Comparison of potentiation post-activation responses in two dominant hip strength exercises

The aim of this study was to evaluate whether an anterior-posterior (hip thrust [HT]) movement generated higher post-activation potentiation (PAP) levels on broad jump (BJ) in comparison to an axial-component exercise (deadlift [DL]). Fifteen resistance-trained rugby players participated in this study (age, 22.7 ± 1.6; body weight, 89.9 ± 10.6; height, 181.8 ± 6.5; BMI, 27.2 ± 2.3; 1RM DL 1RM, 117 ± 20.6; 1RM HT, 133.3 ± 21.5. Subjects attended four sessions to the laboratory with seven days between each session. Anthropometric measurements and one-repetition maximum (1-RM) estimations for HT and DL (grip width 72 cm) were performed in the first and third sessions, while PAP tests for both exercise protocols were performed during the second and fourth sessions (subjects performed two repetitions at 90% 1-RM with 8 minutes of recovery between HT and DL exercises, and then the BJ was performed). Data were analyzed by a general lineal model (GLM) with repeated measures and a delta analysis (Δ = post – pre) was performed to determine the 95% confidence interval (95% CI) for the mean. The GLM showed both significant difference and large effect size on BJ per Time [pre vs. post] (P > 0.001; ηp2 = 0.990), but no differences on Time x Protocol interaction (P = 0.452; ηp2=0.020) or per Group (0.748; ηp2 = 0.004) were found. There were significant changes [X ± SD, CIs 95%] on BJ after HT (5.2±5.6, 2.1–8.3; P = 0.03) and DL (6.9±5.3, 4.0–9.9: P < 0.001); however, no difference was found between the exercise protocols (P = 0.748). Our results suggest that DL and HP provided with large effects on PAP to improve the BJ outcomes, but there was no difference between these exercises.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Identification of novel point mutations in c-kit gene from Leukemia cases: a study from Lucknow, Uttar Pradesh, India

The c-kit gene is a receptor tyrosine kinase (RTK) class III that is expressed in early hematopoietic progenitor cells. Aberrantly activated RTK and related downstream signaling partners have been reported as key elements in the molecular pathogenesis of several malignancies. Within the c-kit gene exon-11 is the most frequent site for mutations in different kinds of tumours. Mutations in c-kit gene may enhance or interfere with the ability of c-kit receptor to initiate the intracellular pathways resulting in cell proliferation. Therefore, we aimed to screen the mutations in c-kit gene at exon-8 and -11 in malignant Leukemias. Ninety Leukemia cases were studied and analyzed by mutation- specific PCR-SSCP followed by DNA sequencing. Twenty point mutations were detected in eight AML (acute myeloid Leukemia) cases within exon-11 which includes Tyr568Ser, Ile571Thr, Thr574Pro, Gln575His, Tyr578Pro, Asp579His, His580Gln, Arg586Thr, Asn587Asp and Arg588Met. The substitutions Lys550Asn, Ile571Leu and Trp582Ser were observed in two independent cases and four novel point mutations at codons Ile563Lys, Val569Leu, Tyr570Ser, and Pro577Ser. Further, six point mutations were detected at exon-8 in six cases (four AML and two CML cases), comprising three novel mutations Asn423Asp, Gln448Thr, and Gln448His. The point mutations Thr417Asp, Tyr418Phe, and Leu421His were observed, but were detected only in three cases. These observations suggest that mutations in c-kit gene might represent a useful molecular genetic marker in Leukemia and incidence of mutation at exon- 8 and -11 is high and might be involve in pathogenesis of AML. Resumen El gen c-kit, que codifica para un receptor tirosina quinasa (RTK) de clase III, se expresa en las primeras células progenitoras hematopoyéticas. La activación de este RTK y su vía de señalización se encuentran involucradas en la patogénesis molecular de varias enfermedades. La mutación del gen c-kit en el exón 11 es una de las mutaciones más frecuentemente reportadas en diferente tipos de tumores. Mutaciones en c-kit podrían incrementar o interferir con la habilidad del receptor c-kit para iniciar la activación de cascadas de señalización intracelulares responsables en la proliferación celular. Por estas razones, estudiamos las mutaciones del gen c-kit en el exon 8 y 11 en casos con Leucemias. Noventa casos de Leucemia en la India fueron estudiados mediante PCR SSCP, seguida por secuenciación de DNA. Veinte mutaciones puntuales fueron detectadas en el exon 11 en tan solo ocho de los casos con AML (leucemia mieloide aguda), entre las que encontraron las mutaciones Tyr568Ser, Ile571Thr, Thr574Pro, Gln575His, Tyr578Pro, Asp579His, His580Gln, Arg586Thr, Asn587Asp y Arg588Met. Las sustituciones Lys550Asn, Ile571Leu y Trp582Ser fueron observadas en tan solo dos casos. Además, cuatro nuevas mutaciones para los codones Ile563Lys, Val569Leu, Tyr570Ser, y Pro577Ser se observaron en este estudio. En el exon 8, seis mutaciones puntuales fueron observadas y en seis de los casos (cuatro en AML y dos en CML) encontramos tres nuevas mutaciones Asn423Asp, Gln448Thr y Gln448His. Sin embargo, las mutaciones puntuales Thr417Asp, Tyr418Phe y Leu421His fueron observadas en varias ocasiones, pero en tan solo tres de los casos estudiados. Estas observaciones sugieren que las mutaciones en c-kit podrían representar un marcador genético para Leucemia. La incidencia en la mutación del exon 8 y 11 es elevada y podría estar relacionada con la patogénesis de la AML. Palabras clave: c-kit, exones 8 y 11, Leucemia, mutación, SSCP PAGE

Cypermethrin, deltamethrin and glyphosate affect the activity of the Ca2 +-ATPase from human erythrocyte

Abstract The extensive use of pesticides can cause many human health problems. However, the effects of pesticides on a biochemical level are still poorly understood. In this study we analyzed the effect of different pesticides on the plasma membrane Ca2+-ATPase (PMCA) and on the sarco/ endoplasmic reticulum Ca2+-ATPase (SERCA) activities. Different amounts of pesticides were added to the Ca2+- dependent ATPase assays in order to determine if they were affecting the ATPase activity. The results showed that PMCA activity was partially inhibited by deltamethrin to 51.85% ± 3.7 when its concentration in the reaction medium was 0.5 mM, while cypermethrin and glyphosate stimulated the PMCA activity at the lowest concentrations tested. The maximum stimulatory effect of cypermethrin was of 155% ± 9.0 at a concentration of 0.2 mM. In addition, the herbicide glyphosate stimulated the activity 111% ± 2.0 at a concentration of 0.2 mM. In conclusion, our results showed that PMCA activity was partially inhibited by deltamethrin, but cypermethrin and glyphosate stimulated their activity. Our findings suggest that cypermethrin and deltamethrin have different structure- activity relationships. However, SERCA was not sensitive to deltamethrin or glyphosate. These differences may be reflected in disturbs over cellular calcium regulation. Resumen Se ha observado que la exposición de personas a plaguicidas puede causar problemas a la salud. Sin embargo, el estudio del efecto de plaguicidas a un nivel bioquímico ha sido pobremente estudiado. En este estudio, analizamos el efecto de cipermetrina, deltametrina y glifosato sobre la actividad enzimática de las Ca2+- ATPasa de membrana plasmática (PMCA) y de retículo sarcoplásmico (SERCA). Diferentes concentraciones de estos pesticidas fueron añadidas a los experimentos de actividad enzimática como estrategia para determinar si estos compuestos eran capaces de afectar la actividad enzimática de PMCA. Nuestros resultados demuestran que la actividad enzimática de PMCA fue parcialmente inhibida por deltametrina en un 51.85% ± 3.7 cuando su concentración fue de 0.5 mM, mientras que cipermetrina y glifosato estimularon la actividad enzimática de PMCA a menores concentraciones. El máximo efecto estimulatorio de cipermetrina fue de 155% ± 9.0 cuando el compuesto alcanzó una concentración de 0.2 mM. Además, el herbicida glifosato fue capaz de estimular la actividad enzimática de PMCA en un 111% ± 2.0 a concentraciones de 0.1-0.2 mM. En conclusión, nuestros resultados demostraron que la actividad enzimática de PMCA fue también parcialmente inhibida con deltametrina, pero al contrario de cipermetrina y glifosato la estimularon. Nuestros resultados sugieren que las diferencias en las estructuras químicas de cipermetrina y deltametrina se ven reflejadas en el efecto provocado sobre PMCA. Sin embargo, una enzima similar, SERCA, no fue afectada ni por deltametrina ni glifosato. Estas diferencias se pudieran ver reflejadas en disturbios sobre la regulación celular del calcio. Palabras clave: PMCA (ATPasa de Ca2+ de membrana plasmática), SERCA (ATPasa de Ca2+ de retículo endo/sarcoplásmico), actividad enzimática, cipermetrina, deltametrina, glifosato

Procesos de Oxidación avanzada en el tratamiento de agua

A lo largo de este libro diversos autores especializados exponen el tema permitiendo al lector encontrar desde principios básicos, hasta aplicaciones de procesos, resultando ser una fuente de consulta con una visión amplia de los procesos de oxidación avanzada y sus aplicaciones dentro del tratamiento de agua.El agua es un líquido vital, sin ella no podemos subsistir. Además de usarla en nuestro hogar, se utiliza en gran variedad de procesos industriales para la transformación de materias primas en productos terminados. El agua usada industrialmente cambia su composición fisicoquímica, ya que agregamos un sinfín de compuestos orgánicos e inorgánicos. Por ello, es necesario desarrollar nuevas metodologías que permitan de manera segura y eficiente recuperar la calidad del agua usada originalmente para poder usarla.Universidad Autónoma del Estado de Méxic

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

American College of Rheumatology Provisional Criteria for Clinically Relevant Improvement in Children and Adolescents With Childhood-Onset Systemic Lupus Erythematosus

10.1002/acr.23834ARTHRITIS CARE & RESEARCH715579-59

Phosphorylation of the glycine transporter 1

The extracellular levels of the neurotransmitter glycine in the brain are tightly regulated by the high-affinity glycine transporter 1 (GlyT1) and the clearance of glycine depends on its rate of transport and the levels of cell surface GlyT1. Over the past years, it has been shown that PKC activation diminishes the activity and promoted phosphorylation of several neurotransmitter transporters including the dopamine, serotonin and norepinephrine transporters however, its role is unknown for the glycine transporter. To get insights into the role of PKC activation on GlyT1 regulation, we used three N-terminus GlyT1 isoforms stably expressed in porcine aortic endothelial (PAE) cells and assaying for [32P]-orthophosphate metabolic labeling. We demonstrated that the isoforms GlyT1a, GlyT1b, and GlyT1c were constitutively phosphorylated, and that phosphorylation was dramatically enhanced, in a time-dependent fashion, after PKC activation by phorbol ester (PMA). The phosphorylation was PKC-dependent, since pre-incubation of the cells with bisindolylmaleimide I (BIM), a selective PKC inhibitor, abolished the phorbol ester-induced phosphorylation. Moreover, blotting of a purified GlyT1 fraction with specific antibodies to phosphorylated tyrosine residues did not yield any signal that could correspond to GlyT1 phosphorylation, suggesting that the phosphorylation occurs at serine and/or threonine residues. In addition, by using more specific inhibitors to the different PKC isoenzymes, we were able to determine the PKC isoenzyme(s) involved in downregulation of glycine uptake and GlyT1 phosphorylation. Specifically, we found that pre-incubation of the cells with the selective PKCα/β inhibitor Go6976 completely abolished the effect of phorbol ester on uptake and phosphorylation. On the other hand, incubation with either selective PKCβ inhibitors (PKCβ inhibitor or LY333531) prevented the PKC-dependent phosphorylation of GlyT1 without affecting the downregulation triggered by PMA. Taken together, this data suggest that conventional PKCα/β regulates the uptake of glycine and probably its efflux, whereas PKCβ is responsible for GlyT1 phosphorylation. In order to determine the sites of phosphorylation, we mutated all the threonine and serine residues found at either, the N- or C-terminus to alanine residues. Incubation of cells stably expressing the GlyT1 N- or C-mutants with PMA triggered phosphorylation at similar levels to those obtained for the wild-type transporter, suggesting that both N- and C-terminal tails in the GlyT1 are phosphorylated. Although the results demonstrate GlyT1 phosphorylation, the role of this modification still remains to be elucidated. Additionally, in order to obtain structural information about the potential glycine-binding site, a model of three-dimensional structure of the GlyT1 was built based on the atomic coordinates of the related bacterial leucine transporter (LeuT). The resulting model showed a helical bundle and a substrate binding-pocket similar to the LeuT. This model was used to analyze the reactivity of cysteine residues in the GlyT1 to fluorescently-labeled maleimides. The reactivity of the cysteine residues was assayed by incubation of cells expressing the wild-type transporter with the hydrophilic fluorescein maleimide resulting in labeling of transporter without any significant effect on glycine uptake. By contrast, incubation with either hydrophobic pyrenyl or coumarin maleimides resulted in rapid inhibition of glycine uptake. These results suggest that inhibition of the activity could be due a modification of a cysteine residue(s) lying in or near to the glycine-binding site. In agreement with this hypothesis, our model suggests the presence of the Cys-152 in the putative substrate-pocket. Further mutagenesis analysis combined with chemical modification should pave the way for studying the glycine-binding site and translocation pathway in the GlyT1

Revisión: Mecanismos moleculares de la neurofibromatosis tipo 2

En este trabajo presentamos una revisión sobre hallazgos más relevantes de la neurofibromatosis tipo 2 (NF2), la cual se conoce por ser un desorden autosómico dominante caracterizado por la presencia de schwannomas vestibulares bilaterales, aunque pueden presentarse otros tumores como meningiomas y ependimomas. Esta enfermedad es causada por diversas mutaciones en el gen NF2, mismo que codifica una proteína conocida como merlina o schwannomina. Merlina está relacionada estructuralmente con la familia de proteínas ERM (Ezrina-Radixina- Moesina), encargadas de acoplar las señales provenientes de las glucoproteínas de la membrana plasmática con el citoesqueleto de actina. El gen NF2 es considerado como un supresor de tumores, y las evidencias indican que merlina funciona regulando la proliferación y el crecimiento celular. Sin embargo, los mecanismos específicos por medio de los cuales merlina cumple con su función siguen siendo un enigma. Se han identificado diversas moléculas que interactúan con merlina, lo que ha proporcionado indicios acerca de los diversos procesos celulares en los cuales esta molécula participa. Entre las proteínas que interactúan con merlina se incluyen proteínas de función estructural, receptores de membrana plasmática, proteínas citosólicas, GTPasas y adaptadores citoesqueléticos. Las mutaciones en el gen NF2 afectan la funcionalidad de merlina, lo que produce alteraciones en los mecanismos de acción de merlina dando como origen a la NF2. Son necesarios más estudios para determinar con certeza el papel de merlina en el control de la proliferación celular. Abstract In this work, we present a review over the most relevant information of the neurofibromatosis type 2 (NF2), which is known as an autosomal dominant disorder characterized by the presence of bilateral vestibular schwannomas. Other tumors such as meningiomas and ependymomas may be present. The disease is caused by mutations in the NF2 gene, which encodes a protein known as merlin or schwannomin. Merlin is structurally related to the ERM (Ezrina-Radixina-Moesina) family of proteins, a group of molecules responsible for linking the signals coming from the plasma membrane glycoproteins to the actin cytoskeleton. The NF2 gene is considered as a tumor suppressor gene, and the evidence indicates that merlin functions by regulating the cell growth and proliferation. However, the specific mechanisms through which merlin fulfill its functions as a tumor suppressor remains enigmatic. Several molecules that interact with merlin have been identified. This has provided clues to determine the cellular processes in which merlin participates. These molecules include structural proteins, plasma membrane receptors, cytosolic proteins, GTPases, and cytoskeletal adapters. Mutations in the NF2 gene affect the functionality of merlin, altering tha mecanisms of action of merlin, giving rise to NF2. Further studies are needed to determine the precise role of merlin on the control of cell proliferation. Keywords: neurofibromatosis type 2, merlin, cytoskeleton, plasma membrane

El plaguicida glifosato incrementa la Vmax de la Ca2 +-ATPasa de membrana plasmática de eritrocito humano sin afectar su afinidad (Km) por el substrato

Los plaguicidas son un amplio grupo de sustancias heterogéneas que producen un beneficio mediante la disminución de vectores que trasmiten enfermedades, y en el control de plagas que afectan la producción agrícola. En el presente trabajo estudiamos el efecto del glifosato (GF) sobre la actividad hidrolítica de la ATPasa dependiente de Ca2+ de membrana (PMCA) de eritrocito humano en su forma nativa (sin calmodulina [CaM]) o en el complejo activo CaM-PMCA. Nuestros resultados evidenciaron que el GF produjo un efecto dual, estimulatorio e inhibitorio, y el efecto fue dosis-dependiente: concentraciones bajas de GF (0.05-0.1 mM) estimularon la hidrólisis de ATP por PMCA en su conformación nativa en un 62.5%, y en el complejo activo por CaM el incremento fue de tan solo del 20% (basados en sus respectivos controles sin GF). Ambas formas conformaciones de PMCA fueron parcialmente inhibidas con GF 0.3-0.5 y 0.4-0.5 mM. El análisis sobre el efecto de GF en la afinidad de la enzima por el substrato (ATP), mostró que GF fue capaz de incrementar significativamente el consumo de ATP (Vmax), pero solo para la conformación nativa de PMCA. Sin embargo, la afinidad de la enzima por el sustrato (Km) no fue afectada en ninguna de las dos conformaciones de PMCA. En conclusión, GF afecta la velocidad del ciclo catalítico de PMCA sin afectar su afinidad por el sustrato, ello puede deberse a una interacción del plaguicida con el C-terminal de PMCA. Este efecto, pudiera ocasionar disturbios sobre la homeostasis celular del calcio desencadenando la activación de procesos celulares tales como la apoptosis. Abstract Pesticides are a broad group of heterogeneous substances with great benefit to the human population, as they are used for decreasing or killing a broad number of pests involved in transmission of diseases or involved in decreasing the levels of food production. In this work we studied the effect of glyphosate (GF) over the hydrolytic activity of plasma membrane Ca2+-ATPase (PMCA) from human erythrocytes, either in their native conformational state (without calmodulin [CaM]) or in the active complex CaM-PMCA. Our results showed that GF produced a dual effect, activation and inhibition, and the effect was a dose-response effect: at low-concentrations of GF (0.05-0.1 mM), ATP hydrolysis mediated by PMCA was stimulated up to 62.5%, however with CaM the increase was only 20% (with respect to the controls without GF). Both conformational forms of PMCA were partially inhibited with GF 0.3-0.5 and 0.4- 0.5 mM respectively. An analysis performed on PMCA affinity for the substrate (ATP) showed that GF was significantly increasing the ATP consumption (Vmax) and it was only significant for PMCA in their native conformation. However, for the two conformations studied, GF was unable to significantly affect the affinity of PMCA for the substrate ATP (Km). In conclusion, GF affects the velocity of the PMCA’s catalytic cycle without affecting its affinity for ATP, and this may be due to an interaction between GF and the C-terminus of PMCA. This effect may disturb calcium homeostasis in the cell, leading to the activation of several cellular processes such as apoptosis. Keywords: Ca2+-ATPase, plasma membrane, glyphosate, calmodulin, human erythrocyte