Trypanosoma evansi contains two auxiliary enzymes of glycolytic metabolism: Phosphoenolpyruvate carboxykinase and pyruvate phosphate dikinase.

Trypanosoma evansi is a monomorphic protist that can infect horses and other animal species of economic importance for man. Like the bloodstream form of the closely related species Trypanosoma brucei, T. evansi depends exclusively on glycolysis for its free-energy generation. In T. evansi as in other kinetoplastid organisms, the enzymes of the major part of the glycolytic pathway are present within organelles called glycosomes, which are authentic but specialized peroxisomes. Since T. evansi does not undergo stage-dependent differentiations, it occurs only as bloodstream forms, it has been assumed that the metabolic pattern of this parasite is identical to that of the bloodstream form of T. brucei. However, we report here the presence of two additional enzymes, phosphoenolpyruvate carboxykinase and PPi-dependent pyruvate phosphate dikinase in T. evansi glycosomes. Their colocalization with glycolytic enzymes within the glycosomes of this parasite has not been reported before. Both enzymes can make use of PEP for contributing to the production of ATP within the organelles. The activity of these enzymes in T. evansi glycosomes drastically changes the model assumed for the oxidation of glucose by this parasite.

The scale of population structure in Arabidopsis thaliana.

The population structure of an organism reflects its evolutionary history and influences its evolutionary trajectory. It constrains the combination of genetic diversity and reveals patterns of past gene flow. Understanding it is a prerequisite for detecting genomic regions under selection, predicting the effect of population disturbances, or modeling gene flow. This paper examines the detailed global population structure of Arabidopsis thaliana. Using a set of 5,707 plants collected from around the globe and genotyped at 149 SNPs, we show that while A. thaliana as a species self-fertilizes 97% of the time, there is considerable variation among local groups. This level of outcrossing greatly limits observed heterozygosity but is sufficient to generate considerable local haplotypic diversity. We also find that in its native Eurasian range A. thaliana exhibits continuous isolation by distance at every geographic scale without natural breaks corresponding to classical notions of populations. By contrast, in North America, where it exists as an exotic species, A. thaliana exhibits little or no population structure at a continental scale but local isolation by distance that extends hundreds of km. This suggests a pattern for the development of isolation by distance that can establish itself shortly after an organism fills a new habitat range. It also raises questions about the general applicability of many standard population genetics models. Any model based on discrete clusters of interchangeable individuals will be an uneasy fit to organisms like A. thaliana which exhibit continuous isolation by distance on many scales.

Diversity of genetic lesions characterizes new Arabidopsis flavonoid pigment mutant alleles from T-DNA collections.

The visual phenotypes afforded by flavonoid pigments have provided invaluable tools for modern genetics. Many Arabidopsis transparent testa (tt) mutants lacking the characteristic proanthocyanidin (PA) seed coat pigmentation and often failing to accumulate anthocyanins in vegetative tissues have been characterized. These mutants have significantly contributed to our understanding of flavonoid biosynthesis, regulation, and transport. A comprehensive screening for tt mutants in available large T-DNA collection lines resulted in the identification of 16 independent lines lacking PAs and anthocyanins, or with seed coat pigmentation clearly distinct from wild type. Segregation analyses and the characterization of second alleles in the genes disrupted by the indexed T-DNA insertions demonstrated that all the lines contained at least one additional mutation responsible for the tt phenotypes. Using a combination of RNA-Seq and whole genome re-sequencing and confirmed through complementation, we show here that these mutations correspond to novel alleles of ttg1 (two alleles), tt3 (two alleles), tt5 (two alleles), ban (two alleles), tt1 (two alleles), and tt8 (six alleles), which harbored additional T-DNA insertions, indels, missense mutations, and large genomic deletion. Several of the identified alleles offer interesting perspectives on flavonoid biosynthesis and regulation.

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds.

Arabidopsis thaliana (Arabidopsis) seedlings often need to be grown on sterile media. This requires prior seed sterilization to prevent the growth of microbial contaminants present on the seed surface. Currently, Arabidopsis seeds are sterilized using two distinct sterilization techniques in conditions that differ slightly between labs and have not been standardized, often resulting in only partially effective sterilization or in excessive seed mortality. Most of these methods are also not easily scalable to a large number of seed lines of diverse genotypes. As technologies for high-throughput analysis of Arabidopsis continue to proliferate, standardized techniques for sterilizing large numbers of seeds of different genotypes are becoming essential for conducting these types of experiments. The response of a number of Arabidopsis lines to two different sterilization techniques was evaluated based on seed germination rate and the level of seed contamination with microbes and other pathogens. The treatments included different concentrations of sterilizing agents and times of exposure, combined to determine optimal conditions for Arabidopsis seed sterilization. Optimized protocols have been developed for two different sterilization methods: bleach (liquid-phase) and chlorine (Cl2) gas (vapor-phase), both resulting in high seed germination rates and minimal microbial contamination. The utility of these protocols was illustrated through the testing of both wild type and mutant seeds with a range of germination potentials. Our results show that seeds can be effectively sterilized using either method without excessive seed mortality, although detrimental effects of sterilization were observed for seeds with lower than optimal germination potential. In addition, an equation was developed to enable researchers to apply the standardized chlorine gas sterilization conditions to airtight containers of different sizes. The protocols described here allow easy, efficient, and inexpensive seed sterilization for a large number of Arabidopsis lines.

[Assessment of curricular contents and perception of competencies of health students related to the integrated health care model in Colombia]. / Evaluación de contenidos curriculares y percepción de competencias de estudiantes del área de la salud respecto del Modelo Integral de Atención en Salud en Colombia.

OBJECTIVE: To identify the perception of the last level students of the professional and technical health training programs regarding their competences to perform in the MIAS, and to evaluate the presence of said competences within the curricular contents of the health programs in Santander. METHODS: A transversal descriptive study was carried out in 451 students and a documentary review of curricula was performed for 26 programs. The content of an online questionnaire on the perception of competences in the areas of knowledge, attitudes and abilities was constructed and validated. RESULTS: In the documentary review, deficiencies were found in the inclusion of competencies such as leadership (80 %) and differential approach in care (84 %). In the perception survey, 36 % of the students stated that they had regular or deficient knowledge about Integral Routes of Health Care, benefit plans and actors of the health system. Additionally, 7.5 % stated they were not competent to carry out intersectoral coordination activities, epidemiological surveillance or public health research. CONCLUSION: The results show the need to include in the curricula of health programs some fundamental aspects for the proper implementation of MIAS.

OBJETIVO: Identificar la percepción de los estudiantes de último nivel de los programas de formación profesional y técnica en salud respecto a sus competencias para desempeñarse en el MIAS, y evaluar la presencia de dichas competencias dentro de los contenidos curriculares de los programas de salud en Santander. MÉTODOS: Se realizó un estudio descriptivo transversal en 451 estudiantes y una revisión documental de currículos de 26 programas. Se construyó y validó el contenido de un cuestionario en línea de percepción de competencias en los ámbitos del saber ser, saber conocer y saber hacer. RESULTADOS: En la revisión documental se encontró deficiencias en la inclusión de competencias tales como las de ejercer liderazgo (80 %) y realizar un enfoque diferencial en la atención (84 %). En la encuesta de percepción, el 36 % de los estudiantes manifestaron tener un conocimiento regular o deficiente sobre Rutas Integrales de Atención en Salud, planes de beneficios y actores del sistema de salud. Adicionalmente, el 7,5 % afirmaron ser poco competentes para realizar actividades de coordinación intersectorial, vigilancia epidemiológica o investigación en salud pública. CONCLUSIONES: Los resultados evidencian la necesidad de incluir en los currículos de programas de salud algunos aspectos fundamentales para la adecuada implementación del MIAS.

Handling Arabidopsis plants: growth, preservation of seeds, transformation, and genetic crosses.

Growing healthy plants is essential for the advancement of Arabidopsis thaliana (Arabidopsis) research. Over the last 20 years, the Arabidopsis Biological Resource Center (ABRC) has collected and developed a series of best-practice protocols, some of which are presented in this chapter. Arabidopsis can be grown in a variety of locations, growth media, and environmental conditions. Most laboratory accessions and their mutant or transgenic derivatives flower after 4-5 weeks and set seeds after 7-8 weeks, under standard growth conditions (soil, long day, 23 ºC). Some mutant genotypes, natural accessions, and Arabidopsis relatives require strict control of growth conditions best provided by growth rooms, chambers, or incubators. Other lines can be grown in less-controlled greenhouse settings. Although the majority of lines can be grown in soil, certain experimental purposes require utilization of sterile solid or liquid growth media. These include the selection of primary transformants, identification of homozygous lethal individuals in a segregating population, or bulking of a large amount of plant material. The importance of controlling, observing, and recording growth conditions is emphasized and appropriate equipment required to perform monitoring of these conditions is listed. Proper conditions for seed harvesting and preservation, as well as seed quality control, are also described. Plant transformation and genetic crosses, two of the methods that revolutionized Arabidopsis genetics, are introduced as well.

Evaluación de contenidos curriculares y percepción de competencias de estudiantes del área de la salud respecto del Modelo Integral de Atención en Salud en Colombia / Assessment of curricular contents and perception of competencies of health students related to the integrated health care model in Colombia

RESUMEN Objetivo Identificar la percepción de los estudiantes de último nivel de los programas de formación profesional y técnica en salud respecto a sus competencias para desempeñarse en el MIAS, y evaluar la presencia de dichas competencias dentro de los contenidos curriculares de los programas de salud en Santander. Métodos Se realizó un estudio descriptivo transversal en 451 estudiantes y una revisión documental de currículos de 26 programas. Se construyó y validó el contenido de un cuestionario en línea de percepción de competencias en los ámbitos del saber ser, saber conocer y saber hacer. Resultados En la revisión documental se encontró deficiencias en la inclusión de competencias tales como las de ejercer liderazgo (80 %) y realizar un enfoque diferencial en la atención (84 %). En la encuesta de percepción, el 36 % de los estudiantes manifestaron tener un conocimiento regular o deficiente sobre Rutas Integrales de Atención en Salud, planes de beneficios y actores del sistema de salud. Adicionalmente, el 7,5 % afirmaron ser poco competentes para realizar actividades de coordinación intersectorial, vigilancia epidemiológica o investigación en salud pública. Conclusiones Los resultados evidencian la necesidad de incluir en los currículos de programas de salud algunos aspectos fundamentales para la adecuada implementación del MIAS.(AU)

ABSTRACT Objective To identify the perception of the last level students of the professional and technical health training programs regarding their competences to perform in the MIAS, and to evaluate the presence of said competences within the curricular contents of the health programs in Santander. Methods A transversal descriptive study was carried out in 451 students and a documentary review of curricula was performed for 26 programs. The content of an online questionnaire on the perception of competences in the areas of knowledge, attitudes and abilities was constructed and validated. Results In the documentary review, deficiencies were found in the inclusion of competencies such as leadership (80 %) and differential approach in care (84 %). In the perception survey, 36 % of the students stated that they had regular or deficient knowledge about Integral Routes of Health Care, benefit plans and actors of the health system. Additionally, 7.5 % stated they were not competent to carry out intersectoral coordination activities, epidemiological surveillance or public health research. Conclusion: The results show the need to include in the curricula of health programs some fundamental aspects for the proper implementation of MIAS.(AU)

First Colombian consensus on congenital Chagas and clinical approach for women of child-bearing age diagnosed with Chagas / Primer consenso colombiano sobre Chagas congénito y orientación clínica a mujeres en edad fértil con diagnóstico de Chagas

Congenital transmission of Chagas disease has not been extensively studied in Colombia, and there are no standardized processes in the health system regarding the specific diagnosis, treatment and follow-up of this disease. To generate recommendations on congenital Chagas disease and Chagas in women of childbearing age in Colombia, a consensus of experts was developed. An extensive literature search through the Medline database was carried out using the MeSH terms: «Chagas disease/congenital¼, «prevention and control¼, «diagnosis¼, «therapeutics¼ and «pregnancy¼. Appropriate abstracts were selected and the full texts were analyzed. The relevant information was synthesized, classified, and organized into tables and figures and was presented to a panel of experts, which was composed of 30 professionals from various fields. Based on the Delphi methodology, three rounds of consultation were conducted. The first and second rounds were based on electronic questionnaires that measured the level of consensus of each question among the participants. The third round was based on a face-to-face discussion focusing on those questions without consensus in the previous consultations. The evidence was adapted to national circumstances on a case-by-case basis, and the content the final document was approved. These recommendations are proposed for use in routine medical practice by health professionals in Colombia.

La transmisión congénita de la enfermedad de Chagas ha sido poco estudiada en Colombia y existen pocos procedimientos rutinarios en el sistema de salud para el manejo de esta enfermedad. Por ello se desarrolló un consenso de expertos dirigido a generar recomendaciones de diagnóstico y tratamiento de Chagas con- génito y orientación a mujeres en edad fértil. Con ese propósito se realizó una búsqueda extensiva de la literatura, empleando una combinación de términos Mes (Chagas, Chagas congénito, prevención, control, diagnóstico, tratamiento y embarazo) para reflejar el estado del arte en cada tema de interés. Después de ello, se leyeron los resúmenes y aquellos seleccionados para análisis del texto completo. La literatura relevante se sintetizo, clasifico y organizo en tablas y se presentó al panel de expertos, el cual estaba constituido por 30 profesionales en diferentes áreas. Mediante la metodología Delphi se realizaron 2 rondas de cuestionarios virtuales y una reunión presencial en los cuales se evaluaron los niveles de acuerdo entre los participantes. Los puntos con falta de consenso durante las 2 rondas virtuales se expusieron durante las mesas de discusión en la ronda presencial. La evidencia utilizada se adaptó a las particularidades nacionales según el caso y se aprobó el contenido del documento final. Se propone que estas recomendaciones sean usadas por profesionales de la salud en Colombia.

Primer consenso colombiano sobre Chagas congénito y orientación clínica a mujeres en edad fértil con diagnóstico de Chagas / First Colombian consensus on congenital Chagas and clinical approach to women of fertile age diagnosed with Chagas

La transmisión congénita de la enfermedad de Chagas ha sido poco estudiada en Colombia y existen pocos procedimientos rutinarios en el sistema de salud para el manejo de esta enfermedad. Por ello se desarrolló un consenso de expertos dirigido a generar recomendaciones de diagnóstico y tratamiento de Chagas congénito y orientación a mujeres en edad fértil. Con ese propósito se realizó una búsqueda extensiva de la literatura, empleando una combinación de términos MeSH (Chagas, Chagas congénito, prevención, control, diagnóstico, tratamiento y embarazo) para reflejar el estado del arte en cada tema de interés. Después de ello, se leyeron los resúmenes y aquellos seleccionados para análisis del texto completo. La literatura relevante se sintetizó, clasificó y organizó en tablas y se presentó al panel de expertos, el cual estaba constituido por 30 profesionales en diferentes áreas. Mediante la metodología Delphi se realizaron 2 rondas de cuestionarios virtuales y una reunión presencial en los cuales se evaluaron los niveles de acuerdo entre los participantes. Los puntos con falta de consenso durante las 2 rondas virtuales se expusieron durante las mesas de discusión en la ronda presencial. La evidencia utilizada se adaptó a las particularidades nacionales según el caso y se aprobó el contenido del documento final. Se propone que estas recomendaciones sean usadas por profesionales de la salud en Colombia.

Congenital transmission of Chagas disease has not been extensively studied in Colombia, and there are no standardized processes in the health system regarding the specific diagnosis, treatment and follow-up of this disease. In order to generate recommendations on congenital Chagas disease and Chagas in women of childbearing age in Colombia, a consensus of experts was developed. An extensive literature search through the Medline database was carried out using the MeSH terms: " Chagas disease/congenital " , " prevention and control " , " diagnosis " , " therapeutics " and " pregnancy " . Appropriate abstracts were selected and the full texts were analyzed. The relevant information was synthesized, classified, and organized into tables and figures and was presented to a panel of experts, which was composed of 30 professionals from various fields. Based on the Delphi methodology, three rounds of consultation were conducted. The first and second rounds were based on electronic questionnaires that measured the level of consensus of each question among the participants. The third round was based on a face-toface discussion focusing on those questions without consensus in the previous consultations. The evidence was adapted to national circumstances on a case-by-case basis, and the content the final document was approved. These recommendations are proposed for use in routine medical practice by health professionals in Colombia.