Disruption of the with no lysine kinase-STE20-proline alanine-rich kinase pathway reduces the hypertension induced by angiotensin II

Objective: The hypertensive effect of angiotensin II (AngII), a peptide hormone, is dependent on its intrarenal actions and the activation of the renal Na–Cl cotransporter (NCC), by AngII requires integrity of the with no lysine kinase/STE20-proline alanine rich kinase (WNK/SPAK) signaling pathway. Here, were analyzed if the integrity of the WNK/SPAK pathway is required for AngII infusion to induce arterial hypertension.Methods: We tested the effect of AngII or aldosterone administration on the blood pressure and on pNCC/NCC ratio in SPAKT243A/243A knock-in mice in which the kinase and thus NCC cannot be activated by WNK kinases. AngII or aldosterone was infused at 1440 or 700 mg/kg/d, respectively, for 14 days using osmotic minipumps. The aldosterone-treated mice were exposed to NaCl drinking water (1%) during the hormone administration. The arterial blood pressure was assessed using radiotelemetry.Results: We observed that in the SPAK knock-in mice, the AngII-induced hypertensive effect was significantly reduced and associated with an absence of AngII-induced NCCphosphorylation. In contrast, the hypertensive effect of aldosterone was enhanced and was related with an increased response to amiloride, but not to thiazide-typediuretics, without a significant increase in NCC phosphorylation.Conclusion: Our data suggest that AngII-induced hypertension requires, at least partly, NCC activation via the WNK/SPAK signaling pathway, whereas aldosterone-induced hypertension depends on ENaC activation in a WNK/SPAKindependent manner. SPAK knock-in mice emerge as a useful model to distinguish between the effects of AngII and aldosterone on distal nephrons

Evaluación de las condiciones predisponentes a enfermedades en granjas porcinas a pequeña escala en un ambiente urbano en el noroeste de la Ciudad de México

The objective of the work was to develop and apply an instrument to identify the predisposing conditions to the occurrence of diseases in 12 small-scale swine farms in an urban environment. The percentage of negative points obtained in general and by type of farm was analyzed according to its production, fattening (T1) or full cycle (T2), where the highest percentage was for T1 (50 %) and for T2 (66.0 %). Likewise, the data were analyzed to compare the farms T1 and T2 in relation to the percentages of each section that makes up the survey, where only a difference in the “health state” section (P<0.0001) was found. The relationship between the population density per m2 with respect to the maximum percentage of points reached by farms showed no difference (R2, 0.03; P=0.854). No correlation was found between the percentage of points obtained with the number of animals (R2, 0.13; P=0.722). In relation to the average percentage per section by population size, only a difference in the “feeding” section (P<0.0006) was detected, indicating that farms with 10 to 40 swine obtained fewer points in this section. It is concluded that the methodology for the evaluation of conditions predisposing to diseases in this type of farms proved to be applicable. It was determined that farm size and population density are not a predisposing factor in these farms, but the predisposing conditions to the occurrence of diseases differ between full-cycle and fattening farms.El objetivo del trabajo fue desarrollar y aplicar un instrumento para identificar las condiciones predisponentes a la presentación de enfermedades en 12 granjas porcinas a pequeña escala en un ambiente urbano. Se analizó el porcentaje de puntos negativos obtenidos de manera general y por tipo de granja en función a su producción, engorda (T1) o ciclo completo (T2), donde el mayor porcentaje fue para T1 (50 %) y para T2 (66.0 %). Del mismo modo se analizaron los datos para comparar a las granjas T1 y las T2 en relación con los porcentajes de cada sección que integra la encuesta, donde solo se encontró diferencia en la sección “estado de salud” (P<0.0001). La relación entre la densidad de población por m2 con respecto al porcentaje máximo de puntos alcanzados por granjas, no mostró diferencia (R2, 0.03; P=0.854). No se encontró correlación entre el porcentaje de puntos obtenidos con el número de animales (R2, 0.13; P=0.722). En relación al porcentaje promedio por sección por tamaño de la población, solo se detectó diferencia en la sección “alimentación” (P<0.0006), indicando que las granjas con 10 a 40 cerdos obtuvieron menos puntos en esta sección. Se concluye que la metodología para la evaluación de las condiciones predisponentes a enfermedades en este tipo de granjas resultó ser aplicable. Se determinó que el tamaño de las granjas y la densidad de población, no son un factor predisponente en estas granjas, pero las condiciones predisponentes a la presentación de enfermedades difieren entre granjas de ciclo completo y engorda

La investigación sobre género, sexualidades y cuerpos en la educación secundaria argentina. Un estado de la cuestión

El artículo desarrolla un estado del arte sobre la investigación educativa en Argentina que aborda el cruce entre género, sexualidades y cuerpo en la escuela secundaria. Se describen las líneas y enfoques predominantes en el campo, los temas con mayor desarrollo, que plantean novedosos de pensar la escuela secundaria desde una perspectiva de género. El artículo pone en diálogo el enfoque de género con algunas de las preocupaciones centrales de la investigación educativa y la agenda pública de discusión sobre la educación secundaria en Argentina, como la construcción de poder entre estudiantes, la producción de la desigualdad, el rol docente y el desarrollo de políticas que buscan ofrecer otra experiencia de convivencia y sociabilidad en el marco de derechos. Se identifican los aportes de la investigación sobre las relaciones de género, las construcciones de las sexualidades y la producción y regulación de las corporalidades en la escuela secundaria, tanto como un enfoque transversal como un subcampo específico en las discusiones y aportes de la investigación. La expansión en la producción científica sobre estos temas confluye con la consolidación del feminismo académico, la articulación con el movimiento de mujeres, disidencias y diversidad sexual, y una agenda política que le brinda cada vez mayor especificidad al tópico, al tiempo que se identifican problemáticas sobre la producción de desigualdades en la escuela secundaria argentina. Se asume el enfoque de género en educación como una categoría para repensar casi todo acerca de los procesos de escolarización en el nivel medio: saberes, vínculos y relaciones pedagógicas, participación y convivencia, currículum, trayectorias, políticas e incluso modalidade

Artrodesis tibiotalocalcanea con placa philos. Experiencia en el Hospital Militar Central. Enero 2007 a diciembre de 2009

Estudio descriptivo observacional, retrospectivo tipo serie de casos, realizado entre enero de 2007 y diciembre de 2009 en el Hospital Militar Central. Fueron tratados 21 pacientes con artrosis de la articulación tibio talar y subtalar de etiología diversa, con seguimiento mínimo de 6 meses. Se evaluó;la función pre operatoria y post operatoria por medio de la escala AOFAS, el dolor mediante la escala visual análoga, el tiempo de consolidación con controles radiográficos, el tipo de injerto utilizado y las complicaciones asociadas. Se encontraron como complicaciones 2 pacientes con infección superficial, que requirió manejo con antibiótico endovenoso, y 2 pacientes con retardo en la consolidación que requirieron cirugía de revisión. Los resultados obtenidos demuestran que la artrodesis tibiotalocalcanea con placa philos es un excelente método de fijación (90,4% consolidación) en pacientes con artrosis del tobillo y sub talar y mala calidad ósea

The European Eel NCCβ Gene Encodes a Thiazide-Resistant Na-Cl Cotransporter

The thiazide-sensitive Na-Cl cotransporter (NCC) is the major pathway for salt reabsorption in the mammalian distal convoluted tubule. NCC plays a key role in the regulation of blood pressure. Its inhibition with thiazides constitutes the primary baseline therapy for arterial hypertension. However, the thiazide-binding site in NCC is unknown. Mammals have only one gene encoding for NCC. The eel, however, contains a duplicate gene. NCCα is an ortholog of mammalian NCC and is expressed in the kidney. NCCβ is present in the apical membrane of the rectum. Here we cloned and functionally characterized NCCβ from the European eel. The cRNA encodes a 1043-amino acid membrane protein that, when expressed in Xenopus oocytes, functions as an Na-Cl cotransporter with two major characteristics, making it different from other known NCCs. First, eel NCCβ is resistant to thiazides. Single-point mutagenesis supports that the absence of thiazide inhibition is, at least in part, due to the substitution of a conserved serine for a cysteine at position 379. Second, NCCβ is not activated by low-chloride hypotonic stress, although the unique Ste20-related proline alanine-rich kinase (SPAK) binding site in the amino-terminal domain is conserved. Thus, NCCβ exhibits significant functional differences from NCCs that could be helpful in defining several aspects of the structure-function relationship of this important cotransporter

Sequence and structural variations determining the recruitment of WNK kinases to the KLHL3 E3 ligase

This is the final version. Available on open access from Portland Press via the DOI in this recordData Availability: Structure factors and co-ordinates have been deposited in the PDB with accession code 5NKP. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [50] partner repository with the dataset identifier PXD031606 and 10.6019/PXD031606.The BTB-Kelch protein KLHL3 is a Cullin3-dependent E3 ligase that mediates the ubiquitin-dependent degradation of kinases WNK1-4 to control blood pressure and cell volume. A crystal structure of KLHL3 has defined its binding to an acidic degron motif containing a PXXP sequence that is strictly conserved in WNK1, WNK2 and WNK4. Mutations in the second proline abrograte the interaction causing the hypertension syndrome pseudohypoaldosteronism type II. WNK3 shows a diverged degron motif containing four amino acid substitutions that remove the PXXP motif raising questions as to the mechanism of its binding. To understand this atypical interaction, we determined the crystal structure of the KLHL3 Kelch domain in complex with a WNK3 peptide. The electron density enabled the complete 11-mer WNK-family degron motif to be traced for the first time revealing several conserved features not captured in previous work, including additional salt bridge and hydrogen bond interactions. Overall, the WNK3 peptide adopted a conserved binding pose except for a subtle shift to accommodate bulkier amino acid substitutions at the binding interface. At the centre, the second proline was substituted by WNK3 Thr541, providing a unique phosphorylatable residue among the WNK-family degrons. Fluorescence polarisation and structural modelling experiments revealed that its phosphorylation would abrogate the KLHL3 interaction similarly to hypertension-causing mutations. Together, these data reveal how the KLHL3 Kelch domain can accommodate the binding of multiple WNK isoforms and highlight a potential regulatory mechanism for the recruitment of WNK3

Role of KLHL3 and dietary K⁺ in regulating KS-WNK1 expression

This is the author accepted manuscript. The final version is available from the American Physiological Society via the DOI in this recordThe physiological role of the shorter isoform of WNK1 that is exclusively expressed in the kidney (KS-WNK1), with particular abundance in the distal convoluted tubule, remains elusive. KS-WNK1 despite lacking the kinase domain, is nevertheless capable of stimulating the NaCl cotransporter (NCC), apparently through activation of WNK4. It has recently been shown that a less severe form of the Familial Hyperkalemic Hypertension featuring only hyperkalemia is caused by missense mutations in the WNK1 acidic domain that preferentially affect CUL3-KLHL3 E3-induced degradation of KS-WNK1, rather than that of the full-length WNK1 (L-WNK1). Here we show that L-WNK1 is indeed less impacted by the CUL3-KLHL3 E3 ligase complex compared to KS-WNK1. We demonstrate that the unique 30 amino acid amino N-terminal fragment of KS-WNK1 is essential for its activating effect on NCC and recognition by KLHL3. We identify specific amino acid residues in this region critical for the functional effect of KS-WNK1 and KLHL3 sensitivity. To further explore this, we generated KLHL3-R528H knock-in mice that mimic human mutations causing Familial Hyperkalemic Hypertension. These mice revealed that the KLHL3 mutation specifically increased expression of KS-WNK1 in the kidney. We also observed that in wild type mice, expression of KS-WNK1 is only detectable after exposure to low potassium diet. These findings provide new insights into the regulation and function of KS-WNK1 by the CUL3-KLHL3 complex in DCT and indicate that this pathway is regulated by dietary K+ levels.National Institutes of Health (NIH)Conacyt MexicoPAPIIT UNAML'OréalMedical Research Council (MRC

NRBP1 and TSC22D proteins affect distal convoluted tubule physiology through modulation of the WNK pathway

The with-no-lysine (K) (WNK) kinases regulate processes such as cell volume and epithelial ion transport through the modulation of cation chloride cotransporters such as the NaCl cotransporter (NCC) present in the distal convoluted tubule (DCT) of the kidney. Recently, the interaction of WNKs with nuclear receptor binding protein 1 (NRBP1) and transforming growth factor–β–stimulated clone 22 domain (TSC22D) proteins was reported. Here, we explored the effect of NRBP1 and TSC22Ds on WNK signaling in vitro and in the DCT. TSC22D1.1, TSC22D2, and NRBP1 are localized in DCT WNK bodies, which are cytoplasmic biomolecular condensates associated with WNK activation. In HEK293 cells, long TSC22D isoforms and NRBP1 increase WNK4 activity. DCT-specific NRBP1-knockout mice have reduced NCC phosphorylation and activate a compensatory response. Thus, NRBP1 and long TSC22D proteins are positive modulators of WNK signaling and modulate Na+ reabsorption in the kidney. NRBP1 and TSC22Ds likely influence WNK signaling in other tissues, affecting various physiological processes.</p

Epidemiology of inflammatory bowel disease in Mexico and Colombia: Analysis of health databases, mathematical modelling and a case-series study

Background and aims Ulcerative Colitis (UC) and Crohn’s Disease (CD) have a major impact on quality of life and medical costs. The aim of the study was to estimate the prevalence, incidence and clinical phenotypes of Inflammatory Bowel Disease (IBD) cases in Mexico and Colombia. Methods We analyzed official administrative and health databases, used mathematical modelling to estimate the incidence and complete prevalence, and performed a case-series of IBD patients at a referral center both in Mexico and Colombia. Results The age-adjusted complete prevalence of UC per 100,000 inhabitants for 2015/2016 ranged from 15.65 to 71.19 in Mexico and from 27.40 to 69.97 in Colombia depending on the model considered. The prevalence of CD per 100,000 inhabitants in Mexico ranged from 15.45 to 18.08 and from 16.75 to 18.43 in Colombia. In Mexico, the age-adjusted incidence of UC per 100,000 inhabitants per year ranged from 0.90 to 2.30, and from 0.55 to 2.33 in Colombia. The incidence for CD in Mexico ranged from 0.35 to 0.66 whereas in Colombia, the age-adjusted incidence of CD ranged from 0.30 to 0.57. The case-series included 200 IBD patients from Mexico and 204 patients from Colombia. The UC/CD prevalence ratio in Mexico and Colombia was 1.50:1 and 4.5:1 respectively. In Mexico, the female/male prevalence ratio for UC was 1.50:1 and 1.28:1 for CD, while in Colombia this ratio was 0.68:1 for UC and 0.8:1 for CD. In Mexico the relapse rate for UC was 63.3% and 72.5% for CD, while those rates in Colombia were 58.2% for UC and 58.3% for CD. Conclusions The estimated burden of disease of IBD in Mexico and Colombia is not negligible. Although these findings need to be confirmed by population-based studies, they are useful for decision-makers, practitioners and patients with this condition.Fil: Ciapponi, Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Epidemiología y Salud Pública. Instituto de Efectividad Clínica y Sanitaria. Centro de Investigaciones en Epidemiología y Salud Pública; ArgentinaFil: Virgilio, Sacha Alexis. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Berrueta, Mabel. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Claire Soto, Natalie. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Ciganda, Álvaro. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Rojas Illanes, Moisés Freddy. Instituto de Efectividad Clínica y Sanitaria; ArgentinaFil: Rubio Martinez, Briseida. Hospital Dr. Bernardo Sepúlveda; MéxicoFil: Gamba, Johana. Fundación Universitaria Sanitas; ColombiaFil: González Salazar, Carlos Arturo. Fundación Universitaria Sanitas; ColombiaFil: Rocha Rodríguez, José Nicolás. Fundación Universitaria Sanitas; ColombiaFil: Scarpellini, Bruno. No especifíca;Fil: Bravo Perdomo, Ana María. No especifíca;Fil: Machnicki, Gerardo. No especifíca;Fil: Aldunate, Leandro. No especifíca;Fil: De Paula, Juan. Hospital Italiano; ArgentinaFil: Bardach, Ariel Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Epidemiología y Salud Pública. Instituto de Efectividad Clínica y Sanitaria. Centro de Investigaciones en Epidemiología y Salud Pública; Argentin