Isolation and functional characterization of a cotton ubiquitination-related promoter and 5'UTR that drives high levels of expression in root and flower tissues

<p>Abstract</p> <p>Background</p> <p>Cotton (<it>Gossypium </it>spp.) is an important crop worldwide that provides raw material to 40% of the textile fiber industry. Important traits have been studied aiming the development of genetically modified crops including resistance to insect and diseases, and tolerance to drought, cold and herbicide. Therefore, the characterization of promoters and regulatory regions is also important to achieve high gene expression and/or a specific expression pattern. Commonly, genes involved in ubiquitination pathways are highly and differentially expressed. In this study, we analyzed the expression of a cotton ubiquitin-conjugating enzyme (E2) family member with no previous characterization.</p> <p>Results</p> <p>Nucleotide analysis revealed high identity with cotton <it>E2 </it>homologues. Multiple alignment showed a premature stop codon, which prevents the encoding of the conserved cysteine residue at the <it>E2 </it>active site, and an intron that is spliced in <it>E2 </it>homologues, but not in <it>GhGDRP85</it>. The <it>GhGDRP85 </it>gene is highly expressed in different organs of cotton plants, and has high transcript levels in roots. Its promoter (uceApro2) and the 5'UTR compose a regulatory region named uceA1.7, and were isolated from cotton and studied in <it>Arabidopsis thaliana</it>. uceA1.7 shows strong expression levels, equaling or surpassing the expression levels of CaMV35S. The uceA1.7 regulatory sequence drives GUS expression 7-fold higher in flowers, 2-fold in roots and at similar levels in leaves and stems. GUS expression levels are decreased 7- to 15-fold when its 5'UTR is absent in uceApro2.</p> <p>Conclusions</p> <p>uceA1.7 is a strong constitutive regulatory sequence composed of a promoter (uceApro2) and its 5'UTR that will be useful in genetic transformation of dicots, having high potential to drive high levels of transgene expression in crops, particularly for traits desirable in flower and root tissues.</p

APLICAÇÃO DE BIOTECNOLOGIA NA PRODUÇÃO IN VITRO DE METABÓLITOS DO ANTI-HIPERTENSIVO LASSBIO 897

Introdu&ccedil;&atilde;o e objetivos: A biotransforma&ccedil;&atilde;o, utilizando-se modelos microbianos, tem sido usada para elucida&ccedil;&atilde;o do metabolismo de f&aacute;rmacos e posterior prepara&ccedil;&atilde;o in vitro de metab&oacute;litos. O composto LASSBio 897(um derivado N-acilidraz&ocirc;nico, o 3,4-metilenodioxbenzoil-3-tienilidrazona) foi sintetizado no Laborat&oacute;rio de Avalia&ccedil;&atilde;o e S&iacute;ntese de Subst&acirc;ncias Bioativas (LASSBio) da UFRJ. Devido potente atividade vasodilatadora, o LASSBio 897, &eacute; um promissor candidato a prot&oacute;tipo de f&aacute;rmaco anti-hipertensivo. Estudos farmacocin&eacute;ticos identificaram dois metab&oacute;litos no soro de c&atilde;es. Este trabalho objetivou a prepara&ccedil;&atilde;o in vitro destes metab&oacute;litos com utiliza&ccedil;&atilde;o de fungos filamentosos. Metodologia: Os microrganismos Aspergillus alliaceus NRRL 315, Beauveria bassiana ATCC 7159, Cunninghamella echinulata ATCC 9244, Mortierela isabelina NRRL 1757 e Muccor plumbeus ATCC4740 foram selecionados como biocatalisadores. Ap&oacute;s o t&eacute;rmino da rea&ccedil;&atilde;o, purifica&ccedil;&atilde;o em cromatografia de coluna, monitorada por HPLC-UV, e placa preparativa foi realizada. V&aacute;rias condi&ccedil;&otilde;es de purifica&ccedil;&atilde;o foram testadas de modo a promover maior rendimento. Resultados e discuss&otilde;es: Beauveria bassiana ATCC 7159 e Cunninghamella echinulata ATCC 9244 foram as cepas mais promissoras, produzindo maior quantidade dos metab&oacute;litos desejados, o que pode ser observado por HPLC-UV. Dificuldades na purifica&ccedil;&atilde;o a impediram a completa elucida&ccedil;&atilde;o estrutural dos metab&oacute;litos por Resson&acirc;ncia Magn&eacute;tica Nuclear (RMN) devido a pequena quantidade obtida. Conclus&atilde;o: Os fungos filamentosos mimetizam a produ&ccedil;&atilde;o dos metab&oacute;litos de mam&iacute;feros do LASSBio 897, entretanto os m&eacute;todos cromatogr&aacute;ficos cl&aacute;ssicos n&atilde;o s&atilde;o eficientes para total purifica&ccedil;&atilde;o dos metab&oacute;litos do meio reacional. Novas metodologias de purifica&ccedil;&atilde;o precisam ser desenvolvidas

A worldwide study of white matter microstructural alterations in people living with Parkinson’s disease

The progression of Parkinson’s disease (PD) is associated with microstructural alterations in neural pathways, contributing to both motor and cognitive decline. However, conflicting findings have emerged due to the use of heterogeneous methods in small studies. Here we performed a large diffusion MRI study in PD, integrating data from 17 cohorts worldwide, to identify stage-specific profiles of white matter differences. Diffusion-weighted MRI data from 1654 participants diagnosed with PD (age: 20–89 years; 33% female) and 885 controls (age: 19–84 years; 47% female) were analyzed using the ENIGMA-DTI protocol to evaluate white matter microstructure. Skeletonized maps of fractional anisotropy (FA) and mean diffusivity (MD) were compared across Hoehn and Yahr (HY) disease groups and controls to reveal the profile of white matter alterations at different stages. We found an enhanced, more widespread pattern of microstructural alterations with each stage of PD, with eventually lower FA and higher MD in almost all regions of interest: Cohen’s d effect sizes reached d = −1.01 for FA differences in the fornix at PD HY Stage 4/5. The early PD signature in HY stage 1 included higher FA and lower MD across the entire white matter skeleton, in a direction opposite to that typical of other neurodegenerative diseases. FA and MD were associated with motor and non-motor clinical dysfunction. While overridden by degenerative changes in the later stages of PD, early PD is associated with paradoxically higher FA and lower MD in PD, consistent with early compensatory changes associated with the disorder

Mechanisms of inactivation of the tumour suppressor gene RHOA in colorectal cancer

Reduced RHOA signalling has been shown to increase the growth/metastatic potential of colorectal tumours. However, the mechanisms of inactivation of RHOA signalling in colon cancer have not been characterised. A panel of colorectal cancer cell lines and large cohorts of primary tumours were used to investigate the expression and activity of RHOA, as well as the presence of RHOA mutations/deletions and promoter methylation affecting RHOA. Changes in RHOA expression were assessed by western blotting and qPCR after modulation of microRNAs, SMAD4 and c-MYC. We show here that RHOA point mutations and promoter hypermethylation do not significantly contribute to the large variability of RHOA expression observed among colorectal tumours. However, RHOA copy number loss was observed in 16% of colorectal tumours and this was associated with reduced RHOA expression. Moreover, we show that miR-200a/b/429 downregulates RHOA in colorectal cancer cells. In addition, we found that TGF- β /SMAD4 upregulates the RHOA promoter. Conversely, RHOA expression is transcriptionally downregulated by canonical Wnt signalling through the Wnt target gene c-MYC that interferes with the binding of SP1 to the RHOA promoter in colon cancer cells. We demonstrate a complex pattern of inactivation of the tumour suppressor gene RHOA in colon cancer cells through genetic, transcriptional and post-transcriptional mechanisms

Multidisciplinary scientific cruise to the Rio Grande Rise

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Measurement of the inclusive isolated-photon cross section in pp collisions at √s = 13 TeV using 36 fb−1 of ATLAS data

The differential cross section for isolated-photon production in pp collisions is measured at a centre-of-mass energy of 13 TeV with the ATLAS detector at the LHC using an integrated luminosity of 36.1 fb. The differential cross section is presented as a function of the photon transverse energy in different regions of photon pseudorapidity. The differential cross section as a function of the absolute value of the photon pseudorapidity is also presented in different regions of photon transverse energy. Next-to-leading-order QCD calculations from Jetphox and Sherpa as well as next-to-next-to-leading-order QCD calculations from Nnlojet are compared with the measurement, using several parameterisations of the proton parton distribution functions. The predictions provide a good description of the data within the experimental and theoretical uncertainties. [Figure not available: see fulltext.

Global prevalence and genotype distribution of hepatitis C virus infection in 2015 : A modelling study

Publisher Copyright: © 2017 Elsevier LtdBackground The 69th World Health Assembly approved the Global Health Sector Strategy to eliminate hepatitis C virus (HCV) infection by 2030, which can become a reality with the recent launch of direct acting antiviral therapies. Reliable disease burden estimates are required for national strategies. This analysis estimates the global prevalence of viraemic HCV at the end of 2015, an update of—and expansion on—the 2014 analysis, which reported 80 million (95% CI 64–103) viraemic infections in 2013. Methods We developed country-level disease burden models following a systematic review of HCV prevalence (number of studies, n=6754) and genotype (n=11 342) studies published after 2013. A Delphi process was used to gain country expert consensus and validate inputs. Published estimates alone were used for countries where expert panel meetings could not be scheduled. Global prevalence was estimated using regional averages for countries without data. Findings Models were built for 100 countries, 59 of which were approved by country experts, with the remaining 41 estimated using published data alone. The remaining countries had insufficient data to create a model. The global prevalence of viraemic HCV is estimated to be 1·0% (95% uncertainty interval 0·8–1·1) in 2015, corresponding to 71·1 million (62·5–79·4) viraemic infections. Genotypes 1 and 3 were the most common cause of infections (44% and 25%, respectively). Interpretation The global estimate of viraemic infections is lower than previous estimates, largely due to more recent (lower) prevalence estimates in Africa. Additionally, increased mortality due to liver-related causes and an ageing population may have contributed to a reduction in infections. Funding John C Martin Foundation.publishersversionPeer reviewe