Energetic study of cardioplegic hearts under ischaemia/reperfusion and [Ca2+] changes in cardiomyocytes of guinea-pig: Mitochondrial role

Abstract Aim: To study the role of mitochondria in the recovery of guinea-pig hearts exposed to high-K+-cardioplegia (CPG) and ischaemia/reperfusion (I/R) Methods: We measured contractility and heat release in perfused guineapig hearts and cytosolic and mitochondrial Ca2+ by epifluorescence and confocal microscopy in isolated cardiomyocytes loaded with Fluo-4 or Rhod-2. Results: In hearts, CPG increased the postischaemic contractile recovery, and this was potentiated by the mNCX blocker clonazepam and the mKATP opener diazoxide, which also prevented the fall in muscle economy. Moreover, CPG prevented the stunning induced by ouabain, which was reduced by clonazepam. In cardiomyocytes, CPG increased fluorescent signals of cytosolic and mitochondrial Ca2+, while the addition of a mNCX blocker (CGP37157) increased cytosolic but reduced mitochondrial [Ca2+]. Ouabain in CPG increased cytosolic Ca2+ and resting heat, but the addition of CGP37157 reduced them, as well as mitochondrial Ca2+. Conclusions: CPG, diazoxide and clonazepam improve postischaemic recovery, respectively, by increasing the Ca2+ cycling and by reducing the mitochondrial Ca2+ uptake either by uniporter or by mNCX. The mitochondria compete with the leaky sarcoplasmic reticulum (SR) as sink of Ca2+ in guinea-pig hearts, affecting the postischaemic contractility. CPG also prevented the ouabain-induced dysfunction by avoiding the Ca2+ overload. Ouabain reduced the synergism between CPG and clonazepam suggesting that [Na+]i and SR load influence the mNCX role.Fil: Ragone, María Inés. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Cátedra de Farmacología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata; ArgentinaFil: Torres, N. S.. Cardiovascular Research And Training Institute; Estados UnidosFil: Consolini, A. E.. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas. Cátedra de Farmacología; Argentin

T Cell Leukemia/Lymphoma 1A is essential for mouse epidermal keratinocytes proliferation promoted by insulin-like growth factor 1

T Cell Leukemia/Lymphoma 1A is expressed during B-cell differentiation and, when overexpressed, acts as an oncogene in mouse (Tcl1a) and human (TCL1A) B-cell chronic lymphocytic leukemia (B-CLL) and T-cell prolymphocytic leukemia (T-PLL). Furthermore, in the murine system Tcl1a is expressed in the ovary, testis and in pre-implantation embryos, where it plays an important role in blastomere proliferation and in embryonic stem cell (ESC) proliferation and self-renewal. We have also observed that Tcl1-/-adult mice exhibit alopecia and deep ulcerations. This finding has led us to investigate the role of TCL1 in mouse skin and hair follicles. We have found that TCL1 is expressed in the proliferative structure (i.e.The secondary hair germ) and in the stem cell niche (i.e.The bulge) of the hair follicle during regeneration phase and it is constitutively expressed in the basal layer of epidermis where it is required for the correct proliferative-differentiation program of the keratinocytes (KCs). Taking advantage of the murine models we have generated, including the Tcl1-/-and the K14-TCL1 transgenic mouse, we have analysed the function of TCL1 in mouse KCs and the molecular pathways involved. We provide evidence that in the epidermal compartment TCL1 has a role in the regulation of KC proliferation, differentiation, and apoptosis. In particular, the colony-forming efficiency (CFE) and the insulin-like growth factor 1 (IGF1)-induced proliferation are dramatically impaired, while apoptosis is increased, in KCs from Tcl1-/-mice when compared to WT. Moreover, the expression of differentiation markers such as cytokeratin 6 (KRT6), filaggrin (FLG) and involucrin (IVL) are profoundly altered in mutant mice (Tcl1-/-). Importantly, by over-expressing TCL1A in basal KCs of the K14-TCL1 transgenic mouse model, we observed a significant rescue of cell proliferation, differentiation and apoptosis of the mutant phenotype. Finally, we found TCL1 to act, at least in part, via increasing phospho-ERK1/2 and decreasing phospho-P38 MAPK. Hence, our data demonstrate that regulated levels of Tcl1a are necessary for the correct proliferation and differentiation of the interfollicular KC

Cool Core Clusters from Cosmological Simulations

We present results obtained from a set of cosmological hydrodynamic simulations of galaxy clusters, aimed at comparing predictions with observational data on the diversity between cool-core (CC) and non-cool-core (NCC) clusters. Our simulations include the effects of stellar and AGN feedback and are based on an improved version of the smoothed particle hydrodynamics code GADGET-3, which ameliorates gas mixing and better captures gas-dynamical instabilities by including a suitable artificial thermal diffusion. In this Letter, we focus our analysis on the entropy profiles, the primary diagnostic we used to classify the degree of cool-coreness of clusters, and on the iron profiles. In keeping with observations, our simulated clusters display a variety of behaviors in entropy profiles: they range from steadily decreasing profiles at small radii, characteristic of cool-core systems, to nearly flat core isentropic profiles, characteristic of non-cool-core systems. Using observational criteria to distinguish between the two classes of objects, we find that they occur in similar proportions in both simulations and in observations. Furthermore, we also find that simulated cool-core clusters have profiles of iron abundance that are steeper than those of NCC clusters, which is also in agreement with observational results. We show that the capability of our simulations to generate a realistic cool-core structure in the cluster population is due to AGN feedback and artificial thermal diffusion: their combined action allows us to naturally distribute the energy extracted from super-massive black holes and to compensate for the radiative losses of low-entropy gas with short cooling time residing in the cluster core.Comment: 6 pages, 4 figures, accepted in ApJL, v2 contains some modifications on the text (results unchanged

High prevalence of anti-hepatitis e virus antibodies among blood donors in central Italy, february to march 2014

Prevalence of anti-hepatitis E virus (HEV) antibodies is highly variable in developed countries, which seems partly due to differences in assay sensitivity. Using validated sensitive assays, we tested 313 blood donors attending a hospital transfusion unit in central Italy in January and February 2014 for anti-HEV IgG and IgM and HEV RNA. Data on HEV exposure were collected from all donors. Overall anti-HEV IgG prevalence was 49% (153/313). Eating raw dried pig-liver sausage was the only independent predictor of HEV infection (adjusted prevalence rate ratio = 2.14; 95% confidence interval: 1.23–3.74). Three donors were positive for either anti-HEV IgM (n = 2; 0.6%) or HEV RNA (n = 2; 0.6%); they were completely asymptomatic, without alanine aminotransferase (ALT) abnormalities. Of the two HEV RNA-positive donors (both harbouring genotype 3), one was anti-HEV IgG- and IgM-positive, the other was anti-HEV IgG- and IgM-negative. The third donor was positive for anti-HEV IgG and IgM but HEV RNA-negative. HEV infection is therefore hyperendemic among blood donors (80% men 18–64 years-old) from central Italy and associated with local dietary habits. Nearly 1% of donors have acute or recent infection, implying potential transmission to blood recipients. Neither ALT nor anti-HEV IgM testing seems useful to prevent transfusion-transmitted HEV infection. © 2016, European Centre for Disease Prevention and Control

Farmacodinamia general e interacciones medicamentosas : Mecanismos de acción de fármacos y metodologías de estudio experimental

Este libro aúna la experiencia docente y de investigación de los autores, que además de enseñar Farmacología ejercemos tareas de tiempo completo en distintos ámbitos de aplicación, como son tres diversos laboratorios de investigación de la Facultad de Ciencias Exactas de UNLP, dos centros de investigación de las facultades de Medicina de la UBA y de UNLP respectivamente, la pertenencia al CONICET y un sector de Farmacología y Toxicología de la industria farmacéutica. Aspiramos a que nuestra experiencia asista al alumno en la construcción de los primeros conceptos farmacológicos, y en la interpretación o realización de experimentos básicos de laboratorio. En este sentido, en la cátedra venimos observando que el proceso de adquisición de los conceptos farmacológicos es algo así como el aprendizaje de la lectura en un niño: al inicio hay gran dificultad en articular los diversos conceptos estudiados separadamente, hasta que se hace una repentina integración de los mismos y se empieza a pensarlos, a trabajarlos, a interpretar algunos resultados experimentales, a hipotetizar otros, y a generar protocolos de evaluación de efectos y mecanismos. Creemos que estos capítulos ayudarán a adquirir estos hábitos junto a la discusión oral en talleres y trabajos prácticos, con el último objetivo de que la Farmacología sea una asignatura adquirible en modo más razonado que memorístico, y que inspire al futuro profesional a ahondar el conocimiento farmacológico mediante la participación en equipos multidisciplinarios para el estudio preclínico de nuevos fármacos en la investigación, la industria o en organismos estatales de investigación y control farmacológico y toxicológico. En el último capítulo, este libro brindará nuestra experiencia en la compilación y comprensión de mecanismos que originan interacciones medicamentosas, tema en el que hemos dado varios cursos de posgrado, y que representa otra dificultad al alumno. Esto es debido a que las mismas se citan en diversos capítulos de los libros de texto y no siempre se describe claramente el origen y significación clínica de los mismos. El futuro profesional participará del equipo de salud que evalúa estudios clínicos junto al médico, en los cuales pueden aparecer o evaluarse las interacciones medicamentosas. Además, deberá detectar o prevenir interacciones medicamentosas durante el seguimiento farmacoterapéutico de sus pacientes o en el consejo en la dispensa de un medicamento.Facultad de Ciencias Exacta

Evidence That Qpx (Quahog Parasite Unknown) Is Not Present In Hatchery-Produced Hard Clam Seed

A protistan parasite known as QPX (Quahog Parasite Unknown) has been recently associated with disease and mortality of adult hard clams, Mercenaria mercenaria, from Canada to Virginia. There is concern that the organism may be transported in hatchery-reared seed. Tissue sections of 2,203 seed clams (\u3c1-20 mm) from 13 different hatcheries in six states, collected from 1995 to 1997 and examined by pathologists in three laboratories, failed to show QPX or QPX-like organisms. Further, QPX was not detected in a total of 756 hatchery-produced clams examined during their first year of field growout. From this, we conclude that hatchery-produced seed clams are an unlikely source of QPX organisms

Cosmological hydrodynamical simulations of galaxy clusters: X-ray scaling relations and their evolution

We analyse cosmological hydrodynamical simulations of galaxy clusters to study the X-ray scaling relations between total masses and observable quantities such as X-ray luminosity, gas mass, X-ray temperature, and $Y_{X}$. Three sets of simulations are performed with an improved version of the smoothed particle hydrodynamics GADGET-3 code. These consider the following: non-radiative gas, star formation and stellar feedback, and the addition of feedback by active galactic nuclei (AGN). We select clusters with $M_{500} > 10^{14} M_{\odot} E(z)^{-1}$, mimicking the typical selection of Sunyaev-Zeldovich samples. This permits to have a mass range large enough to enable robust fitting of the relations even at $z \sim 2$. The results of the analysis show a general agreement with observations. The values of the slope of the mass-gas mass and mass-temperature relations at $z=2$ are 10 per cent lower with respect to $z=0$ due to the applied mass selection, in the former case, and to the effect of early merger in the latter. We investigate the impact of the slope variation on the study of the evolution of the normalization. We conclude that cosmological studies through scaling relations should be limited to the redshift range $z=0-1$, where we find that the slope, the scatter, and the covariance matrix of the relations are stable. The scaling between mass and $Y_X$ is confirmed to be the most robust relation, being almost independent of the gas physics. At higher redshifts, the scaling relations are sensitive to the inclusion of AGNs which influences low-mass systems. The detailed study of these objects will be crucial to evaluate the AGN effect on the ICM.Comment: 24 pages, 11 figures, 5 tables, replaced to match accepted versio

Superclusters of galaxies from the 2dF redshift survey. II. Comparison with simulations

We investigate properties of superclusters of galaxies found on the basis of the 2dF Galaxy Redshift Survey, and compare them with properties of superclusters from the Millennium Simulation. We study the dependence of various characteristics of superclusters on their distance from the observer, on their total luminosity, and on their multiplicity. The multiplicity is defined by the number of Density Field (DF) clusters in superclusters. Using the multiplicity we divide superclusters into four richness classes: poor, medium, rich and extremely rich. We show that superclusters are asymmetrical and have multi-branching filamentary structure, with the degree of asymmetry and filamentarity being higher for the more luminous and richer superclusters. The comparison of real superclusters with Millennium superclusters shows that most properties of simulated superclusters agree very well with real data, the main differences being in the luminosity and multiplicity distributions.Comment: 15 pages, 13 Figures, submitted for Astronomy and Astrophysic

EFECTOS DEL HIPOTIROIDISMO EN LA ISQUEMIA-REPERFUSIÓN MIOCÁRDICA: ESTUDIO MECÁNICO-CALORIMÉTRICO DE LA PARTICIPACIÓN MITOCONDRIAL Y FARMACOLOGÍA

Se estudiarán las consecuencias de las alteraciones tiroideas  en la energética y recuperación de corazones de rata expuestos a isquemia y reperfusión (I/R). Se estudiarán dos modelos: de atontamiento y de I/R severa, integrando la metodología mecánico-calorimétrica de los corazones enteros perfundidos con mediciones bioquímicas de vías celulares cardioprotectoras y estimación de cambios de [Ca2+] en compartimentos celulares de cardiomiocitos aislados. Se generarán modelos de ratas hipotiroideas (HipoT, por administración de metimazol vía oral) y controles no tratados (C). Los corazones aislados y perfundidos por técnica de Langendorff se introducirán en un calorímetro de flujo y se expondrán a uno de los dos modelos de I/R, sin y con herramientas farmacológicas. El objetivo general es caracterizar los efectos mecánico-energéticos de las diversas condiciones en la disfunción por I/R, la interacción funcional entre mitocondrias, retículo sarcoplásmico y citosol (Mit-RS-cit) en la homeostasis de calcio, y los mecanismos implicados en la disfunción, mediante el tratamiento con herramientas farmacológicas selectivas. Se evaluarán las consecuencias energéticas y los mecanismos cardioprotectores directos en el miocardio de fármacos antiarrítmicos amiodarona y dronedarona en HipoT y C