Phospholipase D signaling: orchestration by PIP2 and small GTPases

Hydrolysis of phosphatidylcholine by phospholipase D (PLD) leads to the generation of the versatile lipid second messenger, phosphatidic acid (PA), which is involved in fundamental cellular processes, including membrane trafficking, actin cytoskeleton remodeling, cell proliferation and cell survival. PLD activity can be dramatically stimulated by a large number of cell surface receptors and is elaborately regulated by intracellular factors, including protein kinase C isoforms, small GTPases of the ARF, Rho and Ras families and, particularly, by the phosphoinositide, phosphatidylinositol 4,5-bisphosphate (PIP2). PIP2 is well known as substrate for the generation of second messengers by phospholipase C, but is now also understood to recruit and/or activate a variety of actin regulatory proteins, ion channels and other signaling proteins, including PLD, by direct interaction. The synthesis of PIP2 by phosphoinositide 5-kinase (PIP5K) isoforms is tightly regulated by small GTPases and, interestingly, by PA as well, and the concerted formation of PIP2 and PA has been shown to mediate receptor-regulated cellular events. This review highlights the regulation of PLD by membrane receptors, and describes how the close encounter of PLD and PIP5K isoforms with small GTPases permits the execution of specific cellular functions

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Not AvailableConcurrent occurrence of Leptonema in a Brucella infected vet is presented and discussed. The merits of the preliminary laboratory tests in detecting these multiple infections and confirmation by molecular tests for quick and accurate diagnosis and treatment are emphasized.Not Availabl

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Not AvailableBrucellosis in pigs, caused by the bacterium Brucella suis, is an important zoonotic infection. In the present study, fluorescence polarization assay (FPA) was standardized and compared with indirect enzyme linked immunosorbent assay (iELISA) and competitive ELISA (cELISA) for diagnosis of porcine brucellosis. Test performances were evaluated using representative panel (n = 100), samples from swine brucellosis outbreak (n = 300), samples from brucellosis suspected animals (n = 291) and sera samples from apparently healthy animals (n = 1121). With panel samples, the FPA cut-off ≥11ΔmP was arrived with sensitivity (Se) and specificity (Sp) of 95.00 and 98.75%, respectively. Testing of samples from swine brucellosis outbreak, the diagnostic Se and Sp of 100 and 95.14% by iELISA, 73.91 and 100% by cELISA and 86.96 and 100% by FPA, respectively were recorded. Similarly, in case of swine brucellosis suspected samples, relative performance of FPA with cELISA had revealed higher kappa value of 0.864 with an accuracy of 93.47. Indirect ELISA was found to be highly sensitive but showed cross reactivity mainly for Yersinia enterocolitica O9 antibodies than cELISA and FPA. The high specificity of FPA test recorded in various types of samples in the study indicated that, FPA could serve as confirmatory test for individual animal diagnosis, outbreak confirmation, surveillance and quarantine of swine brucellosis cases.Not Availabl

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Not AvailableIn the present study, PCR amplified products of partial alk B gene (498 bp) and that of 731 bp products of B. abortus and B. melitensis strains respectively were analysed. Further, the 498 bp product of B. abortus was ligated into pGEM-T Easy vector and the recombinant plasmid after RE digestion with EcoRI released the insert and sequenced. Upon comparison of the nucleotide sequence of B. abortus S 99 strain with those of reference sequences from USA, it was found that, there was 100% homology with all the three sequences. Similarly, the PCR amplified product of B. melitensis 16M strain (731 bp) was similarly cloned and the nucleotide sequences were aligned with two reference sequences from USA and one from Belgium. B. abortus and B. melitensis strains showed 100 and 98.8% similarity respectively with the reference sequences available, thus confirming the purity of the strains being used in the diagnostic kits.Not Availabl

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Not AvailableLeptospirosis, considered to be the most widespread zoonoses in the world, is caused by serovars belonging to 7 pathogenic species of the genus Leptospira, viz. L. borgpetersenii, L. inadai, L. interrogans, L. kirschneri, L. noguchii, L. santarosai, and L. weilii (Ramadass et al. 1992). This paper reports the development of molecular technique for detecion of pathogenic Leptospira species by PCR amplification of a partial fragment of the major outer- membrane lipoprotein LipL32 gene, whose protein appears to be an important virulence factor (Haake et al. 2000, Yang et al. 2002), confined to pathogenic strains of all Leptospira spp. The technique could detect majority of serovars of L. interrogans, viz. icterohaemorrhagiae, canicola, autumnalis, javonica, pomona, pyrogenes, australis; L. borgpetersenii serovar Hardjobovis, and L. inadai.Not Availabl

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Not AvailableAIM: To evaluate the virulence determinants and genetic diversity of Staphylococcus aureus from bovine subclinical mastitis milk. METHODS AND RESULTS: PCR detection of virulence genes was performed for 173 Staph. aureus from bovine subclinical mastitis milk. Further, genetic diversity was analysed by agr and spa typing followed by pulsed field gel electrophoresis (PFGE) of selected isolates. Screening of virulence genes (n = 19) showed the adherence genes viz. fnbA, clfA, fnbB and cna in 98·8, 97·1, 68·8 and 28·3 percentage of isolates, respectively, and 80 strains (46·24%) positive for enterotoxin genes were distributed as 23 toxinotypes, of which, 5 genotypes contained a single gene and the rest comprised of multiple toxin genes. Out of agr type-1 (87·3%), 74·2 per cent belonged to the three predominant spa types. Of 27 spa types, 11 were identified for the first time. The predominant spa types were t267 (N =44), t359 (N = 42) and t6877 (N =29), which together accounts to 66·5 per cent of isolates. PFGE analysis of isolates (N = 45) covering all the spa types revealed mostly similar or closely related pulsotypes. Local emergence of spa type t6877 in herd-dependant manner was observed. spa sequence-based phylogenetic analysis suggested t267 as the ancestral clone of t359, t6877 and other spa types except two. CONCLUSION: Heterogenous virulence profile of the isolates had no significant association with the genotype. High prevalence of agr group I reaffirms their association with persistent subclinical mastitis. The spa type t267 appears to be the ancestral clone endemic in the region causing subclinical mastitis. In addition, few new spa types have emerged in the geographic region. SIGNIFICANCE AND IMPACT OF STUDY: Gives an insight into the genetic and evolutionary behaviour of Staph. aureus associated with bovine subclinical mastitis in India. The study would pave the way for devising effective control strategy for bovine mastitis in Indian context.Not Availabl

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Not AvailableEpidemiological mapping shows Staphylococcus aureus to be the leading mastitis causing pathogen in India with diverse genetic lineages circulating in the dairy cattle population. We previously reported that endemic clonal strains of S. aureus isolated from subclinical mastitis lead to specific alteration of epigenetic modulators resulting in deviating immune response in intramammary infection mouse model. However, the extent of transcriptome modulation and associated alternative splicing in S. aureus mastitis is poorly understood. Hence, to gain a deeper insight of the extent of modulation of transcriptome landscape, we expanded the study here using high throughput, paired-end RNA sequencing analysis of the mouse mammary gland inoculated with three strains of S. aureus (SA1, SA2, and SA3) possessing specific genotype, virulence and enterotoxin traits. Overall, we detected 35,878 transcripts in S. aureus inoculated mammary gland, 23% more than those annotated in the reference genome. Expression of 20,756 transcripts was > 1 fragment per kilobase of transcript per million mapped fragments and 25.95% of multi-exonic genes were alternatively spliced. We noted Alternative Splicing (AS) events for > 100 immune-related genes. S. aureus infection quantitatively altered AS events in mice mammary gland. Collectively, the majority of differentially expressed significant genes clustered into immune-associated, cell adhesion and metabolic process categories. We observed AS events for 379 transcripts of genes putatively encoding several splicing associated proteins and transcription factors besides inflammatory mediators. The present analysis provides new insights into global transcriptome landscape and AS events in host-defense related genes in response to S. aureus intramammary infection, suggesting the need for studies focusing on multi-target and/or network therapeutics approach to combat mastitis.Not Availabl

Physiological impact of Zinc nanoparticle on germination of rice (Oryza sativa L) seed

Nanoparticles affects growth and development of Plant. Zinc is an important micronutrient that regulates various physiological responses in plant. Application of nanoparticles for modulating plants physiological response is a recent practice. Zinc nanoparticles has been widely used in industry for several decades. However, no significant work had been made on its potential use in agriculture. Understanding physiological effect of Zn NP on rice seed germination could suggest the basis for its prospective application in agriculture to improve plant growth. In the present experiment effect of Zn NP was studied in Kmj-6-1-1 which is a commonly growing rice cultivar of Karimganj district of Assam, India. An exposure to Zn NP (0 mg/L, 5mg/L,10mg/L, 15mg/L, 20mg/L & 50mg/L) caused significant changes in radicle and plumule length , mass ( fresh & dry mass) and seed moisture content in rice. Antioxidant enzymes like guaiacol peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD) and gluthathione reductase (GR) also increased due to ZnNP treatment. This suggest that Zn NP may significantly alters antioxidant metabolism during rice seed germination. In conclusion, Zn NP protected rice plants from ROS damage by improving levels of antioxidant enzyme activities during germination. As a consequence the Zn NP treated seeds, showed better potential for germination. Further, genomic analysis of germinating rice seeds are needed to elucidate the molecular mechanisms by which Zn NP modulates germination process in rice

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Not AvailableBrucellosis in pigs, caused by the bacterium Brucella suis, is an important zoonotic infection. In the present study, fluorescence polarization assay (FPA) was standardized and compared with indirect enzyme linked immunosorbent assay (iELISA) and competitive ELISA (cELISA) for diagnosis of porcine brucellosis. Test performances were evaluated using representative panel (n = 100), samples from swine brucellosis outbreak (n = 300), samples from brucellosis suspected animals (n = 291) and sera samples from apparently healthy animals (n = 1121). With panel samples, the FPA cut-off ≥11ΔmP was arrived with sensitivity (Se) and specificity (Sp) of 95.00 and 98.75%, respectively. Testing of samples from swine brucellosis outbreak, the diagnostic Se and Sp of 100 and 95.14% by iELISA, 73.91 and 100% by cELISA and 86.96 and 100% by FPA, respectively were recorded. Similarly, in case of swine brucellosis suspected samples, relative performance of FPA with cELISA had revealed higher kappa value of 0.864 with an accuracy of 93.47. Indirect ELISA was found to be highly sensitive but showed cross reactivity mainly for Yersinia enterocolitica O9 antibodies than cELISA and FPA. The high specificity of FPA test recorded in various types of samples in the study indicated that, FPA could serve as confirmatory test for individual animal diagnosis, outbreak confirmation, surveillance and quarantine of swine brucellosis cases.Not Availabl