Molecular characterization of Fusarium oxysporum f. sp. ciceri causing wilt of chickpea

Thirty isolates of Fusarium oxysporum f. sp. ciceri were isolated from rhizosphere soil of chickpea from different locations in Northern India. The amount of genetic variation was evaluated by polymerase chain reaction (PCR) amplification with a set of 40 RAPD primers and 2 IGS primers. Less than 10% of the amplified fragments in each case were polymorphic. Genetic similarity between each of the isolates was calculated and results indicate that there was little genetic variability among the isolates collected from the different locations. At the 0.75 similarity index the isolates divides into three groups. Isolates Foc-A18, Foc-A19, Foc-A20 forming a similar group and far different from other isolates.Key words: Fusarium oxysporum f. sp. ciceri, Fusarium wilt of chickpea, RAPD, ITS, IGS

HnRNPK maintains single strand RNA through controlling double-strand RNA in mammalian cells

Although antisense transcription is a widespread event in the mammalian genome, double-stranded RNA (dsRNA) formation between sense and antisense transcripts is very rare and mechanisms that control dsRNA remain unknown. By characterizing the FGF-2 regulated transcriptome in normal and cancer cells, we identified sense and antisense transcripts IER3 and IER3-AS1 that play a critical role in FGF-2 controlled oncogenic pathways. We show that IER3 and IER3-AS1 regulate each other\u27s transcription through HnRNPK-mediated post-transcriptional regulation. HnRNPK controls the mRNA stability and colocalization of IER3 and IER3-AS1. HnRNPK interaction with IER3 and IER3-AS1 determines their oncogenic functions by maintaining them in a single-stranded form. hnRNPK depletion neutralizes their oncogenic functions through promoting dsRNA formation and cytoplasmic accumulation. Intriguingly, hnRNPK loss-of-function and gain-of-function experiments reveal its role in maintaining global single- and double-stranded RNA. Thus, our data unveil the critical role of HnRNPK in maintaining single-stranded RNAs and their physiological functions by blocking RNA-RNA interactions

<em>In vitro</em> evaluation of antimicrobial activities and antibiotic susceptibility profiling of culturable actinobacteria from fresh water streams

665-673Actinobacteria are major producers of antibiotics, industrially significant enzymes and many pharmaceutically important biologically active compounds. Twenty two actinobacterial strains were isolated from fresh water stream sediment samples of Murlen National Park, Mizoram, India. The actinobacterial strains were screened against antifungal pathogens (Fusarium oxysporum, Fusarium udum, Fusarium proliferatum, Fusarium oxysporum ciceri and Fusarium graminearum), and antibacterial activities against five bacterial pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis and Escherichia coli) and a yeast pathogen Candida albicans. All strains showed antibacterial activity against E. coli and F. proliferatum. Based on the results of antagonistic, antibacterial and anti-yeast, two most potent strains Kocuria sp. and Streptomyces intermidus were further evaluated for their antibiotics susceptibility activity against 21 different antibiotics. Kocuria sp. showed resistance to 10 antibiotics whereas Streptomyces intermidus was resistance to 15 antibiotics. Modular genes Polyketide Synthase (PKS II) and Nonribosomal Peptide Synthetase (NRPS) were also detected in these two strains, which might be responsible for the production of secondary metabolites. Two volatile compounds, Di-N-octyl phthalate and 1-Bromo-3, 7-Dimethyloctane were identified from the extract of Streptomyces intermidus BPSWAC29 strain using Gas chromatography Mass spectrometry (GC-MS). This study highlights the promise of discovering novel actinobacteria with antimicrobial activity from underexplored niche biotopes such as fresh water stream sediments

Ultrasound-guided Glossopharyngeal Nerve Block- A New Paradigm in Pain Management

We report eight cases of oropharyngeal carcinoma in which ultrasound-guided percutaneous distal glossopharyngeal nerve (GPN) block was performed for pain relief. Mean age of the patients was 52 ± 11.5 [SD] years and median baseline pain score was 7 (IQR, 5–8). Under ultrasound guidance, mixture of ropivacaine and dexamethasone was injected into parapharyngeal space. Pain reduced after four weeks in all patients (median [IQR], 4 [2.5–5]). Median quality of life score improved as compared with baseline in physical health (63 [44–69] vs 50 [44–63]) and psychological domains (56 [56–63] vs 50 [50–63]), reduced in social relationships domain (31 [19–44] vs 44 [31–44]), remained same in environment domain (56 [44–69] vs 56 [56–56]). Seven patients showed improvement on Patients' Global Impression of Change scale, while six showed improvement on Clinical Global Impressions scale. These early results show that ultrasound guided distal GPN block can reduce pain intensity in oropharyngeal carcinoma patients

Molecular characterization of Fusarium oxysporum f. sp. ciceri causing wilt of chickpea

Thirty isolates of Fusarium oxysporum f. sp. ciceri were isolated from rhizosphere soil of chickpea from different locations in Northern India. The amount of genetic variation was evaluated by polymerase chain reaction (PCR) amplification with a set of 40 RAPD primers and 2 IGS primers. Less than 10% of the amplified fragments in each case were polymorphic. Genetic similarity between each of the isolates was calculated and results indicate that there was little genetic variability among the isolates collected from the different locations. At the 0.75 similarity index the isolates divides into three groups. Isolates Foc-A18, Foc-A19, Foc-A20 forming a similar group and far different from other isolates

In Vitro and In Vivo Plant Growth Promoting Activities and DNA Fingerprinting of Antagonistic Endophytic Actinomycetes Associates with Medicinal Plants.

Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 μg/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 μg/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 μg/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these plants and were shown to have antagonistic and plant growth promoting abilities. These results clearly suggest the possibility of using endophytic actinomycetes as bioinoculant for plant growth promotion, nutrient mobilization or as biocontrol agent against fungal phytopathogens for sustainable agriculture

In vitro and in vivo plant growth promoting activities and dna fingerprinting of antagonistic endophytic actinomycetes associates with medicinal plants

Endophytic actinomycetes have shown unique plant growth promoting as well as antagonistic activity against fungal phytopathogens. In the present study forty-two endophytic actinomycetes recovered from medicinal plants were evaluated for their antagonistic potential and plant growth-promoting abilities. Twenty-two isolates which showed the inhibitory activity against at least one pathogen were subsequently tested for their plant-growth promoting activities and were compared genotypically using DNA based fingerprinting, including enterobacterial repetitive intergenic consensus (ERIC) and BOX repetitive elements. Genetic relatedness based on both ERIC and BOX-PCR generates specific patterns corresponding to particular genotypes. Exponentially grown antagonistic isolates were used to evaluate phosphate solubilization, siderophores, HCN, ammonia, chitinase, indole-3-acetic acid production, as well as antifungal activities. Out of 22 isolates, the amount of indole-3-acetic acid (IAA) ranging between 10-32 mu g/ml was produced by 20 isolates and all isolates were positive for ammonia production ranging between 5.2 to 54 mg/ml. Among 22 isolates tested, the amount of hydroxamate-type siderophores were produced by 16 isolates ranging between 5.2 to 36.4 mu g/ml, while catechols-type siderophores produced by 5 isolates ranging from 3.2 to 5.4 mu g/ml. Fourteen isolates showed the solubilisation of inorganic phosphorous ranging from 3.2 to 32.6 mg/100ml. Chitinase and HCN production was shown by 19 and 15 different isolates, respectively. In addition, genes of indole acetic acid (iaaM) and chitinase (chiC) were successively amplified from 20 and 19 isolates respectively. The two potential strains Streptomyces sp. (BPSAC34) and Leifsonia xyli (BPSAC24) were tested in vivo and improved a range of growth parameters in chilli (Capsicum annuum L.) under greenhouse conditions. This study is the first published report that actinomycetes can be isolated as endophytes from within these plants and were shown to have antagonistic and plant growth promoting abilities. These results clearly suggest the possibility of using endophytic actinomycetes as bioinoculant for plant growth promotion, nutrient mobilization or as biocontrol agent against fungal phytopathogens for sustainable agriculture

Global distribution of DNA hydroxymethylation and DNA methylation in chronic lymphocytic leukemia

Abstract Background Chronic lymphocytic leukemia (CLL) has been a good model system to understand the functional role of 5-methylcytosine (5-mC) in cancer progression. More recently, an oxidized form of 5-mC, 5-hydroxymethylcytosine (5-hmC) has gained lot of attention as a regulatory epigenetic modification with prognostic and diagnostic implications for several cancers. However, there is no global study exploring the role of 5-hydroxymethylcytosine (5-hmC) levels in CLL. Herein, using mass spectrometry and hMeDIP-sequencing, we analysed the dynamics of 5-hmC during B cell maturation and CLL pathogenesis. Results We show that naïve B-cells had higher levels of 5-hmC and 5-mC compared to non-class switched and class-switched memory B-cells. We found a significant decrease in global 5-mC levels in CLL patients (n = 15) compared to naïve and memory B cells, with no changes detected between the CLL prognostic groups. On the other hand, global 5-hmC levels of CLL patients were similar to memory B cells and reduced compared to naïve B cells. Interestingly, 5-hmC levels were increased at regulatory regions such as gene-body, CpG island shores and shelves and 5-hmC distribution over the gene-body positively correlated with degree of transcriptional activity. Importantly, CLL samples showed aberrant 5-hmC and 5-mC pattern over gene-body compared to well-defined patterns in normal B-cells. Integrated analysis of 5-hmC and RNA-sequencing from CLL datasets identified three novel oncogenic drivers that could have potential roles in CLL development and progression. Conclusions Thus, our study suggests that the global loss of 5-hmC, accompanied by its significant increase at the gene regulatory regions, constitute a novel hallmark of CLL pathogenesis. Our combined analysis of 5-mC and 5-hmC sequencing provided insights into the potential role of 5-hmC in modulating gene expression changes during CLL pathogenesis

Field emission gun-scanning electron microscopy (FEG-SEM) micrographs of (A) <i>Streptomyces</i> sp. (BPSAC34) and (B) <i>Leifsonia xyli</i> (BPSAC24) showing spore chain morphology.

<p>Field emission gun-scanning electron microscopy (FEG-SEM) micrographs of (A) <i>Streptomyces</i> sp. (BPSAC34) and (B) <i>Leifsonia xyli</i> (BPSAC24) showing spore chain morphology.</p

PCR amplification of (A) <i>iaa</i>M gene and (B) <i>chi</i>C gene for endophytic actinomycetes isolates. M: low range (100bp -3 kb) molecular marker; N: negative control; numerical numbers represents different isolates.

<p>PCR amplification of (A) <i>iaa</i>M gene and (B) <i>chi</i>C gene for endophytic actinomycetes isolates. M: low range (100bp -3 kb) molecular marker; N: negative control; numerical numbers represents different isolates.</p