Transgenesis and conditional lethality in Aedes albopictus

The Asian tiger mosquito Aedes albopictus (Skuse) is a vector of several arboviruses including dengue and chikungunya. This highly invasive species originates from Southeast Asia and has spread across the world in the last 30 years. It is now established in Europe, North and South America, Africa, the Middle East and the Caribbean. In the absence of vaccine or antiviral drugs, efficient mosquito control strategies are crucial. Conventional control methods have so far failed to adequately control Ae. albopictus. Using germline transformation technology, a technique known as Release of Insects carrying a Dominant Lethal (RIDL) proposes to enhance the sterile insect technique by replacing irradiation with inherited dominant lethal genes. While this technology has recently shown some success in the field against the yellow fever mosquito, Aedes aegypti (L.), it remains to be implemented against Ae. albopictus. This thesis presents the development and application of gene transfer and site-specific integration technologies in Ae. albopictus, as well as the creation of tetracycline-repressible, female-specific flightless lines for vector control based on the RIDL method. Germline transformation and site-specific integration were performed using the piggyBac transposon and the ФC31 system, respectively. Ae. albopictus RIDL strains showing a conditional female-specific flightless phenotype were created using both the Ae. aegypti and the Ae. albopictus Actin-4 regulatory regions. Conditionality was provided by the ‘Tet-Off’ system, which is suppressed in the presence of tetracycline (and suitable analogues). One of these strains was assessed for attributes relevant to a RIDL control programme. Specific tailoring of the RIDL transgene with alternative transactivator elements was investigated using the ФC31 system. The work presented in this thesis lays the foundations for the application of the RIDL strategy to Ae. albopictus, an innovative vector-control method offering a promising alternative for efficient control of this highly invasive insect

Optimization of somatic cell injection in the perspective of nuclear transfer in goldfish

<p>Abstract</p> <p>Background</p> <p>Nuclear transfer has the potential to become one strategy for fish genetic resources management, by allowing fish reconstruction from cryopreserved somatic cells. Survival rates after nuclear transfer are still low however. The part played by unsuitable handling conditions is often questioned, but the different steps in the procedure are difficult to address separately. In this work led on goldfish (<it>Carassius auratus</it>), the step of somatic cells injection was explored. Non-enucleated metaphase II oocytes were used as a template to explore the toxicity of the injection medium, to estimate the best location where the cell should be injected, and to assess the delay necessary between cell injection and oocyte activation.</p> <p>Results</p> <p>Trout coelomic fluid was the most suitable medium to maintain freshly spawned oocytes at the metaphase II stage during oocyte manipulation. Oocytes were then injected with several media to test their toxicity on embryo development after fertilization. Trout coelomic fluid was the least toxic medium after injection, and the smallest injected volume (10 pL) allowed the same hatching rates as the non injected controls (84.8% ± 23). In somatic cell transfer experiments using non enucleated metaphase II oocytes as recipient, cell plasma membrane was ruptured within one minute after injection. Cell injection at the top of the animal pole in the oocyte allowed higher development rates than cell injection deeper within the oocyte (respectively 59% and 23% at mid-blastula stage). Embryo development rates were also higher when oocyte activation was delayed for 30 min after cell injection than when activation was induced without delay (respectively 72% and 48% at mid-blastula stage).</p> <p>Conclusions</p> <p>The best ability of goldfish oocytes to sustain embryo development was obtained when the carrier medium was trout coelomic fluid, when the cell was injected close to the animal pole, and when oocyte activation was induced 30 min after somatic cell injection. Although the experiments were not designed to produce characterized clones, application of these parameters to somatic cell nuclear transfer experiments in enucleated metaphase II oocytes is expected to improve the quality of the reconstructed embryos.</p

The evolution in the stellar mass of Brightest Cluster Galaxies over the past 10 billion years

Using a sample of 98 galaxy clusters recently imaged in the near infra-red with the ESO NTT, WIYN and WHT telescopes, supplemented with 33 clusters from the ESO archive, we measure how the stellar mass of the most massive galaxies in the universe, namely Brightest Cluster Galaxies (BCG), increases with time. Most of the BCGs in this new sample lie in the redshift range $0.2<z<0.6$, which has been noted in recent works to mark an epoch over which the growth in the stellar mass of BCGs stalls. From this sample of 132 clusters, we create a subsample of 102 systems that includes only those clusters that have estimates of the cluster mass. We combine the BCGs in this subsample with BCGs from the literature, and find that the growth in stellar mass of BCGs from 10 billion years ago to the present epoch is broadly consistent with recent semi-analytic and semi-empirical models. As in other recent studies, tentative evidence indicates that the stellar mass growth rate of BCGs may be slowing in the past 3.5 billion years. Further work in collecting larger samples, and in better comparing observations with theory using mock images is required if a more detailed comparison between the models and the data is to be made.Comment: 15 pages, 8 tables, 7 figures - Accepted for publication in MNRA

Genome wide analysis of gene dosage in 24,092 individuals estimates that 10,000 genes modulate cognitive ability

International audienceGenomic copy number variants (CNVs) are routinely identified and reported back to patients with neuropsychiatric disorders, but their quantitative effects on essential traits such as cognitive ability are poorly documented. We have recently shown that the effect size of deletions on cognitive ability can be statistically predicted using measures of intolerance to haploinsufficiency. However, the effect sizes of duplications remain unknown. It is also unknown if the effect of multigenic CNVs are driven by a few genes intolerant to haploinsufficiency or distributed across tolerant genes as well. Here, we identified all CNVs > 50 kilobases in 24,092 individuals from unselected and autism cohorts with assessments of general intelligence. Statistical models used measures of intolerance to haploinsufficiency of genes included in CNVs to predict their effect size on intelligence. Intolerant genes decrease general intelligence by 0.8 and 2.6 points of intelligence quotient when duplicated or deleted, respectively. Effect sizes showed no heterogeneity across cohorts. Validation analyses demonstrated that models could predict CNV effect sizes with 78% accuracy. Data on the inheritance of 27,766 CNVs showed that deletions and duplications with the same effect size on intelligence occur de novo at the same frequency. We estimated that around 10,000 intolerant and tolerant genes negatively affect intelligence when deleted, and less than 2% have large effect sizes. Genes encompassed in CNVs were not enriched in any GOterms but gene regulation and brain expression were GOterms overrepresented in the intolerant subgroup. Such pervasive effects on cognition may be related to emergent properties of the genome not restricted to a limited number of biological pathways

Effect of Hydrocortisone on Mortality and Organ Support in Patients With Severe COVID-19: The REMAP-CAP COVID-19 Corticosteroid Domain Randomized Clinical Trial.

Importance: Evidence regarding corticosteroid use for severe coronavirus disease 2019 (COVID-19) is limited. Objective: To determine whether hydrocortisone improves outcome for patients with severe COVID-19. Design, Setting, and Participants: An ongoing adaptive platform trial testing multiple interventions within multiple therapeutic domains, for example, antiviral agents, corticosteroids, or immunoglobulin. Between March 9 and June 17, 2020, 614 adult patients with suspected or confirmed COVID-19 were enrolled and randomized within at least 1 domain following admission to an intensive care unit (ICU) for respiratory or cardiovascular organ support at 121 sites in 8 countries. Of these, 403 were randomized to open-label interventions within the corticosteroid domain. The domain was halted after results from another trial were released. Follow-up ended August 12, 2020. Interventions: The corticosteroid domain randomized participants to a fixed 7-day course of intravenous hydrocortisone (50 mg or 100 mg every 6 hours) (n = 143), a shock-dependent course (50 mg every 6 hours when shock was clinically evident) (n = 152), or no hydrocortisone (n = 108). Main Outcomes and Measures: The primary end point was organ support-free days (days alive and free of ICU-based respiratory or cardiovascular support) within 21 days, where patients who died were assigned -1 day. The primary analysis was a bayesian cumulative logistic model that included all patients enrolled with severe COVID-19, adjusting for age, sex, site, region, time, assignment to interventions within other domains, and domain and intervention eligibility. Superiority was defined as the posterior probability of an odds ratio greater than 1 (threshold for trial conclusion of superiority >99%). Results: After excluding 19 participants who withdrew consent, there were 384 patients (mean age, 60 years; 29% female) randomized to the fixed-dose (n = 137), shock-dependent (n = 146), and no (n = 101) hydrocortisone groups; 379 (99%) completed the study and were included in the analysis. The mean age for the 3 groups ranged between 59.5 and 60.4 years; most patients were male (range, 70.6%-71.5%); mean body mass index ranged between 29.7 and 30.9; and patients receiving mechanical ventilation ranged between 50.0% and 63.5%. For the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively, the median organ support-free days were 0 (IQR, -1 to 15), 0 (IQR, -1 to 13), and 0 (-1 to 11) days (composed of 30%, 26%, and 33% mortality rates and 11.5, 9.5, and 6 median organ support-free days among survivors). The median adjusted odds ratio and bayesian probability of superiority were 1.43 (95% credible interval, 0.91-2.27) and 93% for fixed-dose hydrocortisone, respectively, and were 1.22 (95% credible interval, 0.76-1.94) and 80% for shock-dependent hydrocortisone compared with no hydrocortisone. Serious adverse events were reported in 4 (3%), 5 (3%), and 1 (1%) patients in the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively. Conclusions and Relevance: Among patients with severe COVID-19, treatment with a 7-day fixed-dose course of hydrocortisone or shock-dependent dosing of hydrocortisone, compared with no hydrocortisone, resulted in 93% and 80% probabilities of superiority with regard to the odds of improvement in organ support-free days within 21 days. However, the trial was stopped early and no treatment strategy met prespecified criteria for statistical superiority, precluding definitive conclusions. Trial Registration: ClinicalTrials.gov Identifier: NCT02735707

A novel approach for the generation of genetically modified mammary epithelial cell cultures yields new insights into TGF-beta signaling in the mammary gland

Abstract Introduction Molecular dissection of the signaling pathways that underlie complex biological responses in the mammary epithelium is limited by the difficulty of propagating large numbers of mouse mammary epithelial cells, and by the inability of ribonucleic acid interference-based knockdown approaches to fully ablate gene function. Here we describe a method for the generation of conditionally immortalized mammary epithelial cells with defined genetic defects, and we show how such cells can be used to investigate complex signal transduction processes using the transforming growth factor beta (TGF&#946;)/Smad pathway as an example. Methods We intercrossed the previously described H-2Kb-tsA58 transgenic mouse (Immortomouse), which expresses a temperature-sensitive mutant of the simian virus-40 large T-antigen (tsTAg), with mice of differing Smad genotypes. Conditionally immortalized mammary epithelial cell cultures were derived from the virgin mammary glands of offspring of these crosses and were used to assess the Smad dependency of different biological responses to TGF&#946;. Results IMECs could be propagated indefinitely at permissive temperatures and had a stable epithelial phenotype, resembling primary mammary epithelial cells with respect to several criteria, including responsiveness to TGF&#946;. Using this panel of cells, we demonstrated that Smad3, but not Smad2, is necessary for TGF&#946;-induced apoptotic, growth inhibitory and epithelial-to-mesenchymal transition responses, whereas either Smad2 or Smad3 can support TGF&#946;-induced invasion as long as a threshold level of total Smad is exceeded. Conclusions The present work demonstrates the practicality and utility of generating conditionally immortalized mammary epithelial cell lines from genetically modified Immortomice for detailed investigation of complex signaling pathways in the mammary epithelium.http://deepblue.lib.umich.edu/bitstream/2027.42/78285/1/bcr2728.xmlhttp://deepblue.lib.umich.edu/bitstream/2027.42/78285/2/bcr2728.pdfPeer Reviewe

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570

L'apamine, utilisation comme marqueur d'un canal potassium calcium-dependant et relations structure-activite

SIGLEINIST T 75357 / INIST-CNRS - Institut de l'Information Scientifique et TechniqueFRFranc

Existe-t-il une corrélation entre les données radiologiques et anatomopathologiques des berges des pièces opératoires de carcinome canalaire in situ du sein révélés par des microcalcifications dans le cadre d'une chirurgie conservatrice

Le but du traitement chirurgical conservateur du carcinome canalaire in situ du sein (CCIS) est de réaliser une exérèse carcinologique la plus complète possible, afin d'éviter toute récidive locale tumorale. L'état des berges d'exérèse est le facteur pronostic le plus important, c'est pourquoi l'analyse histologique est primordiale pour s'assurer d'une exérèse correcte. Notre étude montre que la radiographie de pièce opératoire permet de juger de l'exérèse complète ou non des microcalcifications, mais ses performances sont insuffisantes pour prévoir l'histologie finale des berges de la pièce de tumorectomie avec fiabilité. Le geste chirurgical doit donc s'assurer de l'exérèse complète des microcalcifications, et être le plus large possible tout en tenant compte du volume mammaire de la patiente. A la vue de la littérature, nous proposons plusieurs possibilités pour améliorer les performances de la pièce opératoire.NANTES-BU Médecine pharmacie (441092101) / SudocPARIS-BIUM (751062103) / SudocSudocFranceF