mu-Opioid inhibition of Ca2+ currents and secretion in isolated terminals of the neurohypophysis occurs via ryanodine-sensitive Ca2+ stores

mu-Opioid agonists have no effect on calcium currents (I(Ca)) in neurohypophysial terminals when recorded using the classic whole-cell patch-clamp configuration. However, mu-opioid receptor (MOR)-mediated inhibition of I(Ca) is reliably demonstrated using the perforated-patch configuration. This suggests that the MOR-signaling pathway is sensitive to intraterminal dialysis and is therefore mediated by a readily diffusible second messenger. Using the perforated patch-clamp technique and ratio-calcium-imaging methods, we describe a diffusible second messenger pathway stimulated by the MOR that inhibits voltage-gated calcium channels in isolated terminals from the rat neurohypophysis (NH). Our results show a rise in basal intracellular calcium ([Ca(2+)]i) in response to application of [D-Ala(2)-N-Me-Phe(4),Gly5-ol]-Enkephalin (DAMGO), a MOR agonist, that is blocked by D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP), a MOR antagonist. Buffering DAMGO-induced changes in [Ca(2+)]i with BAPTA-AM completely blocked the inhibition of both I(Ca) and high-K(+)-induced rises in [Ca(2+)]i due to MOR activation, but had no effect on kappa-opioid receptor (KOR)-mediated inhibition. Given the presence of ryanodine-sensitive stores in isolated terminals, we tested 8-bromo-cyclic adenosine diphosphate ribose (8Br-cADPr), a competitive inhibitor of cyclic ADP-ribose (cADPr) signaling that partially relieves DAMGO inhibition of I(Ca) and completely relieves MOR-mediated inhibition of high-K(+)-induced and DAMGO-induced rises in [Ca(2+)]i. Furthermore, antagonist concentrations of ryanodine completely blocked MOR-induced increases in [Ca(2+)]i and inhibition of I(Ca) and high-K(+)-induced rises in [Ca(2+)]i while not affecting KOR-mediated inhibition. Antagonist concentrations of ryanodine also blocked MOR-mediated inhibition of electrically-evoked increases in capacitance. These results strongly suggest that a key diffusible second messenger mediating the MOR-signaling pathway in NH terminals is [Ca(2+)]i released by cADPr from ryanodine-sensitive stores

Estudio Preliminar Sobre La Decoloración Del Residual Líquido De La Producción De Papel Mediante Ozonización

Preliminarily, the relationship between Color Reduction (%RC), Total Suspended Solids concentration (SST), and as independent variable, the time of Ozone Oxidation Process, was evaluated with reference values of DQO of the black liquor adjusting its concentration from SST to 300 and 50mg/dm3. A batch reactor of 12,60dm3, an effective height of 3,2dm and an ozone generating device from SEFILTRA company (air flow of 2dm3/min, c(O3) of 9,50mg/dm3 with a O3(g) production 19mg/min) was used. The polynomic correlation was determined from 192 data sets with satisfactory adjustment level (R=0,92; p <0,001). Although, preliminarily, the DQO reduction (%RDQO) was not included in the polynomial relationship, it can be concluded that: a) The %RC and %RDQO is reduced when color and DQO increases from the ozonized sample at same time intervals; this reduction is greater when the SST levels of the ozonized sample increases. b) The %RC and %RDQO decrease when the SST level of the ozonized sample increases. c) Depending on the initial color, of the DQO and of the SST level of the sample, values from 10 to 53 %Rc were obtained after 10 min, and of 66 to 94 %Rc after 60 min

Lectura y comprensión de textos en el nivel superior: un desafío compartido entre alumnos y docentes

La comprensión lectora se ha convertido, en las últimas décadas, en una preocupación de quienes tienen la responsabilidad de la educación superior. Así, en el ISFD “Santa María” (San Juan, Argentina) se desarrolló durante los años 2006-2007 la investigación denominada "La Comprensión Lectora en el Nivel Superior: un estudio exploratorio". El objetivo fue indagar los factores que inciden en la comprensión lectora de alumnos de nivel superior. La estrategia metodológica se enmarcó dentro del paradigma interpretativo mediante un abordaje predominantemente cualitativo, con alumnos de dicho Instituto de la cohorte 2005. Se desarrolló en dos etapas: una enfocada a descubrir categorías desde los datos, mediante técnicas de recolección de bajo nivel de preestructuración (entrevistas en profundidad); la otra profundiza y relaciona categorías específicas del abordaje de textos escritos, usando técnicas de obtención de datos más estructuradas (cuestionario semi-estructurado autoadministrado). En la primera etapa —año 2006— la investigación estuvo orientada a recuperar la perspectiva de los alumnos para descubrir factores facilitadores y obstaculizadores de la comprensión lectora en su trayectoria educativa. Mediante un estudio de casos se puso de manifiesto la diversidad e interrelación de factores que inciden en el problema. A modo de conclusión, encontramos que la dimensión afectiva, los condicionantes académicos y las disposiciones de los alumnos se articulan para dar lugar a una determinada forma de encarar la formación de nivel superior. La segunda etapa —año 2007— se centró específicamente en el trabajo con los textos, tanto del docente como de los alumnos. Como resultado de este estudio se puso de relieve la importancia de la mediación del docente para facilitar la comprensión del material de estudio

Self-assembled coumarin- and 5-fluorouracil-PEG micelles as multifunctional drug delivery systems

The copper(I)-catalyzed azide/alkyne cycloaddition is recognized as one of the most successful click reactions to access self-assembling amphiphilic polymer-drug conjugates (PDCs). In this way, poor water-soluble drugs can be linked covalently to hydrophilic poly(ethylene glycol) (PEG) to obtain PEGylated drug micelles that can be used as versatile carriers for the delivery of diverse therapeutic agents. In this work, two novel amphiphilic PDCs that combine PEG with privileged scaffolds well-known for their anticancer properties, such as coumarin and 5-fluorouracil, have been synthesized and characterized. These conjugates were able to self-assemble into micelles at relatively high critical micellar concentration, probably due to the large portion of hydrophilic PEG. The micelles allowed to load other anticancer drugs (paclitaxel, curcumin, and gemcitabine), providing a unique opportunity to develop promising co-delivery carriers for synergistic cancer therapy. The Korsmeyer-Peppas mathematical model was used for describing the in vitro kinetics of drug release from the micelles. Similar sustained and controlled drug release profiles were obtained for paclitaxel and curcumin in both conjugates, which was attributed to the excellent stability driven by the strong interaction between polymeric conjugates and drugs in the micelle core. In contrast, the high instability observed for the gemcitabine-loaded micelles provided an initial uncontrolled burst release of drug. A preliminary in vitro cytotoxicity study of the micelles against human pancreatic cancer cells PANC-1 and BxPC-3 was also carried out, demonstrating that both coumarin and 5-fluorouracil retain their anticancer properties after conjugation with PEG.La cicloadición de azida/alquino catalizada por cobre (I) se reconoce como una de las reacciones de clic más exitosas para acceder a conjugados de polímero-fármaco anfifílicos (PDC) autoensamblados. De esta manera, los fármacos poco solubles en agua se pueden unir covalentemente al poli(etilenglicol) (PEG) hidrofílico para obtener micelas de fármacos PEGilados que se pueden utilizar como vehículos versátiles para la administración de diversos agentes terapéuticos. En este trabajo, dos PDC anfifílicos novedosos que combinan PEG con andamios privilegiados conocidos por sus propiedades anticancerígenas, como la cumarina .y 5-fluorouracilo, han sido sintetizados y caracterizados. Estos conjugados pudieron autoensamblarse en micelas a una concentración micelar crítica relativamente alta, probablemente debido a la gran porción de PEG hidrofílico. Las micelas permitieron cargar otros medicamentos contra el cáncer (paclitaxel, curcumina y gemcitabina), lo que brinda una oportunidad única para desarrollar portadores de administración conjunta prometedores para la terapia sinérgica contra el cáncer. Se utilizó el modelo matemático de Korsmeyer-Peppas para describir la cinética in vitro de la liberación del fármaco desde las micelas. Se obtuvieron perfiles similares de liberación sostenida y controlada del fármaco para paclitaxel.y curcumina en ambos conjugados, lo que se atribuyó a la excelente estabilidad impulsada por la fuerte interacción entre los conjugados poliméricos y los fármacos en el núcleo de la micela. Por el contrario, la alta inestabilidad observada para las micelas cargadas con gemcitabina proporcionó una liberación inicial descontrolada del fármaco. También se llevó a cabo un estudio preliminar de citotoxicidad in vitro de las micelas contra las células de cáncer de páncreas humano PANC-1 y BxPC-3, demostrando que tanto la cumarina como el 5-fluorouracilo conservan sus propiedades anticancerígenas después de la conjugación con PEG

Reconstruction of ancient microbial genomes from the human gut

Loss of gut microbial diversity in industrial populations is associated with chronic diseases, underscoring the importance of studying our ancestral gut microbiome. However, relatively little is known about the composition of pre-industrial gut microbiomes. Here we performed a large-scale de novo assembly of microbial genomes from palaeofaeces. From eight authenticated human palaeofaeces samples (1,000–2,000 years old) with well-preserved DNA from southwestern USA and Mexico, we reconstructed 498 medium- and high-quality microbial genomes. Among the 181 genomes with the strongest evidence of being ancient and of human gut origin, 39% represent previously undescribed species-level genome bins. Tip dating suggests an approximate diversification timeline for the key human symbiont Methanobrevibacter smithii. In comparison to 789 present-day human gut microbiome samples from eight countries, the palaeofaeces samples are more similar to non-industrialized than industrialized human gut microbiomes. Functional profiling of the palaeofaeces samples reveals a markedly lower abundance of antibiotic-resistance and mucin-degrading genes, as well as enrichment of mobile genetic elements relative to industrial gut microbiomes. This study facilitates the discovery and characterization of previously undescribed gut microorganisms from ancient microbiomes and the investigation of the evolutionary history of the human gut microbiota through genome reconstruction from palaeofaeces

Truncating FLNC Mutations Are Associated With High-Risk Dilated and Arrhythmogenic Cardiomyopathies

BACKGROUND: Filamin C (encoded by the FLNC gene) is essential for sarcomere attachment to the plasmatic membrane. FLNC mutations have been associated with myofibrillar myopathies, and cardiac involvement has been reported in some carriers. Accordingly, since 2012, the authors have included FLNC in the genetic screening of patients with inherited cardiomyopathies and sudden death. OBJECTIVES: The aim of this study was to demonstrate the association between truncating mutations in FLNC and the development of high-risk dilated and arrhythmogenic cardiomyopathies. METHODS: FLNC was studied using next-generation sequencing in 2,877 patients with inherited cardiovascular diseases. A characteristic phenotype was identified in probands with truncating mutations in FLNC. Clinical and genetic evaluation of 28 affected families was performed. Localization of filamin C in cardiac tissue was analyzed in patients with truncating FLNC mutations using immunohistochemistry. RESULTS: Twenty-three truncating mutations were identified in 28 probands previously diagnosed with dilated, arrhythmogenic, or restrictive cardiomyopathies. Truncating FLNC mutations were absent in patients with other phenotypes, including 1,078 patients with hypertrophic cardiomyopathy. Fifty-four mutation carriers were identified among 121 screened relatives. The phenotype consisted of left ventricular dilation (68%), systolic dysfunction (46%), and myocardial fibrosis (67%); inferolateral negative T waves and low QRS voltages on electrocardiography (33%); ventricular arrhythmias (82%); and frequent sudden cardiac death (40 cases in 21 of 28 families). Clinical skeletal myopathy was not observed. Penetrance was >97% in carriers older than 40 years. Truncating mutations in FLNC cosegregated with this phenotype with a dominant inheritance pattern (combined logarithm of the odds score: 9.5). Immunohistochemical staining of myocardial tissue showed no abnormal filamin C aggregates in patients with truncating FLNC mutations. CONCLUSIONS: Truncating mutations in FLNC caused an overlapping phenotype of dilated and left-dominant arrhythmogenic cardiomyopathies complicated by frequent premature sudden death. Prompt implantation of a cardiac defibrillator should be considered in affected patients harboring truncating mutations in FLNC.Instituto de Salud Carlos III [PI11/0699, PI14/0967, PI14/01477, RD012/0042/0029, RD012/0042/0049, RD012/0042/0066, RD12/0042/0069]; Spanish Ministry of Economy and Competitiveness [SAF2015-71863-REDT]; Plan Nacional de I+D+I; Plan Estatalde I+D+I, European Regional Development Fund; Health in Code SLS

Epigenetic loss of RNA‑methyltransferase NSUN5 in glioma targets ribosomes to drive stress adaptive translational program

Tumors have aberrant proteomes that often do not match their corresponding transcriptome profiles. One possible cause of this discrepancy is the existence of aberrant RNA modification landscapes in the so-called epitranscriptome. Here, we report that human glioma cells undergo DNA methylation-associated epigenetic silencing of NSUN5, a candidate RNA methyltransferase for 5-methylcytosine. In this setting, NSUN5 exhibits tumor-suppressor characteristics in vivo glioma models. We also found that NSUN5 loss generates an unmethylated status at the C3782 position of 28S rRNA that drives an overall depletion of protein synthesis, and leads to the emergence of an adaptive translational program for survival under conditions of cellular stress. Interestingly, NSUN5 epigenetic inactivation also renders these gliomas sensitive to bioactivatable substrates of the stress-related enzyme NQO1. Most importantly, NSUN5 epigenetic inactivation is a hallmark of glioma patients with long-term survival for this otherwise devastating disease

TNOs are cool: a survey of the transneptunian region

Over one thousand objects have so far been discovered orbiting beyond Neptune. These trans-Neptunian objects (TNOs) represent the primitive remnants of the planetesimal disk from which the planets formed and are perhaps analogous to the unseen dust parent-bodies in debris disks observed around other main-sequence stars. The dynamical and physical properties of these bodies provide unique and important constraints on formation and evolution models of the Solar System. While the dynamical architecture in this region (also known as the Kuiper Belt) is becoming relatively clear, the physical properties of the objects are still largely unexplored. In particular, fundamental parameters such as size, albedo, density and thermal properties are difficult to measure. Measurements of thermal emission, which peaks at far-IR wavelengths, offer the best means available to determine the physical properties. While Spitzer has provided some results, notably revealing a large albedo diversity in this population, the increased sensitivity of Herschel and its superior wavelength coverage should permit profound advances in the field. Within our accepted project we propose to perform radiometric measurements of 139 objects, including 25 known multiple systems. When combined with measurements of the dust population beyond Neptune (e.g. from the New Horizons mission to Pluto), our results will provide a benchmark for understanding the Solar debris disk, and extra-solar ones as well

Impact of a Primary Care Antimicrobial Stewardship Program on Bacterial Resistance Control and Ecological Imprint in Urinary Tract Infections

Antimicrobial stewardship programs (ASPs) are a central component in reducing the overprescription of unnecessary antibiotics, with multiple studies showing benefits in the reduction of bacterial resistance. Less commonly, ASPs have been performed in outpatient settings, but there is a lack of available data in these settings. We implemented an ASP in a large regional outpatient setting to assess its feasibility and effectiveness. Over a 5-year post-implementation period, compared to the pre-intervention period, a significant reduction in antibiotic prescription occurred, with a reduction in resistance in E. coli urinary isolates. ASP activities also were found to be cost-effective, with a reduction in medication prescription

HTLV-1 infection in solid organ transplant donors and recipients in Spain

HTLV-1 infection is a neglected disease, despite infecting 10-15 million people worldwide and severe illnesses develop in 10% of carriers lifelong. Acknowledging a greater risk for developing HTLV-1 associated illnesses due to immunosuppression, screening is being widely considered in the transplantation setting. Herein, we report the experience with universal HTLV testing of donors and recipients of solid organ transplants in a survey conducted in Spain. All hospitals belonging to the Spanish HTLV network were invited to participate in the study. Briefly, HTLV antibody screening was performed retrospectively in all specimens collected from solid organ donors and recipients attended since the year 2008. A total of 5751 individuals were tested for HTLV antibodies at 8 sites. Donors represented 2312 (42.2%), of whom 17 (0.3%) were living kidney donors. The remaining 3439 (59.8%) were recipients. Spaniards represented nearly 80%. Overall, 9 individuals (0.16%) were initially reactive for HTLV antibodies. Six were donors and 3 were recipients. Using confirmatory tests, HTLV-1 could be confirmed in only two donors, one Spaniard and another from Colombia. Both kidneys of the Spaniard were inadvertently transplanted. Subacute myelopathy developed within 1 year in one recipient. The second recipient seroconverted for HTLV-1 but the kidney had to be removed soon due to rejection. Immunosuppression was stopped and 3 years later the patient remains in dialysis but otherwise asymptomatic. The rate of HTLV-1 is low but not negligible in donors/recipients of solid organ transplants in Spain. Universal HTLV screening should be recommended in all donor and recipients of solid organ transplantation in Spain. Evidence is overwhelming for very high virus transmission and increased risk along with the rapid development of subacute myelopathy