Coat Protein Gene based Characterization of Cucumber Mosaic Virus Isolates Infecting Banana in India

Banana plants showing typical yellow stripes on leaves as symptoms, in addition to leaf distortion and stunting of plant were collected from Karnataka (KAR), Maharashtra (MH) and Uttar Pradesh (UP) in India. The causal agent was identified as Cucumber mosaic virus (CMV) on the basis of transmission electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). Complete coat protein (CP) gene of all isolates were amplified using gene specific primers for coat protein (CP), followed by cloning into desired cloning vector for sequencing. Sequenced region were found containing complete single open reading frame of 657 nucleotides, potentially coding 219 amino acids. Sequence analysis of CP gene showed 93%-98% (at nucleotide) and 94%-99% (at amino acid) sequence identity between all three Indian isolates. On comparing CP gene sequences of CMV KAR, CMV MH and CMV UP with CMV P isolate (Physalis minima); we got 94%, 99% and 96% identity respectively. High degree identity at nucleotide level between these isolates of banana and Physalis minima (a weed) suggest that Physalis minima could be an alternate host of CMV banana. Phylogenetic analysis of nucleotide along with amino acid sequence of coat protein gene revealed that all our isolates belong to IB subgroup.  In short, it appears that there occurs a high incidence of CMV infecting banana belonging to IB subgroup in most parts of Indian subcontinent.Key words: Banana, CMV, CP gene, RT-PC

Alanine scanning of transmembrane helix 11 of Cdr1p ABC antifungal efflux pump of Candida albicans: identification of amino acid residues critical for drug efflux

Objectives: To investigate the role of transmembrane segment 11 (TMS11) of Candida albicans drug resistance protein (Cdr1p) in drug extrusion. Methods: We replaced each of the 21 putative residues of TMS11 with alanine by site-directed mutagenesis. The Saccharomyces cerevisiae AD1-8u− strain was used to overexpress the green fluorescent protein tagged wild-type and mutant variants of TMS11 of Cdr1p. The cells expressing mutant variants were functionally characterized. Results: Out of 21 residues of TMS11, substitution of seven residues, i.e. A1346G, A1347G, T1351A, T1355A, L1358A, F1360A and G1362A, affected differentially the substrate specificity of Cdr1p, while 14 mutants had no significant effect on Cdr1p function. TMS11 projection in an α-helical configuration revealed with few exceptions (A1346 and F1360), a distinct segregation of mutation-sensitive residues (A1347, T1351, T1355, L1358 and G1362) towards the more hydrophilic face. Interestingly, mutation-insensitive residues seem to cluster towards the hydrophobic side of the helix. Competition of rhodamine 6G efflux, in the presence of excess of various substrates in the cells expressing native Cdr1p, revealed for the first time the overlapping binding site between azoles (such as ketoconazole, miconazole and itraconazole) and rhodamine 6G. The ability of these azoles to compete with rhodamine 6G was completely lost in mutants F1360A and G1362A, while it was selectively lost in other variants of Cdr1p. We further confirmed that fungicidal synergism of calcineurin inhibitor FK520 with azoles is mediated by Cdr1p; wherein in addition to conserved T1351, substitution of T1355, L1358 and G1362 of TMS11 also resulted in abrogation of synergism. Conclusions: Our study for the first time provides an insight into the possible role of TMS11 of Cdr1p in drug efflux

Synthesis and mechanistic studies of diketo acids and their bioisosteres as potential antibacterial agents

A series of diketo esters and their pertinent bioisosteres were designed and synthesized as potent antibacterial agents by targeting methionine amino peptidases (MetAPs). In the biochemical assay against purified MetAPs from Streptococcus pneumoniae (SpMetAP1a), Mycobacterium tuberculosis (MtMetAP1c), Enterococcus faecalis (EfMetAP1a) and human (HsMetAP1b), compounds 3a, 4a and 5a showed more than 85% inhibition of all the tested MetAPs at 100 μM concentration. Compounds 4a and 5a also exhibited antibacterial potential with MIC values 62.5 μg/mL (S. pneumoniae), 31.25 μg/mL (E. faecalis), 62.5 μg/mL (Escherichia coli) and 62.5 μg/mL (S. pneumoniae), 62.5 μg/mL (E. coli), respectively. Moreover, 5a also significantly inhibited the growth of multidrug resistant E. coli strains at 512 μg/mL conc., while showing no cytotoxic effect towards healthy CHO cells and thus being selected. Growth kinetics study showed significant inhibition of bacterial growth when treated with different conc. of 5a. TEM analysis also displayed vital damage to bacterial cells by 5a at MIC conc. Moreover, significant inhibition of biofilm formation was observed in bacterial cells treated with MIC conc. of 5a as visualized by SEM micrographs. Interestingly, 5a did not cause an alteration in the hemocyte density in Galleria mellonella larvae which is considered in vivo model for antimicrobial studies and was non-toxic up to a conc. of 2.5 mg/mL

Determination of Lead Acetate Effects on Heavy Weight Protein of Musca domestica

So many insects are influenced easily by many pollutants; therefore, the influence of lead (as lead acetate) on Musca domestica was studied,lead is considered to be an important toxic waste which could contaminate the environment, such as soil, air and water, therefore, insects could be influenced by the lead. Musca domestica, was studied at 48 hours post treatment, under the effects of lead acetate, in different concentrations. Thus the lead is found to exert a definite specific physiological and morphological effect on these flies, It was observed that under the effects of lead abnormalities and deformity were developed in the larvae of flies. Thus these flies could present a useful module for the quick transmission of the environmental hazards due to lead contamination, which exerts a specific physiological and morphological effect on these flies. The purpose of the present work was to determin the effects of lead on proteins as a major indicator of physiological features alongwith morphology features of larvae of Diptera flies

Determination of Lead Acetate Effects on Heavy Weight Protein of Musca domestica

So many insects are influenced easily by many pollutants; therefore, the influence of lead (as lead acetate) on Musca domestica was studied,lead is considered to be an important toxic waste which could contaminate the environment, such as soil, air and water, therefore, insects could be influenced by the lead. Musca domestica, was studied at 48 hours post treatment, under the effects of lead acetate, in different concentrations. Thus the lead is found to exert a definite specific physiological and morphological effect on these flies, It was observed that under the effects of lead abnormalities and deformity were developed in the larvae of flies. Thus these flies could present a useful module for the quick transmission of the environmental hazards due to lead contamination, which exerts a specific physiological and morphological effect on these flies. The purpose of the present work was to determin the effects of lead on proteins as a major indicator of physiological features alongwith morphology features of larvae of Diptera flies

Spin-isospin SU(4) symmetry breaking in statistical spectroscopy

Centroid energies and variances of fixed spin-isospin and SU(4) symmetry spectral distributions are calculated without any approximation in d-s shell nuclei by using some previously developed propagation techniques. Intensities of various irreducible representations of SU(4) near the ground state are evaluated and these provide a measure of symmetry breaking. The influence of the residual interaction is studied. The results are compared with those coming from fixed SU(4) symmetry distributions or from fixed spin-isospin and SU(4) symmetry distributions with an approximate calculation of the variances. © 1982.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Shell model and spectral distribution study of SU(4) symmetry mixing in the s-d shell

Ground state intensities of SU(4) irreducible representations are evaluated by shell model diagonalization in various nuclei of the first half of the s-d shell. They are well reproduced by using SU(4) spin-isospin distributions with exactly calculated widths. Such distributions can then serve to predict the intensities in the other nuclei. The SU(4) symmetry is most often badly broken. © 1982.SCOPUS: ar.jinfo:eu-repo/semantics/publishe

Adverse Effects of Lead Acetate on Light Weight Protein of Bactrocera dorsalis

ALarge number of insects are influenced easily by a number of pollutants; such as, the influence of lead (as lead acetate) on Bactrocera dorsalis. Lead is considered to be an important toxic waste which could contaminate the environment, such as soil, air and water. Therefore, insects could be influenced by the lead. Bactrocera dorsalis, was studied at 48 hours post treatment, under the effects of lead acetate, in different concentrations. Lead is found to exert a definite specific physiological and morphological effect on these flies. It was observed that under the effect of lead abnormalities and deformities were developed in the larvae of flies. Thus these flies could present a useful module for the quick transmission of the environmental hazards due to lead contamination, which exerts a specific physiological and morphological effect on these flies. The purpose of the present work was to determine the effects of lead on proteins as a major indicator of physiological features along with morphological features of larvae of Bactrocera dorsalis flies

Heavy Protein Alteration under the Effects of Lead Acetate in Bactrocera cucurbitae

Lead is supposed to be an important poisonous waste which could contaminate the environment, therefore, insects could be influenced easily by the lead., Bactrocera cucurbitae was studied at 48 hours post treatment, under the effects of lead acetate, in different concentrations of 0.125 mg., 0.25 mg., 0.5 mg, 1.0 mg and 2.0 mg. It was observed that under the effects of lead abnormalities and deformity were developed in the larvae of flies. Thus these flies could present a useful module for the quick transmission of the environmental hazards due to lead contamination, which exerts a specific physiological and morphological effect on these flies