the amplitude of the fluctuating temperatures was small and approached their temperature optima for growth. An antibody probe was used to detect the activity of native or an introduced strain of a thermophilic fungus,Thermomyces lanuginosus, under field conditions. The results suggest that although widespread, thermophilic fungi are ordinarily not an active component of soil microflora. Their presence in soil most likely may be the result of the aerial dissemination of propagules from composting plant material.

    • Mercury ions inhibit photosynthetic electron transport at multiple sites in the cyanobacteriumSynechococcus 6301

      S D S Murthy Prasanna Mohanty

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      Inhibition of electron transport activities in the spheroplasts ofSynechococcus 6301 by HgCl2 is dependent on the concentration of mercury ions. The inhibition of whole chain electron transport activity occurs at low concentration of Hg2+ (6 ΜM@#@). This inhibition occurs mostly due to interaction of Hg2+ on plastocyanin. At an elevated concentration (24 ΜM@#@), mercury induces inhibition chiefly in photosystem II catalyzed electron transport. At this concentration it also alters both the absorption and emission characteristics of the phycocyanin. The photosystem I catalyzed electron transport was inhibited by 50% only at high concentrations (36 ΜM@#@) of HgCl2. However, electron transport catalyzed by photosystems I and II from reduced duroquinone to methylviologen which involves intersystem electron transport is extremely sensitive to mercury (low concentration 6–9 ΜM) like that of whole chain assay indicating that the observed inhibition in whole chain electron transport at low concentrations is mostly contributed by the damage involving other intersystem electron transport carrier(s) like plastocyanin. Thus mercury ions depending on the concentration affects the electron transport at multiple sites in the spheroplasts ofSynechococcus.

    • [125I] gonadotropin binding to the ovary of an Indian major carp,Catla catla, at different stages of reproductive cycle

      Pulak Ranjan Manna Samir Bhattacharya

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      At different stages of the annual reproductive cycle ofCatla catla, a major Indian carp, specific binding of gonadotropic hormone to the plasma membrane receptors was demonstrated. Maximum specific binding of [125I]Catla gonadotropic hormone was obtained at 30‡C and pH 7.5 during 2 h of incubation.Catla gonadotropic hormone binding was saturable with high affinity. Competitive inhibition experiment showed that binding site was specifically occupied by piscine gonadotropic hormone,Catla gonadotropic hormone and murrel gonadotropic hormone, human chorionic gonadotropin was a weak competitor while bovine thyroid stimulating hormone, bovine prolactin and ovine follicle stimulating hormone had no effect. Scatchard analysis ofCatla gonadotropic hormone binding to the plasma membrane preparation from the carp oocytes of different reproductive stages showed that the range of dissociation constant(Kd) varied from 0.78 to 0.97 x 10-10 M. However, maximum binding capacity (B-max) varied remarkably between the different stages of reproductive cycle, it was 6.11 ± 0.36 fmol/mg protein in the preparatory stage which increased to about three-fold in prespawning stage of reproductive cycle (17.0 ± 0.29 fmol/mg protein) and spawning (18.7 ± 0.17 fmol/mg protein) and lowest in postspawning stage of reproductive cycle (5.28 ± 0.28 fmol/mg protein). Fluctuation in the number of gonadotropic hormone binding site at different stages of annual reproductive cycle was found to be coincided well with the pattern of ovarian steroidogenesis in response toCatla gonadotropic hormone as determined by the formation of progesterone from pregnenolone.

    • Frequency of HLA antibodies in south India

      R M Pitchappan S Amutha V Mahendran V Brahmajothi U Shankar Kumar K Balakrishnan R Shantha Kumari M Sashidhar SamalRaj May Manuel V Indra Raja

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      Sera from 4088 pregnant women (649 antenatal bleeding and 3439 post-partum bleeding) living in Madurai, were collected and screened for anti-HLA. A, B and DR antibodies. 696 of them were screened for anti-HLA DR antibodies. Ten per cent (65/649) of antenatal sera and 13 4% of post-partum sera (463/3439) were positive for HLA A and B antibodies: nonetheless the percentage of monospecific sera were almost the same in both. Screening for HLA DR antibodies were carried out using platelet absorption in test tray technique: seventy three of 696 (10.5%) were positive. The incidence of anti-HLA A, B antibodies correlates to the allelic frequencies in the population. Thus in India, collection and screening post-partum haemorrhage is the simplest and cost effective method of acquiring polyclonal sera for routine laboratory and diagnostic use.

    • Bioacoustic analysis of frog calls from northeast India

      Debjani Roy Andreas Elepfandt

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      Mating calls of three frog species abundant in northeast IndiaRana tigerina,Rana cyanophlyctis andRana limnocharis were recorded in the fields of Assam and Meghalaya during their breeding season (July-August, 1991). The calls were analysed for their temporal and spectral characters. They were species specific, with distinct call duration and call period, number of pulses per call and interpulse interval, and dominant frequency and frequency domain. A comparison of the mating calls ofRana cyanophlyctis with those of the siblingRana ehrenbergi from Yemen showed differences in their temporal and spectral characters, supporting the suggestion that these two species are distinct species, rather than subspecies of the same species. Differences in the temporal and spectral pattern were found in the mating calls of morphologically alike specimens ofRana limnocharis, indicating that the present morphotypeRana limnocharis in northeast India is composed of several species.

    • Nutrient cycling in an excessively rainfed subtropical grassland at Cherrapunji

      H N Pandey R S Tripathi Uma Shankar

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      Cycling of six mineral elements (N, P, K, Na, Ca and Mg) was studied in a humid subtropical grassland at Cherrapunji, north-eastern India during 1988-1989. Elemental concentrations in the shoot of four dominant grass species,viz., Arundinella khaseana, Chrysopogon gryllus, Eragrostiella leioptera andEulalia trispicata were very low, and none of the species appears suitable for fodder use. Among different vegetation compartments, live root was the largest reservoir of all the nutrients (except Ca) followed by live shoot, dead shoot, litter and dead root. For Ca, live shoot was the major storage compartment. The total annual uptake (kg ha-1) was 137.3, 10.4, 51.1, 5.5, 8.7 and 18.2 for N, P, K, Na, Ca and Mg, respectively. In an annual cycle 98% N, 77% P, 49% K, 109% Na, 87% Ca and 65% Mg returned to the soil through litter and belowground detritus. A major portion of N, P and Na was recycled through the belowground system, whereas nearly half of K, Ca and Mg was recycled through the shoot system. Precipitation acts as the source of N and P input, but at the same time causes loss of cations.

    • Mitochondrial genome organization and cytoplasmic male sterility in plants

      C K K Nair

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      Plant mitochondrial genomes are much larger and more complex than those of other eukaryotic organisms. They contain a very active recombination system and have a multipartite genome organization with a master circle resolving into two or more subgenomic circles by recombination through repeated sequences. Their protein coding capacity is very low and is comparable to that of animal and fungal systems. Several subunits of mitochondrial functional complexes, a complete set of tRNAs and 26S, 18S and 5S rRNAs are coded by the plant mitochondrial genome. The protein coding genes contain group II introns. The organelle genome contains stretches of DNA sequences homologous to chloroplast DNA. It also contains actively transcribed DNA sequences having open reading frames. Plasmid like DNA molecules are found in mitochondria of some plants

      Cytoplasmic male sterility in plants, characterized by failure to produce functional pollen grains, is a maternally inherited trait. This phenomenon has been found in many species of plants and is conveniently used for hybrid plant production. The genetic determinants for cytoplasmic male sterility reside in the mitochondrial genome. Some species of plants exhibit more than one type of cytoplasmic male sterility. Several nuclear genes are known to control expression of cytoplasmic male sterility. Different cytoplasmic male sterility types are distinguished by their specific nuclear genes(rfs) which restore pollen fertility. Cytoplasmic male sterility types are also characterized by mitochondrial DNA restriction fragment length polymorphism patterns, variations in mitochondrial RNAs, differences in protein synthetic profiles, differences in sensitivity to fungal toxins and insecticides, presence of plasmid DNAs or RNAs and also presence of certain unique sequences in the genome. Recently nuclear male sterility systems based on (i) over expression of agrobacterialrol C gene and (ii) anther specific expression of an RNase gene have been developed in tobacco andBrassica by genetic engineering methods.

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