Burnout y factores predictores en profesionales de Enfermería chilenos a un año de pandemia por COVID-19 / Burnout and predicting factors in Chilean nursing professional after one year of COVID-19 pandemic

Objetivo: analizar la asociación de variables sociodemográficas y laborales, estado de salud, inteligencia emocional, apoyo social percibido y espiritualidad en el desarrollo del burnout en profesionales de Enfermería a un año del comienzo de la pandemia COVID-19 en Chile.Método: se llevó a cabo un estudio descriptivo transversal (julio-octubre 2021). La población de estudio fueron enfermeras, que hubieran trabajado durante la pandemia en cualquier centro de atención sanitaria en Chile, atendiendo a pacientes al menos durante tres meses. Se aplicó un cuestionario online que incluía perfil enfermera, Maslach Burnout Inventory, Cuestionario Nórdico Estandarizado, Trait-Meta Mood Scale-24, Escala Multidimensional de Apoyo Social Percibido e Índice Breve de Religiosidad y Espiritualidad. Se llevó a cabo análisis descriptivo, correlaciones y regresión lineal.Resultados: participaron 192 profesionales, 181 (94,3%) eran mujeres. La edad media (DE) fue de 30,8 (6,81) años. La antigüedad laboral media (DE) fue de 5,6 (5,98) años. El 64,1% de las enfermeras presentó burnout. Se evidenció relación inversa y moderada entre las dimensiones de burnout e inteligencia emocional (directas, para el caso de realización personal). Se observó relación directa y moderada entre la realización personal y el apoyo social de amigos. También hubo relación indirecta y cercana a moderada entre cansancio emocional y espiritualidad. En los modelos predictivos, el dolor físico y la inteligencia emocional se asociaron con burnout.Conclusiones: más de la mitad de los profesionales de Enfermería de la muestra presentó burnout, siendo sus principales predictores el dolor físico y la inteligencia emocional.(AU)

Objective: to analyse the impact of sociodemographic and occupational variables, health status, emotional intelligence, perceived social support and spirituality, upon the development of burnout in Nursing professionals at one year after the start of the COVID-19 pandemic in Chile.Methods: a descriptive cross-sectional study was conducted (July to October 2021). The study population were nurses who had worked during the pandemic at any healthcare centre in Chile, seeing patients during at least three months. An online questionnaire was applied, including the nurse profile, the Maslach Burnout Inventory, the Standardized Nordic questionnaire, the Trait-Meta Mood Scale-24, the Multidimensional Scale of Perceived Social Support, and the Brief Spirituality/Religiousness Index. Descriptive analysis, correlations and linear regression were conducted.Results: the study included 192 professionals, 181 (94,3%) were female. Their mean age (SD) was 30.8 (6.81) years. Their mean seniority (SD) was 5.6 (5.98) years. Of these nurses, 64.1% presented burnout. A reverse and moderate relationship was observed between the burnout and emotional intelligence dimensions (direct in the case of personal fulfilment). A direct and moderate relationship was observed between personal fulfilment and social support by friends. There was also an indirect and close to moderate relationship between emotional exhaustion and spirituality. In the predictive models, physical pain and emotional intelligence were associated with burnout.Conclusions: more than half of Nursing professionals presented burnout, and its main predictors were physical pain and emotional intelligence.(AU)

Spef1/CLAMP binds microtubules and actin-based structures and regulates cell migration and epithelia cell polarity.

During migration, cells invade, repair, and create barriers leading to the formation of new cellular contacts in target tissues. Cell migration requires many proteins that collectively form the cytoskeleton. The main cytoskeletal elements are actin filaments, microtubules (MTs), and intermediate filaments. These structures work in concert with a large number of accessory proteins that contribute in a variety of ways to regulate filament assembly and turnover, to alter the configuration or arrangement of filaments by bundling or crosslinking, to link the cytoskeleton to other structures in the cell, such as membranes and junctions, and to transport cargo along the filaments. Sperm flagella protein-1 (Spef1), also designated calponin homology and microtubules-associated protein (CLAMP), is a multifunctional protein that interacts with cytoskeletal structures, including MTs, actin filaments, and focal adhesions in epithelia. In this review, we outline Spef1/CLAMP structure and expression in several cellular models. The function of Spef1/CLAMP in flagellar and ciliary motility, MT-binding and stability, regulation of planar cell polarity, and potential contribution to the maintenance of actin-based structures, such as lamellipodia and filopodia during cell migration, are also discussed.

Experience in a rehabilitation center for mastectomized women at the start of the COVID-19 pandemic.

OBJECTIVE: Describe the experience lived in an interdisciplinary follow-up care center for mastectomized women at a public university in São Paulo during the beginning of the COVID-19 pandemic. METHOD: Experience report on the health care provided in the health center for mastectomized women. RESULTS: The care was provided three times a week by an interdisciplinary health team. The mentioned areas that cover the women care in the center: Physical, Psychological, Social Support and Health Education. CONCLUSIONS: The attention by an interdisciplinary team becomes prevalent in the care of mastectomized women, since cancer and its treatment produce various changes in women's lives in the short and long term, so follow-up and support must be biopsychosocial, covering all areas that may be affected, especially during the pandemic.

Experience in a rehabilitation center for mastectomized women at the start of the COVID-19 pandemic / Experiencia en centro de rehabilitación de mujeres mastectomizadas al inicio de la pandemia por COVID-19 / Experiência em centro de reabilitação de mulheres mastectomizadas no início da pandemia do COVID-19

ABSTRACTObjective: Describe the experience lived in an interdisciplinary follow-up care center for mastectomized women in a public university in Sao Paulo during the onset of the COVID-19 pandemic.Method: Experience report on the health care provided in the health center for mastectomized women.Results: The health care was provided three times a week by an interdisciplinary health team. The areas of care for women at the centre were physical, psychological, social support; health education and support during the pandemic.Conclusions: The attention by an interdisciplinary team becomes prevalent in the care of mastectomized women, since cancer and its treatment produce various changes in women's lives in the short and long term, so follow-up and support must be biopsychosocial, covering all areas that may be affected, especially during the pandemic.Keywords: Women's health. Breast neoplasms. Mastectomy. Comprehensive health care. Coronavirus infections.

RESUMENObjetivo: Describir la experiencia vivida en un centro de atención interdisciplinaria de seguimiento a mujeres mastectomizadas de una universidad pública de Sao Paulo durante el inicio de la pandemia de COVID-19.Método: Relato de experiencia sobre las atenciones de salud brindadas en el centro de salud para mujeres mastectomizadas.Resultados: Las atenciones fueron realizadas tres veces por semana por un equipo interdisciplinario de salud. Se mencionan las áreas que abarcan la atención de las mujeres en el centro: Acompañamiento físico, Psicológico, Social y la Educación en Salud.Conclusión: La atención por un equipo interdisciplinario se torna imperante en el cuidado de mujeres mastectomizadas, ya que el cáncer y su tratamiento producen diversos cambios en la vida de la mujer, a corto y largo plazo, por lo que el seguimiento y acompañamiento debe ser biopsicosocial, abarcando todas las áreas que pueden verse afectadas, especialmente durante la pandemia. Palabras clave: Salud de la mujer. Neoplasias de la mama. Mastectomía. Atención integral de salud. Infecciones por coronavirus.

RESUMOObjetivo: Descrever a experiência vivida em um centro de acompanhamento interdisciplinar de mulheres mastectomizadas de uma universidade pública de São Paulo durante o início da pandemia COVID-19.Método: Relato de experiência sobre cuidados de saúde prestados no centro de saúde para mulheres mastectomizadas.Resultados: Os atendimentos foram realizadas três vezes por semana por uma equipe interdisciplinar de saúde. São mencionadas as áreas que abrangem o atendimento à mulher no centro: Acompanhamento físico, psicológico, social e Educação em saúde.Conclusão: A atenção por uma equipe interdisciplinar torna-se predominante no cuidado de mulheres mastectomizadas, pois o câncer e seu tratamento produzem diversas mudanças na vida das mulheres, a curto e longo prazo. Portanto o acompanhamento devem ser biopsicossociais, cobrindo todas as áreas que podem ser afetadas, especialmente durante a pandemia.Palavras-chave: Saúde da mulher. Neoplasias da mama. Mastectomia. Assistência integral à saúde. Infecções por coronavirus.

Validity and Reliability of the Richmond Agitation-Sedation Scale in Pediatric Intensive Care Patients: A Multicenter Study.

Background: There is limited data about the psychometric properties of the Richmond Agitation-Sedation Scale (RASS) in children. This study aims to analyze the validity and reliability of the RASS in assessing sedation and agitation in critically ill children. Methods: A multicenter prospective study in children admitted to pediatric intensive care, aged between 1 month and 18 years. Twenty-eight observers from 14 PICUs (pediatric intensive care units) participated. Every observation was assessed by 4 observers: 2 nurses and 2 pediatric intensivists. We analyzed RASS inter-rater reliability, construct validity by comparing RASS to the COMFORT behavior (COMFORT-B) scale and the numeric rating scale (NRS), and by its ability to distinguish between levels of sedation, and responsiveness to changes in sedative dose levels. Results: 139 episodes in 55 patients were analyzed, with a median age 3.6 years (interquartile range 0.7-7.8). Inter-rater reliability was excellent, weighted kappa (κw) 0.946 (95% CI, 0.93-0.96; p < 0.001). RASS correlation with COMFORT-B scale, rho = 0.935 (p < 0.001) and NRS, rho = 0.958 (p < 0.001) was excellent. The RASS scores were significantly different (p < 0.001) for the 3 sedation categories (over-sedation, optimum and under-sedation) of the COMFORT-B scale, with a good agreement between both scales, κw 0.827 (95% CI, 0.789-0.865; p < 0.001), κ 0.762 (95% CI, 0.713-0.811, p < 0.001). A significant change in RASS scores (p < 0.001) was recorded with the variance of sedative doses. Conclusions: The RASS showed good measurement properties in PICU, in terms of inter-rater reliability, construct validity, and responsiveness. These properties, including its ability to categorize the patients into deep sedation, moderate-light sedation, and agitation, makes the RASS a useful instrument for monitoring sedation in PICU.

Experience in a rehabilitation center for mastectomized women at the start of the COVID-19 pandemic / Experiência em centro de reabilitação de mulheres mastectomizadas no início da pandemia do COVID-19 / Experiencia en centro de rehabilitación de mujeres mastectomizadas al inicio de la pandemia por COVID-19

ABSTRACT Objective Describe the experience lived in an interdisciplinary follow-up care center for mastectomized women at a public university in São Paulo during the beginning of the COVID-19 pandemic Method Experience report on the health care provided in the health center for mastectomized women. Results The care was provided three times a week by an interdisciplinary health team. The mentioned areas that cover the women care in the center: Physical, Psychological, Social Support and Health Education. Conclusions The attention by an interdisciplinary team becomes prevalent in the care of mastectomized women, since cancer and its treatment produce various changes in women's lives in the short and long term, so follow-up and support must be biopsychosocial, covering all areas that may be affected, especially during the pandemic.

RESUMO Objetivo Descrever a experiência vivida em um centro de acompanhamento interdisciplinar de mulheres mastectomizadas de uma universidade pública de São Paulo durante o início da pandemia COVID-19. Método Relato de experiência sobre cuidados de saúde prestados no centro de saúde para mulheres mastectomizadas. Resultados Os atendimentos foram realizados três vezes por semana por uma equipe interdisciplinar de saúde. São mencionadas as áreas que abrangem o atendimento à mulher no centro: Acompanhamento físico, psicológico, social e Educação em saúde. Conclusão A atenção por uma equipe interdisciplinar torna-se predominante no cuidado de mulheres mastectomizadas, pois o câncer e seu tratamento produzem diversas mudanças na vida das mulheres, a curto e longo prazo. Portanto o acompanhamento deve ser biopsicossocial, cobrindo todas as áreas que podem ser afetadas, especialmente durante a pandemia.

RESUMEN Objetivo Describir la experiencia vivida en un centro de atención interdisciplinaria de seguimiento a mujeres mastectomizadas de una universidad pública de Sao Paulo durante el inicio de la pandemia de COVID-19. Método Relato de experiencia sobre las atenciones de salud brindadas en el centro de salud para mujeres mastectomizadas. Resultados Las atenciones fueron realizadas tres veces por semana por un equipo interdisciplinario de salud. Se mencionan las áreas que abarcan la atención de las mujeres en el centro: Acompañamiento físico, Psicológico, Social y la Educación en Salud. Conclusión La atención por un equipo interdisciplinario se torna imperante en el cuidado de mujeres mastectomizadas, ya que el cáncer y su tratamiento producen diversos cambios en la vida de la mujer, a corto y largo plazo, por lo que el seguimiento y acompañamiento debe ser biopsicosocial, abarcando todas las áreas que pueden verse afectadas, especialmente durante la pandemia.

Enteropathogenic Escherichia coli (EPEC) Recruitment of PAR Polarity Protein Atypical PKCζ to Pedestals and Cell-Cell Contacts Precedes Disruption of Tight Junctions in Intestinal Epithelial Cells.

Enteropathogenic Escherichia coli (EPEC) uses a type three secretion system to inject effector proteins into host intestinal epithelial cells, causing diarrhea. EPEC induces the formation of pedestals underlying attached bacteria, disrupts tight junction (TJ) structure and function, and alters apico-basal polarity by redistributing the polarity proteins Crb3 and Pals1, although the mechanisms are unknown. Here we investigate the temporal relationship of PAR polarity complex and TJ disruption following EPEC infection. EPEC recruits active aPKCζ, a PAR polarity protein, to actin within pedestals and at the plasma membrane prior to disrupting TJ. The EPEC effector EspF binds the endocytic protein sorting nexin 9 (SNX9). This interaction impacts actin pedestal organization, recruitment of active aPKCζ to actin at cell-cell borders, endocytosis of JAM-A S285 and occludin, and TJ barrier function. Collectively, data presented herein support the hypothesis that EPEC-induced perturbation of TJ is a downstream effect of disruption of the PAR complex and that EspF binding to SNX9 contributes to this phenotype. aPKCζ phosphorylates polarity and TJ proteins and participates in actin dynamics. Therefore, the early recruitment of aPKCζ to EPEC pedestals and increased interaction with actin at the membrane may destabilize polarity complexes ultimately resulting in perturbation of TJ.

Sperm Flagellar 1 Binds Actin in Intestinal Epithelial Cells and Contributes to Formation of Filopodia and Lamellipodia.

BACKGROUND & AIMS: Sperm flagellar 1 (also called CLAMP) is a microtubule-associated protein that regulates microtubule dynamics and planar cell polarity in multi-ciliated cells. We investigated the localization and function of sperm flagellar 1, or CLAMP, in human intestinal epithelia cells (IECs). METHODS: We performed studies with SKCO-15 and human intestinal enteroids established from biopsies from different intestinal segments (duodenal, jejunum, ileal, and colon) of a single donor. Enteroids were induced to differentiation after incubation with growth factors. The distribution of endogenous CLAMP in IECs was analyzed by immunofluorescence microscopy using total internal reflection fluorescence-ground state depletion and confocal microscopy. CLAMP localization was followed during the course of intestinal epithelial cell polarization as cells progressed from flat to compact, confluent monolayers. Protein interactions with endogenous CLAMP were determined in SKCO-15 cells using proximity ligation assays and co-immunoprecipitation. CLAMP was knocked down in SKCO-15 monolayers using small hairpin RNAs and cells were analyzed by immunoblot and immunofluorescence microscopy. The impact of CLAMP knock-down in migrating SKCO-15 cells was assessed using scratch-wound assays. RESULTS: CLAMP bound to actin and apical junctional complex proteins but not microtubules in IECs. In silico analysis predicted the calponin-homology domain of CLAMP to contain conserved amino acids required for actin binding. During IEC polarization, CLAMP distribution changed from primarily basal stress fibers and cytoplasm in undifferentiated cells to apical membranes and microvilli in differentiated monolayers. CLAMP accumulated in lamellipodia and filopodia at the leading edge of migrating cells in association with actin. CLAMP knock-down reduced the number of filopodia, perturbed filopodia polarity, and altered the organization of actin filaments within lamellipodia. CONCLUSIONS: CLAMP is an actin-binding protein, rather than a microtubule-binding protein, in IECs. CLAMP distribution changes during intestinal epithelial cell polarization, regulates the formation of filopodia, and appears to assist in the organization of actin bundles within lamellipodia of migrating IECs. Studies are needed to define the CLAMP domains that interact with actin and whether its loss from IECs affects intestinal function.

EPEC NleH1 is significantly more effective in reversing colitis and reducing mortality than NleH2 via differential effects on host signaling pathways.

Enteropathogenic Escherichia coli (EPEC) is a foodborne pathogen that uses a type III secretion system to translocate effector molecules into host intestinal epithelial cells (IECs) subverting several host cell processes and signaling cascades. Interestingly, EPEC infection induces only modest intestinal inflammation in the host. The homologous EPEC effector proteins, NleH1 and NleH2, suppress the nuclear factor-κB (NF-κB) pathway and apoptosis in vitro. Increased apoptosis and activation of NF-κB and MAP kinases (MAPK) contribute to the pathogenesis of inflammatory bowel diseases (IBD). The aim of this study was to determine if NleH1 and NleH2 also block MAPK pathways in vitro and in vivo, and to compare the effects of these bacterial proteins on a murine model of colitis. Cultured IECs were infected with various strains of EPEC expressing NleH1 and NleH2, or not, and the activation of ERK1/2 and p38 was determined. In addition, the impact of infection with various strains of EPEC on murine DSS colitis was assessed by change in body weight, colon length, histology, and survival. Activation of apoptosis and MAPK signaling were also evaluated. Our data show that NleH1, but not NleH2, suppresses ERK1/2 and p38 activation in vitro. Interestingly, NleH1 affords significantly greater protection against and hastens recovery from dextran sodium sulfate (DSS)-induced colitis compared to NleH2. Strikingly, colitis-associated mortality was abolished by infection with EPEC strains expressing NleH1. Interestingly, in vivo NleH1 suppresses activation of ERK1/2 and p38 and blocks apoptosis independent of the kinase domain that inhibits NF-κB. In contrast, NleH2 suppresses only caspase-3 and p38, but not ERK1/2. We conclude that NleH1 affords greater protection against and improves recovery from DSS colitis compared to NleH2 due to its ability to suppress ERK1/2 in addition to NF-κB, p38, and apoptosis. These findings warrant further investigation of anti-inflammatory bacterial proteins as novel treatments for IBD.

EPEC effector EspF promotes Crumbs3 endocytosis and disrupts epithelial cell polarity.

Enteropathogenic Escherichia coli (EPEC) uses a type III secretion system to inject effector proteins into host intestinal epithelial cells causing diarrhoea. EPEC infection redistributes basolateral proteins ß1-integrin and Na+ /K+ ATPase to the apical membrane of host cells. The Crumbs (Crb) polarity complex (Crb3/Pals1/Patj) is essential for epithelial cell polarisation and tight junction (TJ) assembly. Here, we demonstrate that EPEC displaces Crb3 and Pals1 from the apical membrane to the cytoplasm of cultured intestinal epithelial cells and colonocytes of infected mice. In vitro studies show that EspF, but not Map, alters Crb3, whereas both effectors modulate Pals1. EspF perturbs polarity formation in cyst morphogenesis assays and induces endocytosis and apical redistribution of Na+ /K+ ATPase. EspF binds to sorting nexin 9 (SNX9) causing membrane remodelling in host cells. Infection with ΔespF/pespFD3, a mutant strain that ablates EspF binding to SNX9, or inhibition of dynamin, attenuates Crb3 endocytosis caused by EPEC. In addition, infection with ΔespF/pespFD3 has no impact on Na+ /K+ ATPase endocytosis. These data support the hypothesis that EPEC perturbs apical-basal polarity in an EspF-dependent manner, which would contribute to EPEC-associated diarrhoea by disruption of TJ and altering the crucial positioning of membrane transporters involved in the absorption of ions and solutes.

Modulation of epithelial cell polarity by bacterial pathogens.

Epithelial cells constitute a physical barrier that aids in protecting the host from microbial pathogens. Polarized epithelial cells contain distinct apical and basolateral membrane domains separated by intercellular junctions, including tight junctions (TJs), which contribute to the maintenance of apical-basal polarity. Polarity complexes also contribute to the establishment of TJ formation. Several pathogens perturb epithelial TJ barrier function and structure in addition to causing a loss of apical-basal polarity. Here, we review the impact of pathogenic bacteria on the disruption of cell-cell junctions and epithelial polarity.

ZO proteins redundantly regulate the transcription factor DbpA/ZONAB.

The localization and activities of DbpA/ZONAB and YAP transcription factors are in part regulated by the density-dependent assembly of epithelial junctions. DbpA activity and cell proliferation are inhibited by exogenous overexpression of the tight junction (TJ) protein ZO-1, leading to a model whereby ZO-1 acts by sequestering DbpA at the TJ. However, mammary epithelial cells and mouse tissues knock-out for ZO-1 do not show increased proliferation, as predicted by this model. To address this discrepancy, we examined the localization and activity of DbpA and YAP in Madin-Darby canine kidney cells depleted either of ZO-1, or one of the related proteins ZO-2 and ZO-3 (ZO proteins), or all three together. Depletion of only one ZO protein had no effect on DbpA localization and activity, whereas depletion of ZO-1 and ZO-2, which is associated with reduced ZO-3 expression, resulted in increased DbpA localization in the cytoplasm. Only depletion of ZO-2 reduced the nuclear import of YAP. Mammary epithelial (Eph4) cells KO for ZO-1 showed junctional DbpA, demonstrating that ZO-1 is not required to sequester DbpA at junctions. However, further depletion of ZO-2 in Eph4 ZO-1KO cells, which do not express ZO-3, caused decreased junctional localization and expression of DbpA, which were rescued by the proteasome inhibitor MG132. In vitro binding assays showed that full-length ZO-1 does not interact with DbpA. These results show that ZO-2 is implicated in regulating the nuclear shuttling of YAP, whereas ZO proteins redundantly control the junctional retention and stability of DbpA, without affecting its shuttling to the nucleus.

MgcRacGAP interacts with cingulin and paracingulin to regulate Rac1 activation and development of the tight junction barrier during epithelial junction assembly.

The regulation of Rho-family GTPases is crucial to direct the formation of cell-cell junctions and tissue barriers. Cingulin (CGN) and paracingulin (CGNL1) control RhoA activation in epithelial cells by interacting with RhoA guanidine exchange factors. CGNL1 depletion also inhibits Rac1 activation during junction assembly. Here we show that, unexpectedly, Madin-Darby canine kidney epithelial cells depleted of both CGN and CGNL1 (double-KD cells) display normal Rac1 activation and tight junction (TJ) formation, despite decreased junctional recruitment of the Rac1 activator Tiam1. The expression of the Rac1 inhibitor MgcRacGAP is decreased in double-KD cells, and the barrier development and Rac1 activation phenotypes are rescued by exogenous expression of MgcRacGAP. MgcRacGAP colocalizes with CGN and CGNL1 at TJs and forms a complex and interacts directly in vitro with CGN and CGNL1. Depletion of either CGN or CGNL1 in epithelial cells results in decreased junctional localization of MgcRacGAP but not of ECT2, a centralspindlin-interacting Rho GEF. These results provide new insight into coordination of Rho-family GTPase activities at junctions, since apical accumulation of CGN and CGNL1 at TJs during junction maturation provides a mechanism to spatially restrict down-regulation of Rac1 activation through the recruitment of MgcRacGAP.

The control of gene expression and cell proliferation by the epithelial apical junctional complex.

The AJC (apical junctional complex) of vertebrate epithelial cells orchestrates cell-cell adhesion and tissue barrier function. In addition, it plays a pivotal role in signalling. Several protein components of the AJC, e.g. the cytoplasmic proteins ß-catenin, p120-catenin and ZO (Zonula Occludens)-2, can shuttle to the nucleus, where they interact with transcription factors to regulate gene expression and cell proliferation. Other junctional proteins, e.g. angiomotin, α-catenin and cingulin, are believed to act by sequestering either transcription factors, such as YAP (Yes-associated protein), or regulators of small GTPases, such as GEF (guanine-nucleotide-exchange factor)-H1, at junctions. The signalling activities of AJC proteins are triggered by different extracellular and intracellular cues, including cell density, and physiological or pathological activation of developmentally regulated pathways, such as the Wnt pathway. The interplay between junctional protein complexes, the actin cytoskeleton and signalling pathways is of crucial importance in the regulation of gene expression and cell proliferation.

[Some ethical dilemmas in the medical praxis]. / Algunos dilemas éticos en la práctica médica.

We reviewed from a bioethical perspective and attempting prevention of potential conflicts derived communication failure during medical practice, palliative treatments and dignified death in the institutional practice as well as general practice; most of conflicts related to patient-doctor relationship could de prevented. We propose an attitude and aptitude plus in deep knowledge of patient, family, friends and legal representatives in terms fully honest communication to prevent most of conflicts and avoid its consequences against doctors and other health workers. Prevention is better and it depends of knowledge of norms, laws, general beliefs and common sense in this country and maybe others.

Algunos dilemas éticos en la práctica médica / Some ethical dilemmas in the medical praxis

Con la intención de prevenir conflictos en el acto médico y desde el punto de vista bioético, éste, el manejo paliativo y de la muerte digna; situaciones que pueden ser fuente de demandas entre el paciente y el médico. Sostenemos que la actitud y aptitud del médico, del paciente, su familia, amigos y representantes legales, con apertura y honestidad, pueden prevenir la gran mayoría de las causas de conflicto y evitar las consecuencias del mismo entre los profesionales de la salud y los enfermos. La prevención es posible si hay buena voluntad y conocimiento de normas, leyes, usos y sentido común.

We reviewed from a bioethical perspective and attempting prevention of potential conflicts derived communication failure during medical practice, palliative treatments and dignified death in the institutional practice as well as general practice; most of conflicts related to patient-doctor relationship could de prevented. We propose an attitude and aptitude plus in deep knowledge of patient, family, friends and legal representatives in terms fully honest communication to prevent most of conflicts and avoid its consequences against doctors and other health workers. Prevention is better and it depends of knowledge of norms, laws, general beliefs and common sense in this country and maybe others.

Phosphorylation of zona occludens-2 by protein kinase C epsilon regulates its nuclear exportation.

Here, we have analyzed the subcellular destiny of newly synthesized tight junction protein zona occludens (ZO)-2. After transfection in sparse cells, 74% of cells exhibit ZO-2 at the nucleus, and after 18 h the value decreases to 17%. The mutation S369A located within the nuclear exportation signal 1 of ZO-2 impairs the nuclear export of the protein. Because Ser369 represents a putative protein kinase C (PKC) phosphorylation site, we tested the effect of PKC inhibition and stimulation on the nuclear export of ZO-2. Our results strongly suggest that the departure of ZO-2 from the nucleus is regulated by phosphorylation at Ser369 by novel PKCepsilon. To test the route taken by ZO-2 from synthesis to the plasma membrane, we devised a novel nuclear microinjection assay in which the nucleus served as a reservoir for anti-ZO-2 antibody. Through this assay, we demonstrate that a significant amount of newly synthesized ZO-2 goes into the nucleus and is later relocated to the plasma membrane. These results constitute novel information for understanding the mechanisms that regulate the intracellular fate of ZO-2.

The tight junction protein ZO-2 blocks cell cycle progression and inhibits cyclin D1 expression.

ZO-2 is an adaptor protein of the tight junction that belongs to the MAGUK protein family. ZO-2 is a dual localization protein that in sparse cultures is present at the cell borders and the nuclei, whereas in confluent cultures it is concentrated at the cell boundaries. Here we have studied whether ZO-2 is able to regulate the expression of cyclin D1 (CD1) and cell proliferation. We have demonstrated that ZO-2 negatively regulates CD1 transcription by interacting with c-Myc at an E box present in CD1 promoter. We have further found that ZO-2 transfection into epithelial MDCK cells triggers a diminished expression of CD1 protein and decreases the rate of cell proliferation in a wound-healing assay.

Zona occludens-2 inhibits cyclin D1 expression and cell proliferation and exhibits changes in localization along the cell cycle.

Here, we have studied the effect of the tight junction protein zona occludens (ZO)-2 on cyclin D1 (CD1) protein expression. CD1 is essential for cell progression through the G1 phase of the cell cycle. We have found that in cultures of synchronized Madin-Darby canine kidney cells, ZO-2 inhibits cell proliferation at G0/G1 and decreases CD1 protein level. These effects occur in response to a diminished CD1 translation and an augmented CD1 degradation at the proteosome triggered by ZO-2. ZO-2 overexpression decreases the amount of Glycogen synthase kinase-3beta phosphorylated at Ser9 and represses beta-catenin target gene expression. We have also explored the expression of ZO-2 through the cell cycle and demonstrate that ZO-2 enters the nucleus at the late G1 phase and leaves the nucleus when the cell is in mitosis. These results thus explain why in confluent quiescent epithelia ZO-2 is absent from the nucleus and localizes at the cellular borders, whereas in sparse proliferating cultures ZO-2 is conspicuously present at the nucleus.