Effect of SARS-CoV-2 S protein on the proteolytic cleavage of the epithelial Na+ channel ENaC.

Severe cases of COVID-19 are characterized by development of acute respiratory distress syndrome (ARDS). Water accumulation in the lungs is thought to occur as consequence of an exaggerated inflammatory response. A possible mechanism could involve decreased activity of the epithelial Na+ channel, ENaC, expressed in type II pneumocytes. Reduced transepithelial Na+ reabsorption could contribute to lung edema due to reduced alveolar fluid clearance. This hypothesis is based on the observation of the presence of a novel furin cleavage site in the S protein of SARS-CoV-2 that is identical to the furin cleavage site present in the alpha subunit of ENaC. Proteolytic processing of αENaC by furin-like proteases is essential for channel activity. Thus, competition between S protein and αENaC for furin-mediated cleavage in SARS-CoV-2-infected cells may negatively affect channel activity. Here we present experimental evidence showing that coexpression of the S protein with ENaC in a cellular model reduces channel activity. In addition, we show that bidirectional competition for cleavage by furin-like proteases occurs between 〈ENaC and S protein. In transgenic mice sensitive to lethal SARS-CoV-2, however, a significant decrease in gamma ENaC expression was not observed by immunostaining of lungs infected as shown by SARS-CoV2 nucleoprotein staining.

Microencapsulation of Theobroma cacao L polyphenols: A high-value approach with in vitro anti-Trypanosoma cruzi, immunomodulatory and antioxidant activities.

Chagas disease (CHD) is the highest economic burden parasitosis worldwide and the most important cardiac infection, without therapeutic alternatives to halt or reverse its progression. In CHD-experimental models, antioxidant and anti-inflammatory compounds have demonstrated therapeutic potential in cardiac dysfunction. Theobroma cacao polyphenols are potent natural antioxidants with cardioprotective and anti-inflammatory action, which are susceptible to degradation, requiring technological approaches to guarantee their protection, stability, and controlled release. Here, 21 cocoa polyphenol-rich microencapsulates were produced by spray-drying and freeze-drying techniques using two wall materials (maltodextrin and gum arabic). Chemical (total and individual phenolic content and antioxidant activity), structural (morphology), and biological parameters (cytotoxicity, trypanocidal, antioxidant, and immunomodulatory activities) were assessed to determine the most efficient microencapsulation conditions on Trypanosoma cruzi-infected myocardioblast and macrophage cells. Significant antiproliferative properties against infected cells (superior to benznidazole) were found in two microencapsulates which also exhibited cardioprotective properties against oxidative stress, inflammation, and cell death.

DHA Supplementation during Pregnancy in Women with Obesity Normalizes IGF2R Levels in the Placenta of Male Newborns.

Introduction: Insulin-like growth factor receptor 2 (IGF2R) regulates placental nutrient transport, and its soluble form is related to obesity in adults. If the placental expression of IGF2R is altered in women with obesity is unknown. Whether maternal supplementation with docosahexaenoic acid (DHA), a polyunsaturated fatty acid with anti-inflammatory properties, has a modulatory role in IGF2R's function has not been elucidated. We hypothesized that maternal obesity (Ob) would be associated with alterations in placental IGF2R expression, which may be prevented with DHA supplementation during pregnancy. Methods: At delivery, we obtained placentas from women with Ob (BMI ≥ 30 kg/m2, n = 17), Ob supplemented with 800 mg/day of DHA during pregnancy (Ob + DHA, n = 13), and normal-weight women (Nw, BMI ≥ 18.5 ≤ 24.9 kg/m2, n = 14). The IGF2R mRNA and protein were determined by RT-PCR and western blotting, respectively. Moreover, we quantified the gene expression of molecules that modulate the IGF2R function in the extracellular domain, such as TACE/ADAM17, PLAU, and IGF2. Mann-Whitney and Kruskal-Wallis nonparametric tests were used to compare results between two or three groups accordingly. Results: The IGF2R levels in the Ob placentas of the male offspring were higher than in the Nw group. The DHA supplementation prevented this effect, suggesting an unknown relationship between IGF2R-Ob-DHA in placental tissues. Conclusion: We report, for the first time, that DHA supplementation during pregnancy in women with obesity normalizes the increased IGF2R levels in male placentas, reducing the risk of adverse outcomes related to the IGF2/IGF2R system in male newborns.

Metabolic Interaction Between Folate, Vitamin B12, and Polyunsaturated Fatty Acids in Pregnancy.

Fetal growth and development are influenced by maternal nutrition and gestational weight gain. Adequate intake of nutrients such as folate, vitamin B12, and docosahexaenoic acid (DHA) is essential for healthy fetal and placental development. Many countries have a national flour fortification program with folic acid (FA), together with pre-pregnancy supplementation of FA (400 µg/day) during the first trimester of pregnancy. The latter has been recommended by the WHO and adapted to local requirements by perinatal guidelines. On the other hand, in population studies, many women of childbearing age have vitamin B12 deficiency (<148 pmol/L), which can be additionally masked by high FA intake and maternal pregestational obesity. Under these conditions, these patients could be having pregnancies in a folate/vitamin B12 imbalance, which is associated with higher adiposity, insulin resistance, altered lipid metabolism, and low DHA levels in their offspring. However, if these neonatal consequences of maternal pregestational obesity and folate/vitamin B12 imbalance can be reverted by DHA supplementation during pregnancy has not been addressed. This chapter reviews the literature and exposes the current gaps in knowledge and challenges in maternal nutrition with a life-course perspective.

Structure-function relationships in the sodium chloride cotransporter.

The thiazide sensitive Na+:Cl- cotransporter (NCC) is the principal via for salt reabsorption in the apical membrane of the distal convoluted tubule (DCT) in mammals and plays a fundamental role in managing blood pressure. The cotransporter is targeted by thiazide diuretics, a highly prescribed medication that is effective in treating arterial hypertension and edema. NCC was the first member of the electroneutral cation-coupled chloride cotransporter family to be identified at a molecular level. It was cloned from the urinary bladder of the Pseudopleuronectes americanus (winter flounder) 30 years ago. The structural topology, kinetic and pharmacology properties of NCC have been extensively studied, determining that the transmembrane domain (TM) coordinates ion and thiazide binding. Functional and mutational studies have discovered residues involved in the phosphorylation and glycosylation of NCC, particularly on the N-terminal domain, as well as the extracellular loop connected to TM7-8 (EL7-8). In the last decade, single-particle cryogenic electron microscopy (cryo-EM) has permitted the visualization of structures at high atomic resolution for six members of the SLC12 family (NCC, NKCC1, KCC1-KCC4). Cryo-EM insights of NCC confirm an inverted conformation of the TM1-5 and TM6-10 regions, a characteristic also found in the amino acid-polyamine-organocation (APC) superfamily, in which TM1 and TM6 clearly coordinate ion binding. The high-resolution structure also displays two glycosylation sites (N-406 and N-426) in EL7-8 that are essential for NCC expression and function. In this review, we briefly describe the studies related to the structure-function relationship of NCC, beginning with the first biochemical/functional studies up to the recent cryo-EM structure obtained, to acquire an overall view enriched with the structural and functional aspects of the cotransporter.

The European and Japanese eel NaCl cotransporters ß exhibit chloride currents and are resistant to thiazide type diuretics.

The thiazide-sensitive Na+-Cl- cotransporter (NCC) is the major pathway for salt reabsorption in the mammalian distal convoluted tubule, and the inhibition of its function with thiazides is widely used for the treatment of arterial hypertension. In mammals and teleosts, NCC is present as one ortholog that is mainly expressed in the kidney. One exception, however, is the eel, which has two genes encoding NCC. The eNCCα is located in the kidney and eNCCß, which is present in the apical membrane of the rectum. Interestingly, the European eNCCß functions as a Na+-Cl- cotransporter that is nevertheless resistant to thiazides and is not activated by low-chloride hypotonic stress. However, in the Japanese eel rectal sac, a thiazide-sensitive NaCl transport mechanism has been described. The protein sequences between eNCCß and jNCCß are 98% identical. Here, by site-directed mutagenesis, we transformed eNCCß into jNCCß. Our data showed that jNCCß, similar to eNCCß, is resistant to thiazides. In addition, both NCCß proteins have high transport capacity with respect to their renal NCC orthologs and, in contrast to known NCCs, exhibit electrogenic properties that are reduced when residue I172 is substituted by A, G, or M. This is considered a key residue for the chloride ion-binding sites of NKCC and KCC. We conclude that NCCß proteins are not sensitive to thiazides and have electrogenic properties dependent on Cl-, and site I172 is important for the function of NCCß.

Factors Associated with Development of Acute Kidney Injury After Liver Transplantation.

BACKGROUND: Early post-liver transplant (LT) acute kidney injury (AKI) has been associated with worse short-term and long-term outcomes, but the incidence and risk factors in our population are unknown. METHODS: We designed a prospective, singlecenter, longitudinal cohort study to determine the incidence of AKI during the immediate postoperative period of LT, and to identify the risk factors associated with AKI after LT. Pre-operative and intraoperative variables were analyzed to determine if there was any correlation with the development of post-operative AKI. RESULTS: Eighty-six patients were included in the final analysis; from them, 45 (52%) developed AKI in the following 30 days after LT. The presence of hepatic encephalopathy prior to LT was the factor most strongly associated with the development of AKI (Relative Risk 3.67, 95% Confidence Interval 1.08-8.95). Other factors associated with AKI development were male gender and a higher serum lactate during surgery. CONCLUSION: AKI was a frequent complication that significantly worsened the prognosis of LT recipients and was associated with an increased 30-day mortality rate. The presence of hepatic encephalopathy strongly predicted the development of severe AKI.

Immunomodulatory, trypanocide, and antioxidant properties of essential oil fractions of Lippia alba (Verbenaceae).

BACKGROUND: Parasite persistence, exacerbated and sustained immune response, and continuous oxidative stress have been described to contribute to the development of the cardiac manifestations in Chronic Chagas Disease. Nevertheless, there are no efficient therapies to resolve the Trypanosoma cruzi infection and prevent the disease progression. Interestingly, trypanocide, antioxidant, and immunodulatory properties have been reported separately for some major terpenes, as citral (neral plus geranial), limonene, and caryophyllene oxide, presents in essential oils (EO) extracted from two chemotypes (Citral and Carvone) of Lippia alba. The aim of this study was to obtain L. alba essential oil fractions enriched with the aforementioned bioactive terpenes and to evaluate the impact of these therapies on trypanocide, oxidative stress, mitochondrial bioenergetics, genotoxicity, and inflammatory markers on T. cruzi-infected macrophages. METHODS: T. cruzi-infected J774A.1 macrophage were treated with limonene-enriched (ACT1) and citral/caryophyllene oxide-enriched (ACT2) essential oils fractions derived from Carvone and Citral-L. alba chemotypes, respectively. RESULTS: ACT1 (IC50 = 45 ± 1.7 µg/mL) and ACT2 (IC50 = 80 ± 1.9 µg/mL) exhibit similar trypanocidal effects to Benznidazole (BZN) (IC50 = 48 ± 2.5 µg/mL), against amastigotes. Synergistic antiparasitic activity was observed when ACT1 was combined with BZN (∑FIC = 0.52 ± 0.13 µg/mL) or ACT2 (∑FIC = 0.46 ± 1.7 µg/mL). ACT1 also decreased the oxidative stress, mitochondrial metabolism, and genotoxicity of the therapies. The ACT1 + ACT2 and ACT1 + BZN experimental treatments reduced the pro-inflammatory cytokines (IFN-γ, IL-2, and TNF-α) and increased the anti-inflammatory cytokines (IL-4 and IL-10). CONCLUSION: Due to its highly trypanocidal and immunomodulatory properties, ACT1 (whether alone or in combination with BZN or ACT2) represents a promising L. alba essential oil fraction for further studies in drug development towards the Chagas disease control.

Disagreement between PCR and serological diagnosis of Trypanosoma cruzi infection in blood donors from a Colombian endemic region. / Desacuerdo entre la PCR y la serología en el diagnóstico de la infección por Trypanosoma cruzi en donantes de una región endémica de Colombia

INTRODUCTION: Chagas' disease is the leading cause of infectious myocarditis worldwide. This infection caused by Trypanosoma cruzi is usually life-long and asymptomatic; however, the third part of infected people can develop severe or even fatal cardiomyopathy. As the parasitemia in the chronic phase is both low-grade and intermittent, T. cruzi infection is principally detected by serology, although this method has sensitivity and specificity limitations. OBJECTIVE: To determine the level of agreement between serologic and molecular tests in 658 voluntary blood donors from six provinces in the Colombian department of Santander. MATERIALS AND METHODS: We evaluated an array of diagnostic technologies by cross-section sampling performing a serological double diagnostic test for T. cruzi antibody detection (Chagas III ELISA™, BiosChile Group, and ARCHITECT Chagas CMIA™, Abbott), and DNA detection by polymerase chain reaction (PCR). We collected the demographic, clinical, and epidemiological information of participants. The sample size was calculated using Epidat™ and the statistical analysis was done with Stata 12.1™. RESULTS: PCR was six times more sensitive in detecting T. cruzi infection than ELISA/CMIA with prevalence values of 1.8% (12/658) and 0.3% (2/658), respectively, and kappa=0.28 (95%CI: -0.03 - 0.59). In contrast, serology showed a sensitivity of 16.7% (95%CI: 2.09 - 48.4) and a specificity of 100% (95%CI: 99.4 - 100). All seropositive samples were found to be positive by PCR. CONCLUSIONS: The implementation of PCR as a complementary method for screening donors could reduce the probability of false negative and the consequent risk of transfusional-transmission of Chagas' disease, especially in endemic regions.

Introducción. La enfermedad de Chagas constituye la principal causa de miocarditis infecciosa en el mundo. Causada por Trypanosoma cruzi, la infección puede persistir toda la vida de manera asintomática y silenciosa, pero un tercio de los infectados desarrolla cardiomiopatía grave. Debido a que la parasitemia en la fase crónica es baja e intermitente, el diagnóstico se hace principalmente mediante la detección de anticuerpos (serología), método que tiene limitaciones de sensibilidad y especificidad. Objetivo. Determinar la concordancia entre el diagnóstico serológico y molecular de T. cruzi en 658 donantes voluntarios de sangre del departamento de Santander, Colombia. Materiales y métodos. Se hizo un estudio de evaluación de tecnologías diagnósticas con muestreo transversal, utilizando un doble diagnóstico serológico para la detección de anticuerpos anti-T. cruzi (Chagas III ELISA™, BiosChile Group, y ARCHITECT Chagas CMIA™, Abbott) y la de ADN por PCR. Se recolectó la información demográfica, clínica y epidemiológica de los participantes. El tamaño de la muestra se estimó utilizando Epidat™ y el análisis estadístico se hizo mediante Stata 12.1™. Resultados. La sensibilidad de la PCR fue seis veces mayor que la de las pruebas de ELISA/CMIA, con prevalencias de 1,8 % (12/658) y 0,3 % (2/658), respectivamente, y kappa de 0,28 (IC95% -0,03 - 0,59). La sensibilidad serológica fue de 16,7 % (IC95% 2,09 - 48,4) y la especificidad de 100 % (IC95% 99,4 - 100). Todas las muestras seropositivas fueron positivas también en la PCR. Conclusiones. El uso de la PCR como método complementario para la tamización de donantes podría reducir el riesgo de falsos negativos y disminuir los casos de transmisión transfusional de la enfermedad de Chagas, especialmente en regiones endémicas.

Maternal Obesity Is Associated With Higher Cord Blood Adipokines in Offspring Most Notably in Females.

BACKGROUND: Deleterious long-term effects in the offspring from women with pregravid obesity have been described; however, the evidence supporting early metabolic and inflammatory markers in the offspring at birth and gender differences are conflicting. OBJECTIVE: The present study aimed to compare cord blood adipokines and cytokines concentrations and anthropometric characteristics of the offspring of women with maternal obesity (MO) and normal-weight mothers (NWM). Also, maternal and neonatal variables on the association of maternal body mass index (BMI) with cord blood adipokines were evaluated. METHODS: A cross-sectional analysis of a subsample of mother-child dyads participating in a cohort study (n = 221) was assessed. Anthropometrics, cord blood adipokines (leptin and adiponectin) and cytokines (interleukin [IL]-1ß, IL-4, IL-10, IL-12 p40, IL-12p70, IL-13, and tumor necrosis factor α) concentrations in the offspring of normal-weight women (BMI >18.5 and <24.9 kg/m2) and women with pregravid obesity (BMI > 30 kg/m2) without comorbidities was performed. RESULTS: Offspring from mothers with obesity had higher birth weight, a higher proportion of large for gestational age, higher ponderal index, and heavier placentae than offspring from normal-weight mothers (P < 0.05). Within the offspring from women with obesity, males had significantly higher weight, length, the proportion of large-for-gestational-age newborns, higher weight for length ratio. Males had more efficient placentas than females (P < 0.05). Higher adiponectin and leptin in both sexes and higher leptin in female offspring of mothers with obesity after adjusting for birth size (P < 0.05) were found. Higher IL-12p40 in the offspring of women with MO with no other differences in other cytokines among groups were evidenced. CONCLUSIONS: Maternal obesity associates with a higher concentration of adiponectin and leptin in their offspring at birth. There is a relevant effect on anthropometrics in male offspring and on leptin in female newborn. Further studies need to evaluate the extension of these effects in postnatal life. TRAIL IDENTIFICATION NUMBER: NCT02903134.

Disagreement between PCR and serological diagnosis of Trypanosoma cruzi infection in blood donors from a Colombian endemic region / Desacuerdo entre la PCR y la serología en el diagnóstico de la infección por Trypanosoma cruzi en donantes de una región endémica de Colombia

Abstract | Introduction: Chagas' disease is the leading cause of infectious myocarditis worldwide. This infection caused by Trypanosoma cruzi is usually life-long and asymptomatic; however, the third part of infected people can develop severe or even fatal cardiomyopathy. As the parasitemia in the chronic phase is both low-grade and intermittent, T. cruzi infection is principally detected by serology, although this method has sensitivity and specificity limitations. Objective: To determine the level of agreement between serologicand molecular tests in 658 voluntary blood donors from six provinces in the Colombian department of Santander. Materials and methods: We evaluated an array of diagnostic technologies by cross-section sampling performing a serological double diagnostic test for T. cruzi antibody detection (Chagas III ELISA™, BiosChile Group, and ARCHITECT Chagas CMIA™, Abbott) , and DNA detection by polymerase chain reaction (PCR). We collected the demographic, clinical, and epidemiological information of participants. The sample size was calculated using Epidat™ and the statistical analysis was done with Stata 12.1™. Results: PCR was six times more sensitive in detecting T. cruzi infection than ELISA/CMIA with prevalence values of 1.8% (12/658) and 0.3% (2/658), respectively, and kappa=0.28 (95%CI: -0.03 - 0.59). In contrast, serology showed a sensitivity of 16.7% (95%CI: 2.09 - 48.4) and a specificity of 100% (95%CI: 99.4 - 100). All seropositive samples were found to be positive by PCR. Conclusions: The implementation of PCR as a complementary method for screening donors could reduce the probability of false negative and the consequent risk of transfusional-transmission of Chagas' disease, especially in endemic regions.

Resumen | Introducción. La enfermedad de Chagas constituye la principal causa de miocarditis infecciosa en el mundo. Causada por Trypanosoma cruzi,la infección puede persistir toda la vida de manera asintomática y silenciosa, pero un tercio de los infectados desarrolla cardiomiopatía grave. Debido a que la parasitemia en la fase crónica es baja e intermitente, el diagnóstico se hace principalmente mediante la detección de anticuerpos (serología), método que tiene limitaciones de sensibilidad y especificidad. Objetivo. Determinar la concordancia entre el diagnóstico serológico y molecular de T. cruzien 658 donantes voluntarios de sangre del departamento de Santander, Colombia. Materiales y métodos. Se hizo un estudio de evaluación de tecnologías diagnósticas con muestreo transversal, utilizando un doble diagnóstico serológico para la detección de anticuerpos anti-T. cruzi (Chagas III ELISA™, BiosChile Group, y ARCHITECT ChagasCMIA™, Abbott) y la de ADN por PCR. Se recolectó la información demográfica, clínica y epidemiológica de los participantes. El tamaño de la muestra se estimó utilizando Epidat™ y el análisis estadístico se hizo mediante Stata 12.1™. Resultados. La sensibilidad de la PCR fue seis veces mayor que la de las pruebas de ELISA/CMIA, con prevalencias de 1,8 % (12/658) y 0,3 % (2/658), respectivamente, y kappa de 0,28 (IC95% -0,03 - 0,59). La sensibilidad serológica fue de 16,7 % (IC95% 2,09 - 48,4) y la especificidad de 100 % (IC95% 99,4 - 100). Todas las muestras seropositivas fueron positivas también en la PCR. Conclusiones. El uso de la PCR como método complementario para la tamización de donantes podría reducir el riesgo de falsos negativos y disminuir los casos de transmisión transfusional de la enfermedad de Chagas, especialmente en regiones endémicas.

WNK3 and WNK4 exhibit opposite sensitivity with respect to cell volume and intracellular chloride concentration.

Cation-coupled chloride cotransporters (CCC) play a role in modulating intracellular chloride concentration ([Cl-]i) and cell volume. Cell shrinkage and cell swelling are accompanied by an increase or decrease in [Cl-]i, respectively. Cell shrinkage and a decrease in [Cl-]i increase the activity of NKCCs (Na-K-Cl cotransporters: NKCC1, NKCC2, and Na-Cl) and inhibit the activity of KCCs (K-Cl cotransporters: KCC1 to KCC4), wheras cell swelling and an increase in [Cl-]i activate KCCs and inhibit NKCCs; thus, it is unlikely that the same kinase is responsible for both effects. WNK1 and WNK4 are chloride-sensitive kinases that modulate the activity of CCC in response to changes in [Cl-]i. Here, we showed that WNK3, another member of the serine-threonine kinase WNK family with known effects on CCC, is not sensitive to [Cl-]i but can be regulated by changes in extracellular tonicity. In contrast, WNK4 is highly sensitive to [Cl-]i but is not regulated by changes in cell volume. The activity of WNK3 toward NaCl cotransporter is not affected by eliminating the chloride-binding site of WNK3, further confirming that the kinase is not sensitive to chloride. Chimeric WNK3/WNK4 proteins were produced, and analysis of the chimeras suggests that sequences within the WNK's carboxy-terminal end may modulate the chloride affinity. We propose that WNK3 is a cell volume-sensitive kinase that translates changes in cell volume into phosphorylation of CCC.

Fatty acid and lipid metabolism in liver of pregnant mice and their offspring is influenced by unbalanced folates/vitamin B12 diets.

Micronutrients (folates and vitamin B12) and long chain polyunsaturated fatty acids (LC-PUFAs) are linked through the one carbon cycle. We studied the effects of pre and postnatal high FA/low B12 diets (HFLB12) on hepatic fatty acid metabolism. Pregnant C57BL/6 mice were divided in two groups: control (2 mg folic acid: FA/25 µg vitamin B12/Kg food) and HFLB12 diets (8 mg FA/5 µg vitamin B12/Kg food). Offspring continued on the same diets until 60 days old. We determined hepatic fatty acid profile in dams and offspring and the expression of PPARα, Cpt-1, Acox-1 and Fas and the enzymatic activity of desaturases, all involved in lipid metabolism. In liver of dams, the HFHB12 diet decreased total fatty acids and desaturase activities; in offspring, effects were opposite, being more noticeable in females. Prenatal and postnatal unbalanced folic acid/B12 diets play a crucial role in regulating genes and enzymes involved in lipid metabolism in liver of dams and their offspring in adulthood.

Inhibition of C. albicans Dimorphic Switch by Cobalt(II) Complexes with Ligands Derived from Pyrazoles and Dinitrobenzoate: Synthesis, Characterization and Biological Activity.

Seven cobalt(II) complexes of pyrazole derivatives and dinitrobenzoate ligands were synthesized and characterized. The single-crystal X-ray diffraction structure was determined for one of the ligands and one of the complexes. The analysis and spectral data showed that all the cobalt complexes had octahedral geometries, which was supported by DFT calculations. The complexes and their free ligands were evaluated against fungal strains of Candida albicans and emerging non-albicans species and epimastigotes of Trypanosoma cruzi. We obtained antifungal activity with a minimum inhibitory concentration (MIC) ranging from 31.3 to 250 µg mL-1. The complexes were more active against C. krusei, showing MIC values between 31.25 and 62.5 µg mL-1. In addition, some ligands (L1-L6) and complexes (5 and Co(OAc)2 · 4H2O) significantly reduced the yeast to hypha transition of C. albicans at 500 µg mL-1 (inhibition ranging from 30 to 54%). Finally, the complexes and ligands did not present trypanocidal activity and were not toxic to Vero cells. Our results suggest that complexes of cobalt(II) with ligands derived from pyrazoles and dinitrobenzoate may be an attractive alternative for the treatment of diseases caused by fungi, especially because they target one of the most important virulence factors of C. albicans.

Structure-function relationships in the renal NaCl cotransporter (NCC).

The thiazide-sensitive Na+-Cl- cotransporter (NCC) is the major pathway for salt reabsorption in the distal convoluted tubule, serves as a receptor for thiazide-type diuretics, and is involved in inherited diseases associated with abnormal blood pressure. The functional and structural characterization of NCC from different species has led us to gain insights into the structure-function relationships of the cotransporter. Here we present an overview of different studies that had described these properties. Additionally, we report the cloning and characterization of the NCC from the spiny dogfish (Squalus acanthias) kidney (sNCC). The purpose of the present study was to determine the main functional, pharmacological and regulatory properties of sNCC to make a direct comparison with other NCC orthologous. The sNCC cRNA encodes a 1033 amino acid membrane protein, when expressed in Xenopus oocytes, functions as a thiazide-sensitive Na-Cl cotransporter with NCC regulation and thiazide-inhibition properties similar to mammals, rather than to teleosts. However, the Km values for ion transport kinetics are significantly higher than those observed in the mammal species. In summary, we present a review on NCC structure-function relationships with the addition of the sNCC information in order to enrich the NCC cotransporter knowledge.

Entrenamiento de capacidades físicas condicionantes en la calidad de vida relacionada con la salud en personas mayores con epoc estadio I y II. Revisión sistemática de literatura / Training of conditioning physical capabilities in quality of life related to health in older people with copd stadium I and II. Systematic review of literature / Treinamento de capacidades de condicionamento físico na qualidade de vida relacionada à saúde em idosos com epoc estágios I e II. Revisão sistemática da literatura

Introducción. La enfermedad pulmonar obstructiva crónica (EPOC) es una patología sistémica y multifactorial que requiere manejo integral e intervención multidisciplinaria. Estudios evidencian la necesidad de que los adultos mayores con EPOC ingresen a un programa de rehabilitación pulmonar en busca la reducción de la disnea y la sensación de cansancio, además de incrementar la tolerancia al ejercicio y la calidad de vida. El entrenamiento de las capacidades físicas condicionantes reduce los efectos que causan la disminución de la capacidad pulmonar en el adulto mayor con EPOC, de la misma manera, favorece los aspectos fisiológicos, las relaciones interpersonales y la calidad de vida. Objetivo. Determinar el efecto del entrenamiento de las capacidades físicas condicionantes en la calidad de vida relacionada con la salud en personas mayores con EPOC, estadios I y II. Materiales y métodos. Se realizó una revisión bibliográfica de artículos científicos, publicados en español e inglés, en bases de datos y bibliotecas virtuales como PUBMED/MEDLINE, EMBASE, LILACS, SCIELO, Registro Cochrane Central de Ensayos Controlados (CENTRAL); se seleccionaron ECA que relacionaran EPOC en adultos mayores y capacidades condicionantes con calidad de vida. Resultado. Las intervenciones con capacidades físicas condicionantes mejoran la calidad de vida relacionada con la salud, en adultos mayores con EPOC y la percepción de estado de salud, además, se observaron mejoras significativas en los condicionantes físicos de fuerza, resistencia muscular y velocidad de la marcha. Conclusiones. La práctica de las capacidades físicas condicionantes tiene efectos positivos sobre la calidad de vida relacionada con la salud en adultos mayores con EPOC.

Introduction. Chronic obstructive pulmonary disease (COPD) is a systemic and multifactorial pathology that requires comprehensive management and multidisciplinary intervention. Studies show the need for older adults with COPD to go into a pulmonary rehabilitation program that may promote the reduction of dyspnea and tiredness, in addition to increase tolerance to exercise and quality of life. Improve physical capacities decline the decrement in lung capacity in the older adult with COPD, moreover it favors the physiological aspects, the interpersonal relationships and the life's quality. Objective. To determine the training effect of the physical conditioning capacities in the quality of life related to health in elderly people with COPD stages I and II. Materials and Methods. A literature review of scientific articles published in Spanish and English languages, articles was found in databases and virtual libraries as PUBMED / MEDLINE, EMBASE, LILACS, SciELO, Cochrane Central Register of Controlled Trials (CENTRAL) were selected RCTs that relate COPD in older adults, conditioning capacities with life's quality. Result: The interventions with conditioning physical capacities improve life's quality related to health, in older adults with COPD and the perception of health status, significant improvements were observed in the physical conditioning factors of strength, muscular resistance and gait speed. Conclusions. The practice of conditioning physical capacities has positive effects on life's quality related to health in older adults with COPD.

Introdução. A doença pulmonar obstrutiva crônica (DPOC) é uma patologia sistêmica e multifatorial que requer uma gestão abrangente e intervenção multidisciplinar. Estudos mostram a necessidade que idosos com DPOC entram num programa de reabilitação pulmonar em busca da redução da dispnéia e da sensação de cansaço, além de aumentar a tolerância ao exercício e a qualidade de vida. O treinamento de capacidades de condicionamento físico reduz os efeitos que causam diminuição da capacidade pulmonar nos idosos com DPOC, da mesma forma, favorece os aspectos fisiológicos, as relações interpessoais e a qualidade de vida. Objetivo. Determinar o efeito do treinamento de capacidades de condicionamento físico na qualidade de vida relacionada à saúde em idosos com DPOC, estágios I e II. Materiais e métodos. Foi realizada uma revisão bibliográfica de artigos científicos, publicados em espanhol e inglês, em bancos de dados e bibliotecas virtuais como PUBMED / MEDLINE, EMBASE, LILACS, SCIELO, Registro Central de Ensaios Controlados da Cochrane (CENTRAL); ECRs; foram selecionados o ECA que relacionava DPOC em idosos e capacidades condicionantes com qualidade de vida. Resultado. As intervenções com capacidades de condicionamento físico melhoram a qualidade de vida relacionada à saúde em idosos com DPOC e a percepção do estado de saúde, além disso, foram observadas melhorias significativas nas condições físicas de força, resistência muscular e velocidade da marcha. Conclusões. A prática das capacidades de condicionamento físico tem efeitos positivos na qualidade de vida relacionada à saúde em idosos com DPOC.

Audiología y Mareo de Origen Cervical / Audiology and dizziness of cervical origin

En la presente revisión sistemática se discute la relación entre la etiología del mareo de origen cervical, el uso de pruebas vestibulares y el rol del especialista en audiología en el abordaje de esta patología. La búsqueda de artículos se realizó a través de bases de datos electrónicas. Se usaron términos libres como mareo de origen cervical, vértigo cervicogénico, cervicogenic dizziness, dolor cervical, latigazo cervical y espondilosis cervical. La valoración de la calidad de los estudios incluidos se realizó con la guía de análisis crítico de calidad metodológica propuesta por (Carvajal C, 2004). Los resultados de la revisión muestran las evidencias etiológicas del mareo de origen cervical reportadas en la literatura y si el especialista en audiología, por su formación, puede abordar esta condición de acuerdo con los procedimientos hallados para su manejo actualmente. Las evidencias sugieren que la afectación de las estructuras musculo esqueléticas del cuello puede derivar en sensaciones de mareo. Adicionalmente, a partir de la revisión se concluye que, aunque la literatura no reporte el rol específico del especialista en audiología en el abordaje del mareo de origen cervical, es pertinente que este lo aborde desde su fase diagnóstica y de inter vención

This systematic review discusses the relationship between the etiology of cervical dizziness, the use of ear canal testing, and the role of the audiology specialist in approaching this pathology. An electronic database was used in researching articles that utilized open terminologies such as cervical dizziness, cervicogenic vertigo, cervicogenic dizziness, sharp cervical pain, and cervical spondylosis. The studies included were evaluated for quality by using the critical analysis guide of methodological quality. The results of the review show reported etiological evidence of cervicogenic dizziness in literature and whether an audiology specialist, through their training, is able to address and handle this condition in accordance with the most up-to-date procedures. The evidence suggests that the affectation of the musculoskeletal framework in the neck can be derived from dizziness symptoms. Additionally, it could be concluded from the review that although the literature does not report the specific role of the audiology specialist in approaching cervical dizziness, it is pertinent to address this role as early as the diagnostic and intervention phases

Induction of programmed cell death in Trypanosoma cruzi by Lippia alba essential oils and their major and synergistic terpenes (citral, limonene and caryophyllene oxide).

BACKGROUND: Chagas Disease caused by Trypanosoma cruzi infection, is one of the most important neglected tropical diseases (NTD), without an effective therapy for the successful parasite eradication or for the blocking of the disease's progression, in its advanced stages. Due to their low toxicity, wide pharmacologic spectrum, and potential synergies, medicinal plants as Lippia alba, offer a promising reserve of bioactive molecules. The principal goal of this work is to characterize the inhibitory properties and cellular effects of the Citral and Carvone L. alba chemotype essential oils (EOs) and their main bioactive terpenes (and the synergies among them) on T. cruzi forms. METHODS: Twelve L. alba EOs, produced under diverse environmental conditions, were extracted by microwave assisted hydrodistillation, and chemically characterized using gas chromatography coupled mass spectrometry. Trypanocidal activity and cytotoxicity were determined for each oil, and their major compounds, on epimastigotes (Epi), trypomastigotes (Tryp), amastigotes (Amas), and Vero cells. Pharmacologic interactions were defined by a matrix of combinations among the most trypanocidal terpenes (limonene, carvone; citral and caryophyllene oxide). The treated cell phenotype was assessed by fluorescent and optic microscopy, flow cytometry, and DNA electrophoresis assays. RESULTS: The L. alba EOs displayed significant differences in their chemical composition and trypanocidal performance (p = 0.0001). Citral chemotype oils were more trypanocidal than Carvone EOs, with Inhibitory Concentration 50 (IC50) of 14 ± 1.5 µg/mL, 22 ± 1.4 µg/mL and 74 ± 4.4 µg/mL, on Epi, Tryp and Amas, respectively. Limonene exhibited synergistic interaction with citral, caryophyllene oxide and Benznidazole (decreasing by 17 times its IC50) and was the most effective and selective treatment. The cellular analysis suggested that these oils or their bioactive terpenes (citral, caryophyllene oxide and limonene) could be inducing T. cruzi cell death by an apoptotic-like mechanism. CONCLUSIONS: EOs extracted from L. alba Citral chemotype demonstrated significant trypanocidal activity on the three forms of T. cruzi studied, and their composition and trypanocidal performance were influenced by production parameters. Citral, caryophyllene oxide, and limonene showed a possible induction of an apoptotic-like phenotype. The best selective anti-T. cruzi activity was achieved by limonene, the effects of which were also synergic with citral, caryophyllene oxide and benznidazole.

Geraniin is a diuretic by inhibiting the Na+-K+-2Cl- cotransporter NKCC2.

Geranium seemannii Peyr is a perennial plant endemic to central Mexico that has been widely used for its diuretic effect, but the responsible compound of this effect is unknown as well as the mechanism by which the diuretic effect is achieved. Geraniin is one of the compounds isolated from this kind of geranium. This study was designed to determinate whether geraniin possesses diuretic activity and to elucidate the mechanism of action. Geraniin was extracted and purified from Geranium seemannii Peyr. Male Wistar rats were divided into four groups: 1) Control, 2) 75 mg/kg of geraniin, 3) 20 mg/kg of furosemide, and 4) 10 mg/kg of hydrochlorothiazide. Each treatment was administered by gavage every 24 h for 7 days. The urinary excretion of electrolytes and the fractional excretion of sodium (FENa) were determined. To uncover the molecular target of geraniin, Xenopus laevis oocytes were microinjected with cRNAs encoding the Na+-Cl- cotransporter (NCC) and the Na+-K+-2Cl- cotransporter NKCC2 to functionally express these cotransporters. Geraniin significantly increased diuresis, natriuresis, and calciuresis to a similar extent as was observed in the furosemide-treated rats. Consistent with the furosemide-like effect, in X. laevis oocytes, geraniin significantly reduced the activity of NKCC2, with no effect on NCC activity. In contrast to furosemide, the effect of geraniin on NKCC2 was irreversible, apparently due to its inhibitory effect on heat shock protein 90. Our observations suggest that geraniin could have a potential role in the treatment of hypertension or edematous states.

The European Eel NCCß 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. NCCß is present in the apical membrane of the rectum. Here we cloned and functionally characterized NCCß 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 NCCß 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, NCCß 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, NCCß exhibits significant functional differences from NCCs that could be helpful in defining several aspects of the structure-function relationship of this important cotransporter.