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Glutathione-S-transferase (GST) a Phase-II drug detoxification enzyme, was detected in Setaria cervi, a bovine filarial parasite. In vitro effect of diethylcarbamazine, butylated hydroxyanisole and phenobarbitone on the GST of adult female S. cervi was assayed by the addition of these compounds in the maintenance medium. The specific activity of GST towards 1-chloro-2,4-dinitrobenzene was increased progressively 1.2–1.97, 1.3–2.4 and 1.2–2.7 times at 10–100 µM of diethylcarbamazine, butylated hydroxyanisole and phenobarbitone, respectively, after 5 h at 37oC. Substrate specificity studies showed a higher increase in specific activity with ethacrynic acid and no change with cumene hydroperoxide. Although the intensity of GST activity band was more in extract from diethylcarbamazine or butylated hydroxyanisole treated worms extract, an extra band of activity appeared in those worm extracts compared to control worm extract. SDS/PAGE showed increased thickness of the band corresponding to purified GST in extracts from diethylcarbamazine/butylated hydroxyanisole/phenobarbitone treated worms. Purification and quantification of GST from diethylcarbamazine and butylated hydroxyanisole treated worms indicated an increase in enzyme specific activity. The increase in GST protein by these agents was blocked by prior treatment with actinomycin D, indicative of a transcription dependent response. The role of this enzyme in motility and viability of microfilariae and adult female was tested in vitro using a range of known GST inhibitors. Of those tested, ethacrynic acid, ellagic acid, 1-chloro-2,4-dinitrobenzene, cibacron blue and butylated hydroxyanisole reduced the viability and motility of microfilariae and adult female worms at micromolar concentrations. These results suggest that S. cervi GST is inducible in response to the antifilarial drug diethylcarbamazine and may play an important role in parasite’s survival, thus could be a potential drug target.
Filarial nematodes infect more than 150 million people worldwide and are responsible for diseases including elephantiasis, river blindness and tropical pulmonary eosinophilia. Antifílarial agents that can kill all the stages in the life cycle of causative filar- iae have yet to be developed. Very little effort has been made towards rational drug design, employing knowledge gained from studies of the biochemistry and physiology of filarial worms and of their interactions with their specific vertebrate hosts. In this review, we highlight the research and development of rational antifílarial agents and we discuss the pitfalls since the discovery of diethylcarbamazine, the only drug of choice for controlling filariasis, despite its adverse side effects.
RNA silencing is one of the important phenomenon in plant defense mechanism, it actively protect host plants against viral infections. Existing viruses must have developed counter defense strategies to survive this arms race. Such counter defense strategy is the viral silencing suppressor (VSRs) which have been reported to directly interfere with the various steps leading to the interference of viral RNAs. Most identified VSRs are multifunctional, besides being RNA-silencing suppressors, they often perform essential roles by functioning as coat proteins, helper components for viral transmission, replicases and movement proteins, proteases or transcriptional regulators. One such identified VSR is AC4 of Sri Lankan cassava mosaic virus strain. Trivial knowledge about the structure –function relationship of this VSR leads to this work, where we focus on the structure generation by modelling to identify the mode of interactions with the various effector molecules of the silencing pathways. Structural analyses have been performed to screen interacting residues. Results indicate conserved structural features which signify propensity of functional interactions and further shows that this VSR can be a potent tool for the analysis of RNA silencing mechanisms and the relationships between different silencing pathways and VSRs.
Plant RNA interference has been a very well studied phenomenon since its discovery. We are well versed with the types of small noncoding RNAs that are prevalent in the plant systems and their pathways of biogenesis and subsequent actions. However, apart from model plant systems such as Arabidopsis and Oryza, very little information is available regarding the other members of the RNA interference machinery; specially Argonaute proteins which acts as the major stabilizing factor for execution of the interference. This work focuses on the exploration of the sequenced crop genomes available on the web using a hybrid approach of computational protein fishing and genome mining. The results indicate that this hybrid approach was successful in the identification of argonaute proteins in the crop genomes under study.
Significant protease activity has been detected in somatic extract of adults and microfilarial stage of Setaria cervi, using general protease substrates and collagen. The pH optima studies of the somatic extract of adult females showed two peaks at 7.0 and 5.0 for collagenase activity. Both forms were purified using sequential DEAE-sepharose and Sephadex G-100 column chromatography. The purified enzymes had the molecular masses of 175 and 20 kDa and pH optima at 7.0 and 5.0, respectively. The 175 kDa collagenase was more sensitive to metal chelators and serine protease inhibitors. However, 20 kDa collagenase was sensitive to cysteine protease inhibitors. The IgG antibodies from W. bancrofti infected human sera inhibited both enzymes. Further the purified collagenases were used to digest total human IgG at their respective pH and for different lengths of time. The 175 kDa protein was capable of cleaving human IgG. The digestion appeared to be restricted to a single cleavage point of H-chain within the hinge region of IgG molecule and produced fragments of similar molecular mass (27 kDa) indicating cleavage to Fab and Fc fragments. The degree of digestion was found to be proportional to the incubation time at 37°C. No further digestion of either fragments were observed. The L-chains were apparently resistant to collagenase digestion in all cases. Thus, the results suggest that S. cervi collagenase might be involved in the defense mechanisms of the parasite against the immune response of the host.
An experiment was conducted to find out the effect of short-term heat stress on morpho-physiological characters and antioxidants in 10 diverse wheat genotypes. Seed were aseptically grown in test tubes containing filter paper whose lower half was dipped in one-fourth MS media. Heat stress conditions were created by exposing the seedlings at 45 ºC for 2 h after 7 days of their germination. Measurements were taken after 3 days of treatment. Heat stress significantly reduced the shoot dry mass, root dry mass, shoot length and root length in all the genotypes. The chlorophyll content and membrane stability index decreased, whereas proline content increased in heat-treated plants. There was significant increase in the activity of catalase, guaiacol peroxidase and superoxide dismutase under stress conditions. The genotypic variations were also significant. On the basis of a coordinated simulation of all these parameters, wheat genotypes Raj 4037 and PBW 373 were identified as tolerant to high temperature stress. The study provides evidence that the tolerant genotypes were equipped with better management of physiological processes along with an efficient antioxidative defence system, sensitivity of which can be evaluated to a sufficient level of certainty at seedling stage.
This paper aims to discover the genetic parameters underlying the physiological efficiency and panicle characters of hill rice plants. In order to improve these characters, it is crucial to learn the nature and magnitude of gene action involved in inheritance of panicle and physiological traits. With this purpose, we combined a line × tester analysis of crosses involving local cultivars and indigenous and exotic germplasm grown at the northeastern hill region of India. From the study it follows that number of tillers per hill, number of ear bearing tillers per hill, panicle length, panicle weight, number of grains per panicle, number of filled grains per panicle, pollen fertility, spikelet fertility, leaf area index, total chlorophyll content and harvest index were mainly under the control of nonadditive gene action, while total dry matter accumulation and grain yield per plant were under the control of additive gene action. The genotypes DR 92, Ngoba, IET 16451 and UPR 1425-1-1-4 were identified as superior parents with high performance and significant general combining ability effects. For heterosis breeding six cross combinations, viz., DR 92/IET 16471, DR 92/UPR 1425-1-1-4, DR 92/IET 15482, Ngoba/IET 16469, Ngoba/H274-27-24, Manipuri/ IET 16451, and for recombination breeding two cross combinations, viz., DR 92/IET 16470 and Ngoba/UPR 1425- 1-1-4, were found suitable to ameliorate the panicle and physiological traits of hill rice grown in northeastern India.
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