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Endoscopic sleeve gastroplasty (ESG) is an effective medium-term procedure for obesity treatment. There are no consistent studies confirming persistence of maintained gastric tubular configuration. We determined sleeve duration, gastric reduction degree, and suture persistence at 12-month follow-up.This is a prospective, single-center study, including patients with obesity undergoing ESG with at least 1-year follow-up, who underwent the following: (1) Barium X-ray study (BS) prior and at 6 months, assessing degree of gastric reduction (severe, moderate, or mild), and (2) gastroscopy at 12 months, accounting the number of persistent sutures (tense, lax, or absent). Secondary outcomes were weight loss data and procedure safety profile.Thirty-eight patients (30 women), median age of 47.0 [40.0-51.0] years, and average baseline BMI of 37.6 [35.5-41.5] kg/mESG is an effective and safe weight loss strategy at 12-month follow-up with persistence of most sutures and maintenance of notable gastric reduction and remodeling.

Alendronate, a bisphosphonate used to prevent osteoporosis, stimulates osteogenesis but impairs adipogenesis. Different clinical trials suggest that the incidence of diabetes may be lower in patients treated with alendronate. Taking into account the importance of adipocytes and macrophages of adipose tissue in insulin resistance and type 2 diabetes, it is necessary to evaluate the effect of alendronate in both cell types. In this paper, we investigated the effect of alendronate on the differentiation to adipocytes of 3T3-L1 fibroblasts, the cell line most used to study adipogenesis, and also its effect on lipid content and oxidative stress in mature adipocytes as well as on the inflammatory response of macrophages. We found that alendronate inhibits differentiation of 3T3-L1 fibroblasts to adipocytes in keeping with reports in other cell lines. On the other hand, treatment of 3T3-L1 adipocytes with alendronate was able to decrease triglyceride content and to prevent H2O2-induced lipid peroxidation which was evaluated as an indicator of oxidative stress. In addition, it was found that activation of RAW 264.7 macrophages to a pro-inflammatory M1 type is inhibited by this bisphosphonate. These results suggest that alendronate may contribute to prevent adipocyte excessive enlargement and the induction of oxidative stress in 3T3-L1 adipocytes as well as the activation of macrophages to a pro-inflammatory M1 type, which are events associated with adipose tissue dysfunction and insulin resistance. In this study, we unraveled the underlying mechanisms of events that were previously observed in clinical trials.

Aims: Reports regarding the effects of antioxidants in obesity have been contradictory. AntioxidantN-acetylcysteine is usually considered a nutritional supplement. Our aim is to evaluate bioactivity ofN-acetylcysteine (NAC) on mature adipocytes, which is a close model to in vivo condition.Study Design: In vitro study.Place and Duration of Study: Department of Basic Science (Universidad Nacional de Lujan),Department of Chemical Biology (Universidad de Buenos Aires), CONICET ? INEDES andCONICET ? IQUIBICEN, between March 2017 and June 2019.Methodology: We evaluated the bioactivity of different concentrations of NAC for 5 days (0.01 mMto 5 mM) on fully differentiated 3T3-L1 cells (mature adipocytes).Results: We demonstrated that NAC treatment was not toxic to mature adipocytes. Only 5mM NACinhibited reactive oxygen species production. 5 mM NAC treatment resulted in a 60% decrease incellular triglycerides content and inhibited 70% cholesterol accumulation. We also determined themRNA and protein expression levels of Peroxisome Proliferator-Activated Receptor  as well as,mRNA levels of lipid protein Perilipin in NAC treated adipocytes; we observed that 5mM NACtreatment caused nearly 30% decrease in the expression of these parameters.Conclusion: These results suggest that NAC could avoid lipid accumulation in mature adipocytes;the antioxidant NAC could be beneficial in obesity treatment. Fil: Soto, Daniela. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Martini, Claudia Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Frontera, Evelyn Micaela. Universidad Nacional de Luján. Departamento de Ciencias Básicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Montaldo, Laura Andrea. Universidad Nacional de Luján. Departamento de Ciencias Básicas; Argentina Fil: Vila, Maria del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Calvo, Juan Carlos. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina Fil: Guerra, Liliana Noemi. Universidad Nacional de Luján. Instituto de Ecología y Desarrollo Sustentable. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Ecología y Desarrollo Sustentable; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina

En esta entrevista, la filologa y artista grafica Marika Vila analiza la presencia femenina en el discurso del comic espanol, contrastandola con su representacion en el discurso misogino y homofobo que ha dominado el medio desde sus origenes. Su trabajo de investigacion resalta las transformaciones de la narrativa grafica espanola desde los anos de la Transicion – cuando, con el advenimiento de la democracia, se asiste al nacimiento del comic de autor y al Boom del comic para adultos – hasta la epoca actual en la que se aprecia un gran surgimiento de obras de autoria femenina. Asimismo, gracias a un trabajo de investigacion de largo recorrido, Vila recupera la visibilidad y resistencia de las autoras pioneras. A partir de las preguntas de Marina Bettaglio, Trujillo revisa el pasado del comic espanol desde una perspectiva de genero para destacar la necesidad de rescatar el trabajo de las creadoras y entender su aporte a lo largo de su historia.

Lead (Pb) is an environmental and industrial contaminant that still represents a public health problem. Elevated Pb exposure has been inversely correlated with femoral bone density and associated with osteoporosis. In the last years, it has been shown that inhibition of osteogenesis from mesenchymal stem cells activates adipogenesis and vice versa. In this paper, we investigated the effect of Pb on the differentiation of 3T3-L1 fibroblasts to adipocytes which is the cell model most used to study adipogenesis. After induction of differentiation, 2 days post-confluent cells re-enter the cell cycle and undergo mitotic clonal expansion (MCE) followed by expression of genes that produce the adipocyte phenotype. The presence of concentrations of Pb up to 10 μM during differentiation of 3T3-L1 fibroblasts did not interfere with MCE but enhanced the accumulation of cytosolic lipids that occur during adipogenesis, as well as, the induction of PPARγ, the master gene in adipogenesis. It is known that PPARγ upregulation is subsequent to induction of C/EBPβ and ERK activation, which are early events in adipogenesis. We found that both events were enhanced by Pb treatment. Our results support a stimulatory effect of Pb on adipogenesis which involves ERK activation and C/EBPβ upregulation prior to PPARγ and adipogenesis activation. Fil: Martini, Claudia Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Gabrielli, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Bonifacino, Graciela. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Codesido, María Magdalena. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Vila, Maria del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina

espanolAntecedentes: el balon intragastrico Elipse® consiste en una capsula ingerible que se rellena bajo control radiologico. Pasadas 16 semanas, su valvula se degrada, el balon se vacia y se elimina por via natural, sin endoscopia. El objetivo del estudio es valorar su viabilidad, eficacia, duracion, seguridad y requerimientos endoscopicos. Metodos: estudio prospectivo y descriptivo, no aleatorizado, de los primeros pacientes remitidos para Elipse®. Tras colocacion, se realizaba sistematicamente una radiografia para confirmar su correcto rellenado. Se determinaba la duracion del balon en funcion de su visualizacion excretado o por radiografia/ecografia. Se recogieron datos de eficacia, tolerancia, efectos adversos y su resolucion (requerimientos endoscopicos), asi como el grado final de satisfaccion a las 16 semanas. Resultados: participaron en el estudio 30 pacientes con peso e indice de masa corporal (IMC) medios basales de 83,3 ± 10,7 kg y 30,6 ± 2,7 kg/m². Todos pudieron ingerir la capsula con correcto control radiologico. A los cuatro meses presentaban una perdida media de 11,2 ± 5,5 kg (12,1 ± 5,8% de perdida total de peso [PTP], 64,7 ± 25% de perdida del exceso de peso [PEP]), con perdida de peso > 10% en el 80% de pacientes (p EnglishBackground: the Elipse® intragastric balloon (EIGB) is a swallowable capsule that is filled under x-ray control. After 16 weeks, its self-releasing valve is degraded and the balloon is deflated and excreted naturally, without endoscopy. The aim of this study was to assess the feasibility of EIGB and its efficacy, duration, safety and endoscopic requirements. Methods: this is a prospective, descriptive, non-randomized study of the first patients enrolled for EIGB. An x-ray was systematically performed after placement to ensure the correct filling of the balloon. The balloon duration was determined according to its excreted visualization or by x-ray/ultrasound. The efficacy, tolerance, adverse events and their resolution outcome (endoscopic requirements), as well as the final satisfaction degree at 16 weeks, were analyzed. Results: the study included 30 patients with a basal mean weight and body mass index (BMI) of 83.3 ± 10.7 kg and 30.6 ± 2.7 kg/m². All subjects swallowed the capsule with correct x-ray control. The mean weight loss was 11.2 ± 5.5 kg (12.1 ± 5.8% of total weight loss [TWL], 64.7 ± 25% of excess weight loss [EWL]), with a weight loss > 10% in 80% of patients (p

Introduccion: la enfermedad hepatica grasa no alcoholica (EHGNA) es la causa mas frecuente de hepatopatia cronica en nuestro medio. Los beneficios de la endoscopia bariatrica en esta enfermedad estan escasamente documentados. Objetivo: evaluar los cambios de EHGNA, mediante metodos no invasivos, en pacientes obesos sometidos a endoscopia bariatrica restrictiva. Secundariamente, analizamos los cambios ponderales y metabolicos y el grado de seguridad tecnica. Metodos: treinta pacientes con EHGNA y obesidad (indice de masa corporal [IMC] medio de 38,22 ± 6,55 kg/m²) sometidos a endoscopia bariatrica restrictiva: 15 balones de un ano y 15 gastroplastias mediante suturas (ESG-Apollo®). Se realiza un analisis prospectivo no invasivo mediante parametros analiticos (funcion hepatica, insulinorresistencia, scores de esteatosis y fibrosis hepatica) y ecograficos. Ademas, se evalua la evolucion antropometrica y de las principales comorbilidades mayores asociadas, todo ello con seguimiento a un ano. Resultados: treinta pacientes (63% mujeres, edad media 46 ± 13,8 anos). A los 12 meses presentaron disminucion en FLI, HSI, NAFLD-Fibrosis Score, esteatosis hepatica ecografica y grasa subcutanea (p 25% en 27/30 pacientes (PP > 10% en 25/30 pacientes) (p < 0,001). Se objetivo resolucion en 17/30 (57%) de las comorbilidades asociadas: 5/8 (62,5%) hipertension arterial (HTA), 5/12 (41,7%) dislipemia (DLP), 2/4 (50%) diabetes mellitus tipo 2 (DM2), 2/3 (66,7%) sindrome de apnea obstructiva del sueno (SAOS) y 3/3 (100%) artropatia. Comparando ambos procedimientos, la mejoria en HbA1c en el grupo de ESG-Apollo (p = 0,017) fue la unica diferencia. Un balon migrado y expulsado espontaneamente fue la unica incidencia tecnica. Conclusiones: la endoscopia bariatrica puede postularse, en un seguimiento a corto plazo, como una alternativa eficaz y segura en aquellos pacientes obesos con EHGNA. Favorece la perdida de peso y mejora los biomarcadores analiticos y ecograficos de grasa hepatica, la insulinorresistencia, la hipertrigliceridemia y las comorbilidades mayores asociadas.

We investigated for the first time the expression of melanoma cell adhesion molecule (MCAM) and its involvement in the differentiation of 3T3-L1 fibroblasts to adipocytes. We found that MCAM mRNA increased subsequent to the activation of the master regulator of adipogenesis, PPARγ, and this increase was maintained in the mature adipocytes. On the other hand, MCAM knockdown impaired differentiation and induction of PPARγ as well as expression of genes activated by PPARγ. However, events that precede and are necessary for early PPARγ activation, such as C/EBPβ induction, β-catenin downregulation, and ERK activation, were not affected in the MCAM knockdown cells. In keeping with this, the increase in PPARγ mRNA that precedes MCAM induction was not altered in the knockdown cells. In conclusion, our findings suggest that MCAM is a gene upregulated and involved in maintaining PPARγ induction in the late but not in the early stages of 3T3-L1 fibroblasts adipogenesis. Fil: Gabrielli, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Romero, Damián Gastón. University of Mississippi; Estados Unidos Fil: Martini, Claudia Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Raiger Iustman, Laura Judith. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Vila, Maria del Carmen. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Glyphosate-based herbicides (GF) are extensively used for weed control. Thus, it is important to investigate their putative toxic effects. We have reported that GF at subagriculture concentrations inhibits proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts. In this investigation, we evaluated the effect of GF on genes upregulated during adipogenesis. GF was able to inhibit the induction of PPAR gamma, the master gene in adipogenesis but not C/EBP beta, which precedes PPAR gamma activation. GF also inhibited differentiation and proliferation of another model of preadipocyte: mouse embryonic fibroblasts. In exponentially growing 3T3-L1 cells, GF increased lipid peroxidation and the activity of the antioxidant enzyme, superoxide dismutase. We also found that proliferation was inhibited with lower concentrations of GF when time of exposure was extended. Thus, GF was able to inhibit proliferation and differentiation of preadipocytes and to induce oxidative stress, which is indicative of its ability to alter cellular physiology.

Glyphosate-based herbicides are extensively used for weed control all over the world. Thus, it is important to investigate the putative toxic effects of these commercial formulations which contain not only glyphosate but also different adjuvants. 3T3-L1 fibroblasts are a useful tool in the study of adipocyte differentiation. We have previously reported that a commercial glyphosate formulation inhibits proliferation and differentiation in these cells. In the present investigation, we further evaluated the effect of different commercial glyphosate-based herbicides on 3T3-L1 fibroblast proliferation and differentiation and compared their effect with glyphosate itself. After treatment of 3T3-L1 fibroblasts with different concentrations of three glyphosate formulations or glyphosate itself, the increase in cell number or cytosolic lipid accumulation was determined. To evaluate the presence of polyethoxylated adjuvants, matrix-assisted laser desorption/ionization (MALDI) analysis of the three commercial glyphosate-based herbicides was also performed. We found that the three commercial formulations tested were more potent inhibitors of both physiological processes: proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts than glyphosate. We also found by MALDI analysis that the adjuvants were polyethoxylated in two of the formulations but not in the third one. It is well known the toxic effect of polyethoxylated adjuvants, but according to our results, non-polyethoxylated adjuvants may also contribute to the toxic effects. Our results further support the ability of glyphosate-based herbicides to disturb cellular physiology and highlight the importance of toxicological assessment of commercial formulations rather than glyphosate alone. Fil: Martini, Claudia Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Gabrielli, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Codesido, María Magdalena. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Vila, Maria del Carmen. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina

Postnatal overfeeding increases the risk of chronic diseases later in life, including obesity, insulin resistance, hepatic steatosis, and type 2 diabetes. Epigenetic mechanisms might underlie the long-lasting effects associated with early nutrition. Here we aimed to explore the molecular pathways involved in early development of insulin resistance and hepatic steatosis, and we examined the potential contribution of DNA methylation and histone modifications to long-term programming of metabolic disease. We used a well-characterized mouse model of neonatal overfeeding and early adiposity by litter size reduction. Neonatal overfeeding led to hepatic insulin resistance very early in life that persisted throughout adulthood despite normalizing food intake. Up-regulation of monoacylglycerol O-acyltransferase ( Mogat) 1 conceivably mediates hepatic steatosis and insulin resistance through increasing intracellular diacylglycerol content. Early and sustained deregulation of Mogat1 was associated with a combination of histone modifications that might favor Mogat1 expression. In sum, postnatal overfeeding causes extremely rapid derangements of hepatic insulin sensitivity that remain relatively stable until adulthood. Epigenetic mechanisms, particularly histone modifications, could contribute to such long-lasting effects. Our data suggest that targeting hepatic monoacylglycerol acyltransferase activity during early life might provide a novel strategy to improve hepatic insulin sensitivity and prevent late-onset insulin resistance and fatty liver disease.-Ramon-Krauel, M., Pentinat, T., Bloks, V. W., Cebrià, J., Ribo, S., Pérez-Wienese, R., Vilà, M., Palacios-Marin, I., Fernández-Pérez, A., Vallejo, M., Téllez, N., Rodríguez, M. À., Yanes, O., Lerin, C., Díaz, R., Plosch, T., Tietge, U. J. F., Jimenez-Chillaron, J. C. Epigenetic programming at the Mogat1 locus may link neonatal overnutrition with long-term hepatic steatosis and insulin resistance., This work was supported by Instituto de Salud Carlos III (PI14/00035, CP11/00312 to J.J.C.), Ministerio de Economía y Competitividad (BFU2011-29739 to J.J.C.), the Netherlands Organization for Health Research and Development (ZonMW, Grant 91211053 to T.P.), and the Spanish Ministry of Economy and Competitiveness (BFU2014-52149-R to M.V.). I.P.-M. is supported by a National Council for Science and Technology in Mexico (CONACyT) Ph.D. Scholarship. M.R.-K. and T.P. share senior authorship.

RESUMEN Introducción: muchos de los pacientes sometidos a cirugía bariátrica (bypass gástrico en Y-de-Roux [RYGB]), con el tiempo, pueden recuperar parte del peso perdido. La reducción transoral del vaciamiento gástrico (TORe) con sutura endoscópica podría ser una alternativa válida en estos pacientes. Métodos: serie inicial retrospectiva que incluye a 13 pacientes consecutivos remitidos por reganancia ponderal tras RYGB y con anastomosis gastroyeyunal dilatada (> 15 mm). El TORe fue realizado mediante un dispositivo endoscópico de suturas transmurales (OverStitch-Apollo(r)), reduciendo el diámetro de la anastomosis y del reservorio gástrico. Se describen los datos iniciales de viabilidad técnica, seguridad y eficacia, con un seguimiento limitado a seis meses. Resultados: tras el RYGB, existía una pérdida media máxima de 37,69 kg y una posterior reganancia media de 21,62 kg. Se redujo el diámetro medio de la anastomosis de 36 mm (rango 20-45) a 9 mm (rango 5-12) (reducción del 75%) con una media de 2,5 suturas y el del reservorio de 7,2 cm (rango 2-10) a 4,7 cm (rango 4-5) (reducción del 34,72%) con una media de 2,7 suturas. La pérdida media de peso a los seis meses tras el TORe fue de 12,29 kg (pérdida del 56,85% del peso reganado tras RYGB). No se registraron complicaciones relacionadas con el procedimiento. Conclusiones: la reducción mediante sutura endoscópica de la anastomosis gastroyeyunal dilatada y del reservorio gástrico parece una opción viable y segura según nuestra limitada experiencia inicial. Dentro de un abordaje multidisciplinar y en un seguimiento a corto plazo, se presenta como una opción mínimamente invasiva y eficaz para controlar la reganancia ponderal tras RYGB. ABSTRACT Introduction: many patients that undergo bariatric surgery (Roux-en-Y gastric bypass [RYGB]) may regain some of their weight lost over time. A transoral outlet reduction (TORe) with endoscopic suture could be a valid alternative in these patients. Methods: this was a retrospective initial series of 13 consecutive patients with weight regain after RYGB and a dilated gastro-jejunal anastomosis (> 15 mm). TORe was performed using an endoscopic transmural suture device (OverStitch-Apollo(r)), which was used to reduce the anastomosis aperture and also to treat the gastric pouch. The initial data of feasibility, safety and weight loss are described with a limited follow-up of six months. Results: there was a mean maximum weight loss of 37.69 kg after RYGB and a subsequent average regain of 21.62 kg. The mean anastomosis diameter was 36 mm (range 20-45) which was reduced to 9 mm (range 5-12) (75% reduction), with an average of 2.5 sutures. The mean pouch size was 7.2 cm (range 2-10), which decreased to 4.7 cm (range 4-5) (34.72% reduction), with an average of 2.7 sutures. The mean weight loss six months after TORe was 12.29 kg, a weight loss of 56.85% of the weight regained after RYGB. No complications related to the procedure were recorded. Conclusions: endoscopic suture reduction of the dilated gastro-jejunal anastomosis and the gastric pouch seems a feasible and safe option in our limited initial experience. With a multidisciplinary approach and a short term follow-up, this seems to be a minimally invasive and effective option to control weight regain after RYGB.

The undercarboxylated form of osteocalcin (ucOC) is an emerging marker of cardiovascular disease. It is unknown if ucOC in related to common cardiovascular risk markers in children. In offspring of families with and without metabolic syndrome (MetS+ and MetS- families), we assessed whether ucOC was related to a continuous metabolic syndrome score (MetS score) and to carotid intima-media thickness (cIMT).ucOC and total OC, MetS score and cIMT were assessed in 203 asymptomatic prepubertal children (age 7.6 ± 0.1 yr; 49% girls), of whom 99 were from MetS+ families.In children from MetS+ families, percent ucOC was higher than in children from MetS- families (p0.01). In offspring from MetS+ families, higher ucOC and especially higher percent ucOC was independently associated with both the MetS score and cIMT (both p ≤ 0.01).The undercarboxylated form of OC is related to common cardiovascular risk markers in children at risk for cardiovascular disease.

Adipogenesis is stimulated in 3T3-L1 fibroblasts by a combination of insulin, dexamethasone and isobutylmethylxanthine, IBMX, (I+D+M). Two transcription factors are important for the acquisition of the adipocyte phenotype, C/EBP beta (CCAT enhancer-binding protein beta) and PPAR gamma (peroxisome proliferator-activated receptor gamma). IBMX increases cAMP content, which can activate protein kinase A (PKA) and/or EPAC (exchange protein activated by cAMP). To investigate the importance of IBMX in the differentiation mixture, we first evaluated the effect of the addition of IBMX on the increase of C/EBP beta and PPAR gamma and found an enhancement of the amount of both proteins. IBMX addition (I+D+M) or its replacement with a cAMP analogue, dibutyryl-cAMP or 8-(4-chlorophenylthio)-2-O′-methyl-cAMP (8CPT-2-Me-cAMP), the latter activates EPAC and not PKA, remarkably increased PPAR gamma mRNA. However, neither I+D nor any of the inducers alone, increased PPAR gamma mRNA to a similar extent, suggesting the importance of the presence of both IBMX and I+D. It was also found that the addition of IBMX or 8CPT-2-Me-cAMP was able to increase the content of C/EBP beta with respect to I+D. In agreement with these findings, a microarray analysis showed that the presence of either 8CPT-2-Me-cAMP or IBMX in the differentiation mixture was able to upregulate PPAR gamma and PPAR gamma-activated genes as well as other genes involved in lipid metabolism. Our results prove the involvement of IBMX-cAMP-EPAC in the regulation of adipogenic genes during differentiation of 3T3-L1 fibroblasts and therfore contributes to elucidate the role of cyclic AMP in this process. Fil: Gabrielli, Matias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Martini, Claudia Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina Fil: Brandani, Javier Nahuel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Raiger Iustman, Laura Judith. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Romero, Damián Gastón. University Of Mississippi; Estados Unidos Fil: Vila, Maria del Carmen. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Glyphosate-based herbicides are extensively used for weed control all over the world. Therefore, it is important to investigate the putative toxic effects of these formulations which include not only glyphosate itself but also surfactants that may also be toxic. 3T3-L1 fibroblasts are a useful tool to study adipocyte differentiation, this cell line can be induced to differentiate by addition of a differentiation mixture containing insulin, dexamethasone and 3-isobutyl-1-methylxanthine. We used this cell line to investigate the effect of a commercial formulation of glyphosate (GF) on proliferation, survival and differentiation. It was found that treatment of exponentially growing cells with GF for 48 h inhibited proliferation in a dose-dependent manner. In addition, treatment with GF dilution 1:2000 during 24 or 48 h inhibited proliferation and increased cell death, as evaluated by trypan blue-exclusion, in a time-dependent manner. We showed that treatment of 3T3-L1 fibroblasts with GF increased caspase-3 like activity and annexin-V positive cells as evaluated by flow cytometric analysis, which are both indicative of induction of apoptosis. It was also found that after the removal of GF, remaining cells were able to restore proliferation. On the other hand, GF treatment severely inhibited the differentiation of 3T3-L1 fibroblasts to adipocytes. According to our results, a glyphosate-based herbicide inhibits proliferation and differentiation in this mammalian cell line and induces apoptosis suggesting GF-mediated cellular damage. Thus, GF is a potential risk factor for human health and the environment.

Background: in recent years new endoscopic strategies and techniques for the treatment of obesity have emerged and develop ed. Aim of the study: in this article we will review and analyze the current state of the following techniques and the basic differential characteristics between each of them: balloons and prosthesis, injection of substances, systems of sutures, malabsorptives techniques and others currently in research. Methods: we will evaluate the endoscopic technique and their main indications, results, tolerances, complications and adverse effects observed, reporting our personal experience and in relation with an extensive literature review. Results: comparatively with the most widespread technique of the Bioenterics balloon, the Spatz balloon can provide greater weight loss but with worse tolerance and more complications and the Heliosphere Bag gets a similar weight loss but with greater technical difficulty. Other balloons and prosthesis (Ullorex, Semistationary, Silimed, Endogast) still require technical improvements and higher studies. The injection of botulinum toxin, although secure, seems to offer a smaller and more transient efficacy. Suture systems (TOGa, endoluminal vertical gastroplasty and POSE) appear to be effective but are technically more laborious. Malabsorptives procedures (Endobarrier, ValenTX) are somewhat laborious but effective, particularly indicated in obese patients with type 2 diabetes mellitus. Conclusions: the development of new endoscopic techniques and improvement in existing designs, suggest an increasingly important role of the endoscopist in the treatment of obesity. We consider it important to individually select and use the endoscopic technique, depending on the desirable outcomes (efficacy, tolerance, safety, adverse effects and risks) and the experience of each hospital. We believe that these techniques should be applied by specifically trained endoscopists in specialized hospitals.

OBJECTIVES: Protein intake impacts on nutritional status and may determine the recurrence of hepatic encephalopathy (HE). A low-protein diet has been considered the standard treatment after an episode of HE, while branched-chain amino acids (BCAA) have been shown to improve minimal HE. We performed a study to investigate the long-term effects of supplementing a protein-controlled diet with BCAA. METHODS: A randomized, double-blind, multicenter study that included 116 patients with cirrhosis and a previous episode of HE was conducted in four tertiary care hospitals. All patients received a standard diet of 35 kcal/kg per day and 0.7 g of proteins/kg per day and a supplement of 30 g of BCAA (BCAA group) or maltodextrin (MDX group) during 56 weeks. RESULTS: The actuarial risk of remaining free of HE did not differ between groups (BCAA = 47%, MDX = 34%, P = 0.274), but patients in the BCAA group exhibited a better outcome on two neuropsychological tests and an increase in the mid-arm muscle circumference. Recurrence was associated with low plasma albumin at baseline and a decrease in sodium and an increase in creatinine during follow-up. Patients with recurrence of HE exhibited a lack of improvement in global cognitive function. CONCLUSIONS: Diet supplementation with BCAA after an episode of HE does not decrease recurrence of HE. However, supplementation with BCAA improves minimal HE and muscle mass. Identification of risk factors for recurrence of HE may allow the development of new preventive therapies that could decrease the neuropsychological sequelae of repeated episodes of HE.

Adipogenesis is stimulated in 3T3-L1 fibroblast by a combination of insulin, dexamethasone, and methylisobutylxanthine (MIX). Mitotic clonal expansion (MCE) precedes differentiation of 3T3-L1 fibroblast to adipocytes. MIX increases cAMP content, which is the activator of protein kinase A (PKA). However, PKA-independent cAMP signaling has also been described. In this paper, it was found that H89, an inhibitor of PKA, was able to block MCE but not differentiation of 3T3-L1 fibroblast. Consistently, MCE did not occur in the absence of MIX in the differentiation mixture but was recovered by overexpression of a catalytic subunit of PKA. In addition, the transfection of 3T3-L1 fibroblast with a dominant-negative mutant of PKA inhibited MCE. On the other hand, differentiation of 3T3-L1 fibroblast to adipocytes did not occur when MIX was not present in the differentiation mixture and it could not be recovered by overexpression of a catalytic subunit of PKA. Differentiation was restored by addition of either dibutyryl-cAMP (db-cAMP) or 8 CPT-2 Me-cAMP. The latter activates cAMP-EPAC but not PKA signaling. These results indicate that cAMP-PKA-independent signaling, is required for 3T3-L1 fibroblasts differentiation to adipocytes and MIX signaling through cAMP-PKA is necessary for MCE, although MCE is not essential for adipogenesis.

Norfloxacin is highly effective in preventing spontaneous bacterial peritonitis recurrence in cirrhosis, but its role in the primary prevention of this complication is uncertain.Patients with cirrhosis and low protein ascitic levels (15 g/L) with advanced liver failure (Child-Pugh scoreor = 9 points with serum bilirubin levelor = 3 mg/dL) or impaired renal function (serum creatinine levelor = 1.2 mg/dL, blood urea nitrogen levelor = 25 mg/dL, or serum sodium levelor = 130 mEq/L) were included in a randomized controlled trial aimed at comparing norfloxacin (35 patients) vs placebo (33 patients) in the primary prophylaxis of spontaneous bacterial peritonitis. The main end points of the trial were 3-month and 1-year probability of survival. Secondary end points were 1-year probability of development of spontaneous bacterial peritonitis and hepatorenal syndrome.Norfloxacin administration reduced the 1-year probability of developing spontaneous bacterial peritonitis (7% vs 61%, P.001) and hepatorenal syndrome (28% vs 41%, P = .02), and improved the 3-month (94% vs 62%, P = .003) and the 1-year (60% vs 48%, P = .05) probability of survival compared with placebo.Primary prophylaxis with norfloxacin has a great impact in the clinical course of patients with advanced cirrhosis. It reduces the incidence of spontaneous bacterial peritonitis, delays the development of hepatorenal syndrome, and improves survival.

We have previously reported that ACTH activates a phospholipase C that hydrolyzes glycosylphosphatidylinositol (GPI), which would release inositolphosphoglycan (IPG) to the extracellular medium, and that an IPG purified from Trypanosoma cruzi is able to inhibit ACTH-mediated steroid production in adrenocortical cells. In the present paper, it was found that anti-inositolphosphoglycan antibodies (anti-CRD) increased ACTH-mediated corticosterone production, which indicates that an endogenous IPG is a physiological inhibitor of ACTH response. On the other hand, we investigated the release to the extracellular medium of the GPI-anchored enzyme, alkaline phosphatase, by ACTH. We found that: (a) the released enzyme appeared in the aqueous phase after Triton X-114 partitioning, consistent with loss of the GPI, (b) the phospholipase C inhibitor, U73122, impaired the release of the enzyme by the hormone and (c) two inhibitors of IPG uptake, inositol 2-monophosphate and 2 M NaCl, increased the amount of alkaline phosphatase in the extracellular medium. These results suggest that ACTH releases alkaline phosphatase by activation of a phospholipase C. Dibutyryladenosine-3′,5′-cyclic monophosphate (db-cAMP) was able to increase the release of alkaline phosphatase from adrenocortical cells and this effect was inhibited by U73122, suggesting that cAMP is involved in the activation of phospholipase C. In addition, it was found that a pertussis-toxin sensitive G-protein is required for ACTH- and db-cAMP-mediated release of alkaline phosphatase and that incorporation of anti-Gi antibodies in adrenocortical cells inhibited the release of alkaline phosphatase by ACTH. Our results suggest that ACTH increases the release of alkaline phosphatase by activation of a phospholipase C through cAMP and Gi which would contribute to produce IPG. It was also found that the two inhibitors of IPG uptake, inositol-2-monophosphate and 2 M NaCl, increased the amount of alkaline phosphatase in the extracellular medium of ACTH-treated cells more than in control cells, indicating that ACTH also stimulates the uptake of IPG. These data support a role of GPI and the involvement of Gi in ACTH action.

Reports about antioxidant effect in obesity are contradictory. We showed that N-acetylcysteine (NAC) inhibits cellular lipid accumulation during adipocyte differentiation, through the inhibition of adipogenic transcriptor factors expression, such as PPARγ and, MAPKs phosphorylation (Pieralisi et al,Redox Biol 2016; Soto el al,Redox Rep 2016). Here we evaluated NAC on fully differentiated cells (3T3-L1 adipocytes: AC). Treatments with 0.01 to 5 mM NAC, included in culture media for 5 days, were not toxic. 5mM NAC treatment on AC (ACN) provoked a decrease of 60% in cellular Triglycerides (Tg) content (1.22+0.09 gTg/g protein [AC] vs 0.49+0.03 gTg/g protein [ACN], p

Background: Prophylaxis with recombinant factor VIII (rFVIII) is considered the optimal treatment for severe or moderate haemophilia A (HA). Bleeding into joints determines a proportional chronic joint damage to its frequency and severity. Knowledge of the individual's pharmacokinetics (PK) using Bayesian analysis helps to individualize prophylaxis therapy with recombinant factor VIII (rFVIII) in severe or moderate haemophilia A (HA) and minimize the risk of bleeding, extracting only 2-3 samples. Methods Retrospective study in HA patients with rFVIII (Advate®) prophylaxis from January 2014 to May of 2016. Bayesian model (myPKFit®) was employed to perform an individual PK profile using the retrospective data of rFVIII levels. PK parameters analyzed were: clearance (Cl); steady state volume (Vss); plasma half-life (t1/2); and time to reach rFVIII levels Results Nineteen patients were analyzed, with a mean age of 32 years (SD: 11.3; range 11-46) and 86 PK monitoring (4.5 per patient). Two patients with p75 14.4 h), with average age of 19.5; 39.4 & 35.8 years, respectively. We detected significant differences between t1/2 and median values of joint state, but not in ABR and AJBR. The mean values of joint scores were: PS (5.5, 22.1; 17; p=0.028), GS (1, 25.9, 17.6; p= 0,008) and NMR (1.2, 2.6, 2.0; p=0.042) for short, normal and long t1/2, respectively. The limited number of patients only allows observe significant differences in patients with short t1/2, patients that also have significantly lower age (p=0.007). The younger age of these patients also justifies the lower joint damage observed. After excluding two patients Conclusion PK monitoring showed a low intraindividual variability in t1/2, but significant interindividual CV. Age could modify PK parameters, so it should be assessed in an integrated manner with other clinical variables. Bayesian estimate with MyPKFit® allows know the PK profile of each patient and could be a useful tool to individualize prophylaxis adjusting by the physical activity and the bleeding pattern. We are performing a personalized one-year program to identify and treat the specific causes of poor bleeding control in prophylaxis therapy, and these are our preliminary results. Acknowledgments: This study will be supported by Baxalta grant "H15-29403". Disclosures Cid: Baxalta Innovations GmbH, now part of Shire: Other: Investigator Clinical Studies.

Heavy metals contamination has become an important risk factor for public health and the environment. Chromium is a frequent industrial contaminant and is also used in orthopaedic joint replacements made from cobalt–chromium-alloy. Since hexavalent chromium (Cr(VI)) was reported as genotoxic and carcinogenic in different mammals, to further evaluate its cytotoxicity, we investigated the effect of this heavy metal in the proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts. These cells, after the addition of a mixture containing insulin, dexamethasone and methylisobutylxanthine, first proliferate, a process known as mitotic clonal expansion (MCE), and then differentiate to adipocytes. In this differentiation process a key transcription factor is induced: peroxisome proliferator-activated receptor gamma (PPAR gamma). We found that treatment of 3T3-L1 fibroblasts with potassium chromate inhibited proliferation in exponentially growing cells and MCE as well as differentiation. A decrease in PPAR gamma content, evaluated by western blot and immunofluorescence, was found in cells differentiated in the presence of chromium. On the other hand, after inhibition of differentiation with chromium, when the metal was removed, differentiation was recovered, which indicates that this may be a reversible effect. We also found an increase in the number of micronucleated cells after treatment with Cr(VI) which is associated with genotoxic effects. According to our results, Cr(VI) is able to inhibit proliferation and differentiation to adipocytes of 3T3-L1 fibroblasts and to increase micronucleated cells, which are all indicative of alterations in cellular physiology and therefore, contributes to further elucidate the cytotoxic effects of this heavy metal. Fil: Martini, Claudia Noemí. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Brandani, Javier Nahuel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gabrielli, Matias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vila, Maria del Carmen. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Long-term primary antibiotic prophylaxis of spontaneous bacterial peritonitis has been suggested to be useful in cirrhotic patients with low ascitic fluid protein levels. However, it is unlikely that all such patients need prophylactic treatment. The aim of this study was to identify the group of cirrhotic patients with low ascitic fluid protein levels at high risk of developing a first episode of spontaneous bacterial peritonitis during outpatient follow-up.One hundred nine cirrhotic patients with low ascitic fluid protein levels and without previous episodes of spontaneous bacterial peritonitis were followed up in an outpatient clinic.Twenty-eight patients developed a first spontaneous bacterial peritonitis episode. In the multivariate analysis, serum bilirubin level (3.2 mg /dL) and platelet count (98.000/mm(3)) independently correlated with the risk of developing the first spontaneous bacterial peritonitis (P0.01 and P0.05, respectively). According to the median relative risk coefficient, a low-risk group (relative risk1.09) and a high-risk group (relative risk1.09) were established. The probability of developing a first spontaneous bacterial peritonitis episode at 1-year follow-up was significantly higher in the high risk-group (low-risk group, 23.6%; high-risk group, 55%; P0.01) as a consequence of a higher probability of the first community-acquired episode (13.7% vs. 47.6%, respectively, P0.01). One-year probability of survival was significantly lower in the high-risk group (low-risk group, 57.6%; high-risk group, 38%, P0.05).Cirrhotic patients with low ascitic fluid protein levels (/=1 g /dL) and high bilirubin level and/or low platelet count are at high risk of developing a first episode of spontaneous bacterial peritonitis during long-term follow-up.

Interest in extra-adrenal corticosteroid synthesis has been revived by technological advances and the quest for answers to clinical problems. The cytochrome P450 21-hydroxylase converts progester-one to deoxycorticosterone, the obligatory substrate for the production of the main adrenal steroids aldosterone, cortisol and corticosterone. The rat P450 21-hydroxylase was cloned and two constructs, 21OH-5 and 21OH-6, sequenced. The constructs are similar, except that 21OH-6 has three additional major insertions of 64, 70 and 84 bp, a 3 bp deletion, and four extra base pairs immediately before the poly-A sequence. The entire coding region of 21OH-5 has 87 and 71% homology with the mouse and human 21-hydroxylase cDNA, respectively, whereas the encoded protein has 84 and 65% homology. Reverse transcriptase-polymerase chain reaction (RT-PCR) combined with Southern blot demonstrated expression of both transcripts in the kidney, aorta, liver, cerebellum, hypothalamus and brain stem, heart and cerebrum, but not the hippocampus, in addition to the adrenal. The entire coding region of 21OH-5 and the corresponding region of 21OH-6 including the three introns were cloned into pCR3 and the plasmids transiently transfected into COS-7 cells. Only 21OH-5 was translated into active protein, converting approximately 64% of 3H-progesterone to deoxycorticosterone in 2 h.

We describe the effect of an inositol phosphoglycan (IPG) purified from Trypanosoma cruzi on the stimulation of aldosterone and cAMP production by ACTH in calf adrenocortical cells. T. cruzi IPG has two galactofuranose residues (Galf) which are not frequent in other IPGs. The effect of IPG with galactofuranose residues (IPG Galf) and IPG without these residues (IPG) was investigated. It was found that IPG Galf slightly decreased the stimulation of aldosterone and cAMP production by ACTH, whereas IPG significantly inhibited ACTH-mediated accumulation of both aldosterone and cAMP. The inhibition of aldosterone content in ACTH-treated cells by IPG was dose dependent. It was also found that the pretreatment of calf adrenocortical cells with IPG inhibited the accumulation of aldosterone provoked by ACTH and dibutyryladenosine-3′,5′-cyclic monophosphate (db-cAMP). On the other hand, the activation of a GPI (glycosyl phosphatidylinositol)-phospholipase C by ACTH was evaluated. First it was found that the release of ceramide from a GPI-like molecule: a glycoinositol-phosphoceramide (LPPG) purified from T. cruzi is increased in ACTH-treated cells. Second, the release of alkaline phosphatase, a GPI-anchored enzyme, to the extracellular medium was increased in these cells by ACTH. These data suggest that ACTH activates a phospholipase C in calf adrenocortical cells, releasing IPG, which in turn may inhibit, or modulate ACTH action.

Submitted by Joana Lopes Dias (jdias@ubi.pt) on 2014-06-06T18:45:12Z No. of bitstreams: 1 Dissertação de Mestrado em Estudos Ibéricos - O Bilinguismo na Comunidade Autónoma da Catalunha.pdf: 5940589 bytes, checksum: 44c460277a943c2a947740153c1b368b (MD5) Made available in DSpace on 2014-06-06T18:45:12Z (GMT). No. of bitstreams: 1 Dissertação de Mestrado em Estudos Ibéricos - O Bilinguismo na Comunidade Autónoma da Catalunha.pdf: 5940589 bytes, checksum: 44c460277a943c2a947740153c1b368b (MD5) Previous issue date: 2010

We tested the hypothesis that the presence of bacterial DNA (bactDNA) in ascitic fluid and serum is associated with decreased survival in patients with cirrhosis. In a prospective, multicenter study, we analyzed the clinical evolution of 156 patients with cirrhosis and ascites (first or recurrence) with lower than 250 polymorphonuclear cells (PMN)/L, negative ascites bacteriological culture, and absence of other bacterial infections being admitted for evaluation of largevolume paracentesis, according to the presence of bactDNA at admission. Survival, causes of death, and successive hospital admissions were determined during a 12-month follow-up period. BactDNA was detected in 48 patients. The most prevalent identified bactDNA corresponded to Escherichia coli (n 32/48 patients, 66.6%). Patients were followed for 12 months after inclusion and in this period 34 patients died: 16 of 108 (15%) bactDNA negative versus 18 of 48 (38%) bactDNA positive (P 0.003). The most frequent cause of death was acute-on-chronic liver failure in both groups (7/16 and 9/18 in patients without or with bactDNA, respectively), although more prevalent in the first month of follow-up in patients with presence of bactDNA (0 versus 4/7). When considering patients with model for end-stage liver disease (MELD) score less than 15, mortality was significantly higher in those with presence of bactDNA. Spontaneous bacterial peritonitis developed similarly in patients with or without bactDNA at admission. Conclusion: The presence of bactDNA in a patient with cirrhosis during an ascitic episode is an indicator of poor prognosis. This fact may be related to the development of acute-on-chronic liver failure at short term and does not predict the development of spontaneous bacterial peritonitis. (HEPATOLOGY 2008;48:1924-1931.)

The extrapancreatic effects of sulfonylurea drugs include increased glucose uptake by certain peripheral tissues. To study this effect, we used BC3H1 myocytes, which are reported to respond to these drugs. Within 30 min, tolbutamide and glyburide increased [3H]-2-deoxyglucose uptake in a dose-dependent manner. The inactive analogue carboxytolbutamide had no effect on glucose transport. Because increases in glucose transport may be mediated by activation of the diacylglycerol-protein kinase C signaling system, we examined the effects of these drugs on lipid metabolism and protein kinase C activity. Unlike insulin, tolbutamide and glyburide failed to increase [3H]glycerol labeling of diacylglycerol or labeling of phospholipids by 32P. After 30 min of treatment with tolbutamide or glyburide, however, membraneassociated and cytosolic protein kinase C activity were each increased. When cells were treated with 12-O- tetradecanoylphorbol-13-acetate (TPA) for 48 h to deplete certain isoforms of protein kinase C, glyburide, tolbutamide, and acute TPA treatment failed to increase glucose uptake, suggesting that TPA and sulfonylureas operate through activation of a common pathway. The effect of glyburide was additive to TPA in stimulating glucose uptake at low but not high TPA concentrations. As with insulin and TPA, extracellular Ca2+ was not essential for sulfonylurea-stimulated glucose uptake. Staurosporine, a protein kinase C inhibitor, blocked glyburide-, tolbutamide-, and insulinstimulated glucose uptake. In intact cells, glyburide stimulated the phosphorylation of both 80,000-Mr and 40,000-Mr proteins, which are markers for protein kinase C activation. Addition of sulfonylureas directly to the protein kinase C assay system in vitro provoked dioleinlike effects, in that sensitivity of the enzyme to Ca2+ was increased. Our findings suggest that tolbutamide and glyburide increase glucose uptake in BC3H1 myocytes by a postreceptor mechanism, which may involve direct activation of protein kinase C.