Sphingosine-1-phosphate restores endothelial barrier integrity in ovarian hyperstimulation syndrome.

STUDY QUESTION: Are follicular fluid (FF) sphingosine-1-phosphate (S1P) levels in patients at risk of developing ovarian hyperstimulation syndrome (OHSS) altered and in part responsible for the high vascular permeability observed in these patients. STUDY ANSWER: FF S1P levels are lower in FF from patients at risk of OHSS and treatment with S1P may reduce vascular permeability in these patients. WHAT IS KNOWN ALREADY: Although advances have been made in the diagnosis, and management of OHSS and in basic knowledge of its development, complete prevention has proven difficult. STUDY DESIGN, SIZE, DURATION: A total of 40 FF aspirates were collected from patients undergoing ART. The women (aged 25-39 years old) were classified into a control group (n = 20) or a group at risk of OHSS (n = 20). The EA.hy926 endothelial cell line was used to assess the efffects of FF from patients at risk of OHSS with or without the addition of S1P. An animal model that develops OHSS in immature Sprague-Dawley rats were also used. PARTICIPANTS/MATERIALS, SETTING, METHODS: Migration assays, confocal microscopy analysis of actin filaments, immunoblotting and quail chorioallantoic membrane (CAM) assays of in-vivo angiogenesis were performed and statistical comparisons between groups were made. MAIN RESULTS AND THE ROLE OF CHANCE: The S1P concentration was significantly lower in FF from patients at risk of OHSS (P = 0.03). The addition of S1P to this FF decreased cell migration (P < 0.05) and prevented VE-cadherin phosphorylation in endothelial cells (P < 0.05). S1P in the FF from patients at risk of OHSS increased the levels of VE-cadherin (P < 0.05), N-cadherin (P < 0.05) and ß-catenin (P < 0.05), and partially reversed actin redistribution in endothelial cells. The addition of S1P in FF from patients at risk of OHSS also decreased the levels of vascular endothelial growth factor (VEGF121; P < 0.01) and S1P lyase (SPL; P < 0.05) and increased the levels of S1PR1 (P < 0.05) in endothelial cells. In CAMs incubated with FF from patients at risk of OHSS with S1P, the number of vessel branch points decreased while the periendothelial cell coverage increased. Additionally, in a rat OHSS model, we demonstrated that vascular permeability and VEGF121 and its receptor KDR expression were increased in the OHSS group compared to the control group and that S1P administration decreased these parameters. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: The results of this study were generated from an in-vitro system. This model reflects the microvasculature in vivo. Even though the ideal model would be the use of human endothelial cells from the ovary, it is obviously not possible to carry out this kind of approach in ovaries of patients from ART. More studies will be necessary to delineate the effects of S1P in the pathogenesis of OHSS. Hence, clinical studies are needed in order to choose the most appropriate method of prevention and management. WIDER IMPLICATIONS OF THE FINDINGS: The use of bioactive sphingolipid metabolites may contribute to finding better and safer therapeutic strategies for the treatment of OHSS and other human diseases that display aberrant vascular leakage. STUDY FUNDING/COMPETING INTERESTS: This work was supported by grants ANPCyT (PICT 2012-897), CONICET (PIP 5471), Roemmers and Baron Foundation, Argentina. The authors declare no conflict of interest.

A link between Notch and progesterone maintains the functionality of the rat corpus luteum.

In this study, we investigated the interaction between the Notch pathway and progesterone to maintain the functionality of the corpus luteum (CL). When Notch signaling is activated, the γ-secretase complex releases the active intracellular domains (NICD) of their receptors, which exert survival effects. We designed studies to analyze whether the in vitro inhibition of Notch affects progesterone production, steroidogenic regulators, apoptotic parameters, and signaling transduction pathways in the cultures of CL isolated from pregnant and superovulated rats. We detected a decrease in progesterone production when corpora lutea (CL) were incubated with N-(N-(3,5-difluorophenacetyl-l-alanyl))-S-phenylglycine t-butyl ester (DAPT), a γ-secretase inhibitor. This effect could be in part due to the decrease detected in the CL protein levels of P450scc because STAR and 3ß-hydroxysteroid dehydrogenase were not affected by Notch inhibition. Besides, the addition of aminoglutethimide to the CL culture medium decreased NICD of NOTCH1. We observed an increase in the expression of active CASPASE3 (CASP3) after inhibition by Notch, which was reversed by the presence of progesterone. The BAX:BCLXL ratio was increased in CL treated with DAPT and the presence of progesterone reversed this effect. In addition, phosphorylation of AKT was inhibited in CL treated with DAPT, but had no effect on ERK activation. To demonstrate that the action of DAPT is specifically related with the inhibition of Notch, CLs were incubated with DLL4 antibody and a decrease in progesterone production was detected. These results suggest the existence of a novel link between progesterone and the Notch signaling pathway to maintain the functionality of the CL.

Gamma secretase inhibitor impairs epithelial-to-mesenchymal transition induced by TGF-ß in ovarian tumor cell lines.

Ovarian cancer is characterized by being highly metastatic, a feature that represents the main cause of failure of the treatment. This study investigated the effects of γ-secretase inhibition on the TGF-ß-induced epithelial-mesenchymal transition (EMT) process in ovarian cancer cell lines. SKOV3 cells incubated in the presence of TGF-ß showed morphological and biochemical changes related to EMT, which were blocked by co-stimulation with TGF-ß and the γ-secretase inhibitor DAPT. In SKOV3 and IGROV1 cells, the co-stimulation blocked the cadherin switch and the increase in the transcription factors Snail, Slug, Twist and Zeb1 induced by TGF-ß. DAPT impaired the translocation of phospho-ß-catenin to the inner cell compartment observed in TGF-ß-treated cells, but was not able to block the induction at protein level induced by TGF-ß. Moreover, the inhibitor blocked the increased cell migration and invasiveness ability of both cell lines induced by TGF-ß. Notch target genes (Hes1 and Hey1) were induced by TGF-ß, decreased by DAPT treatment and remained low in the presence of both stimuli. However, DAPT alone caused no effects on most of the parameters analyzed. These results demonstrate that the γ-secretase inhibitor used in this study exerted a blockade on TGF-ß-induced EMT in ovarian cancer cells.

Dynamic expression of epoxyeicosatrienoic acid synthesizing and metabolizing enzymes in the primate corpus luteum.

Epoxyeicosatrienoic acids (EpETrEs), produced from arachidonic acid via cytochrome P450 (CYP) epoxygenases, regulate inflammation, angiogenesis, cellular proliferation, ion transport and steroidogenesis. EpETrE actions are regulated through their metabolism to diols (dihydroxyeicosatrienoic acids; DiHETrE) via the enzyme soluble epoxide hydrolase (EPHX2). We set out to determine, therefore, whether EpETrE generating (epoxygenases CYP2C8, 2C9, 2C19, 2J2, 1A2 and 3A4) and metabolizing (EPHX2) enzymes are expressed in the primate corpus luteum (CL). CL were isolated from rhesus macaques during the early (day 3-5 post-LH surge), mid (day 6-8), mid-late (day 10-12), late (day 14-16) and very-late (day 17-19: menses) luteal phase of natural menstrual cycles. EPHX2 mRNA levels peaked in mid-late CL (5-fold when compared with early CL, P<0.05) and remained elevated in the late CL. Ablation of pituitary LH secretion and luteal steroid synthesis significantly reduced (P<0.05) EPHX2 mRNA levels in the mid-late CL, with progestin replacement being insufficient to restore its level of expression to control values. EPHX2 protein was localized to large and small luteal cells, as well as vascular endothelial cells. The EpETrE-generating CYP epoxygenase 2J2, 2C9 and 3A4 genes were also expressed in the macaque CL. While CYP2J2 mRNA levels did not significantly change through the luteal phase, CYP2C9 and CYP3A4 levels were significantly (P<0.05) higher in the mid-late phase when compared with the early phase. CYP2C9, 2J2 and 3A4 proteins were each localized to the large luteal cells, with 2C9 and 2J2 also being present in the small luteal, stromal and endothelial cells. These studies demonstrate for the first time that an EpETrE generating and metabolizing system exists in the primate CL, with the latter being regulated by LH and steroid hormone(s).

Indicación quirúrgica tardía en trauma renal: experiencia de 15 años del Hospital San Juan De Dios / Delayed surgical indication for renal trauma: 15 years of experience in the Hospital San Juan De Dios

Introducción: El trauma renal está involucrado en 10 por ciento de los traumatizados, el estudio de imágenes nos ha permitido manejar más conservadoramente a estos pacientes, sin embargo los parámetros clínicos siguen siendo una valiosa herramienta de vigilancia que nos permitirá tomar una conducta quirúrgica oportuna. Objetivo: Análisis de la indicación quirúrgica tardía en pacientes con trauma renal en los últimos 15años en el Servicio de Urología del Hospital San Juan de Dios. Material y método: Estudio retrospectivo de pacientes hospitalizados en Servicio de Urología entre los años 1994 y 2008, se obtienen 70 fichas clínicas, se registra edad, sexo, mecanismo de lesión, estudio de imágenes y tratamiento. La edad promedio fue de 31 años, sexo masculino (85 por ciento), el mecanismo de lesión fue: agresión de terceros en 39 por ciento, caída de altura (22 por ciento), accidente de tránsito (20 por ciento), deportes 10 por ciento y otros 9 por ciento; traumatismo cerrado en 52 pacientes y penetrante en 18 casos ;Grado I: 40 por ciento; GII: 17 por ciento; GIII: 17 por ciento; GIV: 23 por ciento, GV: 3 por ciento. Se define como tardía a la exploración quirúrgica después de una semana ocurrido el accidente. Resultados: Catorce pacientes (20 por ciento) se intervinieron quirúrgicamente en forma tardía; se operaron8 de los 52 pacientes con trauma renal cerrado (15 por ciento), en 6 de ellos se realizó nefrectomía por riñones patológicos, 2 pacientes presentaron a la semana post accidente una caída del hematocrito asociado a hematuria severa, se realizó en uno de ellos drenaje más nefrorrafia y en otro nefrectomía simple. De los pacientes con trauma penetrante se operaron 4 pacientes (22 por ciento), en dos pacientes por un descenso del hematocrito en menos de 48 horas; un tercer paciente se exploró porque se detecta un crecimiento de la masa palpable al examen físico y un 4º paciente cuya indicación

Introduction: Renal trauma is involved in 10 percent of trauma cases, image studies have allowed us to handle these patients with more conservative criteria, and nevertheless clinical parameters continue being a valuable monitoring tool that allows us to take an opportune surgical conduct. Objective: Analysis of delayed surgical indication in patients with renal trauma in the last 15 years in the Service of Urology of the Hospital San Juan de Dios. Material and methods: Retrospective study of patients hospitalized in the Urology unit, between years 1994 and 2008. 70 clinical charts are obtained, and age, sex, mechanism of injury, image studies and treatment are registered. The age average was of 31 years, masculine sex (85 percent), the injury mechanism was: third party aggression in 39 percent, fallen of height (22 percent), traffic accident (20 percent), sports 10 percent and other 9 percent. Trauma was closed in 52 patients and penetrating in 18 cases. Grade I: 40 percent; GII: 17 percent; GIII: 17 percent; GIV: 23 percent, GV: 3 percent. Delayed surgical exploration is defined as the one performed after one week of the accident. Results: Fourteen patients (20 percent) had delayed surgery; 8 of the 52 patients with closed renal trauma were operated (15 percent), in 6 of them a nephrectomy was performed due to pathological kidneys, 2patients presented to the week post In 2 patients a hematocrit fall and severe hematuria was seen a week after the accident, in one of them a drainage and nephrorrafy was performed and in the other a simple nephrectomy.Of the patients with penetrating trauma 4 patients were operated (22 percent). In two patients because a fall of the hematocrit in less than 48 hours, a third patient was explored because of a growing mass detected at physical examination, and a 4th patient whose surgical indication was determined by clinical parameters of infection, in him an infected urinoma was drained...

Carcinoma de células renales: correlación anátomo-radiológica según clasificación tumoral "T" en la experiencia clínica de 10 meses de seguimiento / Renal cell carcinoma: anatomo-radiologic correlation according to \"T\" tumor classification during a 10 month follow up period

Objetivo: Correlacionar las características tomográficas de los tumores renales, con los hallazgos anatomopatológicos, para determinar si es posible su diferenciación por imágenes en el diagnóstico y clasificación T (TNM).Material y Método: Se realizó un seguimiento por 10 meses entre octubre de 2008 y julio de 2009 de todos los tumores renales estudiados y tratados en el Hospital San Juan de Dios. La cohorte recopiló 34 casos de tumores renales sólidos con informe radiológico por TC, de los cuales 30 se sometieron a cirugía, con su posterior análisis anatomopatológico y grado nuclear de Fuhrman. Hombres 23, mujeres 11, con un promedio de edad de 55 años (32-84 años). Se evalúa por TC, patrón de realce, homogeneidad, calcificaciones y compromiso vascular. Se correlacionó el tamaño T imagenológico por TNM, con el pT por histopatología, así como también la presencia o no de síntomas y la modalidad quirúrgica elegida (radical o parcial, laparoscópica o abierta). Resultados: Un promedio de tamaño tumoral de 6,6 cm (1,5-12 cm), 30 fueron catalogados al TC como carcinoma de células renales (CCR), de los cuales 24 fueron confirmados por histopatología, siendo las características tomográficas más predictivas, el comportamiento hipervascular (23) y la heterogenicidad (22). En pocos casos de los CCR, se presentó necrosis (5) y calcificaciones (1). El resto de los tumores (6) correspondieron a pielonefritis xantogranulomatosa, angiomiolipoma, mielolipoma suprarrenal, oncocitoma (2) y quiste hidatídico, de los cuales 5 impresionaron como CCR. El acierto entre la impresión diagnóstica por TC y la por istopatología, fue de 83 por ciento. La correlación en la clasificación T (TNM) fue con acierto total de 75 por ciento. La certeza en el diagnóstico en etapas m órgano confinadas (T1 y T1-2) fue significativa (p= 0,01 y p= 0,02) al confirmarse a la histología. Existe una tendencia de los tumores incidentales a presentarse en etapas más precoces. Conclusión: Casi el...

Objective: To correlate tomographic characteristics of renal tumors with pathologic findings in order to determine if the diagnosis and TNM classification can be performed by imaging. Material and Method: Between October 2008 and July 2009, all renal tumors diagnosed and treated at Hospital San Juan de Dios were followed. The cohort was composed of 34 solid renal tumors. All cases had a CT scan report. Out of the total, 30 patients underwent surgery with a subsequent pathology report that included Fuhrman’s nuclear grade. There were 23 males and 11 females with an average age of 55 years (32-84). CT scan analysis included enhancement pattern, homogeneity, calcifications and vascular involvement. We correlated the T size by images with pT by histology. Also, the presence of symptoms and the surgical technique were recorded. Results: Average tumor size was 6.6 cm (1.5-12); 30 cases were classified by CT scan as renal cell carcinoma (RCC). Of them, 24 cases were confirmed by histology. The CT scan findings that better predicted histology were hipervascularity (23) and heterogeneity (22). Few RCC showed necrosis (5)and calcifications (1). The rest of the cases (6) corresponded to xantogranulomatous pielonefritis, angiomiolipoma, mielolipoma, oncocytoma and hidatidic cyst. Of them, 5 were diagnosed as RCC by images. The agreement between CT scan diagnosis and histopathology was 83 percent. The correlation with the TNM classification was 75 percent overall. Accuracy in the diagnosis of T1 and T2 cases was significant (p =0.01 and p =0.02). There is a trend for incidental tumors to present at earlier stages. Conclusion: Almost 50 percent of RCC cases were incidental findings. Imaging characteristics and enhancement pattern by CT scan are helpful in the diagnosis of RCC and in the T classification. This method has an adequate correlation with postoperative histopathology especially at early stages (T1 and T2). Better diagnostic elements are required...

Compromiso de la grasa periganglionar como factor de riesgo en linfadenectomía inguinal modificada en cáncer de pene / Involvement of the perinodal fat as a risk factor in modified inguinal lymphnode dissection for penile cancer

Introducción: La linfadenectomía en cáncer de pene cumple un importantísimo rol en la etapificación y tratamiento. Históricamente ha sido subutilizada por la morbilidad asociada al procedimiento quirúrgico. Objetivo: Análisis de la linfadenectomía inguinal modificada, su rol etapificador y el compromiso de la grasa periganglionar como factor de mal pronóstico. Material y método: Revisión retrospectiva de 40 linfadenectomías inguinales modificadas, realizadas en 20 pacientes con diagnóstico de cáncer de pene, entre los años 1995 y 2009. La edad promedio de los pacientes estudiados fue de 63,8 años, con un tiempo de evolución promedio de la lesión de 8,2 meses. La histología en la totalidad de los casos fue carcinoma escamoso, realizándose como primera aproximación quirúrgica 8penectomías parciales y 12 totales. Técnica quirúrgica utilizada: Linfadenectomía inguinal modificada. Resultados: Se realizaron 22 linfadenectomías profilácticas, obteniéndose ganglios inguinales superficiales (+) en 4 oportunidades y ganglios profundos (+) en 1 de ellas. Se realizaron 18 linfadenectomías terapéuticas, obteniéndose ganglios inguinales superficiales (+) en 12 procedimientos y ganglios profundos (+) en 4 de ellos. La sobrevida general a 5 años, fue de 15 por ciento y la específica de 10 por ciento.Los factores de mal pronóstico fueron la indiferenciación lesional, el compromiso de ganglios inguinales profundos y el compromiso de la grasa periganglionar. Discusión: No existiendo actualmente exámenes con adecuada sensibilidad y especificidad para detectar el compromiso ganglionar inguinal en el cáncer de pene, se plantea como rol primario de la Linfadenectomía, la etapificación; sin embargo, su rol terapéutico es el más importante, pudiendo ser curativo especialmente en pacientes con micrometástasis. Detectados factores de mal pronóstico, como el compromiso de la grasa periganglionar, se podría mejorar la sobrevida de la enfermedad en pacientes seleccionados con terapias...

Introduction: Lymphnode dissection (LND) in penile cancer (PC) plays an important role both in staging andtreatment. Historically, LND has been underused due to the morbidity associated to the surgical procedure. Objective: To study modified inguinal LND for staging and to evaluate involvement of the perinodal fat as a factor for poor prognosis. Material and method: A retrospective review of 40 modified inguinal LND performed in 20 patients with a diagnosis of PC between 1995 and 2009 was done. Mean age was 63.8 years. The lesion had been present an average of 8.2 month prior to the diagnosis. In all cases, the biopsy showed squamus cell carcinoma. Surgical procedure was partial penectomy in 8 patients and total penectomy in 12 cases. All patients underwent modified inguinal LND. Results: Twenty two prophylactic LND were performed. In 4 cases, (+) superficial lymphnodes were obtained and in 1 case, (+) deep lymphnodes were found. Eighteen therapeutic LND were done. In 12 LND, (+) superficial nodes were found and in 4 cases, deep (+) lymphnodes were found. Overall survival at 5 years was 15 percent, whereas the specific survival was 10 percent. Poor prognostic factors were the following: high histologic grade, involvement of deep inguinal nodes and involvement of the perinodal fat. Discussion: Currently, there are no tests with enough sensitivity and specificity to detect lymphnode involvement in PC. Therefore, LND has an important role in staging; however, its therapeutic role is even more important; the procedure might be curative in patients with micrometastases. When poor prognostic factors such as involvement of the perinodal fat are found, survival could be improved using adjuvant therapies like radiation therapy or combined chemotherapy.

Quiste hidatídico en el niño: experiencia del Servicio de Cirugía Hospital Roberto del Río 1996-1999 / Hidatid cyst in the child: experience of surgery service at the Hospital Roberto del Río 1996-1999

Hidatidosis es una enfermedad endémica en diversas partes del mundo. Entre marzo de 1996 y diciembre 1999 en el Hospital Roberto del Río fueron sometidos a cirugía por hidatidosis de distintas ubicaciones anatómicas 26 niños, 12 niñas y 14 niños, 22 eran de regiones del norte del país y 4 eran de Santiago (de estos últimos 2 habían vivido en áreas rurales). La hidatidosis fue pulmonar en 18 niños, uno de los cuales también tenía ubicación del parásito en hígado. Hidatidosis hepática se observó en 9 niños, ino de los cuales tenía también ubicación renal. El tiempo transcurrido entre la primera consulta y la cirugía varió de 2 a 103 días (mediana 13 días). Los procedimientos quirúrgicos fueron cistectomía, evacuación, drenaje y obliteración de la cavidad residual mediante capitonaje o marsupialización. Solamente 3 pacientes recibieron albendazol antes de la cirugía y 8 después de la intervención. No hubo fallecidos. 12 niños presentaron complicaciones post operatorias