Low concentrations of nitric oxide delay the differentiation of embryonic stem cells and promote their survival

Nitric oxide (NO) is an intracellular messenger in several cell systems, but its contribution to embryonic stem cell (ESC) biology has not been characterized. Exposure of ESCs to low concentrations (2–20 μM) of the NO donor diethylenetriamine NO adduct confers protection from apoptosis elicited by leukaemia inhibitory factor (LIF) withdrawal. NO blocked caspase 3 activation, PARP degradation, downregulation of the pro-apoptotic genes Casp7, Casp9, Bax and Bak1 and upregulation of the anti-apoptotic genes Bcl-2 111, Bcl-2 and Birc6. These effects were also observed in cells overexpressing eNOS. Exposure of LIF-deprived mESCs to low NO prevented the loss of expression of self-renewal genes (Oct4, Nanog and Sox2) and the SSEA marker. Moreover, NO blocked the differentiation process promoted by the absence of LIF and bFGF in mouse and human ESCs. NO treatment decreased the expression of differentiation markers, such as Brachyury, Gata6 and Gata4. Constitutive overexpression of eNOS in cells exposed to LIF deprivation maintained the expression of self-renewal markers, whereas the differentiation genes were repressed. These effects were reversed by addition of the NOS inhibitor L-NMMA. Altogether, the data suggest that low NO has a role in the regulation of ESC differentiation by delaying the entry into differentiation, arresting the loss of self-renewal markers and promoting cell survival by inhibiting apoptosis

Microarray analysis of mRNA from cumulus cells following in vivo or in vitro maturation of mouse cumulus-oocyte complexes

The IVM of mammalian cumulus–oocyte complexes (COCs) yields reduced oocyte developmental competence compared with oocytes matured in vivo. Altered cumulus cell function during IVM is implicated as one cause for this difference. We have conducted a microarray analysis of cumulus cell mRNA following IVM or in vivo maturation (IVV). Mouse COCs were sourced from ovaries of 21-day-old CBAB6F1 mice 46 h after equine chorionic gonadotrophin (5 IU, i.p.) or from oviducts following treatment with 5 IU eCG (61 h) and 5 IU human chorionic gonadotrophin (13 h). IVM was performed in α-Minimal Essential Medium with 50 mIU FSH for 17 h. Three independent RNA samples were assessed using the Affymetrix Gene Chip Mouse Genome 430 2.0 array (Affymetrix, Santa Clara, CA, USA). In total, 1593 genes were differentially expressed, with 811 genes upregulated and 782 genes downregulated in IVM compared with IVV cumulus cells; selected genes were validated by real-time reverse transcription–polymerase chain reaction (RT-PCR). Surprisingly, haemoglobin α (Hba-a1) was highly expressed in IVV relative to IVM cumulus cells, which was verified by both RT-PCR and western blot analysis. Because haemoglobin regulates O2 and/or nitric oxide availability, we postulate that it may contribute to regulation of these gases during the ovulatory period in vivo. These data will provide a useful resource to determine differences in cumulus cell function that are possibly linked to oocyte competence.Karen L. Kind, Kelly M. Banwell, Kathryn M. Gebhardt, Anne Macpherson, Ashley Gauld, Darryl L. Russell and Jeremy G. Thompso

Regulated release of nitric oxide by nonhematopoietic stroma controls expansion of the activated T cell pool in lymph nodes

Fibroblastic reticular cells (FRCs) and lymphatic endothelial cells (LECs) are nonhematopoietic stromal cells of lymphoid organs. They influence the migration and homeostasis of naive T cells; however, their influence on activated T cells remains undescribed. Here we report that FRCs and LECs inhibited T cell proliferation through a tightly regulated mechanism dependent on nitric oxide synthase 2 (NOS2). Expression of NOS2 and production of nitric oxide paralleled the activation of T cells and required a tripartite synergism of interferon-γ, tumor necrosis factor and direct contact with activated T cells. Notably, in vivo expression of NOS2 by FRCs and LECs regulated the size of the activated T cell pool. Our study elucidates an as-yet-unrecognized role for the lymph node stromal niche in controlling T cell responses

Role of nitric oxide signaling components in differentiation of embryonic stem cells into myocardial cells

Nitric oxide (NO) is involved in number of physiological and pathological events. Our previous studies demonstrated a differential expression of NO signaling components in mouse and human ES cells. Here, we demonstrate the effect of NO donors and soluble guanylyl cyclase (sGC) activators in differentiation of ES cells into myocardial cells. Our results with mouse and human ES cells demonstrate an increase in Nkx2.5 and myosin light chain (MLC2) mRNA expression on exposure of cells to NO donors and a decrease in mRNA expression of both cardiac-specific genes with nonspecific NOS inhibitor and a concomitant increase and decrease in the mRNA levels of sGC α1 subunit. Although sGC activators alone exhibited an increase in mRNA expression of cardiac genes (MLC2 and Nkx2.5), robust inductions of mRNA and protein expression of marker genes were observed when NO donors and sGC activators were combined. Measurement of NO metabolites revealed an increase in the nitrite levels in the conditioned media and cell lysates on exposure of cells to the different concentrations of NO donors. cGMP analysis in undifferentiated stem cells revealed a lack of stimulation with NO donors. Differentiated cells however, acquired the ability to be stimulated by NO donors. Although, 3-(4-amino-5-cyclopropylpyrimidin-2-yl)-1-(2-fluorobenzyl)-1H-pyrazolo [3,4-b]pyridine (BAY 41-2272) alone was able to stimulate cGMP accumulation, the combination of NO donors and BAY 41-2272 stimulated cGMP levels more than either of the agents separately. These studies demonstrate that cGMP-mediated NO signaling plays an important role in the differentiation of ES cells into myocardial cells

Self Reported Lifetime Physical Activity in a Sample of Rural Cancer Survivors: A Pilot Study

Please see the pdf version of the abstract

Overpressure Effects from Propane Detonation : Comparison of Simulation Results with Experimental Data

International audienceAccidental gas explosion is a standard scenario considered in risk assessment for process industries. Blast walls or barriers are an efficient way to reduce the effects of pressure waves originated from an explosion. However, the proper design of a blast wall requires heavy investigation considering real accidental scenarios. CFD-based method can be used for a parametric investigation and for design optimization instead of experimental investigations. Nevertheless, the first CFD codes must be validated versus corresponding experimental data. The purpose of this paper is to present comparison of CFD simulations of propane/oxygen detonation and experimental measurements for small scale explosions

“They’re very passionate about making sure that women stay healthy”: Women’s experiences participating in a community paramedicine program

Community paramedicine programs (i.e., physician-directed preventive care by emergency medical services personnel embedded in communities) offer a novel approach to community-based health care. Project Swaddle, a community paramedicine program for mothers and their infants, seeks to address (directly or through warm referrals) the physical, mental, social, and economic needs of its participants. The objective of this process evaluation was to describe women’s experiences in Project Swaddle. By understanding their experiences, our work begins to build the foundation for similar programs and future examinations of the efficacy and effectiveness of these approaches. We completed 21 interviews with women living in Indiana (July 2019 - February 2020) who were currently participating in or had graduated from Project Swaddle. Interviews were audio-recorded, transcribed, and analyzed using a six-phase approach to thematic analysis. Four emerging themes resulted: 1) Program enrollment was influenced by the community paramedics’ experience and connections; 2) Participating women view the community paramedics as a trusted provider who supplied necessary health information and support; 3) In their role as physician extenders, the community paramedics enhanced patient care through communication with other providers; 4) The program and its community paramedics connected participants to outside resources, which aimed to support their holistic health and wellbeing. The results of our process evaluation suggest Project Swaddle is a promising approach to improving the health and wellbeing of women and infants. Future outcome and impact evaluations are necessary to quantify the effect of Project Swaddle on its participants

Vented explosion of hydrogen/air mixture: An intercomparison benchmark exercise

International audienceExplosion venting is a widely used mitigation solution in the process industry to protect indoor equipment or buildings from excessive internal pressure caused by accidental explosions. However, vented explosions are very complicated to model using computational fluid dynamics (CFD). In the framework of a French working group, the main target of this investigation is to assess the predictive capabilities of five CFD codes used by five different organizations by means of comparison with recent experimental data. On this basis several recommendations for the CFD modelling of vented explosions are suggested

A Mild and Selective Method for the Catalytic Hydrodeoxygenation of Cyanurate Activated Phenols in Multi-Phasic Continuous Flow.

A low energy, high selectivity approach to the catalytic hydrodeoxygenation of phenols is reported using batch or continuous flow methods to react 3 equivalents of phenol with cyanuric chloride then hydrogenolyzing the triarylcyanurate intermediate to give 3 equivalents of de-oxo aromatic. The use of cyanuric chloride compares favorably with existing activation methods, showing improved scalability, atom efficiency and economics. The scope of both the activation and hydrogenolysis stages are explored using lignin-related phenols. Initial development has identified that continuous stir tank reactors (CSTRs) enable a multi-phasic process for converting guaiacol to anisole, and at steady-state overcome the catalyst deactivation issues observed in batch, seemingly caused by the cyanurate by-product. Green chemistry aspects, and the potential for industrial adoption are discussed

Le universit\ue0 e la Riforma protestante. Studi e ricerche nel quinto centenario delle tesi luterane

Nel quinto centenario delle tesi luterane, il Centro interuniversitario per la storia delle universit\ue0 italiane - CISUI ha promosso un\u2019aggiornata riflessione sul rapporto tra Riforma e universit\ue0, approfondendo, attraverso alcuni esempi indicativi, il significato e le ricadute che i movimenti riformatori ebbero sul sistema d\u2019insegnamento superiore tra medioevo ed et\ue0 moderna. Come la proposta religiosa di Lutero s\u2019innest\uf2 in un clima umanistico-rinascimentale gi\ue0 attraversato da fermenti di rinnovamento, cos\uec le riforme provocarono e/o accelerarono un processo di trasformazione dei luoghi d\u2019insegnamento e dei metodi educativi, in parte gi\ue0 in atto. Universitas semper reformanda: se fin dalle sue origini l\u2019istituzione accademica aveva accolto e accompagnato i cambiamenti storici, dal XVI secolo le nuove proposte universitarie s\u2019inserirono nel solco delle riforme religiose e in un contesto culturale, politico, sociale ed economico ormai confessionale. Eredit\ue0 della Riforma fu la gemmazione, intorno a universit\ue0 stricto sensu, di istituzioni pi\uf9 o meno deputate a erogare i gradi accademici, ma provviste di insegnamenti \uabsuperiori\ubb, con finalit\ue0 esclusivamente professionali non soddisfatte in pieno dagli atenei: le accademie calviniste svizzere e francesi per la formazione dei pastori, i seminari cattolici post-tridentini per la preparazione dei sacerdoti, fino ai seminaria nobilium o alle scuole d\u2019ingegneria civile o militare. La Riforma protestante, avviata con Lutero, Zwingli e Calvino, provoc\uf2 un\u2019autentica rivoluzione dell\u2019ambito educativo che scosse le fondamenta delle comunit\ue0 accademiche e l\u2019epistemologia di alcune discipline, come l\u2019astronomia o la medicina