Metformin Treatment Regulates the Expression of Molecules Involved in Adiponectin and Insulin Signaling Pathways in Endometria from Women with Obesity-Associated Insulin Resistance and PCOS.

Polycystic ovary syndrome (PCOS) is an endocrine/metabolic disorder associated with insulin resistance (IR) and obesity. Endometria from women with PCOS present failures in insulin action, glucose uptake and signaling of insulin-sensitizing molecules, such as adiponectin, with consequences for reproduction. Metformin (MTF) treatment improves insulin signaling in endometrial tissues, but its mechanism is not fully understood. This study addresses the MTF effect, as well as adiponectin agonist action, on levels of molecules associated with insulin and adiponectin signaling pathways in endometrial tissue and cells, as assessed by immunohistochemistry and immunocytochemistry, respectively. Endometrial tissues were obtained from women and divided into five groups: Normal Weight (control); Obesity + IR; Obesity + IR + PCOS; Obesity + IR + MTF; Obesity + IR + PCOS + MTF. Endometrial cells stimulated with TNFα (as obesity-marker) were also used to partially emulate an obesity environment. The results showed low levels of insulin/adiponectin signaling in the endometria from women with obesity, IR and PCOS compared with the control group. MTF re-established these levels, independently of PCOS. TNFα-associated molecules were elevated in pathologic endometria, whereas MTF diminished these levels. The low levels of insulin/adiponectin molecules in endometrial cells treated with TNFα were reverted by MTF, similar to what was observed in the case of the adiponectin agonist. Therefore, independently of PCOS, MTF can re-establish levels of molecules involved in insulin/adiponectin signaling in endometrial cells, suggesting an improvement in insulin action and reproductive failures observed in endometria from women with obesity/PCOS.

The Autophagy Protein Pacer Positively Regulates the Therapeutic Potential of Mesenchymal Stem Cells in a Mouse Model of DSS-Induced Colitis.

Mesenchymal stem cells (MSC) have emerged as a promising tool to treat inflammatory diseases, such as inflammatory bowel disease (IBD), due to their immunoregulatory properties. Frequently, IBD is modeled in mice by using dextran sulfate sodium (DSS)-induced colitis. Recently, the modulation of autophagy in MSC has been suggested as a novel strategy to improve MSC-based immunotherapy. Hence, we investigated a possible role of Pacer, a novel autophagy enhancer, in regulating the immunosuppressive function of MSC in the context of DSS-induced colitis. We found that Pacer is upregulated upon stimulation with the pro-inflammatory cytokine TNFα, the main cytokine released in the inflammatory environment of IBD. By modulating Pacer expression in MSC, we found that Pacer plays an important role in regulating the autophagy pathway in this cell type in response to TNFα stimulation, as well as in regulating the immunosuppressive ability of MSC toward T-cell proliferation. Furthermore, increased expression of Pacer in MSC enhanced their ability to ameliorate the symptoms of DSS-induced colitis in mice. Our results support previous findings that autophagy regulates the therapeutic potential of MSC and suggest that the augmentation of autophagic capacity in MSC by increasing Pacer levels may have therapeutic implications for IBD.

Pro-Inflammatory Markers Negatively Regulate IRS1 in Endometrial Cells and Endometrium from Women with Obesity and PCOS.

A pro-inflammatory environment is characteristic of obesity and polycystic ovary syndrome (PCOS). This environment through cytokines secretion negatively affects insulin action. Endometria from women with both conditions (obesity and PCOS) present high TNF-α level and altered insulin signaling. In addition, these patients present reproductive failures that could be associated to an abnormal endometrial function. Here, TNF-α and IL-6 effects on insulin signaling pathway were evaluated. Serum and endometrial IL-6, phospho-IRS1-S270 (inactive form) and phospho-IRS1-Y612 (active form) levels were evaluated in women with: Normal-Weight, Obesity and Obesity-PCOS. In endometrial cells under hyperandrogenic/hyperinsulinic conditions resembling PCOS, it was evaluated IL-6/TNF-α effects on phospho-IRS1-S270, phospho-IRS1-Y612, phospho-AKT-S473 levels, and S6K and JNK activation (IRS1-inactivating molecules). In obesity groups, diminution of IRS1-active form was observed, being more significantly in Obesity-PCOS; whereas, IRS1-inactive form increased in Obesity-PCOS. Serum and endometrial IL-6 were higher in Obesity-groups compared to Normal-Weight. In endometrial cells, TNF-α increases phospho-IRS1-S270, while IL-6 decreases phospho-IRS1-Y612. Importantly, TNF-α and IL-6 promote S6K and JNK activation; TNF-α increases and IL-6 decreases phospho-AKT-S473 levels. Thus, pro-inflammatory cytokines in endometrium could negatively influence insulin signaling by different mechanisms: TNF-α promotes activation of IRS1-inactivating kinases, whereas, IL-6 decreases IRS1 and AKT activation. Moreover, when obesity and PCOS are present the disruption of insulin signaling is aggravated. These effects could explain endometrial abnormal function and reproductive failures observed in women with obesity and PCOS.

Histological comparison of DBBM and platelet rich fibrin for guided bone regeneration in a rabbit model.

PURPOSE: To histologically evaluate the use of bovine derived deproteinized xenograft (DBBM), leukocyte and platelet rich fibrin (L-PRF) and the combination of both in Guided Bone Regeneration (GBR) performed in non-critical size defects in rabbit. METHODS: A prospective experimental study was performed. Four bone defects in the tibiae of 12 rabbits were made and each of them was filled with DBBM, L-PRF, a combination of DBBM + L-PRF or was left to heal as control site. All defects were covered with a collagen membrane. Rabbits were randomly distributed in three groups and euthanatized at 3, 6 or 9 weeks. Samples were obtained and histologically analyzed to determine vital bone, connective tissue and remaining graft particles percentage. Analysis of variance, Kruskal Wallis and non-paired t-test where used to evaluate the significance of the results. RESULTS: At 3 weeks of healing, DBBM showed significantly more vital bone percentage than L-PRF (p = 0,05) and DBBM + L-PRF showed significantly less connective tissue than control (p < 0,05). All other groups showed no statistical difference between them. At 6 weeks of healing, DBBM showed significantly more vital bone percentage than L-PRF (p < 0,05), DBBM + L-PRF (p < 0,05) and control (p < 0,05) and there wasn't any other significant difference regarding to connective tissue or remaining particle percentage between groups. At t 9 weeks healing period, there weren't any significant differences between groups. CONCLUSIONS: DBBM seems to enhance vital bone formation at early healing stages. The use of L-PRF alone or combined with DBBM, didn't show any histological improvement regarding to vital bone formation. The use of DBBM, alone or in conjunction with L-PRF showed a trend to reduce connective tissue percentage. The use of L-PRF combined with DBBM didn't affect the remaining particle percentage.

Photoreduction of oxoisoaporphines by amines: laser flash and steady-state photolysis, pulse radiolysis, and TD-DFT studies.

Photoreduction of oxoisoaporphine (OIA) (1-aza-benzo-[de]anthracen-7-one) and its 5-methoxy (5-MeO-OIA) derivative by selected amines (two non-alpha-hydrogen-donating amines (1,4-diaza[2.2.2]-bicyclooctane (DABCO) and 2,2,6,6-tetramethylpiperidine (TMP)) and three alpha-hydrogen-donating amines (triethylamine (TEA), diethylmethylamine (DEMA), and dimethylethylamine (DMEA))) has been studied in deaerated neat acetonitrile solutions using laser flash and steady-state photolysis. The triplet excited states of OIA and 5-MeO-OIA are characterized by intense absorption maxima located at lambda(max) = 450 nm and lifetimes of 34.7 +/- 0.5 and 44.6 +/- 0.4 micros, respectively. In the presence of tertiary amines, both triplets are quenched with a rate constant that varies from the near diffusion limit (>10(9) M(-1) s(-1)) to a rather low value (approximately 10(7) M(-1) s(-1)) and shows the expected dependence on the reduction potential for one-electron-transfer reactions. The transient absorption spectra observed after quenching of the respective triplet states are characterized by distinct absorption maxima located at lambda(max) = 480 and 490 nm (for OIA and 5-MeO-OIA, respectively) and accompanied by broad shoulders in the range of 510-560 nm. They were assigned to either solvent-separated radical ion pairs and/or isolated radical anions. In the presence of alpha-hydrogen-donating amines these species undergo protonation that leads to the formation of neutral hydrogenated radicals A1H(*)/A2H(*) with two possible sites of protonation, N and O atoms. Pulse radiolysis and molecular modeling together with TD-DFT calculations were used to support the conclusions about the origin of transients.

Prevalencia de la exóstosis del conducto auditivo externo entre practicantes de surf y bodyboard en la costa central de Chile / Prevalence of auditory canal exostosis among surfers and bodyboarding athletes of the central Chilean coast

La exóstosis del conducto auditivo externo (ECAE), también conocida como oído de surfista, es una alteración del tejido óseo del oído externo, principalmente causada debido a la sobrexposición al frío. Considerando que los practicantes de deportes acuáticos como el surf y bodyboard de las costas del pacífico sur se exponen a aguas con temperaturas entre 12-16 °C, se busca determinar, en este estudio, la prevalencia y grado de ECAE en practicantes de surf y bodyboard de las localidades de Reñaca y Concón durante el año 2018. Se realizó una evaluación del CAE mediante video-otoscopía a 67 personas (134 oídos) practicantes de surf y bodyboard de las playas de Reñaca y Concón, y se les aplicó un cuestionario respecto a sus hábitos de práctica. Como resultado, se observó una prevalencia de ECAE del 77,6%, siendo el 61,2% ECAE bilateral y el 16,4% ECAE unilateral. Se pudo determinar, además, que el 62,3%de los participantes no usa protecciones. A partir de estos hallazgos, es posible concluir que existe una alta prevalencia de la ECAE en practicantes de surf y bodyboard en las costas centrales chilenas, lo que debería alertar tanto a la población practicante como a profesionales de la salud auditiva con el fin de promover una vida saludable en esta población.

External auditory canal exostosis (ECAE), also known as surfer's ear, is an alteration of the bone tissue of the external ear, mainly caused due to overexposure to cold. Considering that those who practice water sports such as surfing and bodyboarding on the Pacific coast are exposed to waters with temperatures between 12-16 °C,. We seek to determine, in this study, the prevalence and degree of ECAE in surfers and bodyboarders from the coast of the south of Pacific Ocean, Reñaca and Concón, during the year 2018. An evaluation of the ECAE was carried out by means of video-otoscopy in 67 people (134 ears) who practiced surfing and bodyboarding from the beaches of Reñaca and Concón, and a questionnaire was used to collect information about their practice habits. As a result, a prevalence of ECAE of 77.6% was observed, with 61.2% bilateral ECAE and 16.4% unilateral ECAE. It was also possible to determine that 62.3% of the participants do not use protections. Based on these findings, it is possible to conclude that there is a high prevalence of ECAE in surfers and bodyboarders on the central Chilean coasts, which should alert both the practicing population and hearing health professionals to promote a healthy life in this population.

Hyperandrogenism Decreases GRP78 Protein Level and Glucose Uptake in Human Endometrial Stromal Cells.

BACKGROUND: Women with polycystic ovary syndrome (PCOS) exhibit a low fertility by chronic hyperandrogenemia. Different evidence have shown that androgens could regulate the endoplasmic reticulum (ER) homeostasis and glucose metabolism. However, it is unclear whether androgens can exert these effects on human endometrial stromal cells. Our goal was to study the protein content of GRP78 (an ER homeostasis marker) in endometria from women with PCOS and healthy women and to assess the GRP78 protein levels and its relationship with glucose uptake on a human endometrial stromal cell line stimulated with testosterone. METHODS: Immunohistochemistry assays for GRP78 were performed on endometrial samples obtained from women with PCOS (n = 8) and control women subjected to hysterectomy (n = 8). Western blot analysis for GRP78 and glucose uptake was assessed in a telomerase-immortalized human endometrial stromal cell line (T-HESC) exposed to testosterone for 24 or 48 hours and challenged to an insulin short-term stimulation. Tukey test was performed for human samples comparison. Student t test or ANOVA-Bonferroni test was carried out according to the in vitro experiment. P < .05 was considered as significant. RESULTS: GRP78 stromal immunostaining was reduced in PCOS endometria compared to controls (P < .05). The T-HESC shows a testosterone-dependent downregulation of GRP78 protein content (P < .05), concomitant with half-reduction in glucose uptake compared to controls (P < .05). Moreover, enhanced small interfering RNA against GRP78 messenger RNA leads to a decrease in glucose uptake (P < .05). Such effects were reverted by hydroxyflutamide, an inhibitor of androgen receptor. CONCLUSION: These results suggest that hyperandrogenemic PCOS environment could compromise the endometrial homeostasis confirmed by the decrease in glucose uptake induced by testosterone and exhibited by stromal cells.

Expression of steroid sulfated transporters and 3ß-HSD activity in endometrium of women having polycystic ovary syndrome.

Intracrinology mechanism involves the metabolism of steroids in peripheral tissues, such as DHEA, to molecules with estrogenic or androgenic activity. Proliferation rate of endometria from Polycystic Ovary Syndrome women (PCOS) is increased, favoring hyperplasia development. Besides, in endometria from PCOS-women the synthesis of androst-5-ene-3ß,17ß-diol (androstenediol), an estrogenic molecule, is enhanced concomitantly to increased cellular proliferation. DHEA, the major intracrinological precursor, circulates mainly in its sulfated form and requires transporters for cell intake, that belong to the families of organic anion transporting polypeptides (OATP) and organic anion transporters (OAT). The aim of this study was to determine protein levels and activity of sulfated steroid transporters OATP2B1, OATP3A1, OATP4A1 and OAT4 in endometria from control and PCOS-women and to evaluate the activity of the enzyme 3ß-HSD. Levels of transporters were done by RT-PCR (OAT4 only) and Western-blot (WB). Additionally, in primary culture cells stimulated with steroids, protein levels by WB and uptake of tritiated DHEAS, were evaluated; 3ß-HSD activity was assessed using radiolabel substrate. PCOS-endometrium had higher levels of OATP2B1 and OATP4A1 than CE (p<0.05); decreased OATP4A1 levels were found in androstenediol or testosterone-stimulated cells. Accordingly, the entry of DHEAS to cells was lower in cells stimulated with testosterone (p<0.05); 3ß-HSD-activity was similar in control and PCOS-endometria. Therefore, this study describes that steroids can modulate the expression and activity of transporters of OATPs-family in human endometria and that some transporter levels are increased in PCOS-endometria, suggesting a potential role in the pathogenesis of endometrial hyperplasia of these patients.

Increased SNAIL expression and low syndecan levels are associated with high Gleason grade in prostate cancer.

Prostate cancer (PC) is a leading male oncologic malignancy wideworld. During malignant transformation, normal epithelial cells undergo genetic and morphological changes known as epithelial-mesenchymal transition (EMT). Several regulatory genes and specific marker proteins are involved in PC EMT. Recently, syndecans have been associated with malignancy grade and Gleason score in PC. Considering that SNAIL is mainly a gene repressor increased in PC and that syndecan promoters have putative binding sites for this repressor, we propose that SNAIL might regulate syndecan expression during PC EMT. The aim of this study was to analyze immunochemically the expression of SNAIL, syndecans 1 and 2 and other EMT markers in a tissue microarray (TMA) of PC samples and PC cell lines. The TMAs included PC samples of different Gleason grade and benign prostatic hyperplasia (BPH) samples, as non­malignant controls. PC3 and LNCaP cell lines were used as models of PC representing different tumorigenic capacities. Semi-quantitative immunohistochemistry was performed on TMAs and fluorescence immunocytochemistry and western blot analysis were conducted on cell cultures. Results show that SNAIL exhibits increased expression in high Gleason specimens compared to low histological grade and BPH samples. Accordingly, PC3 cells show higher SNAIL expression levels compared to LNCaP cells. Conversely, syndecan 1, similarly to E-cadherin (a known marker of EMT), shows a decreased expression in high Gleason grades samples and PC3 cells. Interestingly, syndecan 2 shows no changes associated to histological grade. It is concluded that increased SNAIL levels in advanced PC are associated with low expression of syndecan 1. The mechanism by which SNAIL regulates the expression of syndecan 1 remains to be investigated.

The role of androst-5-ene-3ß,17ß-diol (androstenediol) in cell proliferation in endometrium of women with polycystic ovary syndrome.

Women with polycystic ovary syndrome (PCOS) show high prevalence of endometrial hyperplasia and adenocarcinoma. Endometrial proliferation is increased, evaluated by high levels of Ki67 (cell cycle marker) and low levels of p27 (negative regulator of cell cycle). Nevertheless, endometrial changes in cyclin D1 (positive regulator of cell cycle) in PCOS-women are not described. Androst-5-ene-3ß,17ß-diol (androstenediol), steroid with estrogenic activity present in endometria, could be related to increased endometrial cell proliferation. The objective of this study was to determine protein content of cyclin D1 and androstenediol levels in endometria from PCOS and control-women and to evaluate the possible mechanism favoring cell proliferation associated with hormonal characteristics of patients. Therefore, cyclin D1 protein content in PCOS-women and control-endometrial tissue were assessed by western blot and immunohistochemistry. The androstenediol levels were evaluated by ELISA. To further analyze the effect of steroids (androstenediol, 17ß-estradiol, testosterone) in cell proliferation, levels of proteins cyclin D1, p27 and Ki67 were evaluated in an in vitro model of stromal endometrial cells T-HESC and St-T1b. An increase in cyclin D1 and androstenediol was observed in tissues from PCOS-women relative to control group (p<0.05). In the in vitro model, androstenediol exerted increase in cyclin D1 (p<0.05) and a decrease in p27 protein level (p<0.05), while Ki67 in St-T1b cells increased under this stimulus (p<0.05). Testosterone produces opposite effects in the levels of the above markers (p<0.05). Therefore, the hormonal imbalance associated with this syndrome could alter endometrial tissue homeostasis, promoting cell proliferation. Androstenediol is a molecule that could be involved by stimulating proliferation, whereas testosterone elicits a role of cell cycle repressor.

Elaboración y validación de un instrumento para la medición de la violencia laboral externa y sus factores de riesgo en población de trabajadores y trabajadoras chileno/as / Development and validation of an instrument for the measurement of external violence at work and its risk factors in a population of Chilean workers

Resumen: La violencia laboral externa, aquella que ejercen personas que no tienen una relación de trabajo directa con las víctimas (clientes, usuarios, asaltantes), es parte constitutiva de la violencia laboral, un problema de salud pública de creciente magnitud. En Chile la medición de la violen cia laboral externa así como sus vínculos con la salud mental han sido poco estudiados, en parte por la inexistencia de instrumentos de eva luación. Objetivo: Elaborar y validar un instrumento para medir y caracterizar violencia externa, factores de riesgo y efectos en salud mental en trabajadores/as de la salud, educación, retail y administración pública. Metodología: técnicas cualitativas y cuantitativas: grupos focales, panel de expertos y aplicación del instrumento a 554 trabaja dores/as. Resultados: El análisis factorial obtiene 7 factores referidos a violencia física, no física y a causas vinculadas al lugar de trabajo, los/ as usuarios/as y los/as trabajadores/as. La consistencia interna y confia bilidad del instrumento son estadísticamente significativas. Se confirma su capacidad para discriminar la existencia de violencia laboral externa, su intensidad y factores de riesgo en los sectores estudiados. No se observan correlaciones significativas con psicopatología medida a tra vés del GHQ-12. Conclusiones: El instrumento tiene las propiedades para discriminar y medir la violencia laboral externa en los sectores estudiados.

Abstract: External workplace violence, is understood as that perpetrated by persons outside the organizations such us customers, users, assail ants, and it is a form of workplace violence, a problem of public health of growing magnitude. In Chile, the measurement of external labor violence as well as its links with mental health have been little studied, in part by the absence of evaluation tools. Objective: to develop and validate an instrument to measure and characterize external workplace violence, risk factors and effects on mental health in workers from the health, education, retail, and public administration. Methodology: qualitative and quantitative tech niques: focus groups, expert panel and application of the instrument to 554 workers. Results: factor analysis obtained 7 factors relating to physical, not physical violence and causes related to the work place, the users, and workers - men and women. The internal con sistency and reliability of the instrument are statistically significant. Confirms its ability to discriminate the existence of external work place violence, it intensity and risk factors in the studied sectors. Failure to observe significant correlations with psychopathology measured by GHQ-12. Conclusion: The instrument has the properties to discriminate and measure the external workplace violence in the sectors considered.

Validación del inventario de violencia y acoso psicológico en el trabajo -IVAPT-PANDO- en tres ámbitos laborales chilenos / Validation of the violence and bullying at work inventory -IVAPT-PANDO- in three Chilean fields of work

INTRODUCCIÓN: En el contexto de las políticas públicas sobre riesgos psicosociales en Chile, se plantea la necesidad de incluir entre ellos la violencia y el acoso psicológico y disponer de instrumentos válidos y confiables para su medida. OBJETIVOS: se propone validar el Inventario de Violencia y Acoso Psicológico en el Trabajo (IVAPT-Pando) en la población laboral chilena. MÉTODO: El proceso consistió en: a) validación de contenidos; b) adaptación semántica del instrumento e incorporación de nuevos ítems orientados a medir violencia ejercida por actores externos a la organización; c) aplicación de la versión final del instrumento a una muestra de 700 trabajadores/as para evaluar sus propiedades psicométricas; d) análisis de validez convergente con el GHQ; y, e) elaboración de baremos referenciales. RESULTADOS: Se confirma la presencia de tres factores componentes del instrumento original y la emergencia de un cuarto factor relativo a violencia externa. La consistencia interna y confiabilidad de ambas versiones son estadísticamente significativas. Se confirma la capacidad del instrumento para discriminar la existencia de presencia e intensidad de la violencia psicológica y de acoso laboral en los sectores estudiados. Se presentan correlaciones significativas con la presencia de psicopatología medida a través del GHQ. CONCLUSIONES: El instrumento tiene las propiedades para discriminar y medir la violencia laboral en los sectores estudiados, pero debe atenderse a algunas dificultades en su aplicación.

INTRODUCTION: In the context of public policies on psychosocial risks in Chile, the need arises to include amongst these violence and bullying at work and obtain valid and reliable instruments for its measurement. OBJECTIVES: It is proposed to validate the inventory of Violence and Bullying at Work (IVAPT - Pando) in Chilean workforce; METHOD: The process consisted of: a) content validation; b) semantic adaptation of the questionnaire and its complementarity with the addition of items designed to measure violence by actors external to the organization; c) final application to a total sample of 700 subjects to analyze the psychometric properties of the instrument; d) elaboration of reference scales; and, e) study of convergent validity with the GHQ. RESULTS: The presence of three components of the original instrument is confirmed, and in the modified instrument emerges a fourth factor on violence by actors outside the organization. Internal consistency and reliability of both versions remain high. The instrument's capacity to discriminate the presence and intensity of violence and harassment in the sectors studied is confirmed. Significant correlations are presented with the presence of psychopathology as measured by the GHQ. CONCLUSIONS: The instrument is useful for measuring workplace violence but it should be paid attention to some difficulties in its application.