Niveles de burnout y de exposición a conflicto ético de los profesionales de enfermería de cuidados intensivos / Levels of burnout and exposure to ethical conflict and assessment of the practice environment in nursing professionals of intensive care

Introducción: Los profesionales de enfermería que trabajan en unidades de cuidados intensivos (UCI) poseen un alto riesgo de desarrollar respuestas emocionales negativas, así como problemas emocionales y espirituales relacionados con cuestiones éticas. El diseño de estrategias efectivas que mejoren estos aspectos viene determinado por el conocimiento de los niveles de burnout y conflicto ético de dichos profesionales, así como la influencia que el entorno de la práctica puede tener en ellos. Objetivo: Analizar la relación existente entre niveles de burnout, exposición a conflicto ético y la percepción del ambiente de la práctica entre sí y con las variables sociodemográficas de los diferentes profesionales de enfermería de cuidados intensivos. Metodología: Estudio transversal correlacional en una UCI de un hospital universitario de nivel terciario. Se evaluó el nivel de burnout con la escala Maslach Burnout Inventory-Human-Services Survey; el nivel de conflicto ético, con el cuestionario de conflictividad ética para enfermeros, y la percepción del entorno, con la escala Practice-Environment-Scale of the Nursing-Work-Index. La asociación entre variables categóricas ha sido analizada mediante el test exacto de Fisher de chi-cuadrado (χ2). Resultados: Se evaluaron 39 enfermeras y 8 auxiliares, obteniendo una tasa de participación del 82,93%. El 31,10% de los profesionales de enfermería presentaron signos de burnout, el 14,89% consideraron que trabajan en un entorno desfavorable y el 87,23% presentaron un índice de exposición a conflicto ético medio-alto.El nivel educativo (χ2=11,084, p=0,011) y la categoría profesional (χ2=5,007, p=0,025) influyeron en el nivel de burnout, presentando las auxiliares mayores niveles del mismo...(AU)

Background: Nursing professionals working in intensive care units (ICU) are at high risk of developing negative emotional responses as well as emotional and spiritual problems related to ethical issues. The design of effective strategies that improve these aspects is determined by knowing the levels of burnout and ethical conflict of these professionals, as well as the influence that the practice environment might have on them. Objectives: To analyze the relationship between levels of burnout, the exposure to ethical conflicts and the perception of the practice environment among themselves and with sociodemographic variables of the different intensive care nursing professionals. Methods: Descriptive, correlational, cross-sectional, observational study in an ICU of a tertiary level university hospital. The level of burnout was evaluated with the Maslach Burnout Inventory Human Services Survey scale; the level of ethical conflict with the Ethical Conflict Questionnaire for Nurses; and the perception of the environment with the Practice Environment Scale of the Nursing Work Index. Descriptive and inferential statistics were performed. The association between categorical variables was analyzed using Fisher's exact chi-square test (χ2). Results: 31 nurses and 8 nursing assistants were evaluated, which meant a participation rate of 82.93%. 31.10% of the nursing professionals presented signs of burnout, 14.89% considered that they work in an unfavorable environment and 87.23% presented a medium-high index of exposure to ethical conflict. The educational level (χ2=11.084, P=.011) and the professional category (χ2=5.007, P=.025) influenced the level of burnout: nursing assistants presented higher levels of this. When comparing the level of burnout with the environment and the index of ethical conflict, there were no statistically significant differences...(AU)

Levels of burnout and exposure to ethical conflict and assessment of the practice environment in nursing professionals of intensive care.

BACKGROUND: Nursing professionals working in Intensive Care Units (ICU) are at high risk of developing negative emotional responses as well as emotional and spiritual problems related to ethical issues. The design of effective strategies that improve these aspects is determined by knowing the levels of burnout and ethical conflict of these professionals, as well as the influence that the practice environment might have on them. OBJECTIVES: To analyze the relationship between levels of burnout, the exposure to ethical conflicts and the perception of the practice environment among themselves and with sociodemographic variables of the different intensive care nursing professionals. METHODS: Descriptive, correlational, cross-sectional, observational study in an ICU of a tertiary level university hospital. The level of burnout was evaluated with the Maslach Burnout Inventory Human Services Survey scale; the level of ethical conflict with the Ethical Conflict Questionnaire for Nurses and the perception of the environment with the Practice Environment Scale of the Nursing Work Index. Descriptive and inferential statistics were performed. The association between categorical variables was analyzed using Fisher's exact chi-square test (χ2) RESULTS: 31 nurses and 8 nursing assistants were evaluated, which meant a participation rate of 82,93%. 31,10% of the nursing professionals presented signs of burnout, 14,89% considered that they work in an unfavorable environment and 87,23% presented a medium-high index of exposure to ethical conflict. The educational level (χ2=11.084, p=0.011) and the professional category (χ2=5.007, p=0.025) influenced the level of burnout: nursing assistants presented higher levels of this. When comparing the level of burnout with the environment and the index of ethical conflict, there were no statistically significant differences. CONCLUSIONS: The absence of association found in the study between Burnout and ethical conflict with the perception of the practice environment suggests that personal factors may influence its development.

Nurses' knowledge about palliative care in a critical care unit.

INTRODUCTION AND OBJECTIVE: Patients admitted to the Critical Care Unit (CCU) have a high mortality rate due to their complexity. Palliative care (PC) is a key aspect that can improve patient care. Because of the essential role of the nurse in providing this care, training, and including it in daily practice are needed. Our objective was to review the level of knowledge among the nurses in the CCU regarding PC and assess whether there is an association between each of the study variables. METHODOLOGY: We performed a descriptive observational cross-sectional study in the CCU of a tertiary level university hospital. The questionnaire Palliative Care Quiz for Nurses, previously validated and translated into Spanish, was used. This is a self-administered questionnaire consisting of 20 multiple-choice questions (True/False/Do not know-Do not answer) which evaluates three aspects of PC: philosophy, psychosocial and control of pain and other symptoms. In addition, sociodemographic data was collected. Descriptive and inferential statistics were used, a p < .05 was considered statistically significant in all cases. RESULTS: The questionnaire was administered to 68 nursers, with an average age of 34.98 ±â€¯12.12 years, and 13.00 ±â€¯11.75 years of professional experience. Twelve nurses have Master studies and 28 nurses have received training in PC. The percent of correct answers of the questionnaire was 56.98%. There were no statistically significant differences between the total average score and the variables studied. However, looking at each aspect on the scale, an association was found between PC training and control of pain and other symptoms (p = .033). CONCLUSION: Critical care nurses have a basic knowledge of PC, it being insufficient in the psychological sphere. Developing a training programme which identifies misconceptions and training deficits might improve the management of symptom control in palliative care patients, quality of care and its application.

Conocimientos de las enfermeras acerca de los cuidados paliativos en un área de críticos / Nurses’ knowledge about palliative care in a critical care unit

Introducción y objetivos: Los pacientes que ingresan en las áreas de críticos (AC) tienen alta tasa de morbi-mortalidad debido a su complejidad. Los cuidados paliativos (CP) se presentan como un componente clave y pueden ayudar a mejorar el cuidado del paciente. La enfermera es esencial para la administración de estos cuidados; para ello, es necesario tener formación y saber integrarlos en la práctica diaria. El objetivo fue examinar el nivel de conocimientos de las enfermeras del AC acerca de los CP, y evaluar si existe asociación entre cada una de las variables estudiadas. Metodología: Estudio observacional descriptivo transversal en el AC de un hospital universitario de nivel terciario. Se utilizó el cuestionario Palliative Care Quiz for Nurses, traducido y validado al español. Es un cuestionario auto-administrado que consta de 20 ítems de respuesta múltiple (verdadero/falso/no sabe-no contesta) que evalúa tres aspectos de los CP: filosofía, psicosocial, y control del dolor y otros síntomas. Además, se recogieron datos sociodemográficos. Se realizó estadística descriptiva e inferencial, considerándose estadísticamente significativo un valor de p<0,05 en todos los casos. Resultados: El cuestionario se administró a 68 enfermeras, con una edad media de 34,98±12,12 años, y 13,00±11,75 años de experiencia profesional. Doce enfermeras tenían formación de Máster y 28 enfermeras habían recibido formación en CP. El porcentaje de aciertos del cuestionario fue de 56,98%. No hubo diferencias estadísticamente significativas entre la puntuación media total y las variables estudiadas. Al analizar los aspectos de la escala, se encontró asociación entre la formación en paliativos y el control de síntomas (p=0,033). Conclusión: Las enfermeras del AC tienen un conocimiento básico sobre los CP, siendo el aspecto psicosocial del mismo insuficiente.(AU)

Introduction and objetive: Patients admitted to the Critical Care Unit (CCU) have a high mortality rate due to their complexity. Palliative care (PC) is a key aspect that can improve patient care. Because of the essential role of the nurse in providing this care, training, and including it in daily practice are needed. Our objective was to review the level of knowledge among the nurses in the CCU regarding PC and assess whether there is an association between each of the study variables. Methodology: We performed a descriptive observational cross-sectional study in the CCU of a tertiary level university hospital. The questionnaire Palliative Care Quiz for Nurses, previously validated and translated into Spanish, was used. This is a self-administered questionnaire consisting of 20 multiple-choice questions (True/False/Do not Know-Do not answer) which evaluates three aspects of PC: philosophy, psychosocial and control of pain and other symptoms. In addition, sociodemographic data was collected. Descriptive and inferential statistics were used, a p<.05 was considered statistically significant in all cases. Results: The questionnaire was administered to 68 nursers, with an average age of 34.98±12.12 years, and 13.00±11.75 years of professional experience. Twelve nurses have Master studies and 28 nurses have received training in PC. The percent of correct answers of the questionnaire was 56.98%. There were no statistically significant differences between the total average score and the variables studied. However, looking at each aspect on the scale, an association was found between PC training and control of pain and other symptoms (p=.033). Conclusion: Critical care nurses have a basic knowledge of PC, it being insufficient in the psychological sphere. Developing a training programme which identifies misconceptions and training deficits might improve the management of symptom control in palliative care patients, quality of care and its application.(AU)

Impact of biotin supplemented diet on mouse pancreatic islet ß-cell mass expansion and glucose induced electrical activity.

Biotin supplemented diet (BSD) is known to enhance ß-cell replication and insulin secretion in mice. Here, we first describe BSD impact on the islet ß-cell membrane potential (Vm) and glucose-induced electrical activity. BALB/c female mice (n ≥ 20) were fed for nine weeks after weaning with a control diet (CD) or a BSD (100X). In both groups, islet area was compared in pancreatic sections incubated with anti-insulin and anti-glucagon antibodies; Vm was recorded in micro dissected islet ß-cells during perfusion with saline solutions containing 2.8, 5.0, 7.5-, or 11.0 mM glucose. BSD increased the islet and ß-cell area compared with CD. In islet ß-cells of the BSD group, a larger ΔVm/Δ[glucose] was found at sub-stimulatory glucose concentrations and the threshold glucose concentration for generation of action potentials (APs) was increased by 1.23 mM. Moreover, at 11.0 mM glucose, a significant decrease was found in AP amplitude, frequency, ascending and descending slopes as well as in the calculated net charge influx and efflux of islet ß-cells from BSD compared to the CD group, without changes in slow Vm oscillation parameters. A pharmacological dose of biotin in mice increases islet insulin cell mass, shifts islet ß-cell intracellular electrical activity dose response curve toward higher glucose concentrations, very likely by increasing KATP conductance, and decreases voltage gated Ca2+ and K+ conductance at stimulatory glucose concentrations.

Realización y comunicación de resultados de test de autodiagnóstico de antígenos SARS- CoV-2 bajo supervisión en farmacia de Cantabria / Performance and communication of results of self-diagnostic test for SARS-CoV-2 antigens under supervision in pharmacies in Cantabria

JUSTIFICACIÓN: la Consejería de Sanidad del Gobierno de Cantabria y el Colegio Oficial de Farmacéuticos de Cantabria colaboran en estrategias para asegurar la trazabilidad de resultados positivos de test de autodiagnóstico de antígenos SARS-COV-2 a través de las farmacias comunitarias. En enero de 2022, ambas entidades han iniciado un procedimiento mediante el cual la realización del test bajo supervisión por el profesional farmacéutico en farmacia es considerado para el diagnóstico de confirmación de infección activa, y los resultados positivos del test se consideran casos COVID-19 confirmados.OBJETIVOS: •Realizar in situ en la farmacia, o acompañar a la población que lo solicite, test de autodiagnóstico de antígenos SARS-COV-2 •Comunicar el resultado obtenido a la Consejería de Sanidad mediante la Aplicación móvil Plataforma CAÑIA. •Informar a los usuarios sobre la actuación a seguir según el resultado obtenido.MATERIAL Y MÉTODOS: la población susceptible de solicitar el test es toda persona que en riesgo de haber adquirido infección por SARS-COV-2 por presentar síntomas compatibles de COVID-19 o haber tenido contacto con un positivo. Las farmacias que participan voluntariamente se identifican con un cartel informativo, y comunican los resultados al Servicio Cántabro de Salud (SCS) mediante la aplicación móvil Cañía, de uso exclusivo para profesionales sanitarios, que mediante tecnología de inteligencia artificial realiza la lectura del resultado del test y de la identidad de la persona y la carga automáticamente en el sistema del SCS. Si el resultado es positivo, el paciente entra directamente en el circuito de Atención Primaria y de Farmacovigilancia del SCS. Si el resultado es negativo, el paciente puede descargar durante 48 horas un certificado a través del SCS. (AU)

Comunicación de incidencias con la medicación entre farmacias de Cantabria y Servicios de Farmacia de Atención Primaria del Servicio Cántabro de Salud / Communication of medication incidents between pharmacies in Cantabria and the Primary Care Pharmacy Services of the Cantabrian Health Service

INTRODUCCIÓN: el Colegio Oficial de Farmacéuticos de Cantabria (COF Cantabria) ha colaborado con la Consejería de Sanidad del Gobierno de Cantabria para crear un sistema de comunicación directo entre las 279 farmacias comunitarias de Cantabria y los Servicios de Farmacia de Atención Primaria del Servicio Cántabro de Salud (SFAP). El protocolo de comunicación, que comenzó en octubre de 2020 y en marzo de 2022 continúa en activo, constituye una herramienta que las OF han integrado en su práctica diaria para informar y solucionar incidencias relacionadas con la medicación prescrita en receta electrónica.OBJETIVOS: desarrollar e implantar un sistema de comunicación entre las farmacias comunitarias y los SFAP, para resolver incidencias relacionadas con la receta electrónica de los pacientes, asegurando la adherencia al tratamiento farmacológico y contribuyendo a descongestionar la Atención Primaria.MATERIAL Y MÉTODOS: se creó un grupo de trabajo para desarrollar un protocolo de comunicación de incidencias con la medicación, con participantes del COF Cantabria, de la Consejería de Sanidad y de la Gerencia de Atención Primaria del Servicio Cántabro de Salud.RESULTADOS Y DISCUSIÓN: los resultados del seguimiento del protocolo se presentaron a los farmacéuticos/as en febrero de 2021 mediante una conferencia online, durante la cual se debatieron las incidencias más frecuentes y las propuestas para mejorar la comunicación de las mismas. El grado de satisfacción de los farmacéuticos/as con el funcionamiento del protocolo se evaluó mediante una encuesta online de seis preguntas. En las respuestas se observó que un 35,4 % de los farmacéuticos se muestra satisfecho con el funcionamiento y un 29,2 % valora positivamente el tiempo de resolución de incidencias por parte de SFAP. En cuanto a la utilidad para resolver la incidencia respecto al paciente, un 25 % se muestra bastante satisfecho. (AU)

Dispensación colaborativa entre los servicios de farmacia hospitalaria y farmacias comunitarias en el contexto de la COVID-19 / Collaborative dispensing between hospital pharmacy services and community pharmacies in the context of COVID-19

JUSTIFICACIÓN: de marzo a mayo de 2020, se desarrolló una experiencia piloto de continuidad asistencial en la atención farmacéutica de pacientes externos hospitalarios, en la que la medicación hospitalaria se entregaba en la farmacia comunitaria de elección del paciente. En septiembre de 2020 se consolidó dicho procedimiento, según el protocolo “Entrega de Medicación Hospitalaria de Dispensación Ambulatoria -MHDA- por la farmacia comunitaria en el contexto de la COVID-19”, elaborado por la Consejería de Sanidad del Gobierno de Cantabria, el Colegio de Farmacéuticos de Cantabria y la distribución farmacéutica que opera en Cantabria.OBJETIVOS: establecer un protocolo para limitar la presencia en centros hospitalarios de pacientes que recogen sus medicamentos hospitalarios (MH) en los Servicios de Farmacia Hospitalaria (SFH), y sustituirlo por el envío, a través del Distribuidor Farmacéutico (DF), de los MH a Farmacias de Cantabria (FC) que harán la entrega a los pacientes.MATERIAL Y MÉTODOS: las FC reciben, a través del DF, la medicación de cada paciente desde los SFH. Los pacientes seleccionados son previamente contactados e informados por el SFH correspondiente, al que indican la farmacia a la que quieren que se envíe su medicación.RESULTADOS Y DISCUSIÓN: se analizan los datos de febrero a diciembre de 2021. El número total de pacientes a los que se ha entregado MH, a través de las FC es 6832. (El 80 % reside en la provincia y el 20 % en Santander capital).El número de FC que han entregado MH es 222. (El 72 % en la provincia y el 28 % en Santander capital).El número total de entregas de MH realizadas por los SFH ha sido 5644, correspondiendo 5066 al Hospital Universitario Marqués de Valdecilla, 565 al Hospital Sierrallana y 13 al Hospital de Laredo. En cuanto a la DF, Centro Farmacéutico del Norte (CENFARTE) ha realizado 2.447 repartos, Cooperativa Farmacéutica Española (COFARES) 1809 y Cooperativa Farmacéutica Asturiana (COFAS) 1213. (AU)

Connexin 30.2 is expressed in exocrine vascular endothelial and ductal epithelial cells throughout pancreatic postnatal development.

Previously we have demonstrated that the GJ protein connexin 30.2 (Cx30.2) is expressed in pancreatic beta cells and endothelial cells (ECs) of the islet. In the present study, we address whether Cx30.2 is expressed in the exocrine pancreas, including its vascular system. For this, adult mouse pancreatic sections were double labeled with specific antibodies against Cx30.2 and CD31, an endothelial cell marker, or with anti-α-actin smooth muscle, a smooth muscle cell (SMC) marker or anti-mucin-1, a marker of epithelial ductal cells, using immunofluorescence (IF) studies. Cx30.2-IF hot spots were found at junctional membranes of exocrine ECs and SMCs of blood vessels. Furthermore, Cx30.2 was localized in mucin-1 positive cells or epithelial ductal cells. Using immunohistochemistry (IHC) studies, it was found that in vessels and ducts of different diameters, Cx30.2 was also expressed in these cell types. In addition, it was found that Cx30.2 is already expressed in these cell types in pancreatic sections of 3, 14 and 21 days postpartum. Moreover, this cell specific pattern of expression was also found in the adult rat, hamster and guinea pig pancreas. Expression of Cx30.2 mRNA and protein in the pancreas of all these species was confirmed by RT-PCR and Western blot studies. Overall, our results suggest that intercellular coupling mediated by Cx30.2 intercellular channels may synchronize the functional activity of ECs and SMCs of vascular cells, as well as of epithelial ductal cells after birth.

Neurons and satellite glial cells in adult rat lumbar dorsal root ganglia express connexin 36.

Previous studies have shown that following peripheral nerve injury there was a downregulation of the gap junction protein connexin 36 (Cx36) in the spinal cord; however, it is not known whether Cx36 protein is expressed in the dorsal root ganglia (DRGs), nor if its levels are altered following peripheral nerve injuries. Here we address these aspects in the adult rat lumbar DRG. Cx36 mRNA was detected using qRT-PCR, and Cx36 protein was identified in DRG sections using immunohistochemistry (IHC) and immunofluorescence (IF). Double staining revealed that Cx36 co-localizes with both anti-ß-III tubulin, a neuronal marker, and anti-glutamine synthetase, a satellite glial cell (SGC) marker. In neurons, Cx36 staining was mostly uniform in somata and fibers of all sizes and its intensity increased at the cell membranes. This labeling pattern was in contrast with Cx36 IF dots mainly found at junctional membranes in islet beta cells used as a control tissue. Co-staining with anti-Cx43 and anti-Cx36 showed that whereas mostly uniform staining of Cx36 was found throughout neurons and SGCs, Cx43 IF puncta were localized to SGCs. Cx36 mRNA was expressed in normal lumbar DRG, and it was significantly down-regulated in L4 DRG of rats that underwent sciatic nerve injury resulting in persistent hypersensitivity. Collectively, these findings demonstrated that neurons and SGCs express Cx36 protein in normal DRG, and suggested that perturbation of Cx36 levels may contribute to chronic neuropathic pain resulting from a peripheral nerve injury.

Linajes madurativos de las células que regeneran al hígado / Maturative lineages of cells that regenerate the liver

Resumen Diferentes linajes de las células hepáticas participan en la regeneración del hígado y los hepatocitos guiescentes pueden responder ante un daño hepático leve con división celular moderada, mientras gue las células troncales y progenitoras responderán para una amplia restitución del parénquima hepático severamente dañado. Ocho linajes madurativos se sitúan dentro del hígado con diferente grado de madurez. Cuatro de ocho subpoblaciones gue presentan marcadores para células troncales o progenitoras se ubican en nichos intrahepáticos como los canales de Hering y la vena porta, estructuras en las gue pueden continuar su diferenciación si se recibe algún estímulo. Recientemente se describió un nuevo nicho entrahepático con una composición celular heterogénea ubicado entre los grandes duetos biliares y en el páncreas. Las células gue se encuentran en estos compartimentos expresan marcadores tempranos de células troncales y se consideran como precursores de las células troncales hepáticas.

Abstract Different cellular lineages participate in liver regeneration. Quiescent hepatocytes may respond to a mild liver injury with moderate cell division. Meanwhile, stem and progenitor cells are responsible for the large restitution of a severe damaged organ. Eight matu rational lineages are in the liver, each one with a different maturity grade. Four subpopulations out of eight express stem or progenitor cell markers are found within intrahepatic niches such as the canals of Hering and the portal vein. Each structure is able to continue its differentiation if a stimuli is received. Recently, a new extra-hepatic stem niche with a heterogeneous cellular composition was found between the large biliary ducts and the pancreas. Stem or progenitor cells contained in these compartments express early stem markers and are considered as a precursor of hepatic stem cells.

Modulation of spontaneous intracellular Ca²âº fluctuations and spontaneous cholinergic transmission in rat chromaffin cells in situ by endogenous GABA acting on GABAA receptors.

Using fluorescence [Ca(2+)]i imaging in rat adrenal slices, we characterized the effects of agonists and antagonists of the GABAA receptor (GABAA-R) on resting intracellular Ca(2+) ([Ca(2+)]i) and spontaneous [Ca(2+)]i fluctuations (SCFs) in hundreds of individual chromaffin cells (CCs) recorded simultaneously in situ. Muscimol, a GABAA-R agonist (20 µM; 25 s), induced an increase of resting [Ca(2+)]i in 43 ± 3 % of CCs, a decrease in 26 ± 2 %, and no response in 30 ± 5 %. In Ca(2+)-free external medium, SCFs ceased completely and muscimol failed to elicit [Ca(2+)]i rises. All muscimol-induced [Ca(2+)]i changes were blocked by the GABAA-R antagonist bicuculline, suggesting that they result from changes in membrane potential depending on the cell's Cl(-) equilibrium potential. Unexpectedly, bicuculline increased the amplitude and frequency of SCFs in 54 % of CCs, revealing a tonic inhibition of SCFs by ambient GABA acting through GABAA-R. Mecamylamine (a specific nicotinic cholinergic blocker) decreased basal SCF activity in 18 % of CCs and inhibited bicuculline-induced SCF intensification, suggesting that spontaneous acetylcholine (ACh) release from nerve endings contributes to SCF generation in CCs in situ and that blockade of presynaptic GABAA-Rs intensifies SCFs in part through the disinhibition of spontaneous cholinergic transmission. Electrophysiological experiments confirmed that spontaneous excitatory postsynaptic currents recorded from CCs in situ were enhanced by bicuculline. To our knowledge, this is the first description of a regulatory effect of endogenous GABA on synaptic currents and SCFs of adrenal CCs. These findings denote a novel GABA-mediated presynaptic and postsynaptic regulatory mechanism of CC activity which may participate in the control of catecholamine secretion.

CDKN3 mRNA as a Biomarker for Survival and Therapeutic Target in Cervical Cancer.

The cyclin-dependent kinase inhibitor 3 (CDKN3) gene, involved in mitosis, is upregulated in cervical cancer (CC). We investigated CDKN3 mRNA as a survival biomarker and potential therapeutic target for CC. CDKN3 mRNA was measured in 134 CC and 25 controls by quantitative PCR. A 5-year survival study was conducted in 121 of these CC patients. Furthermore, CDKN3-specific siRNAs were used to investigate whether CDKN3 is involved in proliferation, migration, and invasion in CC-derived cell lines (SiHa, CaSki, HeLa). CDKN3 mRNA was on average 6.4-fold higher in tumors than in controls (p = 8 x 10-6, Mann-Whitney). A total of 68.2% of CC patients over expressing CDKN3 gene (fold change ≥ 17) died within two years of diagnosis, independent of the clinical stage and HPV type (Hazard Ratio = 5.0, 95% CI: 2.5-10, p = 3.3 x 10-6, Cox proportional-hazards regression). In contrast, only 19.2% of the patients with lower CDKN3 expression died in the same period. In vitro inactivation of CDKN3 decreased cell proliferation on average 67%, although it had no effect on cell migration and invasion. CDKN3 mRNA may be a good survival biomarker and potential therapeutic target in CC.

Connexin 36, a key element in pancreatic beta cell function.

The prevalence of diabetes at a global scale has markedly increased during the last three decades. Diabetes is a chronic disease that includes a group of metabolic disorders, in which high serum glucose levels is a common factor. Insulin is the only hormone that decreases serum glucose levels. Therefore, it is relevant to deepen our understanding of cell mechanisms that regulate insulin production and release. Insulin is produced in pancreatic islet beta cells. They are excitable cells and most of them are electrically coupled through gap junction channels. Connexin 36 (Cx36) has been identified at junctional membranes of islet beta cells in both rodents and humans. Co-localization of Cx36 with Cx30.2 has been recently identified. Functional studies in Cx36 deficient mice have provided direct evidence that Cx36 gap junction channels are necessary for the synchronization of [Ca(2+)]i oscillations in islet beta cells. The latter allows for the generation of insulin pulses in a single perfused islet. Moreover, Cx36 deficient mice were found to have altered serum insulin pulse dynamics and to be glucose intolerant. In addition, Cx36 has been recently identified as an early gene that is specifically expressed in embryonic beta cells, whose transcript and protein are upregulated in unison with the main wave of beta cell differentiation. In conclusion, Cx36 is critical for endocrine pancreatic function and may represent a molecular target for future prevention and treatment of diabetes. This article is part of the Special Issue Section entitled 'Current Pharmacology of Gap Junction Channels and Hemichannels'.

Connexin 30.2 is expressed in mouse pancreatic beta cells.

Nowadays, connexin (Cx) 36 is considered the sole gap junction protein expressed in pancreatic beta cells. In the present research we investigated the expression of Cx30.2 mRNA and protein in mouse pancreatic islets. Cx30.2 mRNA and protein were identified in isolated islet preparations by qRT-PCR and Western blot, respectively. Immunohistochemical analysis showed that insulin-positive cells were stained for Cx30.2. Confocal images from double-labeled pancreatic sections revealed that Cx30.2 and Cx36 fluorescence co-localize at junctional membranes in islets from most pancreases. Abundant Cx30.2 tiny reactive spots were also found in cell cytoplasms. In beta cells cultured with stimulatory glucose concentrations, Cx30.2 was localized in both cytoplasms and cell membranes. In addition, Cx30.2 reactivity was localized at junctional membranes of endothelial or cluster of differentiation 31 (CD31) positive cells. Moreover, a significant reduction of Cx30.2 mRNA was found in islets preparations incubated for 24h in 22mM as compared with 3.3mM glucose. Therefore, it is concluded that Cx30.2 is expressed in beta and vascular endothelial cells of mouse pancreatic islets.

High glucose concentrations induce TNF-α production through the down-regulation of CD33 in primary human monocytes.

BACKGROUND: CD33 is a membrane receptor containing a lectin domain and a cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM) that is able to inhibit cytokine production. CD33 is expressed by monocytes, and reduced expression of CD33 correlates with augmented production of inflammatory cytokines, such as IL-1ß, TNF-α, and IL-8. However, the role of CD33 in the inflammation associated with hyperglycemia and diabetes is unknown. Therefore, we studied CD33 expression and inflammatory cytokine secretion in freshly isolated monocytes from patients with type 2 diabetes. To evaluate the effects of hyperglycemia, monocytes from healthy donors were cultured with different glucose concentrations (15-50 mmol/l D-glucose), and CD33 expression and inflammatory cytokine production were assessed. The expression of suppressor of cytokine signaling protein-3 (SOCS-3) and the generation of reactive oxygen species (ROS) were also evaluated to address the cellular mechanisms involved in the down-regulation of CD33. RESULTS: CD33 expression was significantly decreased in monocytes from patients with type 2 diabetes, and higher levels of TNF-α, IL-8 and IL-12p70 were detected in the plasma of patients compared to healthy donors. Under high glucose conditions, CD33 protein and mRNA expression was significantly decreased, whereas spontaneous TNF-α secretion and SOCS-3 mRNA expression were increased in monocytes from healthy donors. Furthermore, the down-regulation of CD33 and increase in TNF-α production were prevented when monocytes were treated with the antioxidant α-tocopherol and cultured under high glucose conditions. CONCLUSION: Our results suggest that hyperglycemia down-regulates CD33 expression and triggers the spontaneous secretion of TNF-α by peripheral monocytes. This phenomenon involves the generation of ROS and the up-regulation of SOCS-3. These observations support the importance of blood glucose control for maintaining innate immune function and suggest the participation of CD33 in the inflammatory profile associated with type 2 diabetes.

Biophysical evidence that connexin-36 forms functional gap junction channels between pancreatic mouse beta-cells.

Connexin-36 (Cx36) is the only gap junction protein that has been unambiguously identified in rodent pancreatic beta-cells. However, properties of gap junction channel unitary currents between beta-cells remain unrevealed. To address whether Cx36 forms functional channels in beta-cells, we characterized biophysical properties of macro- and microscopic junctional currents recorded from dual whole cell voltage clamp isolated pairs of dispersed mouse beta-cells. Electrical coupling was recorded in 80% of cell pairs with a junctional conductance (g(j)) of 355 +/- 45 pS (n = 20). Transjunctional voltage dependence was identified in three of seven cell pairs with high-input membrane resistances. Normalized steady-state g(j) (Gj) and transjunctional-voltage relation were well described by a two-state Boltzmann equation [maximal conductance (Gmax) = 1.0, voltage-insensitive conductance (Gmin) = 0.3 and 0.28, voltage gating sensitivity (A) = 0.21 and 0.23, and voltage at which one-half of the initial voltage-dependent conductance was reached (Vo) = -85 and 87 mV for negative and positive potentials, respectively]. Halothane reversibly uncoupled beta-cell pairs, and, during recovery, unitary conductances of 5-10 pS were recorded while using patch pipettes containing mainly CsCl. Although these properties are similar to those previously described for Cx36 channels in mammalian cell systems, we found that beta-cell junctional currents were insensitive to quinine. Cx36 transcript and protein expression in islets and freshly dispersed cell preparations was confirmed by RT-PCR and immunofluorescence. In conclusion, biophysical properties of junctional channels between beta-cells are similar but not identical to those previously described for homomeric Cx36 channels. Cell type-specific mechanisms that may account for these differences are discussed.

Connexin43 is expressed in mouse fetal ovary.

Developmental studies have shown that connexin43 (Cx43) is expressed in the ovary from the first day of life and throughout the rest of postnatal development. In both mouse embryonic ovaries and testes, target-directed deletion of Cx43 gene induces a significant decrease in germinal cells, but the exact mechanism determining this reduction remains unknown. Moreover, recently we found that Cx43 is abundantly expressed in mouse testes from the earliest stages of its fetal development. In the present work we investigate whether Cx43 transcript and protein are expressed in mouse embryonic ovaries. Total RNA was analyzed with specific Cx43 oligonucleotides in RT-PCR studies. A Cx43 PCR product was detected in ovaries at 16.5 and 18.5 days postcoitum (dpc). Bands of 43-45 kDa, characteristic of Cx43, were detected in immunoblots of total homogenates of ovaries at 14.5 and 18.5 dpc. Cell type-specific expression of Cx43 was investigated using double-labeled sections incubated with specific antibodies against Cx43 and the enzyme 3beta-hydroxysteroid dehydrogenase (3betaHSD) or a germ cell nuclear antigen (GCNA1), which are cell markers of steroidogenic and germinal cells, respectively. At 18.5 dpc, Cx43 was found in conglomerates of 3betaHSD-positive cells. Cx43 was also localized at homocellular junctions between parenchyma pregranulosa cells, and at heterocellular junctions between pregranulosa and germinal cells. At these two latter localizations, Cx43 was traced back to 12.5 dpc. In conclusion, this study demonstrates for the first time that from the earliest stages of embryonic ovary development, Cx43 is expressed in principal cell types involved in control of female fertility. These data suggest that the gap junctions formed with Cx43 between somatic and germinal cells may be necessary for prenatal expansion of germinal cells at initial stages of fetal gonadal development.

Developmental regulation of connexin 43 expression in fetal mouse testicular cells.

Multiple connexins have been identified in testicular cells. Several lines of evidences indicate that, among them, connexin 43 (Cx43) may be unique for control of gonad development and spermatogenesis. To date, however, it is not known whether Cx43 is expressed in the fetal testis and what possible types of cellular interactions mediated by this connexin are critical to male fertility. In the present work, expression of Cx43 was investigated at various developmental ages in cryosections from mouse testis by using specific antibodies against Cx43. In serial or double-labeled sections, Cx43 localization was compared with immunocytochemical distribution of steroidogenic enzyme, 3beta-hydroxysteroid dehydrogenase (3betaHSD), Mullerian inhibitory hormone (MIH), and germinal nuclear cell antigen (GCNA1), which are specific markers, respectively, of interstitial Leydig, Sertoli, and germinal cells. Sections were analyzed by fluorescence microscopy. We found that Cx43 immunofluorescence (IF) was uniformly distributed in the undifferentiated gonad at 11.5 days post coitus (dpc) and in cells of the mesonephric tubules. In the undifferentiated gonad, Cx43 was localized between primordial germ cells and somatic cells. At 12.5 dpc, when the gonad has undergone sexual differentiation, in the interstitium Cx43 was localized in Leydig cells and in the seminiferous cord it was localized between adjacent Sertoli cells. In Leydig and Sertoli cells, Cx43 labeling increased at 14.5, 16.5, and 18.5 dpc. From day 12.5 up to 18.5 dpc, Cx43 was also localized in cell borders between germinal and Sertoli cells. In conclusion, this study demonstrates that from the earliest stages of gonadal development, Cx43 is expressed in the principal cell types that participate in the control of male fertility. It also shows that Cx43 expression in Leydig and Sertoli cells increase during fetal life. Finally, it provides evidence that, throughout embryonic life, Cx43 forms gap junctions between Sertoli and germinal cells.

Expression of connexin43 in mouse Leydig, Sertoli, and germinal cells at different stages of postnatal development.

Connexin 43 (Cx43) is the most abundant and ubiquitously distributed gap junction protein in testicular cells. Lack of Cx43 expression results in male infertility. We investigated whether Cx43 is expressed and regulated in Leydig, Sertoli and germinal cells at different stages of postnatal development. Cx43 was detected using three different antibodies shown by immunoblotting to be highly specific. At different postnatal ages Cx43 localization was compared in serial or double labeled testicular cryosections with immunocytochemical distribution of steroidogenic enzyme, 3 betahydroxysteroid-dehydrogenase (3betaHSD), Mullerian inhibitory hormone (MIH), and germinal nuclear cell antigen (GNCA1), which are specific markers of interstitial Leydig, Sertoli and germinal cells, respectively. In the interstitium, round cell clumps (RCC) with lipid droplets positive for 3betaHSD and Cx43 were frequently found at intertubular areas at birth and Cx43 was mainly localized at cell membrane appositions. From day 3, the number and size of 3betaHSD-positive RCC started to decrease, and reached a minimum at 7-14 dpp; Cx43 expressed by them is progressively downregulated. From day 21 an increase in the size and number of RCC positive for Cx43 and 3betaHSD was found that continued at 24, 26 and 28 days and reached a maximum at 35 and 60 dpp. Biphasic expression of interstitial Cx43 and 3betaHSD was also found to be positively and temporally correlated with fluctuations in intratesticular testosterone content at all ages studied. In the seminiferous cord (SC), Cx43 was expressed at birth between adjacent Sertoli cells (MIH positive) localized at the periphery, as well as in their cytoplasm projections that surround centrally localized gonocytes. From days 3 to 7, Cx43 labeling increased in Sertoli cells mainly at their apical border. At day 14, Cx43 distribution in Sertoli cells changed from apical to basal in parallel to migration of germinal (GNCA1-positive) cells from the periphery to the center of the SC. At all these ages, Cx43 was also localized at cell borders between Sertoli and germinal cells. In conclusion, this study demonstrates that Cx43 in Leydig cells is regulated during postnatal development in an age and functional dependent manner. In the tubule, it is demonstrated that Cx43 is modulated in Sertoli cells during the neonatal and prepubertal period. We also provide evidence for the first time that Cx43-gap junctions communicate between Sertoli and germinal cells before and during the first wave of spermatogenesis.