An improved expression and purification protocol enables the structural characterization of Mnt1, an antifungal target from Candida albicans.

BACKGROUND: Candida albicans is one of the most prevalent fungi causing infections in the world. Mnt1 is a mannosyltransferase that participates in both the cell wall biogenesis and biofilm growth of C. albicans. While the cell wall performs crucial functions in pathogenesis, biofilm growth is correlated with sequestration of drugs by the extracellular matrix. Therefore, antifungals targeting CaMnt1 can compromise fungal development and potentially also render Candida susceptible to drug therapy. Despite its importance, CaMnt1 has not yet been purified to high standards and its biophysical properties are lacking. RESULTS: We describe a new protocol to obtain high yield of recombinant CaMnt1 in Komagataella phaffii using methanol induction. The purified protein's identity was confirmed by MALDI-TOF/TOF mass spectroscopy. The Far-UV circular dichroism (CD) spectra demonstrate that the secondary structure of CaMnt1 is compatible with a protein formed by α-helices and ß-sheets at pH 7.0. The fluorescence spectroscopy results show that the tertiary structure of CaMnt1 is pH-dependent, with a greater intensity of fluorescence emission at pH 7.0. Using our molecular modeling protocol, we depict for the first time the ternary complex of CaMnt1 bound to its two substrates, which has enabled the identification of residues involved in substrate specificity and catalytic reaction. Our results corroborate the hypothesis that Tyr209 stabilizes the formation of an oxocarbenium ion-like intermediate during nucleophilic attack of the acceptor sugar, opposing the double displacement mechanism proposed by other reports. CONCLUSIONS: The methodology presented here can substantially improve the yield of recombinant CaMnt1 expressed in flask-grown yeasts. In addition, the structural characterization of the fungal mannosyltransferase presents novelties that can be exploited for new antifungal drug's development.

Evaluación de contenidos de Medicina Natural y Tradicional en Examen Estatal, curso 2013-2014 / Evaluation of contents of Natural and Traditional Medicine in State Exam, course 2013-2014

Se realizó una investigación con el objetivo de describir los resultados de la evaluación de los contenidos de Medicina Natural y Tradicional (MNT) en el examen estatal realizado en el Curso 2013 2014 en la Universidad de Ciencias Médicas de Guantánamo. Para ello se revisaron todos los exámenes estatales realizados en primera convocatoria, donde fue analizada la pregunta relacionada con el tema antes mencionado, la información obtenida se organizó en tablas y gráficos. Se concluyó que el mayor número de estudiantes responden correctamente la planta medicinal a utilizar en el asma bronquial, la mayor dificultad se encuentra en responder correctamente las dos formas de estimulación, un gran número responde una sola forma.

Nivel de conocimiento de los profesores en Medicina Natural y Tradicional en la disciplina de Medicina General Integral (MGl) / Level of knowledge of the natural and traditional medicine professors in the integral general medicine discipline

Se realizó una estudio descriptivo en el campo de la educación médica superior realizada en la Facultad de Ciencias Médicas Enrique Cabrera, en el período comprendido entre septiembre de 2012 y julio de 2013, con el objetivo de identificar el nivel de conocimiento que poseen los profesores sobre Medicina Natural y Tradicional en la disciplina de Medicina General Integral (MGI). El universo de estudio estuvo conformado por los 60 profesores del departamento docente de MGI. Se realizó un análisis documental a las actas de reuniones del colectivo de año 5to. año, actas de reuniones metodológicas, plan de trabajo metodológico y programas de las asignaturas de la carrera de medicina de los cursos 2011-2012 y 2012-2013. Se aplicó un cuestionario estructurado con preguntas cerradas dicotómicas, a los profesores del departamento docente de Medicina General Integral de la facultad. Se evaluaron los resultados del cuestionario según el nivel de conocimiento y se agruparon en dos dimensiones adecuadas y no adecuadas. Como conclusiones se obtuvo que el nivel de conocimientos de Medicina Natural Tradicional de los profesores estudiados no es adecuado, las áreas del conocimiento de la MNT donde más dificultades presentaron están relacionadas con el uso de la acupuntura en el tratamiento de las enfermedades del sistema nervioso central y en algunas afecciones ginecológicas(AU)

A descriptive study was conducted in Enrique Cabrera medical school in the field of higher medical education from September 2012 to July 2013. The objective was to identify the level of knowledge of the traditional and natural herb medicine professors in the Integral General Medicine specialty. The universe of study was made up of 60 professors from the teaching department of integral general medicine discipline. A documentary analysis was made in the 5th year faculty meeting minutes, methodological meeting minutes, the methodological work planning and curricula of the medical subjects in the 2011-2012 and 2012-2013 academic years. The professors answered a structured questionnaire with two-ended closed questions. The results of the questionnaire were evaluated according to the level of knowledge shown and then grouped into two dimensions: adequate and inadequate. It was concluded that the level of knowledge of the traditional and natural herb medicine professors participating in the study was inadequate, being the most difficult areas those related with the use of acupuncture in treating central nervous system diseases and some gynecological diseases(AU)

Nivel de conocimiento de los profesores en medicina natural y tradicional en la disciplina de medicina general integral (MGl) / Level of knowledge of the natural and traditional medicine professors in the integral general medicine discipline

Se realizó una estudio descriptivo en el campo de la educación médica superior realizada en la Facultad de Ciencias Médicas Enrique Cabrera, en el período comprendido entre septiembre de 2012 y julio de 2013, con el objetivo de identificar el nivel de conocimiento que poseen los profesores sobre Medicina Natural y Tradicional en la disciplina de Medicina General Integral (MGI). El universo de estudio estuvo conformado por los 60 profesores del departamento docente de MGI. Se realizó un análisis documental a las actas de reuniones del colectivo de año 5to. año, actas de reuniones metodológicas, plan de trabajo metodológico y programas de las asignaturas de la carrera de medicina de los cursos 2011-2012 y 2012-2013. Se aplicó un cuestionario estructurado con preguntas cerradas dicotómicas, a los profesores del departamento docente de Medicina General Integral de la facultad. Se evaluaron los resultados del cuestionario según el nivel de conocimiento y se agruparon en dos dimensiones adecuadas y no adecuadas. Como conclusiones se obtuvo que el nivel de conocimientos de Medicina Natural Tradicional de los profesores estudiados no es adecuado, las áreas del conocimiento de la MNT donde más dificultades presentaron están relacionadas con el uso de la acupuntura en el tratamiento de las enfermedades del sistema nervioso central y en algunas afecciones ginecológicas.

A descriptive study was conducted in Enrique Cabrera medical school in the field of higher medical education from September 2012 to July 2013. The objective was to identify the level of knowledge of the traditional and natural herb medicine professors in the Integral General Medicine specialty. The universe of study was made up of 60 professors from the teaching department of integral general medicine discipline. A documentary analysis was made in the 5th year faculty meeting minutes, methodological meeting minutes, the methodological work planning and curricula of the medical subjects in the 2011-2012 and 2012-2013 academic years. The professors answered a structured questionnaire with two-ended closed questions. The results of the questionnaire were evaluated according to the level of knowledge shown and then grouped into two dimensions: adequate and inadequate. It was concluded that the level of knowledge of the traditional and natural herb medicine professors participating in the study was inadequate, being the most difficult areas those related with the use of acupuncture in treating central nervous system diseases and some gynecological diseases.

The relationship of cytotoxic and genotoxic damage with blood aluminum levels and oxidative stress induced by this metal in common carp (Cyprinus carpio) erythrocytes.

Aluminum is one of the most abundant elements in nature and is used in diverse industrial processes. As a result, it contaminates aquatic ecosystems, inducing damage on associated biota. In fish, it has been observed to induce hypoxia, hypercapnia, metabolic acidosis and respiratory arrest. Although there is little information on Al-induced cytotoxicity and DNA damage, this type of studies are essential in order to identify the mechanisms of action of this metal. The cytotoxic and genotoxic effects induced by Al on common carp (Cyprinus carpio) erythrocytes were determined in specimens exposed to 0.05, 120 and 239mgAlL(-1) in static exposure systems. Blood samples were taken at 12, 24, 48, 72 and 96h, erythrocytes were separated, and the following were evaluated: frequency of micronuclei and frequency of terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL)-positive cells, blood Al levels, lipid peroxidation, protein carbonyl content, and activity of the antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase. The results show that tested aluminum concentrations produces oxidative stress (increase in lipid peroxidation degree and oxidized proteins content, as well as decrease in antioxidant enzymes activity) and induced higher frequencies of micronuclei and TUNEL-positive cells, so this metal can be considered as a cytotoxic and genotoxic agent for erythrocytes of common carp.

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

Using bacteriological (culture) and molecular (PCR - Restriction Enzyme Analysis, PRA) methods, we investigated the presence of environmental mycobacteria in tap water, antiseptic solutions and surgical gloves, used in carrying out surgical procedures at the Getúlio Vargas University Hospital Surgical Center, in Manaus -Amazonas/Brazil. Samples (105) were collected and analyzed from: tap water (24 - collected from 3 taps in the surgical center); povidone-iodine solution, PVP-I, (8); Chlorhexidine solution (7), that were used to hygienize surgeons' hands; surgical gloves (39); and solutions that were effectively used during the surgical procedure (27). Using bacteriological method 41 mycobacteria were isolated, all from samples of tap water. Using PRA, mycobacterial DNA was detected only in the sample from water that provided over 100 colonies per inoculated culture tubes. The isolated were identified as Mycobacterium celatum pattern II, M. gordonae pattern III, M. gordonae pattern VI, M. intracellulare pattern I, M. lentiflavum pattern III and M. mucogenicum pattern I. The isolating of M. mucogenicum, a species that had already been incriminated for causing post-surgical outbreaks in tap water from the surgical center, indicates that cleaning and monitoring procedures must be carried out in every place of water distribution. This may be necessary to prevent nosological mycobacteriosis outbreaks induced by the use of water in different activities such as handling and hygienizing patients submitted to invasive procedures.

Investigou-se por métodos bacteriológicos (cultivo) e moleculares (PCR - Restriction Enzyme Analysis, PRA), a presença de micobactérias ambientais em águas de torneira, soluções e luvas cirúrgicas, utilizadas nas etapas dos procedimentos cirúrgicos executados no centro cirúrgico do Hospital Universitário Getulio Vargas (HUGV), na cidade de Manaus-Amazonas/Brasil. Foram colhidas e analisadas 105 amostras sendo: 24 de águas (colhidas das 2 torneiras existentes no centro cirúrgico), 8 de solução de Povidine e 7 de solução de Clorhexidina, que servem para a higienização das mãos dos cirurgiões; 39 de luvas cirúrgicas (superfícies internas e externas); e 27 de soluções que foram efetivamente utilizadas durante o ato cirúrgico. Por método bacteriológico obteve-se 41 isolados micobacterianos apenas de águas das torneiras. Pelo PRA obteve-se a detecção de DNA micobacteriano somente na amostra de água que forneceu acima de 100 colônias de micobactérias por tubo semeado. Os isolados foram identificados como sendo Mycobacterium celatum perfil 2, M. gordonae perfil 3, M. gordonae perfil 6, M. intracellulare perfil 1, M. lentiflavum perfil 3 e M. mucogenicum perfil 1. O encontro de M. mucogenicum, espécie já incriminada em surtos pós-cirúrgicos, indica que devem ser efetuados procedimentos de limpeza e monitoramento em todos os pontos de distribuição de águas, visando à prevenção de surtos de micobacterioses nosocomiais induzidos pelo uso das águas nas diferentes atividades de manuseio ou higienização dos pacientes submetidos a procedimentos invasivos.

Pesquisa de Micobactérias Ambientais em água de torneira, luvas e soluções utilizadas em procedimentos cirúrgicos no Hospital Universitário Getúlio Vargas - Manaus/AM

Using bacteriological (culture) and molecular (PCR - Restriction Enzyme Analysis, PRA) methods, we investigated the presence of environmental mycobacteria in tap water, antiseptic solutions and surgical gloves, used in carrying out surgical procedures at the Getúlio Vargas University Hospital Surgical Center, in Manaus -Amazonas/Brazil. Samples (105) were collected and analyzed from: tap water (24 - collected from 3 taps in the surgical center); povidone-iodine solution, PVP-I, (8); Chlorhexidine solution (7), that were used to hygienize surgeons' hands; surgical gloves (39); and solutions that were effectively used during the surgical procedure (27). Using bacteriological method 41 mycobacteria were isolated, all from samples of tap water. Using PRA, mycobacterial DNA was detected only in the sample from water that provided over 100 colonies per inoculated culture tubes. The isolated were identified as Mycobacterium celatum pattern II, M. gordonae pattern III, M. gordonae pattern VI, M. intracellulare pattern I, M. lentiflavum pattern III and M. mucogenicum pattern I. The isolating of M. mucogenicum, a species that had already been incriminated for causing post-surgical outbreaks in tap water from the surgical center, indicates that cleaning and monitoring procedures must be carried out in every place of water distribution. This may be necessary to prevent nosological mycobacteriosis outbreaks induced by the use of water in different activities such as handling and hygienizing patients submitted to invasive procedures.

Investigou-se por métodos bacteriológicos (cultivo) e moleculares (PCR - Restriction Enzyme Analysis, PRA), a presença de micobactérias ambientais em águas de torneira, soluções e luvas cirúrgicas, utilizadas nas etapas dos procedimentos cirúrgicos executados no centro cirúrgico do Hospital Universitário Getulio Vargas (HUGV), na cidade de Manaus-Amazonas/Brasil. Foram colhidas e analisadas 105 amostras sendo: 24 de águas (colhidas das 2 torneiras existentes no centro cirúrgico), 8 de solução de Povidine e 7 de solução de Clorhexidina, que servem para a higienização das mãos dos cirurgiões; 39 de luvas cirúrgicas (superfícies internas e externas); e 27 de soluções que foram efetivamente utilizadas durante o ato cirúrgico. Por método bacteriológico obteve-se 41 isolados micobacterianos apenas de águas das torneiras. Pelo PRA obteve-se a detecção de DNA micobacteriano somente na amostra de água que forneceu acima de 100 colônias de micobactérias por tubo semeado. Os isolados foram identificados como sendo Mycobacterium celatum perfil 2, M. gordonae perfil 3, M. gordonae perfil 6, M. intracellulare perfil 1, M. lentiflavum perfil 3 e M. mucogenicum perfil 1. O encontro de M. mucogenicum, espécie já incriminada em surtos pós-cirúrgicos, indica que devem ser efetuados procedimentos de limpeza e monitoramento em todos os pontos de distribuição de águas, visando à prevenção de surtos de micobacterioses nosocomiais induzidos pelo uso das águas nas diferentes atividades de manuseio ou higienização dos pacientes submetidos a procedimentos invasivos.

Digital soil mapping using multiple logistic regression on terrain parameters in southern Brazil

Soil surveys are necessary sources of information for land use planning, but they are not always available. This study proposes the use of multiple logistic regressions on the prediction of occurrence of soil types based on reference areas. From a digitalized soil map and terrain parameters derived from the digital elevation model in ArcView environment, several sets of multiple logistic regressions were defined using statistical software Minitab, establishing relationship between explanatory terrain variables and soil types, using either the original legend or a simplified legend, and using or not stratification of the study area by drainage classes. Terrain parameters, such as elevation, distance to stream, flow accumulation, and topographic wetness index, were the variables that best explained soil distribution. Stratification by drainage classes did not have significant effect. Simplification of the original legend increased the accuracy of the method on predicting soil distribution.

Os levantamentos de solos são fontes de informação necessárias para o planejamento de uso das terras, entretanto eles nem sempre estão disponíveis. Este estudo propõe o uso de regressões logísticas múltiplas na predição de ocorrência de classes de solos a partir de áreas de referência. Baseado no mapa original de solos em formato digital e parâmetros do terreno derivados do modelo numérico do terreno em ambiente ArcView, vários conjuntos de regressões logísticas múltiplas foram definidas usando o programa estatístico Minitab, estabelecendo relações entre as variáveis do terreno independentes e tipos de solos, usando tanto a legenda original como uma legenda simplificada, e usando ou não estratificação da área de estudo por classes de drenagem. Os parâmetros do terreno como elevação, distância dos rios, acúmulo de fluxo e índice de umidade topográfica foram as variáveis que melhor explicaram a distribuição das classes de solos. A estratificação por classes de drenagem não teve efeito significativo. A simplificação da legenda aumentou a precisão do método na predição da distribuição dos solos.

Digital soil mapping using multiple logistic regression on terrain parameters in southern Brazil

Soil surveys are necessary sources of information for land use planning, but they are not always available. This study proposes the use of multiple logistic regressions on the prediction of occurrence of soil types based on reference areas. From a digitalized soil map and terrain parameters derived from the digital elevation model in ArcView environment, several sets of multiple logistic regressions were defined using statistical software Minitab, establishing relationship between explanatory terrain variables and soil types, using either the original legend or a simplified legend, and using or not stratification of the study area by drainage classes. Terrain parameters, such as elevation, distance to stream, flow accumulation, and topographic wetness index, were the variables that best explained soil distribution. Stratification by drainage classes did not have significant effect. Simplification of the original legend increased the accuracy of the method on predicting soil distribution.

Os levantamentos de solos são fontes de informação necessárias para o planejamento de uso das terras, entretanto eles nem sempre estão disponíveis. Este estudo propõe o uso de regressões logísticas múltiplas na predição de ocorrência de classes de solos a partir de áreas de referência. Baseado no mapa original de solos em formato digital e parâmetros do terreno derivados do modelo numérico do terreno em ambiente ArcView, vários conjuntos de regressões logísticas múltiplas foram definidas usando o programa estatístico Minitab, estabelecendo relações entre as variáveis do terreno independentes e tipos de solos, usando tanto a legenda original como uma legenda simplificada, e usando ou não estratificação da área de estudo por classes de drenagem. Os parâmetros do terreno como elevação, distância dos rios, acúmulo de fluxo e índice de umidade topográfica foram as variáveis que melhor explicaram a distribuição das classes de solos. A estratificação por classes de drenagem não teve efeito significativo. A simplificação da legenda aumentou a precisão do método na predição da distribuição dos solos.