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Biomedical subjects

D K Ghosh

Publications and source records attributed to D K Ghosh.

At least 73 records · Page 4Linked to original sources
PubMed1988

Excreted factors and membrane-associated carbohydrates of Indian leishmaniae.

Antigenic identity between the cell surface carbohydrate ligands and their excreted factor (EF) in all forms of Indian leishmaniasis were assessed by 11 carbohydrate-specific lectins. Our results demonstrated that 3 lectins, viz. PNA, SBA and WFA showed unique selectivity towards skin-dwelling parasites. The EFs of visceral leishmaniasis (VL), post-kala-azar dermal leishmaniasis (PKDL) and cutaneous leishmaniasis (CL) formed band against ConA at 1000, and 500 micrograms/ml concentration, respectively. RCA-120 at 3 mg/ml concentration was positive to both Leishmania tropica and L. major, whereas SBA was specific to L. tropica, L. major and PKDL strains at the same concentration. Antisera directed against the EFs (EF-As) induced agglutination to homologous promastigotes suspension. Cross-reactivity of agglutination was observed in different strains but highest was found among PKDL and CL strains. Although PKDL cases are normally found in patients with the history of visceral leishmaniasis (kala-azar), surprisingly in this study, the surface saccharides of PKDL strains had close affinity to CL type instead of VL.

Animals↗
PubMed1987

Induction of high-density-lipoprotein receptors in rat corpus luteum by human choriogonadotropin. Evidence of protein synthesis de novo.

The present studies investigated the specific binding of 125I-labelled high-density lipoprotein (125I-HDL) to plasma membranes. Golgi, rough endoplasmic reticulum and mitochondria/lysosomes, prepared from ovaries of rats injected with human choriogonadotropin (hCG) or 0.9% NaCl. Treatment in vivo with hCG resulted in 2-3-fold induction of 125I-HDL binding activity in all the subcellular organelles. The specific binding of HDL to various subcellular organelles was dependent on the amount of protein, lipoprotein concentration and incubation time. Equilibrium-binding studies revealed comparable Kd values (13-22 micrograms of HDL protein/ml) for HDL binding in all the subcellular organelles tested. Treatment with cycloheximide (2.0 mg/kg body wt.) before hCG administration abolished the induction of HDL receptors, suggesting the involvement of a protein-synthesis-dependent process in receptor induction. Analysis of equilibrium dissociation constants (Kd) for 125I-HDL binding in membranes from hCG-, cycloheximide-and saline-treated animals suggests that the increase in binding was due to an increase in the number of binding sites rather than a change in the affinity. Additionally, pretreatment with tunicamycin, an inhibitor of N-linked glycosylation, had no effect on hCG-mediated receptor induction, suggesting that glycosylation of the receptor may not be necessary for the interaction of HDL with its receptors.

Animals↗
PubMed1987

Characterization of progesterone 11 alpha-hydroxylase of Aspergillus ochraceus TS: a cytochrome P-450 linked monooxygenase.

The monooxygenase of Aspergillus ochraceus TS capable of 11 alpha-hydroxylation of progesterone has been resolved into three components and characterized as (i) cytochrome P450, (ii) NADPH-cytochrome P450-reductase and (iii) phosphatidyl choline. The 11 alpha-hydroxylase was observed to be NADPH dependent, and hydroxylation was enhanced by a NADPH regenerating system. This fungal monooxygenase has many features in common with that of mammalian liver microsomes. The role of mammalian cytochrome P450 inducers were tested for induction of 11 alpha-hydroxylase in Aspergillus ochraceus TS. The reductase has been partially purified.

Aspergillus↗
PubMed1987

Urea stibamine: an improved method of preparation and its antileishmanial activity.

An improved method for preparation of urea stibamine was developed. The crude p-acetylaminophenyl stibonic acid (II) prepared was purified by dissolving it in Na2CO3 solution, whererin acid (II) dissolved leaving behind the impurities. Acid (II) was directly combined with urea without hydrolyzing the acetyl group to give urea stibamine. Biological activity of the drug in vitro as well as in vivo was also studied. The drug had no inhibitory effect on growth, respiration, incorporation of radiolabeled precursors into promastigotes of L. donovani, and on transformation of amastigotes to promastigotes. In infected hamster, the effect of the drug was highly significant in vivo., as it removed the parasitic burden completely.

Animals↗
PubMed1987

Regulation of cholesterol side-chain cleavage enzyme activity by gonadotropin in rat corpus luteum.

The locus of gonadotropin-induced acute stimulation of pregnenolone production by cholesterol side-chain cleavage (CSCC) enzyme containing cytochrome P450 (cytP450scc) was examined in rat corpus luteum. Mitochondria were isolated from pseudopregnant rat ovaries after treatment with different doses of human CG (hCG) (25-200 IU) for 30 min; Electron Paramagnetic Resonance (EPR) spectra of high spin cholesterol complex of cyt P450scc (type I high spin EPR signal) and the cyt P450scc activity were determined. hCG treatment increased the formation of type I EPR spectra compared to that obtained with saline-treated controls, and pretreatment with cycloheximide (30 mg/kg BW) before hCG abolished this increase. The magnitude of type I EPR signal diminished with increasing pH over the range of 6.2-7.3. The type I EPR signal increased with doses of hCG and correlated well with the pregnenolone production. Aminoglutethimide treatment (competitive inhibitor of CSCC) before hCG injection led to an increased accumulation of cholesterol in inner mitochondrial membranes with a corresponding decrease in the outer membrane cholesterol, and cycloheximide treatment inhibited this accumulation. This suggests that the transport of cholesterol to inner mitochondrial membranes from outer membranes is regulated by hCG. In addition, gonadotropin also regulates the redistribution of cholesterol within the inner mitochondrial membranes.

Animals↗
PubMed1987

A comparison of five selective media for the primary isolation of Leishmania strains in India.

Leishmania parasites of the three distinct pathogenic forms of Indian origin (Cutaneous, visceral and post Kala-azar dermal leishmaniasis) were assessed with five selective media for their rate of primary isolation. Leishmania donovani strains showed the higher rate of isolation in modified Tobie's medium, and Leishmania tropica in Tobie's with FCS. However, strains of post Kala-azar dermal leishmaniasis (PKDL) showed 100% isolation in Schneider's with FCS medium. But as a whole modified Tobie's medium showed a higher rate of parasitic isolation for all the three pathogenic forms. In cutaneous leishmaniasis, the dry type and early lesion are also considered significant in the successful isolation of parasite. The growth study of L. donovani strain in the five selective media showed the modified Tobie's medium was best for the multiplication of the parasite.

Animals↗
PubMed1987

Differentiation of Leishmania strains of Indian origin by lectin-mediated agglutination.

Characterisation of surface saccharides was carried out in 3 different strains of Indian leishmaniae by 7 lectins. Energy dependent agglutination and role of cytoskeletal disintegrator were also analysed. Wisteria floribunda, peanut agglutinin and soybean agglutinin can be used to differentiate clearly the leishmanial strains causing disease like visceral leishmaniasis, post Kala-azar dermal leishmaniasis and cutaneous leishmaniasis. The remaining lectins have mixed agglutination behaviour towards these strains. The effective concentration of dinitrophenol, sodium azide and colchicine is 10 mM, 25 mM and 1.25 x 10(-2) M, respectively.

Agglutination Tests↗
PubMed1986

Identification of gonadotropin inducible, high density lipoprotein receptors in the solubilized membranes from rat ovary.

Specific receptors for high density lipoproteins (HDL3) were solubilized from membranes of rat corpus luteum using different detergents. Among the detergents tested, octyl-beta-D-glucoside (40 mM) was most effective with respect to recovery of binding activity. The receptor activity released into 105,000 X g supernatant, can be assayed directly or with the precipitate obtained after dilution of the soluble supernatant. The 125I-HDL3 binding activity in the precipitated extract was linear with time, proportional to the amount of protein in the incubation mixture and saturable with increasing concentrations of 125I-HDL3. The solubilized receptor has an equilibrium dissociation constant (Kd) of 21.2 micrograms/ml and the binding activity was insensitive to Ca+2, EDTA and NaCl. These properties are similar to the membrane associated receptor. Administration of gonadotropin induced the HDL3 receptor in the solubilized membranes, suggesting that this receptor represents the physiologic receptor in the ovary.

Animals↗
PubMed1985

Leishmania donovani: amastigote inhibition and mode of action of berberine.

Berberine, an alkaloid from Berberis aristata Linnaeus, may be a useful drug for the treatment of visceral leishmaniasis. In both the 8-day and long-term models of Leishmania donovani infection in hamsters, it markedly diminished the parasitic load and proved to be less toxic than pentamidine. It rapidly improved the hematological picture of infected animals. Like pentamidine, it inhibited in vitro multiplication of amastigotes in macrophage culture and their transformation to promastigotes in cell free culture. Manometric studies showed that both drugs had inhibitory action on both the endogenous and the glucose-stimulated respiration of amastigotes. They inhibited incorporation of [14C]adenine, [14C]uracil, and [3H]thymidine into nucleic acids, and of [14C]leucine into the protein of amastigotes, indicating an inhibitory action on macromolecular biosynthesis. They also decreased deoxyglucose uptake. Using spectrophotometric, spectrofluorimetric, and circular dichroism techniques, berberine was found to interact in vitro with nuclear DNA from L. donovani promastigotes.

Animals↗
PubMed1983

Induction of benzo(a)pyrene hydroxylase in Aspergillus ochraceus TS: evidences of multiple forms of cytochrome P-450.

The filamentous fungus Aspergillus ochraceus TS produces an inducible microsomal cytochrome P-450 linked monooxygenase which is capable of hydroxylating benzo(a)pyrene in presence of O2 and NADPH. The addition of Benzo(a)pyrene, 3-Methyl cholanthrene, beta-Naphthoflavone and other aryl hydrocarbons during the induction period causes dramatic improvement in the kinetics of benzo(a) pyrene hydroxylation as was evidenced by large decrease in Km and increase in Vmax values. On the other hand, treatment with Phenobarbital, Polychlorinated biphenyl and Progesterone has no significant effect on the kinetics of benzo(a)pyrene hydroxylation although a significant induction of NADPH-Cyt C reductase activity was observed in all the three cases. Again, both Phenobarbital and 3-Methyl cholanthrene induced microsomes exhibit the characteristic reduced metyrapone difference spectra. These findings together with the results obtained with flavone on the metabolism of benzo(a)pyrene by various microsomal preparations suggest a parallel induction of multiple forms of cytochrome P-450 as observed in mammalian liver under identical condition.

Aryl Hydrocarbon Hydroxylases↗
PubMed1983

Microsomal benzo(a)pyrene hydroxylase in Aspergillus ochraceus TS: assay and characterization of the enzyme system.

Microsomal preparations of Aspergillus ochraceus TS oxidised benzo(a)pyrene very efficiently in the presence of NADPH and 02 and exhibits a pH optimum of 8.0-8.2. The hydroxylation is also effected in presence of NaI04. Hydroxylation was inhibited by metyrapone, SKF-525A, PCMB, imidazole, carbon monoxide and flavone but not by cyanide, azide and antimycin A indicating thereby the involvement of cytochrome P-450 in this reaction. Inhibition by cytochrome C is consistent with the participation of NADPH-cytochrome C reductase in this hydroxylation. Reduced microsomes and its solubilized preparation, when treated with carbon monoxide, showed absorption maxima at 453 and 449 respectively. Different classical inducers of cytochrome P-450 induce the benzo(a)-pyrene hydroxylase activity to varying degree and as such suggests the existence of multiple forms of cytochrome P-450 in this fungus.

Aryl Hydrocarbon Hydroxylases↗