Conformational Stability of Hepatitis C Virus NS3 Protease

AbstractThe hepatitis C virus NS3 protease is responsible for the processing of the nonstructural region of viral precursor polyprotein in infected hepatic cells. NS3 has been considered a target for drug discovery for a long time. NS3 is a zinc-dependent serine protease. However, the zinc ion is not involved in the catalytic mechanism, because it is bound far away from the active site. Thus, zinc is essential for the structural integrity of the protein and it is considered to have a structural role. The first thermodynamic study on the conformational equilibrium and stability of NS3 and the effect of zinc on such equilibrium is presented here. In agreement with a previous calorimetric study on the binding of zinc to NS3, the global unfolding heat capacity is dominated by the zinc dissociation step, suggesting that the binding of zinc induces a significant structural rearrangement of the protein. In addition, contrary to other homologous zinc-dependent proteases, the zinc-free NS3 protease is not completely unstructured. It is apparent that the conformational landscape of hepatitis C virus NS3 protease is fairly complex due to its intrinsic plasticity, and to the interactions with its different effectors (zinc and the accessory viral protein NS4A) and their modulation of the population of the different conformational states

Biophysical analysis of the MHR motif in folding and domain swapping of the HIV capsid protein C-terminal domain

© 2015 Biophysical Society. Infection by human immunodeficiency virus (HIV) depends on the function, in virion morphogenesis and other stages of the viral cycle, of a highly conserved structural element, the major homology region (MHR), within the carboxyterminal domain (CTD) of the capsid protein. In a modified CTD dimer, MHR is swapped between monomers. While no evidence for MHR swapping has been provided by structural models of retroviral capsids, it is unknown whether it may occur transiently along the virus assembly pathway. Whatever the case, the MHR-swapped dimer does provide a novel target for the development of anti-HIV drugs based on the concept of trapping a nonnative capsid protein conformation. We have carried out a thermodynamic and kinetic characterization of the domain-swapped CTD dimer in solution. The analysis includes a dissection of the role of conserved MHR residues and other amino acids at the dimerization interface in CTD folding, stability, and dimerization by domain swapping. The results revealed some energetic hotspots at the domain-swapped interface. In addition, many MHR residues that are not in the protein hydrophobic core were nevertheless found to be critical for folding and stability of the CTD monomer, which may dramatically slow down the swapping reaction. Conservation of MHR residues in retroviruses did not correlate with their contribution to domain swapping, but it did correlate with their importance for stable CTD folding. Because folding is required for capsid protein function, this remarkable MHR-mediated conformational stabilization of CTD may help to explain the functional roles of MHR not only during immature capsid assembly but in other processes associated with retrovirus infection. This energetic dissection of the dimerization interface in MHR-swapped CTD may also facilitate the design of anti-HIV compounds that inhibit capsid assembly by conformational trapping of swapped CTD dimers.Spanish Government (BIO2012-37649) and Comunidad de Madrid (S-2009/MAT/1467) and by an institutional grant from Fundación Ramón Areces.Peer Reviewe

Actividad física y calidad de vida en estudiantes de las instituciones educativas del convenio CIRCA del distrito de Yura, provincia de Arequipa, 2021

La investigación actividad física y la calidad de vida en instituciones de en las instituciones educativas del Consorcio Circa del distrito de Yura provincia de Arequipa surge de la necesidad de establecer en qué medida la actividad física influye en la calidad de vida de los estudiantes. Asimismo, se considera que son variables esenciales de la salud de una población o de una persona. Por esto es importante evaluar las dos variables que son percibidas en todas las etapas de la vida, especialmente en la población infantil. La investigación fue de enfoque cuantitativo, de nivel correlacional, diseño no experimental de tipo transversal. La muestra representada por 146 estudiantes de las Instituciones Educativas del Consorcio Circa del distrito de Yura provincia de Arequipa, los instrumentos aplicados fueron un cuestionario para la actividad física el cual fue validado por un juicio de expertos obteniendo la aprobación del 98%. y un cuestionario de calidad de vida en el cual su Alfa de Cronbach obtuvo un índice de confiabilidad de 0,901, el cual indica que es altamente confiable. Se concluyó que a través de la prueba estadística no paramétrica de Rho de Spearman, se observó que el valor de significancia asintótica tiene un valor de 0,381 el cual es mayor al establecido de 0,05 y al 95% de confianza, este valor indica que se debe de aceptar la hipótesis nula y rechazar la hipótesis alterna. En base a este resultado se concluye que “La actividad física NO influye significativamente en la calidad de vida de los estudiantes en las instituciones educativas del Consorcio Circa del distrito de Yura, provincia de Arequipa, 2021”

Functional Characterization of Nupr1L, A Novel p53-Regulated Isoform of the High-Mobility Group (HMG)-Related Protumoral Protein Nupr1

We have previously demonstrated a crucial role of nuclear protein 1 (NUPR1) in tumor development and progression. In this work, we report the functional characterization of a novel Nupr1-like isoform (NUPR1L) and its functional interaction with the protumoral factor NUPR1. Through the use of primary sequence analysis, threading, and homology-based molecular modeling, as well as expression and immunolocalization, studies reveal that NUPR1L displays properties, which are similar to member of the HMG-like family of chromatin regulators, including its ability to translocate to the cell nucleus and bind to DNA. Analysis of the NUPR1L promoter showed the presence of two p53-response elements at positions -37 and -7, respectively. Experiments using reporter assays combined with site-directed mutagenesis and using cells with controllable p53 expression demonstrate that both of these sequences are responsible for the regulation of NUPR1L expression by p53. Congruently, NUPR1L gene expression is activated in response to DNA damage induced by oxaliplatin treatment or cell cycle arrest induced by serum starvation, two well-validated methods to achieve p53 activation. Interestingly, expression of NUPR1L downregulates the expression of NUPR1, its closely related protumoral isoform, by a mechanism that involves the inhibition of its promoter activity. At the cellular level, overexpression of NUPR1L induces G1 cell cycle arrest and a decrease in their cell viability, an effect that is mediated, at least in part, by downregulating NUPR1 expression. Combined, these experiments constitute the first functional characterization of NUPR1L as a new p53-induced gene, which negatively regulates the protumoral factor NUPR1.Fil: Lopez, Maria Belen. Centre de Recherche En Cancerologie de Marseille; FranciaFil: Garcia, Maria Noé. Centre de Recherche En Cancerologie de Marseille; FranciaFil: Grasso, Daniel Hector. Centre de Recherche En Cancerologie de Marseille; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Bintz, Jennifer. Centre de Recherche En Cancerologie de Marseille; FranciaFil: Molejon, Maria Ines. Centre de Recherche En Cancerologie de Marseille; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Velez, Gabriel. Mayo Clinic; Estados UnidosFil: Lomberk, Gwen. Mayo Clinic; Estados UnidosFil: Neira, Jose Luis. Universidad de Miguel Hernández; EspañaFil: Urrutia, Raul. Mayo Clinic; Estados UnidosFil: Iovanna, Juan. Centre de Recherche En Cancerologie de Marseille; Franci

Spatial characterization of climatic variables for Arica-Parinacota and Tarapacá, Chile using topoclimatic analysis

In the present study, models were developed to determine the monthly and annual spatio-temporal variation of temperature, precipitation, and solar radiation based on topoclimatic analysis of Arica-Parinacota and Tarapacá in northern Chile. To construct the equations of the topoclimatic model, the data from meteorological stations and physiographic factors (latitude, longitude, altitude, and distance to bodies of water) obtained from a digital terrain model with a resolution of 90 m were compiled in a database. The equations of the topoclimatic model were generated by a stepwise regression with a backward selection technique. The equations for average monthly temperature, precipitation, and solar radiation were determined by linear combinations. The results were statistically significant with coefficients of determination greater than 90%, in addition to being greater than the existing climate databases for this area

Caracterización multi-ambiental de la fecha de floración en cultivares de Raigrás anual (L. multiflorum Lam.)

Publicado dentro de la Revista Argentina de Producción Animal, v 39 Suplemento 1 (2019).El importante número de cultivares de raigrás anual (RGA) presentes en el mercado, genera la necesidad de conocer tanto su potencial productivo como el comportamiento fenológico de los mismos, de manera de poder determinar que material se adapta mejor a cada sistema productivo. El objetivo del presente trabajo fue caracterizar el ciclo a floración de cultivares de Raigrás anual en diferentes localidades de la Argentina.EEA PergaminoFil: Ré, Alejo Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concepción del Uruguay; UruguayFil: Lavandera, Javier Eduardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Forrajeras; ArgentinaFil: Barbera, Pablo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes (Corrientes); ArgentinaFil: Romero, Luis Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Gallego, Juan José. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Valle Inferior de Río Negro; ArgentinaFil: Neira Zilli, Fernanda. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Valle Inferior de Río Negro; ArgentinaFil: Méndez, Daniel Gustavo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria General Villegas; Argentin

Genomic characterization of individuals presenting extreme phenotypes of high and low risk to develop tobacco-induced lung cancer

Single nucleotide polymorphisms (SNPs) may modulate individual susceptibility to carcinogens. We designed a genome-wide association study to characterize individuals presenting extreme phenotypes of high and low risk to develop tobacco-induced non-small cell lung cancer (NSCLC), and we validated our results. We hypothesized that this strategy would enrich the frequencies of the alleles that contribute to the observed traits. We genotyped 2.37 million SNPs in 95 extreme phenotype individuals, that is: heavy smokers that either developed NSCLC at an early age (extreme cases); or did not present NSCLC at an advanced age (extreme controls), selected from a discovery set (n = 3631). We validated significant SNPs in 133 additional subjects with extreme phenotypes selected from databases including >39,000 individuals. Two SNPs were validated: rs12660420 (pcombined  = 5.66 × 10-5 ; ORcombined  = 2.80), mapping to a noncoding transcript exon of PDE10A; and rs6835978 (pcombined  = 1.02 × 10-4 ; ORcombined  = 2.57), an intronic variant in ATP10D. We assessed the relevance of both proteins in early-stage NSCLC. PDE10A and ATP10DmRNA expressions correlated with survival in 821 stage I-II NSCLC patients (p = 0.01 and p < 0.0001). PDE10A protein expression correlated with survival in 149 patients with stage I-II NSCLC (p = 0.002). In conclusion, we validated two variants associated with extreme phenotypes of high and low risk of developing tobacco-induced NSCLC. Our findings may allow to identify individuals presenting high and low risk to develop tobacco-induced NSCLC and to characterize molecular mechanisms of carcinogenesis and resistance to develop NSCLC.This work was supported by the Spanish Society of Medical Oncology; Fundación SEOM and Fundación Salud 2000; and Government of Navarra.S

Genomic characterization of individuals presenting extreme phenotypes of high and low risk to develop tobacco-induced lung cancer

Single nucleotide polymorphisms (SNPs) may modulate individual susceptibility to carcinogens. We designed a genome-wide association study to characterize individuals presenting extreme phenotypes of high and low risk to develop tobacco-induced non-small cell lung cancer (NSCLC), and we validated our results. We hypothesized that this strategy would enrich the frequencies of the alleles that contribute to the observed traits. We genotyped 2.37 million SNPs in 95 extreme phenotype individuals, that is: heavy smokers that either developed NSCLC at an early age (extreme cases); or did not present NSCLC at an advanced age (extreme controls), selected from a discovery set (n=3631). We validated significant SNPs in 133 additional subjects with extreme phenotypes selected from databases including >39,000 individuals. Two SNPs were validated: rs12660420 (p(combined)=5.66x10(-5); ORcombined=2.80), mapping to a noncoding transcript exon of PDE10A; and rs6835978 (p(combined)=1.02x10(-4); ORcombined=2.57), an intronic variant in ATP10D. We assessed the relevance of both proteins in early-stage NSCLC. PDE10A and ATP10D mRNA expressions correlated with survival in 821 stage I-II NSCLC patients (p=0.01 and p<0.0001). PDE10A protein expression correlated with survival in 149 patients with stage I-II NSCLC (p=0.002). In conclusion, we validated two variants associated with extreme phenotypes of high and low risk of developing tobacco-induced NSCLC. Our findings may allow to identify individuals presenting high and low risk to develop tobacco-induced NSCLC and to characterize molecular mechanisms of carcinogenesis and resistance to develop NSCLC

Interacción genotipo*localidad de tres ciclos de producción de la red de Raigrás anual en Argentina.

El objetivo del trabajo fue evaluar la producción acumulada anual de materia seca de materiales de raigrás en diez (10) localidades durante el período 2018-20.Estación Experimental Agropecuaria General VillegasFil: Méndez, Daniel. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria General Villegas; ArgentinaFil: Frigerio, Karina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria San Luis; ArgentinaFil: Ruiz, María de los Angeles. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Anguil; ArgentinaFil: Fontana, Laura María Celia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Anguil; ArgentinaFil: Romero, Luis Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; ArgentinaFil: Barbera, Pablo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Mercedes; Argentina.Fil: Ré, Alejo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Concepción del Uruguay; Argentina.Fil: Gallego, Juan José. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Valle Inferior; ArgentinaFil: Neira Zilli, Fernanda. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Valle Inferior; ArgentinaFil: Otondo, José. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Cuenca del Salado. Agencia de Extensión Rural Chascomús; ArgentinaFil: Cicchino, Mariano Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Cuenca del Salado. Agencia de Extensión Rural Chascomús; ArgentinaFil: Bailleres, Matias Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Cuenca del Salado. Agencia de Extensión Rural Chascomús; ArgentinaFil: Melani, Esteban. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Cuenca del Salado. Agencia de Extensión Rural Chascomús; ArgentinaFil: Lavandera, Javier. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Forrajes; Argentina