Transcription factors MYOCD, SRF, Mesp1 and SMARCD3 enhance the cardio-inducing effect of GATA4, TBX5, and MEF2C during direct cellular reprogramming.

Transient overexpression of defined combinations of master regulator genes can effectively induce cellular reprogramming: the acquisition of an alternative predicted phenotype from a differentiated cell lineage. This can be of particular importance in cardiac regenerative medicine wherein the heart lacks the capacity to heal itself, but simultaneously contains a large pool of fibroblasts. In this study we determined the cardio-inducing capacity of ten transcription factors to actuate cellular reprogramming of mouse embryonic fibroblasts into cardiomyocyte-like cells. Overexpression of transcription factors MYOCD and SRF alone or in conjunction with Mesp1 and SMARCD3 enhanced the basal but necessary cardio-inducing effect of the previously reported GATA4, TBX5, and MEF2C. In particular, combinations of five or seven transcription factors enhanced the activation of cardiac reporter vectors, and induced an upregulation of cardiac-specific genes. Global gene expression analysis also demonstrated a significantly greater cardio-inducing effect when the transcription factors MYOCD and SRF were used. Detection of cross-striated cells was highly dependent on the cell culture conditions and was enhanced by the addition of valproic acid and JAK inhibitor. Although we detected Ca(2+) transient oscillations in the reprogrammed cells, we did not detect significant changes in resting membrane potential or spontaneously contracting cells. This study further elucidates the cardio-inducing effect of the transcriptional networks involved in cardiac cellular reprogramming, contributing to the ongoing rational design of a robust protocol required for cardiac regenerative therapies.

In vivo luminescence imaging of NF-κB activity and serum cytokine levels predict pain sensitivities in a rodent model of osteoarthritis.

OBJECTIVE: To investigate the relationship between NF-κB activity, cytokine levels, and pain sensitivities in a rodent model of osteoarthritis (OA). METHODS: OA was induced in transgenic NF-κB-luciferase reporter mice via intraarticular injection of monosodium iodoacetate (MIA). Using luminescence imaging we evaluated the temporal kinetics of NF-κB activity and its relationship to the development of pain sensitivities and serum cytokine levels in this model. RESULTS: MIA induced a transient increase in joint-related NF-κB activity at early time points (day 3 after injection) and an associated biphasic pain response (mechanical allodynia). NF-κB activity, serum interleukin-6 (IL-6), IL-1β, and IL-10 levels accounted for ∼75% of the variability in pain-related mechanical sensitivities in this model. Specifically, NF-κB activity was strongly correlated with mechanical allodynia and serum IL-6 levels in the inflammatory pain phase of this model (day 3), while serum IL-1β was strongly correlated with pain sensitivities in the chronic pain phase of the model (day 28). CONCLUSION: Our findings suggest that NF-κB activity, IL-6, and IL-1β may play distinct roles in pain sensitivity development in this model of arthritis and may distinguish the acute pain phase from the chronic pain phase. This study establishes luminescence imaging of NF-κB activity as a novel imaging biomarker of pain sensitivities in this model of OA.

Molecular characterization of chordoma xenografts generated from a novel primary chordoma cell source and two chordoma cell lines.

OBJECT: Chordoma cells can generate solid-like tumors in xenograft models that express some molecular characteristics of the parent tumor, including positivity for brachyury and cytokeratins. However, there is a dearth of molecular markers that relate to chordoma tumor growth, as well as the cell lines needed to advance treatment. The objective in this study was to isolate a novel primary chordoma cell source and analyze the characteristics of tumor growth in a mouse xenograft model for comparison with the established U-CH1 and U-CH2b cell lines. METHODS: Primary cells from a sacral chordoma, called "DVC-4," were cultured alongside U-CH1 and U-CH2b cells for more than 20 passages and characterized for expression of CD24 and brachyury. While brachyury is believed essential for driving tumor formation, CD24 is associated with healthy nucleus pulposus cells. Each cell type was subcutaneously implanted in NOD/SCID/IL2Rγ(null) mice. The percentage of solid tumors formed, time to maximum tumor size, and immunostaining scores for CD24 and brachyury (intensity scores of 0-3, heterogeneity scores of 0-1) were reported and evaluated to test differences across groups. RESULTS: The DVC-4 cells retained chordoma-like morphology in culture and exhibited CD24 and brachyury expression profiles in vitro that were similar to those for U-CH1 and U-CH2b. Both U-CH1 and DVC-4 cells grew tumors at rates that were faster than those for U-CH2b cells. Gross tumor developed at nearly every site (95%) injected with U-CH1 and at most sites (75%) injected with DVC-4. In contrast, U-CH2b cells produced grossly visible tumors in less than 50% of injected sites. Brachyury staining was similar among tumors derived from all 3 cell types and was intensely positive (scores of 2-3) in a majority of tissue sections. In contrast, differences in the pattern and intensity of staining for CD24 were noted among the 3 types of cell-derived tumors (p < 0.05, chi-square test), with evidence of intense and uniform staining in a majority of U-CH1 tumor sections (score of 3) and more than half of the DVC-4 tumor sections (scores of 2-3). In contrast, a majority of sections from U-CH2b cells stained modestly for CD24 (scores of 1-2) with a predominantly heterogeneous staining pattern. CONCLUSIONS: This is the first report on xenografts generated from U-CH2b cells in which a low tumorigenicity was discovered despite evidence of chordoma-like characteristics in vitro. For tumors derived from a primary chordoma cell and U-CH1 cell line, similarly intense staining for CD24 was observed, which may correspond to their similar potential to grow tumors. In contrast, U-CH2b tumors stained less intensely for CD24. These results emphasize that many markers, including CD24, may be useful in distinguishing among chordoma cell types and their tumorigenicity in vivo.

Rapid behavioral and genomic responses to social opportunity.

From primates to bees, social status regulates reproduction. In the cichlid fish Astatotilapia (Haplochromis) burtoni, subordinate males have reduced fertility and must become dominant to reproduce. This increase in sexual capacity is orchestrated by neurons in the preoptic area, which enlarge in response to dominance and increase expression of gonadotropin-releasing hormone 1 (GnRH1), a peptide critical for reproduction. Using a novel behavioral paradigm, we show for the first time that subordinate males can become dominant within minutes of an opportunity to do so, displaying dramatic changes in body coloration and behavior. We also found that social opportunity induced expression of the immediate-early gene egr-1 in the anterior preoptic area, peaking in regions with high densities of GnRH1 neurons, and not in brain regions that express the related peptides GnRH2 and GnRH3. This genomic response did not occur in stable subordinate or stable dominant males even though stable dominants, like ascending males, displayed dominance behaviors. Moreover, egr-1 in the optic tectum and the cerebellum was similarly induced in all experimental groups, showing that egr-1 induction in the anterior preoptic area of ascending males was specific to this brain region. Because egr-1 codes for a transcription factor important in neural plasticity, induction of egr-1 in the anterior preoptic area by social opportunity could be an early trigger in the molecular cascade that culminates in enhanced fertility and other long-term physiological changes associated with dominance.

Metabolic regulation of T lymphocytes.

T cell activation leads to dramatic shifts in cell metabolism to protect against pathogens and to orchestrate the action of other immune cells. Quiescent T cells require predominantly ATP-generating processes, whereas proliferating effector T cells require high metabolic flux through growth-promoting pathways. Further, functionally distinct T cell subsets require distinct energetic and biosynthetic pathways to support their specific functional needs. Pathways that control immune cell function and metabolism are intimately linked, and changes in cell metabolism at both the cell and system levels have been shown to enhance or suppress specific T cell functions. As a result of these findings, cell metabolism is now appreciated as a key regulator of T cell function specification and fate. This review discusses the role of cellular metabolism in T cell development, activation, differentiation, and function to highlight the clinical relevance and opportunities for therapeutic interventions that may be used to disrupt immune pathogenesis.

Cutting edge: distinct glycolytic and lipid oxidative metabolic programs are essential for effector and regulatory CD4+ T cell subsets.

Stimulated CD4(+) T lymphocytes can differentiate into effector T cell (Teff) or inducible regulatory T cell (Treg) subsets with specific immunological roles. We show that Teff and Treg require distinct metabolic programs to support these functions. Th1, Th2, and Th17 cells expressed high surface levels of the glucose transporter Glut1 and were highly glycolytic. Treg, in contrast, expressed low levels of Glut1 and had high lipid oxidation rates. Consistent with glycolysis and lipid oxidation promoting Teff and Treg, respectively, Teff were selectively increased in Glut1 transgenic mice and reliant on glucose metabolism, whereas Treg had activated AMP-activated protein kinase and were dependent on lipid oxidation. Importantly, AMP-activated protein kinase stimulation was sufficient to decrease Glut1 and increase Treg generation in an asthma model. These data demonstrate that CD4(+) T cell subsets require distinct metabolic programs that can be manipulated in vivo to control Treg and Teff development in inflammatory diseases.

Two Cases of Late Shone Syndrome With Pulmonary Hypertension: Heart-Lung Transplant or Valve Surgery?

Two cases of Shone syndrome with severe mitral and aortic valve problems and pulmonary hypertension were referred for heart-lung transplantation. Severely elevated pulmonary vascular resistance (PVR) was confirmed as was severe periprosthetic mitral and aortic regurgitation. Based on the severity of the valve lesions in both patients, surgery was decided upon and undertaken. Both experienced early pulmonary hypertensive crises, one more than the other, that gradually subsided, followed by excellent recovery and reversal of pulmonary hypertension and PVR. These cases illustrate Braunwald's concept that pulmonary hypertension secondary to left-sided valve disease is reversible.

Estudio del sistema de agronegocios del jabalí en Argentina : antecedentes y situación actual desde la perspectiva de los negocios agroalimentarios

La carne de jabalí es apreciada por sus características organolépticas y nutricionales. El principal consumidor es Europa (Italia, Alemania y Francia) , seguido por Japón. En Argentina, esta especie se adaptó en gran parte del territorio y a diferentes situaciones ecológicas. Asimismo, la crianza en cautiverio, aprovechamiento, elaboración, distribución, precios y características nutricionales no son de amplio conocimiento. El propósito de este trabajo es realizar un diagnóstico del sistema de agronegocios del jabalí en Argentina a fin de identificar los puntos fuertes y débiles y a partir de allí, proponer acciones para dar soluciones a las problemáticas actuales. Se utiliza como metodología el abordaje de la epistemología fenomenológica y el método EPESA, lo cual permite contar con abordaje amplio y sistémico del agronegocio bajo análisis. También, se utiliza el marco teórico de la nueva economía institucional, siguiendo el abordaje de Joskow y sus tres vías de aproximación. Si bien es una producción que posee ventajas comparativas (clima, suelo, extensión, alimento) que permitirían el desarrollo productivo y comercial del sistema, se observa que la informalidad limita la implementación de planteos productivos con mayores niveles de tecnología, registros y datos genéticos, lo que imposibilita la exportación. Por otra parte, existe un problema de reglamentaciones con respecto a la faena, dado que debe ser aprobada por FAUNA y por SENASA, teniendo distintos requerimientos y tiempos de carga y transporte. Esto genera incompatibilidades para la comercialización formal, incentivando a transacciones ilegales, baja transparencia, oportunismo ex ante y ex post de las transacciones y altos costos de transacción. Dado este ambiente de negocios descripto, las empresas bajo estudio en general trabajan con nichos de mercado y no tienen un panorama de ampliación productiva y comercial. Por tanto, la mejora del ambiente institucional es la primera acción a seguir a fin de vencer las limitaciones mencionadas.

Estudio del sistema de agronegocios del jabalí en Argentina : antecedentes y situación actual desde la perspectiva de los negocios agroalimentarios

La carne de jabalí es apreciada por sus características organolépticas y nutricionales. El principal consumidor es Europa (Italia, Alemania y Francia), seguido por Japón. En Argentina, esta especie se adaptó en gran parte del territorio y a diferentes situaciones ecológicas. Asimismo, la crianza en cautiverio, aprovechamiento, elaboración, distribución, precios y características nutricionales no son de amplio conocimiento. El propósito de este trabajo es realizar un diagnóstico del sistema de agronegocios del jabalí en Argentina a fin de identificar los puntos fuertes y débiles y a partir de allí, proponer acciones para dar soluciones a las problemáticas actuales. Se utiliza como metodología el abordaje de la epistemología fenomenológica y el método EPESA, lo cual permite contar con abordaje amplio y sistémico del agronegocio bajo análisis. También, se utiliza el marco teórico de la nueva economía institucional, siguiendo el abordaje de Joskow y sus tres vías de aproximación. Si bien es una producción que posee ventajas comparativas (clima, suelo, extensión, alimento)que permitirían el desarrollo productivo y comercial del sistema, se observa que la informalidad limita la implementación de planteos productivos con mayores niveles de tecnología, registros y datos genéticos, lo que imposibilita la exportación. Por otra parte, existe un problema de reglamentaciones con respecto a la faena, dado que debe ser aprobada por FAUNA y por SENASA, teniendo distintos requerimientos y tiempos de carga y transporte. Esto genera incompatibilidades para la comercialización formal, incentivando a transacciones ilegales, baja transparencia, oportunismo ex ante y ex post de las transacciones y altos costos de transacción. Dado este ambiente de negocios descripto, las empresas bajo estudio en general trabajan con nichos de mercado y no tienen un panorama de ampliación productiva y comercial. Por tanto, la mejora del ambiente institucional es la primera acción a seguir a fin de vencer las limitaciones mencionadas.

La matemática como idioma y su importancia en la enseñanza y aprendizaje del Cálculo

La matemática es un idioma como varios autores han mencionado en diferentes trabajos científicos. En este artículo se analizan y comparan cuatro componentes del lenguaje y la matemática. Por otra parte, la matemática no es exactamente como otros idiomas. De hecho, la matemática parece ser más precisa y más limitada que los otros idiomas y esto tiene varias consecuencias en lo que se refiere a la enseñanza de dicha disciplina. En este artículo comentaremos nuestras experiencias, desarrolladas en Argentina, Alemania y Uruguay, teniendo en cuenta este enfoque de la enseñanza de la matemática como una extensión de la enseñanza de la lengua, y veremos cómo este enfoque ayudó a los estudiantes de los cursos de Cálculo, en diferentes formas.

Publicidad: prensa del corazón y otros temas

Se han publicado en estos días de nuevo las estadísticas sobre el consumo de prensa en nuestro país: seguimos estando en 108 ejemplares por 1000 habitantes. Superamos ligeramente por arriba el nivel del subdesarrollo marcado por la UNESCO: 100 ejemplares. Y lejos, por supuesto, de los niveles de los países de nuestro entorno, que se suele decir ahora. Pero hay un elemento distorsionador. En esos países (Inglaterra, Alemania...) suben las estadísticas las tiradas de la prensa «amarilla», de los periódicos de cotilleos, suplidos aquí por las tiradas inmensas de las revistas del «corazón». A pesar de que suponen un capítulo importante de los medios, y por tanto de la conformación de la opinión pública, no siempre se tienen en cuenta, se les suele despreciar como poco importantes. Algo así ha pasado en esta sección... ¡hasta este momento!

Advanced scrub practitioner toolkit

Toolkit for operating department staff and operating departments for role development NAASP (National Association Of Assistants In Surgical Practice) advanced practitioner toolkit provides outline strategies and a framework within the advanced scrub practitioner (ASP) role can be developed. this document defines the range of knowledge and skills required and sets the standards of practice for the ASP.

Long-Term Oceanographic and Ecological Research in the Western English Channel

Long-term research in the western English Channel, undertaken by the marine laboratories in Plymouth, is described and details of survey methods, sites, and time series given in this chapter. Major findings are summarized and their limitations outlined. Current research, with recent reestablishment and expansion of many sampling programmes, is presented, and possible future approaches are indicated. These unique long-term data sets provide an environmental baseline for predicting complex ecological responses to local, regional, and global environmental change. Between 1888 and the present, investigations have been carried out into the physical, chemical, and biological components (ranging from plankton and fish to benthic and intertidal assemblages) of the western English Channel ecosystem. The Marine Biological Association of the United Kingdom has performed the main body of these observations. More recent contributions come from the Continuous Plankton Recorder Survey, now the Sir Alister Hardy Foundation for Ocean Science, dating from 1957; the Institute for Marine Environmental Research, from 1974 to 1987; and the Plymouth Marine Laboratory, which was formed by amalgamation of the Institute for Marine Environmental Research and part of the Marine Biological Association, from 1988. Together, these contributions constitute a unique data series; one of the longest and most comprehensive samplings of environmental and marine biological variables in the world. Since the termination of many of these time series in 1987-1988 during a reorganisation of UK marine research, there has been a resurgence of interest in long-term environmental change. Many programmes have been restarted and expanded with support from several agencies. The observations span significant periods of warming (1921-1961; 1985-present) and cooling (1962-1980). During these periods of change, the abundance of key species underwent dramatic shifts. The first period of warming saw changes in zooplankton, pelagic fish, and larval fish, including the collapse of an important herring fishery. During later periods of change, shifts in species abundances have been reflected in other assemblages, such as the intertidal zone and the benthic fauna. Many of these changes appear to be related to climate, manifested as temperature changes, acting directly or indirectly. The hypothesis that climate is a forcing factor is widely supported today and has been reinforced by recent studies that show responses of marine organisms to climatic attributes such as the strength of the North Atlantic Oscillation. The long-term data also yield important insights into the effects of anthropogenic disturbances such as fisheries exploitation and pollution. Comparison of demersal fish hauls over time highlights fisheries effects not only on commercially important species but also on the entire demersal community. The effects of acute ("Torrey Canyon" oil spill) and chronic (tributyltin [TBT] antifoulants) pollution are clearly seen in the intertidal records. Significant advances in diverse scientific disciplines have been generated from research undertaken alongside the long-term data series.