El derecho humano al agua y al saneamiento: un tema clave en la intersección Ecología-Derechos Humanos

El articulo pretende contribuir a la reflexión acerca de estos dos derechos (agua y saneamiento), de reciente reconocimiento jurídico internacional

Torulaspora delbrueckii: un modelo para el estudio del estrés por NaCl en levadura de panaderia.

RESUMEN Los hábitos alimenticios del hombre han variado en estos últimos años, aumentando la demanda de productos congelados de panadería y bollería por lo que se requieren levaduras con gran resistencia a diferentes tipos de estrés. En la masa panaria, la presencia de NaCl provoca por una parte un estrés osmótico debido a la pérdida masiva de agua de la célula y por otra parte un estrés iónico debido a las altas concentraciones de iones Na+ y Cl- en el citoplasma, lo que tiene efectos dañinos para la levadura. Torulaspora delbrueckii es una levadura capaz de fermentar una masa panaria y más resistente a estrés que la habitualmente utilizada en la industria, Saccharomyces cerevisiae. A pesar de cumplir con las demandas actuales de la industria, los productores son muy reacios a la introducción de un nuevo microorganismo, ya que esto significaría tener que modificar sus ya bien establecidos procesos de producción. Por ello, hemos utilizado a la levadura T. delbrueckii en este trabajo, como organismo modelo de estudio del estrés por NaCl en levadura de panadería. Nos encontramos ante un microorganismo muy poco estudiado, sobretodo a nivel molecular, por lo que hemos desarrollado herramientas para su utilización en el laboratorio. Hemos conseguido aislar los genes HIS3 y LEU2, que son habitualmente utilizados como marcadores auxotróficos y además hemos conseguido una cepa de T. delbrueckii auxótrofa para el gen LEU2. Gracias a la secuenciación de estos genes y de sus regiones flanqueantes en varias levaduras, hemos podido añadir más información para esclarecer las relaciones filogenéticas entre el grupo de levaduras Saccharomyces sensu stricto y sensu lato. Centrándonos en T. delbrueckii como modelo de reistencia a estrés salino, se han aislado genes cuya sobreexpresión confiere resistencia a NaCl en S. cerevisiae. Así hemos identificado una pauta de lectura abierta que codifica una proteína, con una alta homología a Ena1, Ena2 y Ena5 de S. cerevisiae. El aislamiento de TdENA1 en este rastreo, indicó que la estrategia empleada era correcta ya que este gen codifica la principal ATPasa tipo P implicadas en el bombeo de iones Na+ al exterior de la célula cuya función determina resistencia a estrés iónico. La sobreexpresión de TdDED1 confirió resistencia a sal en Saccharomyces. Ded1p es una RNA helicasa de la denominada familia DEAD-box. Hemos comprobado que este gen se induce tanto en condiciones de estrés salino como en condiciones de baja temperatura, estableciéndose su implicación en resistencia a estrés. Por último la sobreexpresión del extremo 3´ de un gen homólogo a SIP1 de S. cerevisiae, aumentó la tolerancia de la cepa salvaje a NaCl. Sip1p junto a Sip2p y Gal83p son las subunidades de la proteína quinasa Snf1p que juega un papel fundamental en la regulación de genes reprimidos por glucosa. Análisis del fenotipo de un mutante nulo en SNF1 en medios con sal, indicó que esta proteína quinasa tiene un papel funcional como represor de genes que se inducen en estas condiciones de estrés, en concreto genes cuya expresión está regulada por la ruta de la calcineurina. Profundizando en esta línea de trabajo, nuestros resultados nos han permitido establecer una regulación, directa o indirecta de la ruta de la calcineurina por Snf1p en condiciones de normales de crecimiento, es decir en ausencia de estrés salino. ____________________________________________________________________________________________________ SUMMARY Saccharomyces cerevisiae has probably been the first microorganism exploited by men. In baker industry, human food habits have changed in the past few years, increasing frozen products demand. This is why more resistant yeast is required by producers. In our lab, we have focused our attention on improving osmotolerance in baker yeast. The presence of NaCl reduces water activity of the dough and there are also further effects due to the toxicity of sodium and chloride ions. Thus, this stressful environment causes loss of fermentation capacity, leading to longer proofing times and reduced product volume. Torulaspora delbrueckii, a non-conventional yeast, is often isolated from home-made bread-dough. Some strains display a great baking ability and unlike Saccharomyces, they are able to cope with several stresses, including osmotic stress. This trait made this organism a potential model to understand the mechanisms underlying stress resistance in bakers yeast. Otherwise, very little is known about this yeast, including the tools for its manipulation. In the present work we have isolated and characterised the HIS3 and LEU2 gene from T. delbrueckii and the adjacent genes. Analysis of gene order and transcriptional orientation of these regions helped in the study of the phylogenetic relationships among yeasts. We have also isolated T. delbrueckii genes which conferred salt resistance when overexpressed in S. cerevisiae. These genes are the homologous of the RNA helicase DED1, the ATPase ENA1, and the C-terminal part of Sip1p. Finally, we have studied the implications of Snf1p in the salt stress response. Our results suggested that this protein kinase play a role as a repressor of genes induced by saline stress, concretely genes whose expression is regulated by the calcineurin pathway. Snf1p, directly or indirectly control the activation of the calcineurin pathway under non-stress conditions

Tratados bilaterales de inversión :Las regulaciones del comercio y su influencia en el derecho humano al agua y al saneamiento

El derecho humano al agua y al saneamiento está seriamente amenazado. Además del ámbito del Derecho internacional de los derechos humanos, este derecho se ve afectado por otras normas, como las del Comercio Internacional, entre las que se encuentran los tratados multilaterales y bilaterales de inversión -TBI. Estos últimos contienen cláusulas que muy frecuentemente colisionan con las normas internacionales de protección del derecho al agua y al saneamiento, como el Pacto Internacional de Derechos Económicos, Sociales y Culturales. Los conflictos que surgen en la práctica entre empresas afectadas por TBI y estados se resuelven ante tribunales arbitrales, que no incorporan en sus decisiones las categorías y principios del derecho humano al agua potable y al saneamiento. Este texto aborda estos problemas, y propone ciertas recomendacione

Chronic prehepatic portal hypertension in the rat: is it a type of Metabolic Inflammatory Syndrome?

<p>Abstract</p> <p>Background</p> <p>A progressive development of hepatic steatosis with an increase in the lipid hepatocyte content and the formation of megamitochondria have been demonstrated in rats with prehepatic portal hypertension. The aim of this study is to verify the existence of liver and serum lipid metabolism impairments in rats with long-term (2 years) portal hypertension.</p> <p>Methods</p> <p>Male Wistar rats: Control (n = 10) and with prehepatic portal hypertension by triple partial portal vein ligation (n = 9) were used. Liver content of Triglycerides (TG), phospholipids (PL) and cholesterol and serum cholesterol, lipoproteins (HDL and LDL), TG, glucose and Lipid Binding Protein (LBP) were assayed with specific colorimetric commercial kits. Serum levels of insulin and somatostatin were assayed by RIA.</p> <p>Results</p> <p>The liver content of TG (6.30 ± 1.95 <it>vs</it>. 4.17 ± 0.59 μg/ml; p < 0.01) and cholesterol (1.48 ± 0.15 <it>vs</it>. 1.10 ± 0.13 μg/ml; p < 0.001) increased in rats with portal hypertension. The serum levels of cholesterol (97.00+26.02 <it>vs</it>. 114.78 ± 37.72 mg/dl), TG (153.41 ± 80.39 <it>vs</it>. 324.39 ± 134.9 mg/dl; p < 0.01), HDL (20.45 ± 5.14 <it>vs</it>. 55.15 ± 17.47 mg/dl; p < 0.001) and somatostatin (1.32 ± 0.31 <it>vs</it>. 1.59 +0.37 mg/dl) decreased, whereas LDL (37.83 ± 15.39 <it>vs</it>. 16.77 ± 6.81 mg/dl; p < 0.001) and LBP (308.47 ± 194.53 <it>vs</it>. 60.27 ± 42.96 ng/ml; p < 0.001) increased.</p> <p>Conclusion</p> <p>Portal hypertension in the rat presents changes in the lipid and carbohydrate metabolisms similar to those produced in chronic inflammatory conditions and sepsis in humans. These underlying alterations could be involved in the development of hepatic steatosis and, therefore, in those described in the metabolic syndrome in humans.</p

Proinflammatory Liver and Antiinflammatory Intestinal Mediators Involved in Portal Hypertensive Rats

Proinflammatory (TNF-α, IL-1β, and NO) and antiinflammatory (IL-10, CO) levels were assayed in serum, liver, and small bowel in order to verify a hypothetic inflammatory etiopathogeny of portal hypertension that could be the cause of its evolutive heterogeneity. Male Wistar rats were divided into one control group (n = 11) and one group with a triple stenosing ligation of the portal vein (n = 23) after 28 days of evolution. In one subgroup of portal hypertensive rats, portal pressure, collateral venous circulation, mesenteric vasculopathy, and liver and spleen weights were determined. In the remaining rats with portal hypertension TNF-α, IL-1β, and IL-10 were quantified in liver and ileum by enzyme-linked immunosorbent assay. NO synthase activity was studied in liver and ileum. CO and NO were measured in portal and systemic blood by spectrophotometry and Griess reaction, respectively. Portal hypertensive rats with mayor spleen weight show hepatomegaly and mayor development of collateral circulation. Ileum release of IL-10 (0.30 ± 0.12 versus 0.14 ± 0.02 pmol/mg protein; P < .01) is associated with a liver production of both proinflammatory mediators (TNF-α: 2 ± 0.21 versus 1.32 ± 0.60 pmol/mg protein; P < .05, IL-1β: 19.17 ± 2.87 versus 5.96 ± 1.84 pmol/mg protein; P = .005, and NO: 132.10 ± 34.72 versus 61.05 ± 8.30 nmol/mL; P = .005) and an antiinflammatory mediator (CO: 6.49 ± 2.99 versus 3.03 ± 1.59 pmol/mL; P = .005). In short-term prehepatic portal hypertension a gut-liver inflammatory loop, which could be fundamental in the regulation both of the portal pressure and of its complications, could be proposed

Clinical aspects of usher syndrome and the USH2A gene in a cohort of 433 patients

IMPORTANCE A new statistical approach is needed to describe the clinical differences between type I and type II Usher syndrome and between the 2 most frequent mutations in the USH2A gene. OBJECTIVES To describe the primary phenotypic characteristics and differences between type I and type II Usher syndrome and to establish a phenotype-genotype correlation for the 2 most frequent mutations in the USH2A gene. DESIGN, SETTING, AND PARTICIPANTS Cross-sectional study at a genetics department, in which clinical evaluations were performed for 433 patients (297 unrelated families) who were classified as having type I, II, III, atypical, or unclassified Usher syndrome according to their clinical history, pedigree data, results from ophthalmological studies, and audiological, neurophysiological, and vestibular test results. Molecular studies were performed for 304 patients (256 unrelated families). The Mann-Whitney U test or the χ2 test was used for calculating the differences between mean values for the analyzed parameters. MAIN OUTCOMES AND MEASURES Age at diagnosis; age at onset of night blindness, visual field loss, visual acuity loss, and cataracts; and severity and age at diagnosis of hearing loss. RESULTS The comparison between patients with type I Usher syndrome and those with type II Usher syndrome revealed P < .001 for most items analyzed. The most frequent mutations in the USH2A gene were the p.Glu767Serfs*21 and p.Cys759Phe mutations, with an allelic frequency of 23.2%(63 of 272 alleles) and 8.1% (22 of 272 alleles), respectively. The phenotypic analysis for patients carrying p.Cys759Phe showed P < .001 for most items analyzed when compared with patients carrying p.Glu767Serfs*21 and when compared with patients carrying other mutations in the USH2A gene. None of the p.Cys759Phe patients exhibited a severe hearing loss phenotype, and more than 60%had only mild hearing loss. Most patients carrying the p.Glu767Serfs*21 mutation (72.1%) were moderately deaf. CONCLUSIONS AND RELEVANCE Our study presents the clinical differences between type I and type II Usher syndrome and between the 2 most frequent mutations in the USH2A gene. Detailed genotype-phenotype correlations, as presented in our study, allow for a better correlation of clinical signs with a known genotype and can improve the clinical management, genetic counseling, and risk assessment of patients with Usher syndrome because an estimated prognosis of their disease can be madeThis work was supported by grant PI13/00226 (to Servicio de Genética, Instituto de Investigación–Fundación Jiménez Díaz, Madrid, Spain), by grant PI13/00638 (to Unidad de Genética y Diagnóstico Prenatal, Hospital Universitario y Politécnico La Fe, Valencia, Spain), and by grant 06/07/0036 (to Centro de Investigación Biomédica en Red de Enfermedades Raras, Madrid, Spain) from Fundaluce and Organización Nacional de Ciegos Españole

Mutation analysis of 272 Spanish families affected by autosomal recessive retinitis pigmentosa using a genotyping microarray

Contains fulltext : 89342.pdf (publisher's version ) (Open Access)PURPOSE: Retinitis pigmentosa (RP) is a genetically heterogeneous disorder characterized by progressive loss of vision. The aim of this study was to identify the causative mutations in 272 Spanish families using a genotyping microarray. METHODS: 272 unrelated Spanish families, 107 with autosomal recessive RP (arRP) and 165 with sporadic RP (sRP), were studied using the APEX genotyping microarray. The families were also classified by clinical criteria: 86 juveniles and 186 typical RP families. Haplotype and sequence analysis were performed to identify the second mutated allele. RESULTS: At least one-gene variant was found in 14% and 16% of the juvenile and typical RP groups respectively. Further study identified four new mutations, providing both causative changes in 11% of the families. Retinol Dehydrogenase 12 (RDH12) was the most frequently mutated gene in the juvenile RP group, and Usher Syndrome 2A (USH2A) and Ceramide Kinase-Like (CERKL) were the most frequently mutated genes in the typical RP group. The only variant found in CERKL was p.Arg257Stop, the most frequent mutation. CONCLUSIONS: The genotyping microarray combined with segregation and sequence analysis allowed us to identify the causative mutations in 11% of the families. Due to the low number of characterized families, this approach should be used in tandem with other techniques

Probing the Inner Jet of the Quasar PKS 1510-089 with Multi-waveband Monitoring during Strong Gamma-ray Activity

We present results from monitoring the multi-waveband flux, linear polarization, and parsec-scale structure of the quasar PKS 1510-089, concentrating on eight major gamma-ray flares that occurred during the interval 2009.0-2009.5. The gamma-ray peaks were essentially simultaneous with maxima at optical wavelengths, although the flux ratio of the two wavebands varied by an order of magnitude. The optical polarization vector rotated by 720 degrees during a 5-day period encompassing six of these flares. This culminated in a very bright, roughly 1 day, optical and gamma-ray flare as a bright knot of emission passed through the highest-intensity, stationary feature (the "core") seen in 43 GHz Very Long Baseline Array images. The knot continued to propagate down the jet at an apparent speed of 22c and emit strongly at gamma-ray energies as a months-long X-ray/radio outburst intensified. We interpret these events as the result of the knot following a spiral path through a mainly toroidal magnetic field pattern in the acceleration and collimation zone of the jet, after which it passes through a standing shock in the 43 GHz core and then continues downstream. In this picture, the rapid gamma-ray flares result from scattering of infrared seed photons from a relatively slow sheath of the jet as well as from optical synchrotron radiation in the faster spine. The 2006-2009.7 radio and X-ray flux variations are correlated at very high significance; we conclude that the X-rays are mainly from inverse Compton scattering of infrared seed photons by 20-40 MeV electrons.Comment: 10 pages of text + 5 figures, to be published in Astrophysical Journal Letters in 201

Expanding the Clinical and Molecular Heterogeneity of Nonsyndromic Inherited Retinal Dystrophies

A cohort of 172 patients diagnosed clinically with nonsyndromic retinal dystrophies, from 110 families underwent full ophthalmologic examination, including retinal imaging, electrophysiology, and optical coherence tomography, when feasible. Molecular analysis was performed using targeted next-generation sequencing (NGS). Variants were filtered and prioritized according to the minimum allele frequency, and finally classified according to the American College of Medical Genetics and Genomics guidelines. Multiplex ligation-dependent probe amplification and array comparative genomic hybridization were performed to validate copy number variations identified by NGS. The diagnostic yield of this study was 62% of studied families. Thirty novel mutations were identified. The study found phenotypic intra- and interfamilial variability in families with mutations in C1QTNF5, CERKL, and PROM1; biallelic mutations in PDE6B in a unilateral retinitis pigmentosa patient; interocular asymmetry RP in 50% of the symptomatic RPGR-mutated females; the first case with possible digenism between CNGA1 and CNGB1; and a ROM1 duplication in two unrelated retinitis pigmentosa families. Ten unrelated cases were reclassified. This study highlights the clinical utility of targeted NGS for nonsyndromic inherited retinal dystrophy cases and the importance of full ophthalmologic examination, which allows new genotypeephenotype associations and expands the knowledge of this group of disorders. Identifying the cause of disease is essential to improve patient management, provide accurate genetic counseling, and take advantage of gene therapyebased treatments. (J Mol Diagn 2020, 22: 532e543; https:// doi.org/10.1016/j.jmoldx.2020.01.003)Supported by grants from the Instituto de Salud Carlos III (ISCIII) of the Spanish Ministry of Health, including the Center for Biomedical Research Network on Rare Diseases (CIBERER), Fondo de Investigacion Sanitaria grant PI16/00539; the Spanish National Organization for the Blind (Fundación ONCE); and Fundación Mutua Madrileña. G.G.-G. is sponsored by the CIBERER, and A.R.-M. is supported by the Río Hortega program from ISCIII.Medicin

Genetic Screening of the Usher Syndrome in Cuba

BackgroundUsher syndrome (USH) is a recessive inherited disease characterized by sensorineural hearing loss, retinitis pigmentosa, and sometimes, vestibular dysfunction. Although the molecular epidemiology of Usher syndrome has been well studied in Europe and United States, there is a lack of studies in other regions like Africa or Central and South America.MethodsWe designed a NGS panel that included the 10 USH causative genes (MYO7A, USH1C, CDH23, PCDH15, USH1G, CIB2, USH2A, ADGRV1, WHRN, and CLRN1), four USH associated genes (HARS, PDZD7, CEP250, and C2orf71), and the region comprising the deep-intronic c.7595-2144A&gt;G mutation in USH2A.ResultsNGS sequencing was performed in 11 USH patients from Cuba. All the cases were solved. We found the responsible mutations in the USH2A, ADGRV1, CDH23, PCDH15, and CLRN1 genes. Four mutations have not been previously reported. Two mutations are recurrent in this study: c.619C&gt;T (p.Arg207∗) in CLRN1, previously reported in two unrelated Spanish families of Basque origin, and c.4488G&gt;C (p.Gln1496His) in CDH23, first described in a large Cuban family. Additionally, c.4488G&gt;C has been reported two more times in the literature in two unrelated families of Spanish origin.ConclusionAlthough the sample size is very small, it is tempting to speculate that the gene frequencies in Cuba are distinct from other populations mainly due to an “island effect” and genetic drift. The two recurrent mutations appear to be of Spanish origin. Further studies with a larger cohort are needed to elucidate the real genetic landscape of Usher syndrome in the Cuban population