MLKL overexpression leads to Ca2+ and metabolic dyshomeostasis in a neuronal cell model.

The necroptotic effector molecule MLKL accumulates in neurons over the lifespan of mice, and its downregulation has the potential to improve cognition through neuroinflammation, and changes in the abundance of synaptic proteins and enzymes in the central nervous system. Notwithstanding, direct evidence of cell-autonomous effects of MLKL expression on neuronal physiology and metabolism are lacking. Here, we tested whether the overexpression of MLKL in the absence of cell death in the neuronal cell line Neuro-2a recapitulates some of the hallmarks of aging at the cellular level. Using genetically-encoded fluorescent biosensors, we monitored the cytosolic and mitochondrial Ca2+ levels, along with the cytosolic concentrations of several metabolites involved in energy metabolism (lactate, glucose, ATP) and oxidative stress (oxidized/reduced glutathione). We found that MLKL overexpression marginally decreased cell viability, however, it led to reduced cytosolic and mitochondrial Ca2+ elevations in response to Ca2+ influx from the extracellular space. On the contrary, Ca2+ signals were elevated after mobilizing Ca2+ from the endoplasmic reticulum. Transient elevations in cytosolic Ca2+, mimicking neuronal stimulation, lead to higher lactate levels and lower glucose concentrations in Neuro-2a cells when overexpressing MLKL, which suggest enhanced neuronal glycolysis. Despite these alterations, energy levels and glutathione redox state in the cell bodies remained largely preserved after inducing MLKL overexpression for 24-48 h. Taken together, our proof-of-concept experiments are consistent with the hypothesis that MLKL overexpression in the absence of cell death contributes to both Ca2+ and metabolic dyshomeostasis, which are cellular hallmarks of brain aging.

Resistencia a la insulina en mujeres con síndrome de ovario poliquístico / Insulin resistance in women with polycystic ovary syndrome

RESUMEN Introducción: La resistencia a la insulina es frecuente en el síndrome de ovario poliquístico, con diferencias entre fenotipos y discrepancias sobre cómo medirla. Objetivo: Identificar trastornos de la sensibilidad y resistencia a la insulina en mujeres con síndrome de ovario poliquístico, y determinar si es mayor en el fenotipo clásico. Métodos: Incluyó 152 mujeres: 45 sin síndrome de ovario poliquístico (Grupo I); 46 con síndrome de ovario poliquístico clínico (Grupo II); 61 con síndrome de ovario poliquístico clásico (Grupo III). Se realizó prueba de tolerancia a la glucosa oral, se calcularon índices de sensibilidad o resistencia a la insulina en ayunas (HOMA-IR, I0/G0, FIRI, ISI, Belfiore, Bennet, Quicki, Raynaud) y en la prueba de tolerancia a la glucosa oral (Belfiore2, Ribel, Ins2glu2, ATI, IITotal, DATI/DATG, Matsuda, BetaHOMA). Se emplearon las pruebas de Kruskal-Wallis, Mann-Whitney y Chi cuadrado. Resultados: Las mujeres con síndrome de ovario poliquístico tenían más obesidad global y central (p / 0,05), más nivel de glucemia a los 30, 120 y 180 minutos de la prueba de tolerancia a la glucosa oral (p / 0,05) y de insulinemia a los 30, 60 y 120 (p / 0,0001), lo que fue mayor en el grupo III. Se diagnosticó intolerancia en ayunas en una mujer de cada grupo y tolerancia alterada en una del II y III. No hubo diferencias significativas entre grupos para los índices de sensibilidad o resistencia a la insulina en ayunas; ni del HOMA entre mujeres normopeso vs. sobrepeso-obesidad (p / 0,05). La mediana de los índices de la prueba de tolerancia a la glucosa oral fue menor para los de sensibilidad (Belfiore2, Ribel) y mayor para los de resistencia a la insulina (Ins2glu2, ATI, IITotal) en el Grupo III. El DATI/DATG, Matsuda y BetaHOMA no tuvieron diferencias significativas. Conclusiones: Las mujeres con síndrome de ovario poliquístico tienen mayor respuesta glucémica, resistencia a la insulina e hiperinsulinismo postsobrecarga de glucosa que las mujeres con función ovárica normal, más manifiesta en el fenotipo clásico. Los índices de ayuno son menos sensibles, independientemente del peso corporal. Tienen mayor utilidad: insulinemia a los 60 minutos de la prueba de tolerancia a la glucosa oral, Belfiore2, ATI e IITotal(AU)

ABSTRACT Introduction: Insulin resistance is common in polycystic ovary syndrome, with differences between phenotypes and discrepancies on how to measure it. Objective: To identify disorders of insulin sensitivity and resistance in women with polycystic ovarian syndrome and determine if the latter is greater in the classic phenotype. Methods: The study included 152 women. 45 of them had no polycystic ovary syndrome (Group I), 46 had clinical polycystic ovary syndrome (Group II) and 61 had classic polycystic ovary syndrome (Group III). Oral glucose tolerance test was performed, fasting insulin sensitivity or resistance indices (HOMA-IR, I0 / G0, FIRI, ISI, Belfiore, Bennet, Quicki, Raynaud) were calculated and the tolerance test to oral glucose (Belfiore2, Ribel, Ins2glu2, ATI, IITotal, DATI / DATG, Matsuda, BetaHOMA) was also assessed. Kruskal-Wallis, Mann-Whitney and Chi square tests were used. Results: Women with polycystic ovarian syndrome had more global and central obesity (p /0.05), more blood glucose level at 30, 120 and 180 minutes of the oral glucose tolerance test (p /0.05 ) and insulinemia at 30, 60 and 120 (p /0.0001), which was higher in group III. Fasting intolerance was diagnosed in one woman in each group and altered tolerance in one of group II and group III, respectively. There were no significant differences between groups for fasting insulin sensitivity or resistance indices, nor for HOMA among normal weight women vs. overweight-obesity (p / 0.05). The median indexes of the oral glucose tolerance test were lower for those of sensitivity (Belfiore2, Ribel) and higher for those of insulin resistance (Ins2glu2, ATI, IITotal) in Group III. The DATI / DATG, Matsuda and BetaHOMA had no significant differences. Conclusions: Women with polycystic ovarian syndrome have higher glycemic response, insulin resistance and post-overload glucose hyperinsulinism than women with normal ovarian function, which is more evident in the classical phenotype. Fasting rates are less sensitive, regardless of body weight. Tests such as insulinemia 60 minutes after the oral glucose tolerance, Belfiore 2, ATI and IITotal are most useful(AU)

Preparation of Palladium-Supported Periodic Mesoporous Organosilicas and their Use as Catalysts in the Suzuki Cross-Coupling Reaction.

Three periodic mesoporous materials, i.e., two organosilicas with either ethylene or phenylene bridges and one silica, have been used as supports for Pd nanoparticles. All Pd-supported samples (1.0 wt%) were prepared by the incipient wetness method and subsequently reduced in an H2 stream at 200 °C. Both hydrogen chemisorption and temperature programmed reduction experiments revealed significant differences depending on the support. Pd2+ species were more reducible on the mesoporous organosilicas than on their silica counterpart. Also, remarkable differences on the particle morphology were observed by transmission electron microscopy. All Pd-supported samples were active in the Suzuki cross-coupling reaction between bromobenzene and phenylboronic acid.

Once-daily atazanavir/ritonavir compared with twice-daily lopinavir/ritonavir, each in combination with tenofovir and emtricitabine, for management of antiretroviral-naive HIV-1-infected patients: 96-week efficacy and safety results of the CASTLE study.

BACKGROUND: Once-daily atazanavir/ritonavir demonstrated similar antiviral efficacy to twice-daily lopinavir/ritonavir over 48 weeks, with less gastrointestinal disturbance and a better lipid profile, in treatment-naive patients. METHODS: International, multicenter, open-label, 96-week noninferiority randomized trial of atazanavir/ritonavir 300/100 mg once daily vs lopinavir/ritonavir 400/100 mg twice daily, each in combination with fixed-dose tenofovir/emtricitabine 300/200 mg once daily, in antiretroviral-naive, HIV-1-infected patients. The primary end point was the proportion of patients with HIV RNA <50 copies/mL at 48 weeks. Results through 96 weeks are reported. RESULTS: Of 883 patients enrolled, 440 were randomized to atazanavir/ritonavir and 443 to lopinavir/ritonavir. At week 96, more patients receiving atazanavir/ritonavir achieved HIV RNA <50 copies/mL (74% vs 68%, P < 0.05) in the intent-to-treat analysis. On both regimens, 7% of subjects were virologic failures by 96 weeks. Bilirubin-associated disorders were greater in patients taking atazanavir/ritonavir. Treatment-related gastrointestinal adverse events were greater in patients taking lopinavir/ritonavir. Mean changes from baseline in fasting total cholesterol, non-high-density lipoprotein cholesterol, and triglycerides at week 96 were significantly higher with lopinavir/ritonavir (P < 0.0001). CONCLUSIONS: Noninferiority of atazanavir/ritonavir to lopinavir/ritonavir was confirmed at 96 weeks. Atazanavir/ritonavir had a better lipid profile and fewer gastrointestinal adverse events than lopinavir/ritonavir.

Tuberculosis pediátrica, MDR-TB y XDR-TB en la provincia de Buenos Aires durante el período 2002-2007 / Paediatric tuberculosis, MDR-TB and XDR-TB in Buenos Aires province during the period 2002-2007

Los objetivos de este estudio fueron: describir la incidencia de tuberculosis (TB), MDR-TB Y XDR-TB en niños y adultos, y analizar la transmisión entre contactos comprobados. Fue realizado un análisis retrospectivo de los casos notificados al Programa de Control de Tuberculosis de Buenos Aires (PCTB). Durante este lapso se notificaron 30784 casos nuevos de TB, 4276 (14.0%) niños y 26508 adultos. El 4.9% (14/284) y el 71 % (146/ 2042) de los niños y adultos tenían MDR-TB, 5.1 % (13/256) de los niños y 5.3% (86/ 1616) de los adultos eran vírgenes de tratamiento. Se registraron 12 casos de XDR-TB en adultos, 3 sin tratamiento previo. Además 25.0% (40/160) de los casos MDR pertenecían a 9 familias o eran trabajadores de salud. La transmisión en estos casos fue verificada por pruebas moleculares. La evaluación del tratamiento para casos con MDR-TB o XDR-TB pudo realizarse en 47.5% de los casos ya que de 76 adultos y 6 niños no se obtuvo información. Asimismo, se pone de manifiesto la necesidad de contar con una notificación exhaustiva y un riguroso estudio de cohortes que permitan conocer y evaluar la situación real de la TB en la provincia. Debería asimismo recomendarse el incremento en la realización de cultivos y pruebas de sensibilidad que permitan determinar el perfil de resistencia bacteriana para tratar apropiadamente la TBP y la MDR- TB, indicadores de transmisión activa y del impacto de las actividades del PCTB en la comunidad.(AU)

The aims of the study were to describe the incidence of tuberculosis (TB), multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) in children and adults and to analyze the transmission of infection among proved contacts. The method was a retrospective analysis of notified cases to the TB Control Programme of Buenos Aires province. During the period 2002-2007, 30784 TB cases were notified; 4276 (14.0%) were children and 26508 adults. The prevalence of MDR-TB was 4.9% (14/ 284) among children and 7.1 % (146/2042) among adults. The proportions of MDR-TB cases who had not had any previous TB treatment were 5.1 % (13/256) in children and 5.3% (86/1616) in adults. A total of 12 XDR-TB cases were notified in adults; 3 of them without any previous TB treatment. Among the 160 MDR-TB cases, 40 belonged to 9 families or were health workers. Transmission in these cases was traced by molecular epidemiology tests. Data on evaluation of treatment outcomes were only available for 47.5% of MDR-TB cases; no information was registered in 76 adults and 6 children. Intensive notification and strict follow-up of cohort of cases are needed in arder to know and assess the actual TB situation in the province. It is also recommended to increase the use of cultures and drug susceptibility tests to have better estimates of the prevalence of drug-resistant TB, prescribe more effective treatment of TB and MDR- TB, improve indicators of infection transmission and better assess the impact of the TB control activities in the community.(AU)

Tuberculosis pediátrica, MDR-TB y XDR-TB en la provincia de Buenos Aires durante el período 2002-2007 / Paediatric tuberculosis, MDR-TB and XDR-TB in Buenos Aires province during the period 2002-2007

Los objetivos de este estudio fueron: describir la incidencia de tuberculosis (TB), MDR-TB Y XDR-TB en niños y adultos, y analizar la transmisión entre contactos comprobados. Fue realizado un análisis retrospectivo de los casos notificados al Programa de Control de Tuberculosis de Buenos Aires (PCTB). Durante este lapso se notificaron 30784 casos nuevos de TB, 4276 (14.0%) niños y 26508 adultos. El 4.9% (14/284) y el 71 % (146/ 2042) de los niños y adultos tenían MDR-TB, 5.1 % (13/256) de los niños y 5.3% (86/ 1616) de los adultos eran vírgenes de tratamiento. Se registraron 12 casos de XDR-TB en adultos, 3 sin tratamiento previo. Además 25.0% (40/160) de los casos MDR pertenecían a 9 familias o eran trabajadores de salud. La transmisión en estos casos fue verificada por pruebas moleculares. La evaluación del tratamiento para casos con MDR-TB o XDR-TB pudo realizarse en 47.5% de los casos ya que de 76 adultos y 6 niños no se obtuvo información. Asimismo, se pone de manifiesto la necesidad de contar con una notificación exhaustiva y un riguroso estudio de cohortes que permitan conocer y evaluar la situación real de la TB en la provincia. Debería asimismo recomendarse el incremento en la realización de cultivos y pruebas de sensibilidad que permitan determinar el perfil de resistencia bacteriana para tratar apropiadamente la TBP y la MDR- TB, indicadores de transmisión activa y del impacto de las actividades del PCTB en la comunidad.

The aims of the study were to describe the incidence of tuberculosis (TB), multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) in children and adults and to analyze the transmission of infection among proved contacts. The method was a retrospective analysis of notified cases to the TB Control Programme of Buenos Aires province. During the period 2002-2007, 30784 TB cases were notified; 4276 (14.0%) were children and 26508 adults. The prevalence of MDR-TB was 4.9% (14/ 284) among children and 7.1 % (146/2042) among adults. The proportions of MDR-TB cases who had not had any previous TB treatment were 5.1 % (13/256) in children and 5.3% (86/1616) in adults. A total of 12 XDR-TB cases were notified in adults; 3 of them without any previous TB treatment. Among the 160 MDR-TB cases, 40 belonged to 9 families or were health workers. Transmission in these cases was traced by molecular epidemiology tests. Data on evaluation of treatment outcomes were only available for 47.5% of MDR-TB cases; no information was registered in 76 adults and 6 children. Intensive notification and strict follow-up of cohort of cases are needed in arder to know and assess the actual TB situation in the province. It is also recommended to increase the use of cultures and drug susceptibility tests to have better estimates of the prevalence of drug-resistant TB, prescribe more effective treatment of TB and MDR- TB, improve indicators of infection transmission and better assess the impact of the TB control activities in the community.

Once-daily atazanavir/ritonavir versus twice-daily lopinavir/ritonavir, each in combination with tenofovir and emtricitabine, for management of antiretroviral-naive HIV-1-infected patients: 48 week efficacy and safety results of the CASTLE study.

BACKGROUND: Atazanavir/ritonavir is as effective as lopinavir/ritonavir, with a more favourable lipid profile and less gastrointestinal toxicity, in treatment-experienced HIV-1-infected patients. We compared these two combinations directly in treatment-naive patients. METHODS: In this open-label, international non-inferiority study, 883 antiretroviral-naive, HIV-1-infected patients were randomly assigned to receive atazanavir/ritonavir 300/100 mg once daily (n=440) or lopinavir/ritonavir 400/100 mg twice daily (n=443), in combination with fixed-dose tenofovir/emtricitabine 300/200 mg once daily. Randomisation was done with a computer-generated centralised randomisation schedule and was stratified by baseline levels of HIV RNA (viral load) and geographic region. The primary endpoint was the proportion of patients with viral load less than 50 copies per mL at week 48. The main efficacy analysis was done by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00272779. FINDINGS: At week 48, 343 (78%) of 440 patients receiving atazanavir/ritonavir and 338 (76%) of 443 patients receiving lopinavir/ritonavir had achieved a viral load of less than 50 copies per mL (difference 1.7%, 95% CI -3.8 to 7.1). Mean increases from baseline in CD4 cell count were similar (203 cells per muL in the atazanavir/ritonavir group vs 219 cells per muL in the lopinavir/ritonavir group). 25 (6%) patients in the atazanavir/ritonavir group and 26 (6%) in the lopinavir/ritonavir group were virological failures by week 48. Only two patients, both in the atazanavir/ritonavir group, had non-polymorphic protease inhibitor resistance mutations emerge on treatment, which conferred phenotypic resistance to atazanavir in one patient. Serious adverse events were noted in 51 (12%) of 441 patients in the atazanavir/ritonavir group and in 42 (10%) of 437 patients in the lopinavir/ritonavir group. Fewer patients in the atazanavir/ritonavir group than in the lopinavir/ritonavir group experienced grade 2-4 treatment-related diarrhoea (10 [2%] vs 50 [11%]) and nausea (17 [4%] vs 33 [8%]). Grade 2-4 jaundice was seen in 16 (4%) of 441 patients in the atazanavir/ritonavir group versus none of 437 patients in the lopinavir/ritonavir group; grade 3-4 increases in total bilirubin were seen in 146 (34%) of 435 patients on atazanavir/ritonavir and in one (<1%) of 431 patients on lopinavir/ritonavir. INTERPRETATION: In treatment-naive patients, atazanavir/ritonavir once-daily demonstrated similar antiviral efficacy to lopinavir/ritonavir twice-daily, with less gastrointestinal toxicity but with a higher rate of hyperbilirubinaemia.

Cumplimiento con las guías nacionales en pacientes hospitalizados con neumonía adquirida en la comunidad: Resultados del estudio capo en Argentina / Adherence with National Guidelines in hospitalized patients with community-acquired pneumonia: Results of CAPO study in Argentina

Un comité de seis sociedades científicas en Argentina desarrolló guías para la atención de pacientes con neumonía adquirida en la comunidad (NAC). El objetivo de este estudio fue evaluar el nivel de cumplimiento con el cuidado recomendado en las guías nacionales en pacientes hospitalizados con NAC en Argentina. Usando indicadores de calidad, la atención de 436 pacientes hospitalizados en 12 centros de Argentina fue evaluada en las áreas de hospitalización, oxigenoterapia, terapia antibiótica empírica, terapia switch, alta hospitalaria y prevención. El nivel de cumplimiento fue clasificado como óptimo (>90%), intermedio (60% a 90%), y bajo (<60%). La selección de antibióticos empíricos y la administración dentro de las primeras ocho horas de admisión tuvieron una adherencia superior al 80%. Se identificó un nivel de cumplimiento bajo en las áreas del pasaje de antibióticos de la vía endovenosa a la vía oral (53%), la prevención con vacunación antineumocóccica (51%) y el ofrecimiento para dejar de fumar (29%). Usando indicadores de calidad se pueden identificar áreas específicas de atención en pacientes con NAC con un bajo nivel de cumplimiento con las guías nacionales. En Argentina se deben desarrollar inicialmente intervenciones enfocadas a mejorar el manejo del pasaje de antibióticos de la vía endovenosa a la vía oral y prevención de la NAC.

A committee of six scientific organizations from Argentina developed guidelines for the management of patients with community-acquired pneumonia (CAP).The objective of this study was to evaluate the level of adherence with the recommended care suggested by the guidelines in patients hospitalized with CAP in Argentina. Using quality indicators the management of 436 patients who were hospitalized in 12 Argentinean institutions was evaluated. The care given among the following areas was reviewed: need for hospitalization, need for oxygen therapy, empiric antibiotic therapy, switch therapy, hospital discharge, and prevention. The level of adherence was classified as optimal (>90%), intermediate (60% to 90%), and low (<60%).The selection of the empiric antibiotic therapy and the administration of antibiotics between the first 8 hours after arrival had an adherence greater to 80%. A low level of adherence was found in the areas of switch therapy (53%), prevention of CAP with pneumococcal vaccine (51%) and smoking cessation offered (29%). Using quality indicators it is possible to identify specific areas of management in patients with CAP to a low level of adherence with national guidelines. In Argentina interventions to improve switch therapy and prevention of CAP should be developed.

[Adherence with national guidelines in hospitalized patients with community-acquired pneumonia. Results of CAPO study in Argentina]. / Cumplimiento con las guías nacionales en pacientes hospitalizados con neumonía adquirida en la comunidad. Resultados del estudio CAPO en Argentina.

A committee of six scientific organizations from Argentina developed guidelines for the management of patients with community-acquired pneumonia (CAP).The objective of this study was to evaluate the level of adherence with the recommended care suggested by the guidelines in patients hospitalized with CAP in Argentina. Using quality indicators the management of 436 patients who were hospitalized in 12 Argentinean institutions was evaluated. The care given among the following areas was reviewed: need for hospitalization, need for oxygen therapy, empiric antibiotic therapy, switch therapy, hospital discharge, and prevention. The level of adherence was classified as optimal (>90%), intermediate (60% to 90%), and low (<60%).The selection of the empiric antibiotic therapy and the administration of antibiotics between the first 8 hours after arrival had an adherence greater to 80%. A low level of adherence was found in the areas of switch therapy (53%), prevention of CAP with pneumococcal vaccine (51%) and smoking cessation offered (29%). Using quality indicators it is possible to identify specific areas of management in patients with CAP to a low level of adherence with national guidelines. In Argentina interventions to improve switch therapy and prevention of CAP should be developed.