186 research outputs found

    Chemical composition of the essential oil of Ocimum tenuiflorum L. (Krishna Tulsi) from North West Karnataka, India

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    The chemical composition of the essential oil of flowering aerial parts of Ocimum tenuiflorum L. growing in the North West Karnataka, India, was investigated. The hydro-distilled essential oil was analyzed by gas chromatography equipped with flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC/MS). Results demonstrated that the oil was found to be rich in phenyl derivative compounds (83.8%). The major compound was identified as methyl eugenol (82.9%) among twenty-six compounds, comprising 98.9% of the total oil

    Polymorphisms in the WNK1 gene are asociated with blood pressure variation and urinary potassium excretion

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    WNK1 - a serine/threonine kinase involved in electrolyte homeostasis and blood pressure (BP) control - is an excellent candidate gene for essential hypertension (EH). We and others have previously reported association between WNK1 and BP variation. Using tag SNPs (tSNPs) that capture 100% of common WNK1 variation in HapMap, we aimed to replicate our findings with BP and to test for association with phenotypes relating to WNK1 function in the British Genetics of Hypertension (BRIGHT) study case-control resource (1700 hypertensive cases and 1700 normotensive controls). We found multiple variants to be associated with systolic blood pressure, SBP (7/28 tSNPs min-p = 0.0005), diastolic blood pressure, DBP (7/28 tSNPs min-p = 0.002) and 24 hour urinary potassium excretion (10/28 tSNPs min-p = 0.0004). Associations with SBP and urine potassium remained significant after correction for multiple testing (p = 0.02 and p = 0.01 respectively). The major allele (A) of rs765250, located in intron 1, demonstrated the strongest evidence for association with SBP, effect size 3.14 mmHg (95%CI:1.23–4.9), DBP 1.9 mmHg (95%CI:0.7–3.2) and hypertension, odds ratio (OR: 1.3 [95%CI: 1.0–1.7]).We genotyped this variant in six independent populations (n = 14,451) and replicated the association between rs765250 and SBP in a meta-analysis (p = 7×10−3, combined with BRIGHT data-set p = 2×10−4, n = 17,851). The associations of WNK1 with DBP and EH were not confirmed. Haplotype analysis revealed striking associations with hypertension and BP variation (global permutation p10 mmHg reduction) and risk for hypertension (OR<0.60). Our data indicates that multiple rare and common WNK1 variants contribute to BP variation and hypertension, and provide compelling evidence to initiate further genetic and functional studies to explore the role of WNK1 in BP regulation and EH

    A CRISPR/Cas9-based method and primer design tool for seamless genome editing in fission yeast [version 1; peer review: 1 approved, 1 approved with reservations]

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    In the fission yeast Schizosaccharomyces pombe the prevailing approach for gene manipulations is based on homologous recombination of a PCR product that contains genomic target sequences and a selectable marker. The CRISPR/Cas9 system has recently been implemented in fission yeast, which allows for seamless genome editing without integration of a selection marker or leaving any other genomic ‘scars’. The published method involves manual design of the single guide RNA (sgRNA), and digestion of a large plasmid with a problematic restriction enzyme to clone the sgRNA. To increase the efficiency of this approach, we have established and optimized a PCR-based system to clone the sgRNA without restriction enzymes into a plasmid with a dominant natMX6 (nourseothricin) selection marker. We also provide a web-tool, CRISPR4P, to support the design of the sgRNAs and the primers required for the entire process of seamless DNA deletion. Moreover, we report the preparation of G1-synchronized and cryopreserved S. pombe cells, which greatly increases the efficiency and speed for transformations, and may also facilitate standard gene manipulations. Applying this optimized CRISPR/Cas9-based approach, we have successfully deleted over 80 different non-coding RNA genes, which are generally lowly expressed, and have inserted 7 point mutations in 4 different genomic regions

    A CRISPR/Cas9-based method and primer design tool for seamless genome editing in fission yeast [version 2; peer review: 2 approved]

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    In the fission yeast Schizosaccharomyces pombe the prevailing approach for gene manipulations is based on homologous recombination of a PCR product that contains genomic target sequences and a selectable marker. The CRISPR/Cas9 system has recently been implemented in fission yeast, which allows for seamless genome editing without integration of a selection marker or leaving any other genomic ‘scars’. The published method involves manual design of the single guide RNA (sgRNA), and digestion of a large plasmid with a problematic restriction enzyme to clone the sgRNA. To increase the efficiency of this approach, we have established and optimized a PCR-based system to clone the sgRNA without restriction enzymes into a plasmid with a dominant natMX6 (nourseothricin) selection marker. We also provide a web-tool, CRISPR4P, to support the design of the sgRNAs and the primers required for the entire process of seamless DNA deletion. Moreover, we report the preparation of G1-synchronized and cryopreserved S. pombe cells, which greatly increases the efficiency and speed for transformations, and may also facilitate standard gene manipulations. Applying this optimized CRISPR/Cas9-based approach, we have successfully deleted over 80 different non-coding RNA genes, which are generally lowly expressed, and have inserted 7 point mutations in 4 different genomic regions

    Antimicrobial activity of leaf and root methanolic extracts from Vinca pusilla Murr.

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    Vinca pusilla Murr. is a traditional medicinal plant used to treat several diseases. To substantiate the traditional medicinal utility of the plant, the present study aims at screening the antimicrobial activity of methanolic extracts of leaves and roots against five Gram positive, five Gram negative bacterial and four fungal strains. The minimum inhibitory concentration (MIC) were determined by two fold dilution assay. The results indicated that, leaf and root extracts were more effective on Bacillus subtilis and Staphylococcus aureus strains (MIC &lt; 1 mg/mL). The tested organisms were sensitive to root extract compared to leaf extract. Fungal strains were resistant than the bacterial strains to both the extracts. Thus the present study illustrates the antimicrobial potential of the plant.

    Bayesian lasso binary quantile regression

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    In this paper, a Bayesian hierarchical model for variable selection and estimation in the context of binary quantile regression is proposed. Existing approaches to variable selection in a binary classification context are sensitive to outliers, heteroskedasticity or other anomalies of the latent response. The method proposed in this study overcomes these problems in an attractive and straightforward way. A Laplace likelihood and Laplace priors for the regression parameters are proposed and estimated with Bayesian Markov Chain Monte Carlo. The resulting model is equivalent to the frequentist lasso procedure. A conceptional result is that by doing so, the binary regression model is moved from a Gaussian to a full Laplacian framework without sacrificing much computational efficiency. In addition, an efficient Gibbs sampler to estimate the model parameters is proposed that is superior to the Metropolis algorithm that is used in previous studies on Bayesian binary quantile regression. Both the simulation studies and the real data analysis indicate that the proposed method performs well in comparison to the other methods. Moreover, as the base model is binary quantile regression, a much more detailed insight in the effects of the covariates is provided by the approach. An implementation of the lasso procedure for binary quantile regression models is available in the R-package bayesQR

    Fast empirical Bayesian LASSO for multiple quantitative trait locus mapping

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    <p>Abstract</p> <p>Background</p> <p>The Bayesian shrinkage technique has been applied to multiple quantitative trait loci (QTLs) mapping to estimate the genetic effects of QTLs on quantitative traits from a very large set of possible effects including the main and epistatic effects of QTLs. Although the recently developed empirical Bayes (EB) method significantly reduced computation comparing with the fully Bayesian approach, its speed and accuracy are limited by the fact that numerical optimization is required to estimate the variance components in the QTL model.</p> <p>Results</p> <p>We developed a fast empirical Bayesian LASSO (EBLASSO) method for multiple QTL mapping. The fact that the EBLASSO can estimate the variance components in a closed form along with other algorithmic techniques render the EBLASSO method more efficient and accurate. Comparing with the EB method, our simulation study demonstrated that the EBLASSO method could substantially improve the computational speed and detect more QTL effects without increasing the false positive rate. Particularly, the EBLASSO algorithm running on a personal computer could easily handle a linear QTL model with more than 100,000 variables in our simulation study. Real data analysis also demonstrated that the EBLASSO method detected more reasonable effects than the EB method. Comparing with the LASSO, our simulation showed that the current version of the EBLASSO implemented in Matlab had similar speed as the LASSO implemented in Fortran, and that the EBLASSO detected the same number of true effects as the LASSO but a much smaller number of false positive effects.</p> <p>Conclusions</p> <p>The EBLASSO method can handle a large number of effects possibly including both the main and epistatic QTL effects, environmental effects and the effects of gene-environment interactions. It will be a very useful tool for multiple QTL mapping.</p
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