Genetic transformation of tobacco plants with a cacao pathogenesis-related protein 4 for tolerance to water stress study : S03P06

Drought is an important environmental factor limiting the productivity of various crops worldwide. The development of crop cultivars with improved adaptation to drought is a major goal in many crop breeding programs. In addition to classical breeding approaches, genetic transformation to introduce candidate genes into plants for better tolerance to water deficit has been successfully developed. Pathogenesis-related proteins (PR proteins) are defined as plant proteins induced in response to pathogen attack. However, it is known that these proteins may also be involved in response to abiotic stresses. The objective of this study was to transform tobacco plants (as plant model for subsequent analysis of cultivated plant such as citrus) with a PR-4b protein from Theobroma cacao (TcPR-4b) and to select and test the tolerance of such transformed plants to water/osmotic stress. First, an in silico analysis of the TcPR-4b using the BLAST, Pfam,InterProScan, ORF-Finder programs, as well as a search on Cocoa GenDB databank were performed. The TcPR-4b belongs to a small family of PR-4 proteins whose members were mainly located on the chromosomes 5 (five genes) and 10 (one gene). The complete TcPR-4b sequence is 802 bp in length and contains two exons (171 and 258 bp), and one intron (82 bp); the corresponding protein is 142 amino acids in length. For plant transformation experiment, the TcPR-4b cDNA (from cacao-M. perniciosa interaction library) was cloned into the pGem- T Easy vector then subcloned on the pCambia binary vector 1390. Then, Agrobacterium tumefaciens strain EHA 105 was transformed with the35S::TcPR-4b::pCambia construction, and the transformed A. tumefaciens used for Nicotiana tabacum cv.Havana transformation by co-cultivation of leaf segments in selection medium. Transformed shoots from three transformation events are under selection for subsequent in vitro water/osmotic stress, using, among others, mannitol and NaCl. Funding Agency: FAPESB, CNPq, CAPES, EMBRAPA, FINEP/RENORBIO and CIRAD. (Texte intégral

Challenges and opportunities for healthcare workers in a rural district of Chad

Trained healthcare workers are an essential resource for effective health systems. However, healthcare workers' perspective on healthcare, the challenges they face to provide quality health services, and opportunities to improve motivation and providing adequate care are rarely investigated in resource-constrained settings of sub-Saharan Africa.; All reachable nurses of Abou Deia, a primarily rural district in the south-eastern part of Chad, were invited to participate. In-depth interviews were conducted to assess current challenges and opportunities faced in daily work, including factors that influence motivation and social wellbeing. Particular emphasis was placed on paediatric care.; Eight nurses were interviewed. Main work challenges pertained to overall workload, a lack of training and support regarding a serious case mix to be managed on their own, adverse working conditions, issues related to the local communities, and the impact of postings on nurses' private life. Poor working conditions and perceived lack of recognition emerged as the main demotivating factors. Motivation to improve nurses' skills so that they can provide good care, coupled with small, suggested changes in working conditions and health care organisation provide opportunities worth exploring to improve health workers' satisfaction, motivation and the care they can provide.; Health workers in a predominantly rural district in Chad face a wide variety of challenges, and hence their perspectives need to be taken into account to improve health services interventions that aim at enhancing quality of care. Nurses' willingness to further develop skills and knowledge, proactive search of solutions to remedy stock-outs of drugs and other medical devices, and motivational factors to improve the quality of care represent important opportunities for improving health services for all

Hypothesis and theory : a pathophysiological concept of stroke-induced acute phase response and increased intestinal permeability leading to secondary brain damage

Gut integrity impairment leading to increased intestinal permeability (IP) is hypothesized to be a trigger of critically illness. Approximately 15–20% of human ischemic stroke (IS) victims require intensive care, including patients with impaired level of consciousness or a high risk for developing life-threatening cerebral edema. Local and systemic inflammatory reactions are a major component of the IS pathophysiology and can significantly aggravate brain tissue damage. Intracerebral inflammatory processes following IS have been well studied. Until now, less is known about systemic inflammatory responses and IS consequences apart from a frequently observed post-IS immunosuppression. Here, we provide a hypothesis of a crosstalk between systemic acute phase response (APR), IP and potential secondary brain damage during acute and subacute IS stages supported by preliminary experimental data. Alterations of the acute phase proteins (APPs) C-reactive protein and lipopolysaccharide-binding protein and serum level changes of antibodies directed against Escherichia coli-cell extract antigen (IgA-, IgM-, and IgG-anti-E. coli) were investigated at 1, 2, and 7 days following IS in ten male sheep. We found an increase of both APPs as well as a decrease of all anti-E. coli antibodies within 48 h following IS. This may indicate an early systemic APR and increased IP, and underlines the importance of the increasingly recognized gut-brain axis and of intestinal antigen release for systemic immune responses in acute and subacute stroke stages

Safety and tolerability of sirolimus treatment in patients with autosomal dominant polycystic kidney disease

Background. We initiated a randomized controlled clinical trial to assess the effect of sirolimus on disease progression in patients affected by autosomal dominant polycystic kidney disease (ADPKD). Here we report the preliminary safety results of the first 6 months of treatment. Method. A total of 25 patients were randomized to sirolimus 2 mg/day and 25 patients to no treatment except standard care. Treatment adherence was monitored electronically. At baseline and at Month 6, laboratory parameters were analysed and the urinary protein profile in 24-h urine collections was determined. Results. Both treatment groups were well balanced for age, sex and renal function. In 94.1 ± 11.4% of the study days, patients in the sirolimus group were exposed to the drug when assuming a therapeutic efficacy duration of 30 h. At Month 6, the mean sirolimus dose and trough level were 1.28 ± 0.71 mg/day and 3.8 ± 1.9 μg/l, respectively. Glomerular (albumin, transferrin, IgG) and tubular (retinol-binding protein, α1-microglobulin) protein excretion remained unchanged. Glomerular filtration rate also did not change significantly. Haematological parameters were similar in both groups, except for a mild reduction of the mean corpuscular volume of erythrocytes in patients receiving sirolimus. Lipid levels were similar in both groups. Adverse events were transient and mild, and no grade 3 or 4 events occurred. The incidence of infections was similar in the sirolimus group (80%) and the standard group (88%). The most common gastrointestinal adverse events were mucositis (72% in the sirolimus group versus 16% in the standard group, P = 0.0001) and diarrhoea (36% in the sirolimus versus 20% in the standard group, P = 0.345). Conclusion. Treatment of ADPKD patients with sirolimus with a dose of 1-2 mg/day is safe and does not cause proteinuria or impairment of GFR. Treatment adherence was excellent. (ClinicalTrials.gov number, NCT00346918.

Common variation in EMSY and risk of breast and ovarian cancer: a case-control study using HapMap tagging SNPs.

BACKGROUND: EMSY could be involved in low-level susceptibility to breast and ovarian cancer. Gene amplification is seen in a proportion of breast and ovarian tumours and correlates with poor prognosis in breast cancer patients. Furthermore, the EMSY protein silences a transcription activation domain in BRCA2 exon 3. METHODS: We used a genetic association study design to determine if common genetic variation (frequency > or = 5%) in EMSY was associated with breast or ovarian cancer risk in the British population. Haplotype tagging single-nucleotide polymorphisms (htSNPs) were selected from the HapMap database and genotyped using Taqman in two large study sets of white British women (n [breast set] = 2343 cases and 2284 controls, n [ovarian set] = 864 cases and 864 controls). HapMap data might be insufficient to tag genetic variation in EMSY comprehensively. We therefore screened the gene promoter and coding sequences with denaturing high performance liquid chromatography in order to identify additional SNPs that are most likely to be functional. RESULTS: HapMap data on 22 SNPs show that 4 htSNPs tag 4 common haplotypes: rs2282611 (5'up t > g), rs4245443 (IVS7 g > a), rs2513511 (IVS16 a > g), rs2155220 (3'down c > t). We observed no association between any of the genotypes or associated haplotypes and breast or ovarian cancer risk. Seventeen out of the 18 remaining HapMap polymorphisms (94%) were well tagged by the 4 selected htSNPs (r2s > 0.8). Genotype frequencies for two further SNPs identified by screening and located near exon-intron boundaries, rs2508740 (IVS9 a > g) and rs11600501 (IVS10 c > t), were also similar in cases and controls. In order to simulate unidentified SNPs, we performed the leave-one-out cross-validation procedure on the HapMap data; over 95% of the common genetic variation was well represented by tagging polymorphisms. We are therefore likely to have tagged any common, functional variants present in our population. CONCLUSION: We found no association between common genetic variation in EMSY and risk of breast or ovarian cancer in two large study sets of white British women

Common ERBB2 polymorphisms and risk of breast cancer in a white British population: a case-control study.

INTRODUCTION: About two-thirds of the excess familial risk associated with breast cancer is still unaccounted for and may be explained by multiple weakly predisposing alleles. A gene thought to be involved in low-level predisposition to the disease is ERBB2 (HER2). This gene is involved in cell division, differentiation, and apoptosis and is frequently amplified in breast tumours. Its amplification correlates with poor prognosis. Moreover, the coding polymorphism I655V has previously been associated with an increased risk of breast cancer. METHODS: We aimed to determine if common polymorphisms (frequency >or= 5%) in ERBB2 were associated with breast cancer risk in a white British population. Five single-nucleotide polymorphisms (SNPs) were selected for study: SNP 1 near the promoter, SNP 2 in intron 1, SNP 3 in intron 4, SNP 4 in exon 17 (I655V), and SNP 5 in exon 27 (A1170P). We tested their association with breast cancer in a large case-control study (n = 2192 cases and 2257 controls). RESULTS: There were no differences in genotype frequencies between cases and controls for any of the SNPs examined. To investigate the possibility that a common polymorphism not included in our study might be involved in breast cancer predisposition, we also constructed multilocus haplotypes. Our set of SNPs generated all existing (n = 6) common haplotypes and no differences were seen in haplotype frequencies between cases and controls (P = 0.44). CONCLUSIONS: In our population, common ERBB2 polymorphisms are not involved in predisposition to breast cancer

Revised Stellar Properties of Kepler Targets for the Q1-17 (DR25) Transit Detection Run

The determination of exoplanet properties and occurrence rates using Kepler data critically depends on our knowledge of the fundamental properties (such as temperature, radius and mass) of the observed stars. We present revised stellar properties for 197,096 Kepler targets observed between Quarters 1-17 (Q1-17), which were used for the final transiting planet search run by the Kepler Mission (Data Release 25, DR25). Similar to the Q1--16 catalog by Huber et al. the classifications are based on conditioning published atmospheric parameters on a grid of Dartmouth isochrones, with significant improvements in the adopted methodology and over 29,000 new sources for temperatures, surface gravities or metallicities. In addition to fundamental stellar properties the new catalog also includes distances and extinctions, and we provide posterior samples for each stellar parameter of each star. Typical uncertainties are ~27% in radius, ~17% in mass, and ~51% in density, which is somewhat smaller than previous catalogs due to the larger number of improved logg constraints and the inclusion of isochrone weighting when deriving stellar posterior distributions. On average, the catalog includes a significantly larger number of evolved solar-type stars, with an increase of 43.5% in the number of subgiants. We discuss the overall changes of radii and masses of Kepler targets as a function of spectral type, with particular focus on exoplanet host stars.Comment: 19 pages, 13 figures. ApJS in pres