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Biomedical subjects

D K Sharma

Publications and source records attributed to D K Sharma.

At least 19 recordsLinked to original sources
PubMed2006

The release dynamics of model drugs from the psyllium and N-hydroxymethylacrylamide based hydrogels.

In order to utilize the psyllium husk, a medicinally important natural polysaccharide, for developing the novel hydrogels for the controlled drug delivery device, we have prepared psyllium and N-hydroxymethylacrylamide based polymeric networks by using N,N'-methylenebisacrylamide (N,N'-MBAAm) as crosslinker. The polymeric networks thus formed were characterized with scanning electron micrography (SEM), FTIR and thermogravimetric analysis (TGA) techniques to study various structural aspects of the networks and also with the swelling response of the polymeric networks as a function of time, temperature, pH and [NaCl]. Equilibrium swelling has been observed to depend on both structural aspects of the polymers and environmental factors. Maximum P(s) 748.3 was observed at 13.0 x 10(-3)mol/L of [N,N'-MBAAm] in 0.5M NaOH solution. The release dynamics of model drugs (salicylic acid and tetracycline hydrochloride) from hydrogels has also been discussed, for the evaluation of the release mechanism and diffusion coefficients. The effect of pH on the release pattern of tetracycline has been studied by varying the pH of the release medium. In release medium of pH 7.4 buffer the release pattern of tetracycline drastically changes to the extent that mechanism of drug diffusion shifted from non-Fickian diffusion to Fickian diffusion. It has been observed that diffusion exponent "n" have 0.71, 0.67 and 0.52 values and gel characteristic constant 'k' have 1.552 x 10(-2), 2.291 x 10(-2) and 5.309 x 10(-2) values in distilled water, pH 2.2 buffer and pH 7.4 buffer, respectively, for tetracycline release. In solution of pH 7.4 buffer, the rate of polymer chain relaxation was more as compare to the rate of drug diffusion from these hydrogels and it follows Fick's law of diffusion. The value of the initial diffusion coefficient for the release of tetracycline hydrochloride was higher than the value of late time diffusion coefficient in each release medium indicating that in the start, the diffusion of drug from the polymeric matrix was fast as compare to the latter stages.

Acrylamides↗
PubMed2006

Long-term follow-up (11 years plus) results of JRI (Furlong) total hip arthroplasty in young patients: cause for concern regarding acetabular cup?

We describe the clinical and radiological long-term outcomes of 77 primary total hip replacements in 69 patients using the fully hydroxyapatite-coated JRI (Furlong) total hip replacement. The total cases followed up were 77 hips, performed at a mean duration of 11 years and 2 months. Twelve hips could not be followed up for various reasons, which are discussed in the results section. The mean Harris hip score was 89. Seventeen acetabular cups were revised for aseptic loosening. Only one femoral stem was revised, for fracture. By Engh's criteria there were a further two unstable cups with no symptoms, and all femoral stems were stable. Kaplan-Meier survivorship analysis revealed a survival of 98.8% for the femoral stem, 78.7% for the acetabular cup, and a combined survival of 77.8% for both components. Our findings suggest that the JRI (Furlong) hip gives a durable femoral stem implant fixation, whereas the prosthesis-bone interface achieved with the acetabular component is questionable.

Acetabulum↗
PubMed2005

Adsorption of phenols from wastewater.

The present work involves an investigation of the possible use of coal, residual coal, and residual coal treated with H3PO4 as a means of removal of phenol from wastewater. The study was realized using batch experiments, with synthetic wastewater having phenol concentration of 1000 ppm. Other low-cost adsorbents such as petroleum coke, coke breeze, rice husk, and rice husk char have also been used. The effect of system variables such as pH, contact time, and temperature has been investigated. The suitability of the Freundlich, Langmuir, and Redlich-Peterson adsorption models to the equilibrium data was investigated for each phenol-adsorbent system. The results showed that the equilibrium data for all the phenol-sorbent systems fitted the Redlich-Peterson model best. Kinetic modeling of removal of phenols was done using the Lagergren first-order rate expression. A series of column experiments were performed to determine the breakthrough curves.

Journal Article↗
PubMed2005

Anatomical study on true hermaphroditism in an Indian pig (Sus scrofa domesticus).

A pig was confirmed to be a true hermaphrodite on the basis of gross and histomorphological studies of the genital organs. The genitalia was consisted of left ovary, oviduct, two coiled uterine horns, body of uterus along with right testis and an epididymis. Vagina and vulva were absent but male urethra with prostate gland was present. Grossly the size of all the genital organs appeared to be normal. Histomorphologically, testis and epididymis were underdeveloped as there was no clearcut spermatogenia and sertoli cells but Leydig cells were normal. The ovary presents normal histological features with some portion of testicular tissue. Degeneration of uterine epithelium was observed along with normal endometrial glands.

Animals↗
PubMed2004

Biodegradation of acid blue-15, a textile dye, by an up-flow immobilized cell bioreactor.

Acid blue-15, a complex and resonance-stabilized triphenylmethane (TPM) textile dye, resistant to transformation, was decolorized/degraded in an up-flow immobilized cell bioreactor. A consortium comprised of isolates belonging to Bacillus sp., Alcaligenes sp. and Aeromonas sp. formed a multispecies biofilm on refractory brick pieces used as support material. The TPM dye was degraded to simple metabolic intermediates in the bioreactor with 94% decolorization at a flow rate of 4 ml h(-1).

Bacteria↗
PubMed2004

Hypolipidemic effect of different extracts of Clerodendron colebrookianum Walp in normal and high-fat diet fed rats.

Clerodendron colebrookianum Walp (family, Verbenaceae) has been used as a home remedy by the Mizo people of north eastern (NE) region of India in their folklore medicine as a cardioprotective (mainly against high blood pressure) agent. The scientific basis for their use has yet to be established. Therefore, in this investigation an attempt has been made to study the effect of different extracts of the leaves of Clerodendron colebrookianum Walp (CC) on serum lipid profile which is closely associated with many cardiovascular diseases. It was found that after administration of both crude and organic extracts the serum lipid profile, i.e. total cholesterol (TC), triglycerides (TG) and low density lipoprotein (LDL) lowered significantly (P<0.001) from their respective control except TG in methanol extract (ME)-treated rats. The decreased percentage of TC, TG and LDL after administration of crude extract (CE) and ethylacetate extract (EE) with normal diet were found to be 35, 39 and 48%, and 33, 18 and 91%, respectively, after 7 days of treatment. In case of ME the lowering of TC and LDL were noted to be 41 and 85%, respectively. Both TC and TG were lowered significantly (P<0.001) even after the administration of crude extract against high-fat diet. Significant increase of cardioprotective lipid, i.e high density lipoprotein (HDL) after administration of EE (46%) and ME (38%) was another interesting finding of this experiment. Total cholesterol:HDL and LDL:HDL ratios were significantly lower in the experimental group. Therefore, it was concluded that the leaves of CC have definite cardioprotective potentiality, and there is a valid scientific basis for consuming it for better health in NE region of India.

Animals↗
PubMed2004

Biological treatment of textile dye acid violet-17 by bacterial consortium in an up-flow immobilized cell bioreactor.

AIMS: To develop a cost effective and efficient biological treatment process for small scale textile processing industries (TPI) releasing untreated effluents containing intense coloured Acid violet-17 (AV-17), a triphenyl methane (TPM) group textile dye. METHODS AND RESULTS: The samples collected from effluent disposal sites of TPI were used for selective enrichment of microbial populations capable of degrading/decolourizing AV-17. A consortium of five bacterial isolates was used to develop an up-flow immobilized cell bioreactor for treatment of feed containing AV-17. The bioreactor, operating at a flow rate of 6 ml x h(-1), resulted in 91% decolourization of 30 mg AV-17/l with 94.3 and 95.7% removal of biochemical oxygen demand and chemical oxygen demand of the feed. Comparison of the input and output of the bioreactor by UV-visible, thin layer chromatography and (1)H-nuclear magnetic resonance spectroscopy indicates conversion of the parent dye into unrelated metabolic intermediates. SIGNIFICANCE: These results will form a basis for developing 'on-site' treatment system for TPI effluents to achieve decolourization and degradation of residual dyes.

Bacteria↗
PubMed2001

Gall bladder perforation and biliary peritonitis in a young pregnant lady.

A rare event of acute free perforation of gall bladder with biliary peritonitis in a case of calculous cholecystitis in a 28 years old pregnant lady occurring in the absence of the usual factors associated with gall stone disease is reported. The clinical features resembled acute appendicitis and a pre-operative diagnosis could not be made. It is suggested that a thorough attempt should be made to exclude conditions mimicking appendicitis, including those of the biliary system, on finding a normal appendix at emergency appendicectomy without hesitating to convert to full laparotomy if required.

Adult↗
PubMed2000

Soluble GPI8 restores glycosylphosphatidylinositol anchoring in a trypanosome cell-free system depleted of lumenal endoplasmic reticulum proteins.

We previously established an in vitro assay for glycosylphosphatidylinositol (GPI) anchoring of proteins using trypanosome membranes. We now show that GPI anchoring is lost when the membranes are washed at high pH and restored to physiological pH prior to assay. We show that soluble component(s) of the endoplasmic reticulum that are lost in the high-pH wash are required for GPI anchoring. We reconstituted the high-pH extract with high-pH-treated membranes and demonstrated restoration of activity. Size fractionation of the high-pH extract indicated that the active component(s) was 30-50 kDa in size and was inactivated by iodoacetamide. Activity could also be restored by reconstituting the inactivated membranes with Escherichia coli-expressed, polyhistidine-tagged Leishmania mexicana GPI8 (GPI8-His; L. mexicana GPI8 is a soluble homologue of yeast and mammalian Gpi8p). No activity was seen when iodoacetamide-treated GPI8-His was used; however, GPI8-His could restore activity to iodoacetamide-treated membranes. Antibodies raised against L. mexicana GPI8 detected a protein of approx. 38 kDa in an immunoblot of the high-pH extract of trypanosome membranes. Our data indicate (1) that trypanosome GPI8 is a soluble lumenal protein, (2) that the interaction between GPI8 and other putative components of the transamidase may be dynamic, and (3) that GPI anchoring can be biochemically reconstituted using an isolated transamidase component.

Aminoacyltransferases↗
PubMed2000

Interaction between Trypanosoma evansi and Haemonchus contortus infection in goats.

An experimental study on interaction between Trypanosoma evansi and Haemonchus contortus infections was conducted in 42 male Barbari goats of 6-9 months age. Parasitological observations like prepatent period, faecal egg count, worm burden and mortality were made in animals infected with H. contortus followed 1 week later with T. evansi and vice versa. These parameters were also examined in animals with single infection with each of these parasites. It was revealed that T. evansi infection in goats lowered the normal resistance to H. contortus. The prepatent period was markedly reduced to 16-18 days in cases where T. evansi preceded H. contortus as compared to 21-25 and 21-23 days in single H. contortus and in a H. contortus preceding T. evansi infection, respectively. The rate of L(3) establishment was significantly enhanced in infection where T. evansi preceded H. contortus. A higher rate of mortality and more pronounced pathological effects were observed in combined infections than in single ones.

Animals↗
PubMed2000

Fasciolopsiasis--a persisting problem in eastern U.P.--a case report.

Fasciolopsiasis, or infection by the intestinal fluke, Fascilopsis buski, is endemic in the eastern states of our country. While it is by no means a rarity, especially in the rural set up, awareness regarding this common parasitic infestation is still a much-needed entity. The importance of a strong degree of suspicion and early diagnosis cannot be over emphasised, if a successful campaign is to be launched in its control. With this as our central theme, we proceed to report a case of and unsually heavy Fasciolopsis buski infection in our hospital, which had failed to be diagnosed in a semi urban setup in UP.

Adult↗
PubMed1999

Selective inhibitors of the glycosylphosphatidylinositol biosynthetic pathway of Trypanosoma brucei.

Synthetic analogues of D-GlcNalpha1-6D-myo-inositol-1-HPO(4)-3(sn-1, 2-diacylglycerol) (GlcN-PI), with the 2-position of the inositol residue substituted with an O-octyl ether [D-GlcNalpha1-6D-(2-O-octyl)myo-inositol-1-HPO(4)-3-sn-1, 2-dipalmitoylglycerol; GlcN-(2-O-octyl) PI] or O-hexadecyl ether [D-GlcNalpha1-6D-(2-O-hexadecyl)myo-inositol-1-HPO(4)-3-sn-1, 2-dipalmitoylglycerol; GlcN-(2-O-hexadecyl)PI], were tested as substrates or inhibitors of glycosylphosphatidylinositol (GPI) biosynthetic pathways using cell-free systems of the protozoan parasite Trypanosoma brucei (the causative agent of human African sleeping sickness) and human HeLa cells. Neither these compounds nor their N-acetyl derivatives are substrates or inhibitors of GPI biosynthetic enzymes in the HeLa cell-free system but are potent inhibitors of GPI biosynthesis in the T.brucei cell-free system. GlcN-(2-O-hexadecyl)PI was shown to inhibit the first alpha-mannosyltransferase of the trypanosomal GPI pathway. The N-acetylated derivative GlcNAc-(2-O-octyl)PI is a substrate for the trypanosomal GlcNAc-PI de-N-acetylase and this compound, like GlcN-(2-O-octyl)PI, is processed predominantly to Man(2)GlcN-(2-O-octyl)PI by the T.brucei cell-free system. Both GlcN-(2-O-octyl)PI and GlcNAc(2-O-octyl)PI also inhibit inositol acylation of Man(1-3)GlcN-PI and, consequently, the addition of the ethanolamine phosphate bridge in the T.brucei cell-free system. The data establish these substrate analogues as the first generation of in vitro parasite GPI pathway-specific inhibitors.

Acylation↗
PubMed1999

The GPI biosynthetic pathway as a therapeutic target for African sleeping sickness.

African sleeping sickness is a debilitating and often fatal disease caused by tsetse fly transmitted African trypanosomes. These extracellular protozoan parasites survive in the human bloodstream by virtue of a dense cell surface coat made of variant surface glycoprotein. The parasites have a repertoire of several hundred immunologically distinct variant surface glycoproteins and they evade the host immune response by antigenic variation. All variant surface glycoproteins are anchored to the plasma membrane via glycosylphosphatidylinositol membrane anchors and compounds that inhibit the assembly or transfer of these anchors could have trypanocidal potential. This article compares glycosylphosphatidylinositol biosynthesis in African trypanosomes and mammalian cells and identifies several steps that could be targets for the development of parasite-specific therapeutic agents.

Aminoacyltransferases↗
PubMed1999

A cell-free assay for glycosylphosphatidylinositol anchoring in African trypanosomes. Demonstration of a transamidation reaction mechanism.

We established an in vitro assay for the addition of glycosyl-phosphatidylinositol (GPI) anchors to proteins using procyclic trypanosomes engineered to express GPI-anchored variant surface glycoprotein (VSG). The assay is based on the premise that small nucleophiles, such as hydrazine, can substitute for the GPI moiety and effect displacement of the membrane anchor of a GPI-anchored protein or pro-protein causing release of the protein into the aqueous medium. Cell membranes containing pulse-radiolabeled VSG were incubated with hydrazine, and the VSG released from the membranes was measured by carbonate extraction, immunoprecipitation, and SDS-polyacrylamide gel electrophoresis/fluorography. Release of VSG was time- and temperature-dependent, was stimulated by hydrazine, and occurred only for VSG molecules situated in early compartments of the secretory pathway. No nucleophile-induced VSG release was seen in membranes prepared from cells expressing a VSG variant with a conventional transmembrane anchor (i.e. a nonfunctional GPI signal sequence). Pro-VSG was shown to be a substrate in the reaction by assaying membranes prepared from cells treated with mannosamine, a GPI biosynthesis inhibitor. When a biotinylated derivative of hydrazine was used instead of hydrazine, the released VSG could be precipitated with streptavidin-agarose, indicating that the biotin moiety was covalently incorporated into the protein. Hydrazine was shown to block the C terminus of the released VSG hydrazide because the released material, unlike a truncated form of VSG lacking a GPI signal sequence, was not susceptible to proteolysis by carboxypeptidases. These results firmly establish that the released material in our assay is VSG hydrazide and strengthen the proof that GPI anchoring proceeds via a transamidation reaction mechanism. The reaction could be inhibited with sulfhydryl alkylating reagents, suggesting that the transamidase enzyme contains a functionally important sulfhydryl residue.

Amino Acid Sequence↗
PubMed1999

Segregation of glycosylphosphatidylinositol biosynthetic reactions in a subcompartment of the endoplasmic reticulum.

Glycosylphosphatidylinositols (GPIs) are synthesized in the endoplasmic reticulum (ER) via the sequential addition of monosaccharides, fatty acid, and phosphoethanolamine(s) to phosphatidylinositol (PI). While attempting to establish a mammalian cell-free system for GPI biosynthesis, we found that the assembly of mannosylated GPI species was impaired when purified ER preparations were substituted for unfractionated cell lysates as the enzyme source. To explore this problem we analyzed the distribution of the various GPI biosynthetic reactions in subcellular fractions prepared from homogenates of mammalian cells. The results indicate the following: (i) the initial reaction of GPI assembly, i.e. the transfer of GlcNAc to PI to form GlcNAc-PI, is uniformly distributed in the ER; (ii) the second step of the pathway, i.e. de-N-acetylation of GlcNAc-PI to yield GlcN-PI, is largely confined to a subcompartment of the ER that appears to be associated with mitochondria; (iii) the mitochondria-associated ER subcompartment is enriched in enzymatic activities involved in the conversion of GlcN-PI to H5 (a singly mannosylated GPI structure containing one phosphoethanolamine side chain; and (iv) the mitochondria-associated ER subcompartment, unlike bulk ER, is capable of the de novo synthesis of H5 from UDP-GlcNAc and PI. The confinement of these GPI biosynthetic reactions to a domain of the ER provides another example of the compositional and functional heterogeneity of the ER. The implications of these findings for GPI assembly are discussed.

Acetylglucosamine↗
PubMed1999

Cytokinins in pathogenesis and disease resistance of Pyrenophora teres-barley and Dreschslera maydis-maize interactions during early stages of infection.

Infection of Hordeum vulgare L. by Pyrenophora teres and of Zea mays by Dreschslera maydis were characterized by 'green island' formation, higher cytokinin levels and accumulation of metabolites in the infected areas. Higher cytokinin concentrations of the order 6-Y,Y-dimethylallylaminopurine > zeatinriboside > zeatin > dihydrozeatinriboside were detected at infection sites of susceptible hosts. By virtue of these cytokinins, infection sites may be acting as metabolic sinks helping proliferation of the pathogen. Existence of translocatory sinks at infection zones was confirmed from autoradiographic studies, where, accumulation of labeled metabolites was prominent at infection sites of susceptible hosts. Upon infection the lower cytokinin levels of resistant hosts decreased further with progress of infection. In the infected resistant hosts the concentrations of zeatin/zeatinriboside were the maximum among the four identified cytokinins. The pathogen is also capable of secreting cytokinins as evident from quantification of cytokinins in culture filtrate extracts using HPLC. Since detached leaves were used in the experiments the increase/decrease of various cytokinin levels may be attributed to pathogen influence. The increase in cytokinin levels in the susceptible host may be aiding the growth of the pathogen on one hand, while the decrease in the infected resistant host may signal the host to activate defenses against a potential pathogen at the early stage of infection.

Autoradiography↗