Molecular cloning and sequence determination of four different cDNA species coding for α-subunits of G proteins from Xenopus laevis oocytes

AbstractA cDNA library preprared from Xenopus laevis oocytes in λgt10 was screened with a mixture of three oligonucleotide probes designed to detect sequences found in different mammalian genes coding for a-subunits of G-proteins. In addition to a clone coding for a Gαo-type subunit previously reported [(1989) FEBS Lett. 244, 188-192] four additional clones have been found coding for different Gα protein subunits. By comparison with mammalian α-subunits, these oocyte cDNAs correspond to two closely related Gas-la, to a Gαi-1 and to a Gαi-3 species. The derived amino acid sequences showed that both Gαs species contain 379 residues, corresponding to the short species without the serine residue and with a calculated Mr of 42720. The Gαi-1 gene encodes a 354 amino acid protein with an Mr, of 39000 and the Gαi-3 encodes an incomplete open reading frame of 345 residues, lacking the first 9 amino acid residues at the NH2, terminus. All these Gα-subunits showed high identity with their respective mammalian counterparts (75–80%), indicating a great degree of conservation through the evolution and the important cellular regulatory function that they play

Drosophila melanogaster Scramblases modulate synaptic transmission

Scramblases are a family of single-pass plasma membrane proteins, identified by their purported ability to scramble phospholipids across the two layers of plasma membrane isolated from platelets and red blood cells. However, their true in vivo role has yet to be elucidated. We report the generation and isolation of null mutants of two Scramblases identified in Drosophila melanogaster. We demonstrate that flies lacking either or both of these Scramblases are not compromised in vivo in processes requiring scrambling of phospholipids. Instead, we show that D. melanogaster lacking both Scramblases have more vesicles and display enhanced recruitment from a reserve pool of vesicles and increased neurotransmitter secretion at the larval neuromuscular synapses. These defects are corrected by the introduction of a genomic copy of the Scramb 1 gene. The lack of phenotypes related to failure of scrambling and the neurophysiological analysis lead us to propose that Scramblases play a modulatory role in the process of neurotransmission

Sewershed surveillance as a tool for smart management of a pandemic in threshold countries. Case study: Tracking SARS-CoV-2 during COVID-19 pandemic in a major urban metropolis in northwestern Argentina

Wastewater-based epidemiology is an economical and effective tool for monitoring the COVID-19 pandemic. In this study we proposed sampling campaigns that addressed spatial-temporal trends within a metropolitan area. This is a local study of detection and quantification of SARS-CoV-2 in wastewater during the onset, rise, and decline of COVID-19 cases in Salta city (Argentina) over the course of a twenty-one-week period (13 Aug to 30 Dec) in 2020. Wastewater samples were gathered from 13 sewer manholes specific to each sewershed catchment, prior to convergence or mixing with other sewer lines, resulting in samples specific to individual catchments with defined areas. The 13 sewershed catchments selected comprise 118,832 connections to the network throughout the city, representing 84.7 % (534,747 individuals) of the total population. The number of COVID19-related exposure and symptoms cases in each area were registered using an application developed for smartphones by the provincial government. Geographical coordinates provided by the devices were recorded, and consequently, it was possible to geolocalise all app-cases and track them down to which of the 13 sampling catchments belonged. RNA fragments of SARS-CoV-2 were detected in every site since the beginning of the monitoring, anticipating viral circulation in the population. Over the course of the 21-week study, the concentrations of SARS-CoV-2 ranged between 1.77 × 104 and 4.35 × 107 genome copies/L. There was a correspondence with the highest viral load in wastewater and the peak number of cases reported by the app for each catchment. The associations were evaluated with correlation analysis. The viral loads of SARS-CoV-2 in wastewater were a feasible means to describe the trends of COVID-19 infections. Surveillance at sewershed scale, provided reliable and strategic information that could be used by local health stakeholders to manage the COVID-19 pandemic.Fil: Cruz, Mercedes Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; ArgentinaFil: Sanguino Jorquera, Diego Gastón. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; ArgentinaFil: Aparicio González, Mónica Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; ArgentinaFil: Irazusta, Verónica Patricia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; Argentina. Universidad Nacional de Salta. Facultad de Ciencias Naturales; ArgentinaFil: Poma, Hugo Ramiro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; ArgentinaFil: Cristobal, Hector Antonio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; Argentina. Universidad Nacional de Salta. Facultad de Ciencias Naturales; ArgentinaFil: Rajal, Verónica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Salta. Instituto de Investigaciones para la Industria Química. Universidad Nacional de Salta. Facultad de Ingeniería. Instituto de Investigaciones para la Industria Química; Argentina. Nanyang Technological University; Singapu

Productive and reproductive performance and metabolic profiles of ewes supplemented with hydroponically grown green wheat (Triticum aestivum L.)

Inclusion of high levels of concentrate in the diets of late gestation and lactating ewes to improve productive and reproductive performance is a common practice. However cost-effective alternate feeding strategies for small ruminants must be developed and evaluated in order to counteract sustainability issues of feeding them concentrate feeds (Alexandre and Mandonnet, 2005). Hydroponically grown green forages are a potential high feed quality feedstuff in arid and semiarid regions of the world (Al-Faraki and Al-Hashimi, 2012). The nutritive value and fermentative characteristics of hydroponically grown forages positively influenced the performance of late gestation and lactating ewes (Herrera et al., 2010; Gebremedhin, 2015). Earlier investigations emphasized effects of dietary quality on endocrine and metabolic profiles in ewes during pregnancy and lactation (Lemley et al., 2014; Vonnahme et al., 2013). However adequate nutritional status of ewes is associated with favorable productive and reproductive performance whereby blood glucose, non-esterified fatty acids (NEFA) and blood urea nitrogen (BUN) are utilized to sustain a desirable protein and energy balance in ewes during gestation and lactation (Hatfield et al., 1999). Changes in metabolic hormones, such as insulin, play an important role in metabolic adaptation to changes in body weight (BW) and body condition while providing diagnostic information to evaluate ewe nutritional status (Caldeira et al., 2007). Cortisol may be particularly important in this regard as it is the predominant glucocorticoid in sheep blood and has been used as a reliable physiological endpoint to determine ewe responses to a variety of physiological, physical and environmental stress (Moolchandani et al., 2008). A paucity of information is available with respect to the metabolic profile and performance during mating, gestation and lactation of ewes fed diets containing hydroponically grown green wheat (HGW). Thus this experiment was conducted to determine effects of replacement of dry-rolled corn (DRC) and cottonseed meal (CSM) by HGW in an oat hay-based diet on the metabolic profile as well as the productive and reproductive performance of Katahdin female lambs.Twenty six Katahdin ewes (i.e., female lambs from breeding to 2 mo of their 1st lactation) were used in a completely randomized design (13/treatment)to evaluate effects of replacement of dietary dry-rolled corn grain (DRC) and cottonseed meal(CSM) with hydroponically grown whole plant green wheat (HGW; Triticum aestivum L.) on productive parameters and blood metabolites during mating, gestation and lactation, and on body weight (BW) gain of their lambs in their 1st 60 days of age. The gestation diet contained 70% oat hay, 20% rolled corn grain and 10% cottonseed meal, while the lactation diet contained 50% oat hay, 20% DRC and 30% CSM. Treatments consisted of total replacement of DRC and CSM with HGW in the gestation diet, while in the lactation diet HGW replaced 100% of the DRC and 33% of the CSM. There were no diet effects on reproductive parameters, and substitution of DRC and CSM with HGW did not affect dry matter intake during gestation and lactation. The BW gain of the lambs that were fed HGW did not differ from controls in the first 2 months of gestation, while it was lower (P < 0.05) at the last 3 months of gestation. Feeding HGW did not affect birth BW of lambs or subsequent BW gains through 60 days of age. Plasma non-esterified fatty acids (NEFA) were not affected by the diets fed during gestation, but were 56% lower (P < 0.05) at day 60 of lactation. Plasma glucose was only lower (P < 0.05) at day 90 of gestation, and blood urea nitrogen was only lower (P < 0.05) at day 30 of lactation. There were no effects of diets on plasma insulin, cortisol or progesterone during gestation and lactation. Hydroponically grown green wheat is a suitable substitute for a portion of the DRC and CSM in ewes diets during gestation and lactation without negative effects

Results of the first European Source Apportionment intercomparison for Receptor and Chemical Transport Models

In this study, the performance of the source apportionment model applications were evaluated by comparing the model results provided by 44 participants adopting a methodology based on performance indicators: z-scores and RMSEu, with pre-established acceptability criteria. Involving models based on completely different and independent input data, such as receptor models (RMs) and chemical transport models (CTMs), provided a unique opportunity to cross-validate them. In addition, comparing the modelled source chemical profiles, with those measured directly at the source contributed to corroborate the chemical profile of the tested model results. The most used RM was EPA- PMF5. RMs showed very good performance for the overall dataset (91% of z-scores accepted) and more difficulties are observed with SCE time series (72% of RMSEu accepted). Industry resulted the most problematic source for RMs due to the high variability among participants. Also the results obtained with CTMs were quite comparable to their ensemble reference using all models for the overall average (>92% of successful z-scores) while the comparability of the time series is more problematic (between 58% and 77% of the candidates’ RMSEu are accepted). In the CTM models a gap was observed between the sum of source contributions and the gravimetric PM10 mass likely due to PM underestimation in the base case. Interestingly, when only the tagged species CTM results were used in the reference, the differences between the two CTM approaches (brute force and tagged species) were evident. In this case the percentage of candidates passing the z-score and RMSEu tests were only 50% and 86%, respectively. CTMs showed good comparability with RMs for the overall dataset (83% of the z-scores accepted), more differences were observed when dealing with the time series of the single source categories. In this case the share of successful RMSEu was in the range 25% - 34%.JRC.C.5-Air and Climat

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to &lt;90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], &gt;300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of &lt;15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P&lt;0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P&lt;0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Kinship determination using DNA markers Estudios de parentesco mediante marcadores del ADN: Experiencia en resolución de casos en los últimos seis años

Background: Autosomal and Y chromosome short tandem repeats (STRs) and mitochondrial DNA polymorphisms are the most commonly used molecular tools for determination of kinship. Aim: To report a revision of 1,120 kinship cases (paternity and others) analyzed in our laboratory. Material and methods: Revision of all kinship cases analyzed between years 2001-2006. Autosomal and Y chromosome STRs and mitochondrial DNA polymorphisms were analyzed in DNA extracted from blood samples. Results: Paternity was excluded in 27.2% of cases. This figure did not change significantly along years. Most paternity exclusions were confirmed by the discordance in 5 genetic markers (30.5%), followed by exclusion of 4 and 6 genetic markers (20.3 and 20% respectively). Two studied cases were paternal and maternal exclusions, corresponding to a change of children between two families. In one case, the paternal line was assessed through Y chromosome markers, studying 16 STRs of this chromosome, positively confir

La Red Protierra-Chile y sus proyecciones

En el 2013, se constituyó la Red PROTIERRA-Chile de carácter voluntario. Ésta es una red de integración, cooperación técnica de ámbito nacional, que actúa en el desarrollo de la arquitectura y construcción con tierra. La principal misión de la Red es promover y generar conocimiento para el uso del material tierra en la construcción y la arquitectura, en las áreas de “Asentamientos humanos y vivienda en zonas rurales y urbanas” y “Preservación de la diversidad cultural y del patrimonio”. PROTIERRA-Chile contribuye a la promoción del desarrollo sustentable, desde un enfoque integral, considerando los ámbitos social, ambiental, técnico, cultural, económico con la participación de la sociedad civil contribuyendo al desarrollo sustentable en Chile. Las actividades más relevantes realizadas por la red, son las siguientes: la celebración del primer 1º Encuentro Nacional de Arquitectura y Construcción con Tierra-ENACOT; el desarrollo de un registro nacional de intervenciones patrimoniales, de obra nueva y obras menores construidas con tierra; el desarrollo del Prototipo de Vivienda de Emergencia con técnicas de tierra; un plan de viviendas para comunidad rural de la etnia Pehuenche, localizado en el lago Icalma en la región de la Araucanía al sur del país. Todos estas iniciativas y proyectos son congruentes con los objetivos de la red PROTIERRA-Chile y la Red Iberoamericana PROTERRA. Éstos presentan distintos grados de avance y desarrollo y han logrado activar las actividades de la Red en el corto periodo de funcionamiento de ésta. Con lo anterior, se busca diseminar y potenciar la construcción con tierra en Chile, sus prestaciones y potencialidades

Integración, inclusión y justicia social: Reflexiones desde las normativas inclusivas en la educación chilena

Desde 2009, el sistema educacional chileno ha vivido una serie de ajustes y reformas estructurales tendientes a construir escuelas inclusivas que responden a las demandas sociales de equidad y calidad. Este proceso, con continuidades y rupturas, es abordado en este estudio para comprender los sentidos y alcances de lo que se ha comprendido por sistema educativo inclusivo orientado a la justicia social. En este contexto presentamos el despliegue analítico de una investigación cualitativa desarrollada entre los años 2017-2019 que cuestiona sobre los sentidos discursivos latentes en las políticas educativas en Chile en torno a la diversidad e inclusión en el sistema educacional. Se desarrolla una investigación política documental cualitativa, con un enfoque teórico hermenéutico con aspectos críticos. Este proceso investigativo ha encontrado que las configuraciones y reestructuraciones -a nivel paradigmático y social- que han emergido desde las políticas educativas responden a una fuente integradora y no a una matriz inclusiva; esto bajo una declaración discursiva relativa a la intención de entregar servicios educacionales de calidad y equitativos a la población escolar, que presenten o no condición de discapacidad. Otro resultado destacado en este artículo hace referencia a los cambios  emergentes que se observaron durante el tránsito desde el enfoque integrativo al inclusivo; aspectos como los modos de materialización del discurso inclusivo en las aulas regulares, aquellos que proponen la diversificación de enseñanza y la eliminación de barreras frente al aprendizaje. Finalmente, el artículo reflexiona sobre el sustento y el análisis del recorrido de las políticas públicas educativas de carácter inclusivo que se han promulgado en la última década en Chile, declarando los silencios y contradicciones entre ellas