Rapid and precise detection of cryptic tea pathogen Exobasidium vexans: RealAmp validation of LAMP approach.

This work embodies the development of a real time loop mediated isothermal amplification (RealAmp) assay for the rapid detection of the cryptic tea phytopathogen, Exobasidium vexans, the causal organism of blister blight disease. Due to the widespread popularity of tea as a beverage and the associated agro-economy, the rapid detection and management of the fast-spreading blister blight disease have been a longstanding necessity. Loop-mediated isothermal amplification (LAMP) primers were designed targeting the E. vexans ITS rDNA region and the reaction temperature was optimized at 62 °C with a 60 min reaction time. Amplification of the E. vexans isolates in the initial LAMP reactions was confirmed by both agarose gel electrophoresis and SYBR Green I dye based colour change visualization. The specificity of the LAMP primers for E. vexans was validated by negative testing of seven different phytopathogenic test fungi using LAMP and RealAmp assay. The positive findings in RealAmp assay for E. vexans strain were corroborated via detecting fluorescence signals in real-time. Further, the LAMP assays performed with gDNA isolated from infected tea leaves revealed positive amplification for the presence of E. vexans. The results demonstrate that this rapid and precise RealAmp assay has the potential to be applied for field-based detection of E. vexans in real-time.

Prevalence of micronutrient deficiency and its impact on the outcome of childhood cancer: A prospective cohort study.

BACKGROUND & AIMS: Impact of micronutrient deficiency on childhood malignancy is unexplored. We estimated the prevalence of baseline micronutrient deficiency in children with cancer and its impact on event-free survival (EFS) and overall survival (OS). METHODS: A longitudinal cohort study was conducted at a tertiary cancer centre in India. Children (≤18 years) with de novo malignancy were enrolled between October 2012 and May 2014. Baseline levels of vitamin B12, folate, zinc, selenium, copper, and iron were measured and values below 150 pmol/L, 6 ng/mL, International Zinc Nutrition Collaborative Group cut-off, 0.5 µmol/L, 10 µmol/L, and 50 µg/dL, respectively, indicated deficiency. RESULTS: Total 535 children [326 (60.9%) haematological and 209 (39.1%) solid malignancies] were enrolled with median follow-up of 66 months. Vitamin B12, folate, zinc, selenium, copper and iron deficiencies were found in 209 (39.1%), 89 (16.6%), 173 (32.3%), 39 (7.3%), 12 (2.2%), and 231 (43.2%) children, respectively. Selenium deficiency independently predicted poor EFS (hazard ratio [HR] = 1.56; p = 0.038) and OS (HR = 1.65; p = 0.027) in the cohort. In haematological malignancies, selenium deficiency predicted poor EFS (HR = 1.81; p = 0.023) and OS (HR = 2.12; p = 0.004). In solid malignancies, vitamin B12 (HR = 1.55; p = 0.028) and zinc (HR = 1.74; p = 0.009) deficiencies predicted poor EFS, and zinc deficiency predicted poor OS (HR = 1.77; p = 0.009). Multiple micronutrient (≥3) deficiencies also predicted poor EFS (HR = 1.69; p = 0.001) and OS (HR = 1.83; p < 0.001) in the whole cohort. CONCLUSIONS: Selenium deficiency was independently predictive of adverse outcomes in childhood cancer, particularly in haematological malignancies. Zinc deficiency adversely affected solid tumours. The adjunct use of micronutrient supplementation in paediatric malignancies should be explored.

Bioremediation of municipal sludge by vermitechnology and toxicity assessment by Allium cepa.

The aim of this study was to evaluate municipal sludge (MS) for its toxic potential by Allium cepa and also to understand the effect of vermicomposting on the reduction of toxicity, if any. Municipal sludge (MS) and vermicomposted sludge (VS) were evaluated. Elemental analysis of MS showed the presence of heavy metals. Morphological studies of A. cepa roots indicated coiled and wavy roots on exposure to MS but no root abnormality was reported in VS. Under genotoxic studies, inhibition in mitotic index was concentration dependent and the control values of 11.76 gradually reduced to 5.40 at 10% MS leachate whereas mitotic index was increased to 9.48 at 10% VS leachate. Exposure of leachate induced chromosomal aberrations, micronucleus formation and binucleate cells in a dose dependent manner. However, mitotic aberrations were observed significant at 10% MS leachate but they were insignificant at 10% VS leachate. The wet and dry weight of roots, root elongation and chlorophyll contents were reduced as the concentration of leachate increased but VS leachate did not produce considerable reduction. The wet and dry weight of A. cepa roots were 20.312 g and 3.250 g respectively and they were reduced to 10.82 g and 1.68 g respectively at 10% MS leachate but VS leachate showed an increase to 18.127 g and 2.53 g respectively. Total chlorophyll in control, 10% MS leachate and 10% VS leachate were 0.245 g, 0.162 g and 0.214 g respectively. It could be concluded that the MS was toxic to a remarkable extent but vermicomposting of sludge might be beneficial for bioremediation and recommended before land filling.

Synthesis and antiviral properties of arabino and ribonucleosides of 1,3-dideazaadenine, 4-nitro-1,3-dideazapurine and diketopiperazine.

Different arabinosides and ribosides, viz. Ara-DDA or 9(1-beta-D-arabinofuranosyl) 1,3-dideazaadenine (6), Ara-NDDP or 9(1-beta-D-arabinofuranosyl) 4-nitro-1,3-dideazapurine (7), Ara-DKP or 1(1-beta-D-arabinofuranosyl) diketopiperazine (8), Ribo-DDA or 9(1-beta-D-ribofuranosyl) 1,3-dideazaadenine (9) and Ribo-NDDP or 9(1-beta-D-ribofuranosyl) 4-nitro-1,3-dideazapurine (10) have been synthesized as probable antiviral agents. The arabinosides have been synthesized using the catalyst TDA-1 that causes stereospecific formation of beta-nucleosides while a one-pot synthesis procedure was adopted for the synthesis of the ribonucleosides where beta-anomers were obtained in higher yields. All the five nucleoside analogs have been screened for antiviral property against HIV-1 (IIIB), HSV-1 and 2, parainfluenza-3, reovirus-1 and many others. It was observed that arabinosides had greater inhibitory action than ribosides. The compound 7 or Ara-NDDP has shown maximum inhibition of HIV-1 replication than the rest of the molecules with an IC50 of 79.4 microg/mL.