Hexavalent chromium induced changes in growth and biochemical responses of chromate-resistant bacterial strains isolated from tannery effluent.
Explore the source record for details and available documents.
Bacteria↗
Biomedical subjects
O P Shukla
Publications and source records attributed to O P Shukla.
Algal biomass: an economical method for removal of chromium from tannery effluent.
Explore the source record for details and available documents.
Biomass↗
Methaemoglobinaemia in epidemic proportions.
Five cases of methaemoglobinaemia, following exposure to industrial waste, are examined, investigated and managed successfully. The condition is discussed in detail.
Adult↗
Filarial DNA and its interaction with polyamines and antifilarial drugs.
The interaction of DNA from filarial parasite Setaria cervi with polyamines was monitored by melting temperature (Tm) profile, condensation and B to Z DNA transition and compared with DNA of Escherichia coli, Micrococcus luteus and calf thymus having different GC content. Polyamines, viz. spermine and spermidine, stabilized the secondary structure of all DNAs as indicated by increase in Tm value. UV absorption studies indicated B to Z DNA transition in the presence of polyamines. The amount of polyamines required for B to Z transition was dependent upon base composition of DNA and charge of the polyamine. Filarial DNA (AT rich) required six times higher concentration of spermine as compared to GC rich DNA for B to Z DNA transition. Spermidine was not effective in causing transition of S. cervi DNA even at Spd:DNA-P ratio of 20. The antifilarial compound suramin significantly decreased melting temperature of filarial DNA as compared to GC rich DNAs of other parasites. Suramin adversely affected condensation and B to Z DNA transition of various DNAs but prior addition of polyamines protected the DNAs from the destabilizing effect of suramin.
Animals↗
Studies of the hepatic mitochondrial and microsomal mixed-function oxidase system during Plasmodium yoelii infection and inducer treatment in Swiss albino mice.
Plasmodium yoelii infection resulted in depression of hepatic mitochondrial and microsomal mixed-function oxidase system indices, e.g. cytochrome P-450, cytochrome b5 and phase II detoxification enzyme glutathione-S-transferase, while heam and haemozoin registered a marked increase in Swiss albino mice. Phenobarbitone (inducer) treatment showed induced levels of hepatic mitochondrial and microsomal cytochrome P-450 and glutathione-S-transferase in normal as well as in infected mice. The induced cytochrome P-450 and glutathione-S-transferase activities were similar in normal and infected mice. The findings were further supported by the isoenzymic profile and drug-binding properties of the terminal monoxygenase, cytochrome P-450.
Animals↗
Identification of the polyamine N8-acetyltransferase involved in the pathway of 1,3-diaminopropane production in Acanthamoeba culbertsoni.
A cytosolic polyamine N-acetyltransferase that preferentially catalyzes the acetylation of spermidine in the N8-position was identified in the free-living pathogenic amoeba Acanthamoeba culbertsoni. In addition to spermidine, the enzyme also catalyzed the acetylation of spermine and putrescine with Michaelis constants (Km values) of 97, 12, and 10 microM, respectively. The Km value for acetylcoenzyme A (acetyl-CoA) was estimated to be 11 microM, whereas CoA had an inhibitory constant of 6 microM. The N-acetylase has a molecular mass of approximately 45 kDa. That the enzyme preferentially catalyzed the acetylation of spermidine at the N8-position, resulting in N8-acetylspermidine, the preferred substrate of the polyamine oxidase found in A. culbertsoni, indicates a role for the enzyme in the production of 1,3-diaminopropane, the major polyamine found in the Acanthamoeba.
Acanthamoeba↗
Polyamine metabolism in Acanthamoeba culbertsoni.
1,3-Diaminopropane has been identified as the major polyamine of Acanthamoeba culbertsoni. N-acetylputrescine and spermidine were present in appreciable amounts and putrescine as well as N-acetylspermidine were also detected, but spermine was absent. Changes in polyamine levels were observed during the growth of amoebae. Ornithine decarboxylase activity was detected in cell-free extracts but there was very low activity of arginine and lysine decarboxylases. A potent polyamine oxidase was demonstrated which preferentially acted on N8-acetyl-spermidine as the substrate while N1-acetylspermidine was a poor substrate; free polyamines did not serve as a good substrate for this enzyme. Active uptake of polyamines by the amoebae was also demonstrated.
Acanthamoeba↗
Characterization of Ca(2+)-ATPase of Setaria cervi (Nematoda: Filarioidea): effect of phenothiazines and anthelmintics.
1. A Mg2+ independent, Ca(2+)-ATPase requiring high concentrations of Ca2+ (5 mM) for the activation, equally distributed in cuticle-muscular-hypodermis, genital organs and gastrointestinal tissues and mainly localized in 10,000 g pellet fraction, was identified in Setaria cervi, a bovine filarial parasite. 2. Filarial enzyme showed Km value of 3.33 mM for ATP as computed from the double reciprocal Lineweaver-Burk plot. 3. The enzyme could be completely solubilized by sonication with about 4-fold increase in specific activity of the enzyme. 4. The enzyme showed about 2-fold activation by the calmodulin fractions isolated from S. cervi and rat brain homogenates. 5. The enzyme was highly sensitive to inhibition with some phenothiazine derivatives. Trifluoperazine was observed to be the most potent inhibitor followed by promethazine and chlorpromazine. 6. Some anthelmintics viz. diethycarbamazine and centperazine were found to be highly potent inhibitors of the enzyme, significant inhibition of filarial Ca(2+)-ATPase was also observed with levamisole and suramine. 7. Studies indicate Ca(2+)-ATPase of S. cervi as a potential chemotherapeutic target.
Animals↗
Polyamine oxidase from Acanthamoeba culbertsoni specific for N8-acetylspermidine.
Polyamine oxidase plays a key role in the catabolism of polyamines and regeneration of spermidine and putrescine. The mammalian enzyme utilises N1-acetylspermidine, and N8-acetylspermidine, although formed in the mammals, is not catabolised further. We have characterised an enzyme from Acanthamoeba culbertsoni which acts preferentially on N8-acetylspermidine. The highly unstable enzyme was stabilised in the presence of glycerol or dimethylsulphoxide together with spermine and purified 400-fold by a combination of DEAE-cellulose, CM-cellulose, spermine-Sepharose and Sephacryl S-300 chromatography. The enzyme has a pH optimum of 8 and a temperature optimum of 45 degrees C. The relative activities on different substrates are: N8-acetylspermidine 100%, N1-acetylspermine 40%, N1-acetylspermidine 1%, N1,8-diacetylspermidine 1% and N1,12-diacetylspermine 15%. Free polyamines and substrates of monoamine oxidase were not attacked. The enzyme yielded diaminopropane as an end product of catabolism and could be involved in the biosynthesis of this unusual polyamine present in large amounts in this organism.
Acanthamoeba↗
Polyamine metabolism in some helminth parasites.
Polyamine levels of some helminth parasites were analyzed by reverse phase HPLC of benzoyl derivatives. Setaria cervi, Acanthocheilonema viteae, Hymenolepis nana, H. diminuta, and Ascaridia galli contained higher levels of spermine than spermidine while in Ancylostoma ceylanicum and Nippostrongylus brasiliensis the spermidine levels were higher than spermine; putrescine was either absent or present in minor quantities. The enzymes of polyamine biosynthesis viz., ornithine decarboxylase, S-adenosyl methionine (SAM)-decarboxylase, and arginine decarboxylase were present in very low to negligible amounts in all the parasites examined. A. ceylanicum exhibited high activity of ornithine amino transferase (OAT) and catalyzed appreciable decarboxylation of ornithine. The ornithine decarboxylating activity of A. ceylanicum was localized in the particulate fraction containing mitochondria, not inhibited by alpha-difluoromethyl ornithine, the specific inhibitor of ornithine decarboxylase (ODC), but inhibited in the presence of glutamate, suggesting the involvement of mitochondrial OAT rather than a true ODC in ornithine decarboxylation in this parasite. Significant activity of polyamine oxidase was also detected in helminth parasites. The absence of polyamine biosynthesizing enzymes in helminth parasites suggests their dependence on hosts for uptake and interconversion of polyamines, providing a potential target for chemotherapy.
Ancylostoma↗
Action of DL-alpha-difluoromethyl ornithine on Acanthamoeba culbertsoni.
The multiplication of A. culbertsoni in the peptone medium was not inhibited by 10-20 mM concentration of alpha-difluoromethyl ornithine (DMFO) while a partial and transient inhibition of cell multiplication was observed by 10-20 mM DFMO in proteose peptone, yeast extract, glucose (PYG) medium. Ornithine decarboxylase (ODC) activity in the cells and cell free extracts was strongly inhibited by DFMO, excluding enzyme refractoriness and impermeability of cells for DFMO as the possible causes of DFMO resistance. The presence of polyamines in the peptone and PYG media as well as uptake of polyamines by the amoebae has been demonstrated. The growth and multiplication of A. culbertsoni in chemically defined medium was not affected by 1-5 mM DFMO while 10-20 mM DMFO yielded partial inhibition. A lowering of diaminopropane levels and enhancement of spermidine levels was observed in DFMO inhibited cells and level of ODC was drastically reduced in the inhibited cultures. Uptake of polyamines from the growth media may partly account for DFMO resistance of A. culbertsoni. Alternative mechanisms for DFMO resistance are indicated.
Acanthamoeba↗
Hepatic microsomal cytochrome P450 system during experimental hookworm infection.
Experimental infection of golden hamsters with the hookworm, Ancylostoma ceylanicum, caused a profound decline in the hepatic microsomal cytochrome P450 content. Concomitant decrease was also noticed in aminopyrine N-demethylase and benzo[a]pyrene hydroxylase activities. However, aniline hydroxylase activity was only marginally elevated during the infection. Microsomal markers, viz., cytochrome b5, NADH-cytochrome-c reductase, and glucose-6-phosphatase, were not significantly altered. Hepatic tissue exhibited an accumulation of lipids, especially phospholipids, triglycerides, and cholesterol, resulting in fatty necrosis around the central vein region. Isolated hepatic microsomes showed a decrease in phosphatidylcholine content. Impairment in hepatic mixed function oxidase (MFO) activities was further confirmed by prolongation in hexobarbital sleeping time and zoxazolamine-induced paralysis. The hepatic MFO system of A. ceylanicum-infected hamsters responded qualitatively and quantitatively in a manner similar to that of control hamsters, upon stimulation with selective chemical inducers like phenobarbitone and 3-methylcholanthrene. Kinetic and in vitro substrate binding studies revealed that for aminopyrine the substrate affinity and the maximum enzyme activity (Vmax) were decreased, while for aniline the binding affinity was decreased and the binding capacity was enhanced. Results indicate specific/selective impairment of the hepatic microsomal cytochrome P450 system during hookworm infection and may have many practical implications in toxicology and pharmacology.
Ancylostomiasis↗
Biogenic amines, metabolites and monoamine oxidase in the filarial worm Setaria cervi.
Analysis of biogenic monoamines and their metabolites in Setaria cervi adults by reverse phase high performance liquid chromatography (HPLC) revealed dopamine as the major monoamine followed by norepinephrine and 5-hydroxytryptamine (5-HT). 5-Hydroxy indole acetic acid and tryptophan were also detected in significant amounts. A particulate-bound monoamine oxidase (MAO, EC 1.4.3.4.) catalysing the oxidative deamination of several amines was also demonstrated in both microfilariae and adults. The enzyme from the parasites exhibited unusually high Km values for various monoamines. Dopamine was oxidized at the maximum rate while putrescine was not utilized as the substrate. MAO was predominantly associated with the mitochondrial fraction and concentrated mainly in the cuticle-muscle-hypodermis layer of the filariid. The enzyme was most active around pH 7.5 and 37 +/- 2 degrees C, relatively stable in the frozen state but was thermolabile. The specific MAO inhibitors, clorgyline and deprenyl, inhibited the enzyme with Ki values of 2 x 10(-7) M and 5 x 10(-6) M, respectively. Diethylcarbamazine, suramin, levamisole and centperazine significantly inhibited MAO activity. (The characteristics of the enzyme indicated that it may have a role in host-parasite interactions).
Animals↗
Molecular basis of hyperlipidemia in golden hamsters during experimental infection with Ancylostoma ceylanicum (Nematoda:Strongylidae).
An infection of golden hamsters with Ancylostoma ceylanicum, a hookworm parasite, induced profound hyperlipidemia, particularly hypertriglyceridemia, and the effect was directly related to the degree of infection. A significant increase was also noticed in serum cholesterol and phospholipid levels. The appearance of lipoprotein-X, an abnormal low density lipoprotein, was detected in the serum of hookworm-infected animals. The hyperlipidemia was further characterized by an increase in very low density lipoproteins (VLDL) and low density lipoproteins (LDL) with a concomitant decline in high density lipoproteins (HDL). Decreased lipolytic activities, especially triglyceride lipase, in hepatic tissue and induction of lipolytic activities in intestine and adipose tissues indicated mobilization of fats from adipose and jejunum with a defective removal of triglyceride-rich lipoproteins in hepatic tissues. Accumulation of lipids in liver and depletion in adipose tissue supported these results. The derangement may have a significant effect on host parasite interaction and is an important pathophysiological feature occurring during experimental ancylostomiasis.
Ancylostomiasis↗
Action of methylglyoxal bis (guanyl hydrazone) and related antiprotozoals on Acanthamoeba culbertsoni.
Methylglyoxal bis(guanyl hydrazone) (MGBG) and the related diamidine compounds berenil and pentamidine inhibited multiplication of A. culbertsoni. The growth inhibition by MGBG (2.5 mM) in the peptone medium was accompanied by the disappearance of spermidine and a marked reduction in the level of diaminopropane. MGBG and berenil completely inhibited growth in a chemically defined medium at 1 mM and 1-2 microM concentration, respectively. However, there was no decrease in the polyamine levels in the early stages of growth inhibition by these agents. Uptake of putrescine, spermidine and spermine by A. culbertsoni has been demonstrated but addition of exogenous polyamines did not reverse the growth inhibitory action of MGBG and berenil. Inhibition of S-adenosylmethionine decarboxylase and decrease in polyamine synthesis do not seem to be the primary targets for the antiamoebic action of MGBG and berenil.
Acanthamoeba↗
Effect of anthelmintics and phenothiazines on adenosine 5'-triphosphatases of filarial parasite Setaria cervi.
S. cervi showed particulate bound Ca2+ ATPase and Na+,K(+)-ATPase activities while Mg2+ ATPase was detected in traces. ATPase of S. cervi was also differentiated from the nonspecific p-nitrophenyl phosphatase activity. Female parasite and microfilariae exhibited higher Ca2+ ATPase and Na+,K(+)-ATPase activities than the male adults and the enzyme Na+,K(+)-ATPase was mainly concentrated in the gastrointestinal tract of the filarial parasite. Na+,K(+)-ATPase of the filariid was ouabain-sensitive while Ca2(+)-ATPase activity was regulated by concentration of Ca2+ ions and inhibited by EGTA. Phenothiazines, viz. trifluoperazine, promethazine and chlorpromazine caused significant inhibition of Ca2+ ATPase and Na+,K(+)-ATPase. Diethylcarbamazine was a potent inhibitor of these ATPases. Mebendazole, levamisole and centperazine also caused significant inhibition of the ATPases indicating this enzyme system as a common target for the action of anthelmintic drugs.
Adenosine Triphosphatases↗
Enzymes of PEP-succinate pathway in Setaria cervi and effect of anthelmintic drugs.
Localization of different enzymes of PEP-succinate pathway has been done in Setaria cervi, a bovine filarial worm. Succinate dehydrogenase and fumarate reductase were localized in mitochondria rich particulate fraction while all other enzymes were cytosolic. The in vitro effect of certain antifilarial/anthelmintic agents on these enzymes was also investigated. Sumarmin, at low concentration, could cause a marked inhibition of most of the enzymes of this pathway. Centperazine, an antifilarial drug being developed by CDRI showed significant inhibitory action on pyruvate kinase, lactate dehydrogenase, fumarase and succinate dehydrogenase while CDRI compound 72/70 showed significant inhibition of PEP-carboxykinase activity. Diethylcarbamazine and levamisole, however, were found to be more or less ineffective at lower concentrations against all the enzymes of this pathway.
Animals↗
Polyamine metabolism in Setaria cervi, the bovine filarial worm.
Spermine and spermidine were found to be the principal polyamines in the bovine filarial parasite Setaria cervi, whereas putrescine was observed in very low amounts. Studies conducted on the enzymes of polyamine biosynthesis revealed low activity for S-adenosyl-methionine decarboxylase, questionable and negligible activities for the decarboxylation of ornithine and arginine, and appreciable activity for ornithine aminotransferase. Uptake studies with radiolabeled putrescine, spermidine and spermine showed that these amines are rapidly taken up from the medium by an active uptake process. The uptake was temperature-sensitive and abolished at 0-4 degrees C. The questionable presence of biosynthetic enzymes such as ornithine and arginine decarboxylase and, on the other hand, an effective uptake mechanism indicate that the parasite may depend on the host for its polyamine requirement, thereby indicating a possible target for chemotherapy.
Adenosylmethionine Decarboxylase↗