Simple yet functional phosphate-loop proteins.

Abundant and essential motifs, such as phosphate-binding loops (P-loops), are presumed to be the seeds of modern enzymes. The Walker-A P-loop is absolutely essential in modern NTPase enzymes, in mediating binding, and transfer of the terminal phosphate groups of NTPs. However, NTPase function depends on many additional active-site residues placed throughout the protein's scaffold. Can motifs such as P-loops confer function in a simpler context? We applied a phylogenetic analysis that yielded a sequence logo of the putative ancestral Walker-A P-loop element: a ß-strand connected to an α-helix via the P-loop. Computational design incorporated this element into de novo designed ß-α repeat proteins with relatively few sequence modifications. We obtained soluble, stable proteins that unlike modern P-loop NTPases bound ATP in a magnesium-independent manner. Foremost, these simple P-loop proteins avidly bound polynucleotides, RNA, and single-strand DNA, and mutations in the P-loop's key residues abolished binding. Binding appears to be facilitated by the structural plasticity of these proteins, including quaternary structure polymorphism that promotes a combined action of multiple P-loops. Accordingly, oligomerization enabled a 55-aa protein carrying a single P-loop to confer avid polynucleotide binding. Overall, our results show that the P-loop Walker-A motif can be implemented in small and simple ß-α repeat proteins, primarily as a polynucleotide binding motif.

Bacilli glutamate dehydrogenases diverged via coevolution of transcription and enzyme regulation.

The linkage between regulatory elements of transcription, such as promoters, and their protein products is central to gene function. Promoter-protein coevolution is therefore expected, but rarely observed, and the manner by which these two regulatory levels are linked remains largely unknown. We study glutamate dehydrogenase-a hub of carbon and nitrogen metabolism. In Bacillus subtilis, two paralogues exist: GudB is constitutively transcribed whereas RocG is tightly regulated. In their active, oligomeric states, both enzymes show similar enzymatic rates. However, swaps of enzymes and promoters cause severe fitness losses, thus indicating promoter-enzyme coevolution. Characterization of the proteins shows that, compared to RocG, GudB's enzymatic activity is highly dependent on glutamate and pH Promoter-enzyme swaps therefore result in excessive glutamate degradation when expressing a constitutive enzyme under a constitutive promoter, or insufficient activity when both the enzyme and its promoter are tightly regulated. Coevolution of transcriptional and enzymatic regulation therefore underlies paralogue-specific spatio-temporal control, especially under diverse growth conditions.

Environment modulates protein heterogeneity through transcriptional and translational stop codon readthrough.

Stop codon readthrough events give rise to longer proteins, which may alter the protein's function, thereby generating short-lasting phenotypic variability from a single gene. In order to systematically assess the frequency and origin of stop codon readthrough events, we designed a library of reporters. We introduced premature stop codons into mScarlet, which enabled high-throughput quantification of protein synthesis termination errors in E. coli using fluorescent microscopy. We found that under stress conditions, stop codon readthrough may occur at rates as high as 80%, depending on the nucleotide context, suggesting that evolution frequently samples stop codon readthrough events. The analysis of selected reporters by mass spectrometry and RNA-seq showed that not only translation but also transcription errors contribute to stop codon readthrough. The RNA polymerase was more likely to misincorporate a nucleotide at premature stop codons. Proteome-wide detection of stop codon readthrough by mass spectrometry revealed that temperature regulated the expression of cryptic sequences generated by stop codon readthrough in E. coli. Overall, our findings suggest that the environment affects the accuracy of protein production, which increases protein heterogeneity when the organisms need to adapt to new conditions.

Active site center redesign increases protein stability preserving catalysis in thioredoxin.

The stabilization of natural proteins is a long-standing desired goal in protein engineering. Optimizing the hydrophobicity of the protein core often results in extensive stability enhancements. However, the presence of totally or partially buried catalytic charged residues, essential for protein function, has limited the applicability of this strategy. Here, focusing on the thioredoxin, we aimed to augment protein stability by removing buried charged residues in the active site without loss of catalytic activity. To this end, we performed a charged-to-hydrophobic substitution of a buried and functional group, resulting in a significant stability increase yet abolishing catalytic activity. Then, to simulate the catalytic role of the buried ionizable group, we designed a combinatorial library of variants targeting a set of seven surface residues adjacent to the active site. Notably, more than 50% of the library variants restored, to some extent, the catalytic activity. The combination of experimental study of 2% of the library with the prediction of the whole mutational space by partial least squares regression revealed that a single point mutation at the protein surface is sufficient to fully restore the catalytic activity without thermostability cost. As a result, we engineered one of the highest thermal stabilities reported for a protein with a natural occurring fold (137°C). Further, our hyperstable variant preserves the catalytic activity both in vitro and in vivo.

Phenotypic mutations contribute to protein diversity and shape protein evolution.

Errors in DNA replication generate genetic mutations, while errors in transcription and translation lead to phenotypic mutations. Phenotypic mutations are orders of magnitude more frequent than genetic ones, yet they are less understood. Here, we review the types of phenotypic mutations, their quantifications, and their role in protein evolution and disease. The diversity generated by phenotypic mutation can facilitate adaptive evolution. Indeed, phenotypic mutations, such as ribosomal frameshift and stop codon readthrough, sometimes serve to regulate protein expression and function. Phenotypic mutations have often been linked to fitness decrease and diseases. Thus, understanding the protein heterogeneity and phenotypic diversity caused by phenotypic mutations will advance our understanding of protein evolution and have implications on human health and diseases.

Scientific evidence on heated-tobacco products. A narrative review / Evidencia científica sobre productos de tabaco calentado. Una revisión narrativa

Given the impact of smoking on individuals and National Healthcare Systems, it has become necessary for the continuous evaluation of tobacco products, especially when they are claimed or perceived as being less harmful than conventional cigarettes (CCs), and coexisting conflict of interests. The objective of the present manuscript was to perform a complete and updated narrative review of scientific literature on the assessment of harmful and potentially harmful constituents (HPHCs) and the biological and clinical impact from the newest available heated tobacco products (HTPs), in comparison with CCs. A total of 52 studies (46 from the review and 6 manually found) were finally analyzed. Despite some existing differences among the studies, most of them point to a reduction of the emissions of HPHCs as well as exposure to toxicants, and thus in the biological and clinical impact with HTPs when compared with CCs. In the scenario where individuals decide to continue smoking, HTPs seem to be a better option than CCs. Further prospective, independent studies should replicate the existing experiments in order to corroborate the conclusions raised in the original studies. Moreover, long-term investigations (decades) are also needed to obtain more compelling safety information of HTPs. (AU)

Dado el impacto del tabaquismo en las personas y en los Sistemas Nacionales de Salud, se ha vuelto una necesidad la evaluación continua de los nuevos productos de tabaco, especialmente cuando se afirma o se percibe que son menos dañinos que los cigarrillos convencionales (CC). El objetivo del presente manuscrito es presentar una revisión narrativa completa y actualizada de la literatura científica sobre la evaluación de los componentes dañinos y potencialmente dañinos (HPHC) y el impacto biológico y clínico de los productos de tabaco calentado (PTC), más novedosos, en comparación con los CC. Se analizaron un total de 52 estudios (46 de la revisión y 6 encontrados manualmente). A pesar de algunas diferencias existentes entre los estudios, la mayoría de ellos apuntan a una reducción de las emisiones de HPHC, así como a la exposición a sustancias tóxicas y, por lo tanto, en el impacto biológico y clínico con los PTC en comparación con los CC. En un posible escenario de personas que deciden seguir fumando, los PTC parecen ser una mejor opción que los CC. Estudios independientes prospectivos deberían replicar los experimentos existentes para corroborar las conclusiones planteadas en los estudios originales aquí presentados. De la misma manera, investigaciones a largo plazo (décadas) son necesarias para obtener información más completa sobre la seguridad de los PTC. (AU)

Evidencia científica sobre productos de tabaco calentado. Una revisión narrativa / Scientific evidence on heated-tobacco products. A narrative review

Dado el impacto del tabaquismo en las personas y en los Sistemas Nacionales de Salud, se ha vuelto una necesidad la evaluación continua de los nuevos productos de tabaco, especialmente cuando se afirma o se percibe que son menos dañinos que los cigarrillos convencionales (CC). El objetivo del presente manuscrito es presentar una revisión narrativa completa y actualizada de la literatura científica sobre la evaluación de los componentes dañinos y potencialmente dañinos (HPHC) y el impacto biológico y clínico de los productos de tabaco calentado (PTC), más novedosos, en comparación con los CC. Se analizaron un total de 52 estudios (46 de la revisión y 6 encontrados manualmente). A pesar de algunas diferencias existentes entre los estudios, la mayoría de ellos apuntan a una reducción de las emisiones de HPHC, así como a la exposición a sustancias tóxicas y, por lo tanto, en el impacto biológico y clínico con los PTC en comparación con los CC. En un posible escenario de personas que deciden seguir fumando, los PTC parecen ser una mejor opción que los CC. Estudios independientes prospectivos deberían replicar los experimentos existentes para corroborar las conclusiones planteadas en los estudios originales aquí presentados. De la misma manera, investigaciones a largo plazo (décadas) son necesarias para obtener información más completa sobre la seguridad de los PTC. (AU)

Given the impact of smoking on individuals and National Healthcare Systems, it has become necessary for the continuous evaluation of tobacco products, especially when they are claimed or perceived as being less harmful than conventional cigarettes (CCs), and coexisting conflict of interests. The objective of the present manuscript was to perform a complete and updated narrative review of scientific literature on the assessment of harmful and potentially harmful constituents (HPHCs) and the biological and clinical impact from the newest available heated tobacco products (HTPs), in comparison with CCs. A total of 52 studies (46 from the review and 6 manually found) were finally analyzed. Despite some existing differences among the studies, most of them point to a reduction of the emissions of HPHCs as well as exposure to toxicants, and thus in the biological and clinical impact with HTPs when compared with CCs. In the scenario where individuals decide to continue smoking, HTPs seem to be a better option than CCs. Further prospective, independent studies should replicate the existing experiments in order to corroborate the conclusions raised in the original studies. Moreover, long-term investigations (decades) are also needed to obtain more compelling safety information of HTPs. (AU)

Estomatitis Asociada a Metotrexato: Un Reto Diagnóstico / Methotrexate Associated Stomatitis: A Diagnostic Challenge

RESUMEN: Las úlceras orales (UO) son uno de los signos de toxicidad por metotrexato (Mtx) aún en pacientes con esquemas de dosis bajas para el control de artritis reumatoide (AR). En estos casos establecer un diagnóstico correlacionando UO con el medicamento puede ser un reto. Presentamos 2 casos clínicos de pacientes con AR en tratamiento con Mtx, las cuales desarrollaron UO. En los dos casos, interesantemente los pacientes fueron evaluados tanto por especialistas del área médica y oral sin tener un resultado satisfactorio después de múltiples tratamientos. Las UO resolvieron posterior a la suspensión del medicamento. Se estableció el diagnóstico de Estomatitis por Metotrexato (EMtx) por un especialista en medicina oral. El manejo multidisciplinario en estos casos es clave para el establecimiento de un diagnóstico y tratamiento oportuno.

ABSTRACT: Oral ulcers (OU) are a sign of methotrexate (Mtx) toxicity, even in patients with rheumatoid arthritis (RA) that are under a low-dose regime. In those cases, establishing a diagnosis correlating OU with the medication can be quite a challenge. Here we present 2 clinical cases of RA patients under Mtx treatment that developed OU. Interestingly, in both cases the patients were evaluated by two specialists in the medical and dentistry area, and following multiple treatments there was no satisfactory result. However, oral ulcers resolved after stopping the treatment. A diagnosis of Metotrexato stomatitis was established (SMtx) by a specialist in oral medicine. Multidisciplinary management in these cases is key for the establishment of an opportune diagnosis and treatment.

Perspectiva de profesionales de salud sobre la violencia de la pareja en el embarazo / Perspective of the Health Professionals on the Partner Violence during Pregnancy / Perspectiva dos profissionais da saúde sobre a violência do casal na gravidez

Introducción: la violencia de pareja (VP) durante el embarazo afecta los derechos de las gestantes y causa serias complicaciones para la madre o su hijo por nacer. El personal de salud puede enfrentar esta realidad mediante un adecuado proceso de detección y atención. Objetivo: explorar la perspectiva de profesionales de salud de la Unidad de Servicios de Salud Usaquén, de Bogotá, sobre la VP en el embarazo. Método: estudio cualitativo, entre marzo y diciembre de 2016. Se realizaron cuatro grupos focales con la participación de 38 profesionales. El análisis se desarrolló con apoyo del software ATLAS.ti 7. Resultados: la corta edad y la carencia de recursos y redes de apoyo perpetúa la VP. Los participantes sugieren la intervención de equipos multidisciplinarios coordinados en la detección y atención biopsicosocial de la problemática, construir ambientes de contención emocional a las usuarias, empoderarlas para el ejercicio de derechos y capacitar al personal de salud en el manejo de la problemática. Discusión: para el personal de salud es imperioso saber enfrentar esta realidad, de manera que las mujeres salgan fortalecidas. La inclusión de una mirada de género y derechos es una necesidad a la hora de abordar la VP en los contextos de la salud. Conclusiones: para un cambio de condiciones se requiere: a) trabajo multidisciplinario coordinado, b) ambientes amigables, c) atención humanizada a la gestante, d) reducción de la revictimización y e) capacitación al personal de salud para la intervención en VP.

Introduction: Intimate partner violence (IPV) during pregnancy undermines the pregnant woman's rights and brings about serious complications both to the mother and the unborn child. Health professionals will have to cope with this reality through an appropriate process of detection and care. Objective: To explore the perspective by the health professionals in the health center 'Unidad de Servicios de Salud Usaquén' in Bogotá, regarding the IPV during pregnancy. Method: A qualitative study was conducted between March and December 2016. Four focus groups were carried out with 38 health professional taking part in them. The analysis was developed by using the software ATLASti 7. Results: Being young, financial shortfalls and the lack of support feed into the IPV. The participants propose the intervention by coordinated multidisciplinary teams for the detection and bio-psychosocial care to people with this problem, to build emotional containment environments for the users, to empower the women so that they enforce their rights, and to train the health professionals for managing this problem. Discussion: It is a must that the health professionals become able to cope with this reality, so that they can strengthen the women in this situation. There is a need to provide them with a gender-oriented view and rights awareness when it comes to face the IPV in the contexts of health issues. Conclusions: To do a change in the conditions requires: a) coordinated multidisciplinary work; b) friendly environments; c) humanized care to pregnant women; d) to reduce the re-victimization; and e) to train the health workers in IPV intervention.

Introdução: A violência de casal (VC) durante a gravidez afeta os direitos das gestantes e ocasiona complicações fortes para mãe e filho ainda não nascido. O pessoal de saúde pode enfrentar essa realidade mediante um adequado processo de triagem e atendimento. Objetivo: Explorar a perspectiva dos profissionais da saúde da Unidade de Serviços de Saúde Usaquén, de Bogotá, sobre a VC na gravidez. Método: foi realizado um estudo qualitativo entre marco e dezembro de 2016. Foram realizados quatro grupos focais com participação de 38 profissionais. A análise desenvolveu-se com suporte do software ATLASti 7. Resultados: A pouca idade e a carência de recursos e redes de apoio perpetuam a VC. Os participantes sugerem a intervenção de equipes multidisciplinares coordenadas na detecção e atendimento biopsicossocial da problemática, construir ambientes de contenção emocional às usuárias, empoeirá-las para o exercício de direitos e treinar o pessoal de saúde na gestão da problemática. Discussão: Para o pessoal de saúde é imperativo saber enfrentar essa realidade, de jeito tal que as mulheres saírem fortalecidas. A inclusão de um olhar de gênero e direitos é uma necessidade à hora de abordar a VC nos contextos da saúde. Conclusões: Para uma mudança de condições é preciso: a) trabalho multidisciplinar coordenado b) ambientes amigáveis c) atendimento humanizado à gestante, d) redução da revitimização e e) treinamento ao pessoal de saúde para a intervenção em VC.

Conocimiento sobre redes sociales de apoyo para víctimas de bullying cibernético en adolescentes. Parroquia de Yaruquíes, provincia de Chimborazo / Knowledge about social support networks for cyberbullying victms among adolescents. Yaruquíes parish, province of Chimborazo

Introducción: existen nuevas formas de acoso como el daño reiterado e intencionado a través de un medio electrónico, lo que se conoce como bullying cibernético, se puede brindar apoyo a las víctimas mediante redes sociales de apoyo. Objetivo: determinar el conocimiento sobre la importancia las redes sociales de apoyo para víctimas de bullying cibernético en adolescentes. Métodos: se realizó un estudio descriptivo, transversal en adolescentes de 12 a 17 años de las unidades educativas públicas y privadas de la parroquia Yaruquíes, durante el período julio - diciembre 2017. La información se recolectó a través de un cuestionario que se aplicó al universo de 458 adolescentes. La variable respuesta fue el nivel de conocimiento sobre la importancia de las redes sociales de apoyo. Entre las variables independientes se analizó, sexo y autoidentificación como mestizo o indígena. Resultados: 26% (n = 55) de hombres vs. 27% (n = 72) de mujeres y 35% (n = 11) de indígenas vs. 65% (n = 114) de mestizos han reportado haber sufrido bullying cibernético. Solo 26% (n = 120) de los adolescentes entrevistados respondieron tener conocimiento satisfactorio sobre redes sociales de apoyo para víctimas de bullying cibernético. Conclusión: a pesar de que un alto porcentanje de asolescentes ha reportado haber sufrido bullying cibernético, solo pocos conocen sobre la importancia de redes sociales de apoyo. Es necesario implementar invervenciones para dar a conocer alternativas de apoyo para adolescentes

Introduction: there are new forms of harassment such as repeated and intentional damage through electronic means including cyberbullying, is it possible to offer help through social networks. Objective: to determine the knowledge about the importance of social support networks for cyber bullying vitims in adolescents. Methods: a descriptive, cross-sectional study was conducted among adolescents aged 12 to 17 years of public and private educational institutions of Yaruquíes parish, during July - December 2017. Informations was collected through a questionnaire that was applied to the universe of 458 teenagers. Main outcome variable was level of knowledge about the importance of social support networks. Independent variables included sex and self-identification as mestizo or indigenous. Results: 26% (n = 55) of males vs. 27% (n = 72) of females, and 35% (n = 11) of indigenous vs. 65% (n = 114) of mestizos have reported to have suffered cyber bullying. Only 26% (n = 120) of respondents had a satisfactory level of knowledge about the importance of social support networks for cyberbullying victims. Conclusions: although a high percentage of adolscentes haver reported to have endured cyberbullying, only few of them know about social support networks. It is mandatory to implement interventions to making know about support alternatives for adolescts.

Simultaneous determination of multiple constituents in real beer samples of different origins by capillary zone electrophoresis.

Simultaneous determination of alcohols, amines, amino acids, flavonoids, and purine and pyrimidine bases in bottled beer samples directly without any pre-treatment was carried out by capillary zone electrophoresis with diode-array detection. Electrolyte conditions such as pH, composition and concentration of the buffer, working voltage and type and time of injection were checked. The best separation of the cited analytes was achieved in 70 mM sodium borate solution and pH 10.25. The detection limits were from 2.1 to 5.6 mg L(-1) for the 18 compounds studied. The developed method is rapid, sensitive and quantitative and has been applied to seven types of international bottled beers of different origins bought locally.

El delegado judicial de medidas penales alternativas: Intervención y seguimiento / No disponible

No disponible

Conflictos conyugales prevalecientes en mujeres de 45 a 55 años / Marital conflicts prevalents in women between 45 and 55 years of age

Antecedentes. El climaterio es una época de cambios que frecuentemente afecta el comportamiento de una mujer en su matrimonio. Materiales y Métodos. Se realizó un estudio prospectivo, transversar, con recolección de información mediante un cuestionario preparado para tales fines en una muestra de 100 mujeres seleccionadas al azar entre los 45 y 55 años de edad que convivian con sus maridos, que asistian a la consulta de ginecología en la maternidad del Instituto dominicano de Seguros Sociales (IDSS) Dr. Manuel Emilio Perdomo en Santo Domingo, República Dominicana en Santo Domingo, República Dominicana durante el período julio-septiembre 1997. Resultados. La mayoría de las mujeres (69//) estaban casadas con hombres mayores que ellas y 61// tenían conflictos familiares por causas económicas. Un 50// de ellas piensan que en ocasiones conflictivas el hombre comprende lo que ellas piensan que en ocasiones conflictivas el hombre no comprende lo que ella le está diciendo, un 44// de ellas despues de una discusión tiene problemas de comunicación con su esposo y 53// de ellas, despues de una discusión admite estar equivocada. En relación a las actividades sociales un 52// de ellas dijo que el esposos se negaba a salir con ella, el 68// de ellas afirmó que el esposo prefería divertirse sin ella y 45// de las encuestadas se divertían mas en compañia de sus amigos que con sus esposos que con sus esposos. Un 64// de las encuestadas tenían problemas de celos por atenciones de sus maridos hacia otras mujeres. En relación a la actividad sexual el 30// de ellas se quejaron de dispareunia, el 38// de falta de juego sexual antes del coito, el 39// de insatisfacción por la frecuencia de las relaciones sexuales, el 57// tenía dificultad en alcanzar el orgasmo y 68// se quejaba de eyaculación precoz por parte de sus maridos. En otras areas de compartimiento familiar el 52// de las encuestadas pensaba su esposo era muy debil con los hijos y el 47// que era muy rígido, el 48// pensaba su esposo ingería demasiado alcohol y el 23// que por esta causa faltaba a sus deberes en la casa. El 24// de las entrevistadas admitía que sus esposos las golpeaba y ella respondían con golpes también y el 28// admitió ser ellas la agresoras. Comentario. Este estudio revela que existe serios problemas en el matrimonio en mujeres durante el climaterio y varias de ellas se relacionan con la actividad sexual