Local-scale virome depiction in Medellín, Colombia, supports significant differences between Aedes aegypti and Aedes albopictus.

Aedes spp. comprise the primary group of mosquitoes that transmit arboviruses such as dengue, Zika, and chikungunya viruses to humans, and thus these insects pose a significant burden on public health worldwide. Advancements in next-generation sequencing and metagenomics have expanded our knowledge on the richness of RNA viruses harbored by arthropods such as Ae. aegypti and Ae. albopictus. Increasing evidence suggests that vector competence can be modified by the microbiome (comprising both bacteriome and virome) of mosquitoes present in endemic zones. Using an RNA-seq-based metataxonomic approach, this study determined the virome structure, Wolbachia presence and mitochondrial diversity of field-caught Ae. aegypti and Ae. albopictus mosquitoes in Medellín, Colombia, a municipality with a high incidence of mosquito-transmitted arboviruses. The two species are sympatric, but their core viromes differed considerably in richness, diversity, and abundance; although the community of viral species identified was large and complex, the viromes were dominated by few virus species. BLAST searches of assembled contigs suggested that at least 17 virus species (16 of which are insect-specific viruses [ISVs]) infect the Ae. aegypti population. Dengue virus 3 was detected in one sample and it was the only pathogenic virus detected. In Ae. albopictus, up to 11 ISVs and one plant virus were detected. Therefore, the virome composition appears to be species-specific. The bacterial endosymbiont Wolbachia was identified in all Ae. albopictus samples and in some Ae. aegypti samples collected after 2017. The presence of Wolbachia sp. in Ae. aegypti was not related to significant changes in the richness, diversity, or abundance of this mosquito's virome, although it was related to an increase in the abundance of Aedes aegypti To virus 2 (Metaviridae). The mitochondrial diversity of these mosquitoes suggested that the Ae. aegypti population underwent a change that started in the second half of 2017, which coincides with the release of Wolbachia-infected mosquitoes in Medellín, indicating that the population of wMel-infected mosquitoes released has introduced new alleles into the wild Ae. aegypti population of Medellín. However, additional studies are required on the dispersal speed and intergenerational stability of wMel in Medellín and nearby areas as well as on the introgression of genetic variants in the native mosquito population.

Bacteriome depiction and the trophic status of the largest Northern highland lake from Andes system: Lago de Tota, Boyacá, Colombia.

Lago de Tota is the largest highland lake in Colombia and one of the most remarkable of Northern Andean Mountain range. This lake is under an anthropogenic-based eutrophication process as a consequence of non-sustainable agriculture practices developing nearby. Notable relationship between the trophic status and Bacteriome loop dynamics has been increasingly disclosed in lakes worldwide. We performed a 16S sequencing analysis to depict the bacterial community present and we inferred its potential gene function in Lago de Tota. Parameters for determining current trophic condition such as total nitrogen (TN), dissolved carbon (DOC), particulate organic matter (POM), and chlorophyll-a (chl-a) were measured. A total of 440 Operational Taxonomic Units (OTUs) arranged into 50 classes were identified based on V3-V4 regions of the 16S rRNA gene, harboring high-frequent likely found environmental classes such as Actinobacteria, Gammaproteobacteria, Bacteroidia, Acidimicrobia, and Verrucomicrobiae. A total of 26 bacterial classes configure most abundant predicted functional processes involved in organic matter decomposition (i.e., carbohydrate metabolism, amino acid metabolism, xenobiotic biodegradation, and energy metabolism). In general, Actinobacteria, Alphaproteobacteria, and Gammaproteobacteria show the highest potential gene functional contributors, although other low-frequent classes OTUs are also relevant in processes of carbohydrate metabolism, xenobiotic biodegradation, and energy metabolism. The Trophic State Index indicates an oligo-mesotrophic status, and additional variables measured (i.e., POM, DOC) suggest the increasing carbon accumulation. Results provide preliminary evidence for several bacteria groups related to eutrophication of Lago de Tota. Under this picture, we suggest that further studies for Bacteriome loop spatial-temporal description are essential to inform local water quality monitoring strategies.

The absence of the aryl hydrocarbon receptor in the R6/1 transgenic mouse model of Huntington's disease improves the neurological phenotype.

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by an abnormal CAG repeat expansion in the huntingtin gene coding for a protein with an elongated polyglutamine sequence. HD patients present choreiform movements, which are caused by the loss of neurons in the striatum and cerebral cortex. Previous reports indicate that the absence of the aryl hydrocarbon receptor (AhR) protects mice from excitotoxic insults and increases the transcription of neurotrophic factors. Based on these data, we evaluated the effects of the lack of the AhR on a mice model of HD, generating a double transgenic mouse, expressing human mutated huntingtin (R6/1 mice) and knockout for the AhR. Our results show that the body weight of 30-week-old double transgenic mice is similar to that of R6/1 mice; however, feet clasping, an indicative of neuronal damage in the R6/1 animals, was not observed. In addition, motor coordination and ambulatory behavior in double transgenic mice did not deteriorate over time as occur in the R6/1 mice. Moreover, the anxiety behavior of double transgenic mice was similar to wild type mice. Interestingly, astrogliosis is also reduced in the double transgenic mice. The present data demonstrate that the complete loss of the AhR reduces the motor and behavioral deterioration observed in R6/1 mice, suggesting that the pharmacological modulation of the AhR could be a therapeutic target in HD.

Expression of Gas1 in Mouse Brain: Release and Role in Neuronal Differentiation.

Growth arrest-specific 1 (Gas1) is a pleiotropic protein that induces apoptosis of tumor cells and has important roles during development. Recently, the presence of two forms of Gas1 was reported: one attached to the cell membrane by a GPI anchor; and a soluble extracellular form shed by cells. Previously, we showed that Gas1 is expressed in different areas of the adult mouse CNS. Here, we report the levels of Gas1 mRNA protein in different regions and analyzed its expressions in glutamatergic, GABAergic, and dopaminergic neurons. We found that Gas1 is expressed in GABAergic and glutamatergic neurons in the Purkinje-molecular layer of the cerebellum, hippocampus, thalamus, and fastigial nucleus, as well as in dopaminergic neurons of the substantia nigra. In all cases, Gas1 was found in the cell bodies, but not in the neuropil. The Purkinje and the molecular layers show the highest levels of Gas1, whereas the granule cell layer has low levels. Moreover, we detected the expression and release of Gas1 from primary cultures of Purkinje cells and from hippocampal neurons as well as from neuronal cell lines, but not from cerebellar granular cells. In addition, using SH-SY5Y cells differentiated with retinoic acid as a neuronal model, we found that extracellular Gas1 promotes neurite outgrowth, increases the levels of tyrosine hydroxylase, and stimulates the inhibition of GSK3ß. These findings demonstrate that Gas1 is expressed and released by neurons and promotes differentiation, suggesting an important role for Gas1 in cellular signaling in the CNS.

Variantes genéticas de susceptibilidad en epilepsia genética generalizada en familias colombianas / Susceptibility gene variants in genetic generalized epilepsy in Colombian families

INTRODUCCIÓN: las epilepsias genéticas generalizadas (EGG) siguen patrones de herencia compleja. Este fenotipo es producto de la interacción de diferentes genes con factores ambientales. Los genes/loci más consistentemente asociados con este grupo de epilepsias son ECA1, ECA2-GABRG2, ECA3-CLCN2 (también conocido como JME6-CLCN2), JME1-EFHC1 y JME5-GABRA1. En Colombia poco se sabe sobre la contribución de las variantes genéticas en estos genes a la susceptibilidad para ser afectado por cualquiera de las formas de EGG. Nuestro propósito fue evaluar el papel de los cinco genes/loci más consistentemente asociados en otros estudios en un grupo de familias colombianas con EGG. MÉTODOS: se evaluaron dos marcadores para cada locus/gen. Los genotipos se obtuvieron mediante las técnicas de PCR-RFLP y ARMS-Tetraprimer. Los análisis estadísticos incluyeron pruebas de asociación alélica y haplotípica, además de pruebas de interacción gen-gen. RESULTADOS: se incluyeron 98 familias, de las cuales 51 fueron epilepsia de ausencias, mientras que 47 fueron epilepsia mioclónica juvenil. Se identificó una interacción significativa entre el alelo G del marcador rs4428455 (valor P=0,0008; gen GABRA1) y el alelo G de marcador rs719395 (valor P=0,002; gen EFHC1). CONCLUSIÓN: estos dos marcadores parecen incrementar el riesgo de EGG en población colombiana. Otros genes no analizados aquí podrían estudiarse con una muestra de mayor tamaño

INTRODUCTION: Generalized genetic epilepsies (GGE) follow complex inheritance patterns. This phenotype is due to interaction of several genes with environmental factors. The genes/loci most consistently associated with this group of epilepsies are ECA1, ECA2-GABRG2, ECA3-CLCN2 (also known as JME6-CLCN2), JME1-EFHC1 and JME5-GABRA1. In Colombia, little is known about the contribution of gene variants to susceptibility to GGE forms. Our purpose was to evaluate the role of the five most consistently associated genes /loci in other studies, in Colombian families set with GGE. METHODS: Genotypes were obtained by means of PCR-RFLP and ARMS-Tetraprimer. Statistical analyses included both allelic and haplotypic association tests, in addition to gene-gene interaction tests. Two genetic markers were tested for each gene/locus. RESULTS: Ninety-eight families were included, from which 51 had absence epilepsy, and 47 had juvenile myoclonic epilepsy. A significant interaction was identified between allele G at marker rs4428455 (P-value= 0.0008; gene GABRA1) and allele G at marker rs719395 (P-value= 0.002; gene EFHC1). CONCLUSION: Our results suggest that these two markers are associated with GGE in the Colombian population. Other genes not analyzed could be tested using a larger sample size.

Curated Collections for Educators: Five Key Papers about Program Evaluation.

The evaluation of educational programs has become an expected part of medical education. At some point, all medical educators will need to critically evaluate the programs that they deliver. However, the evaluation of educational programs requires a very different skillset than teaching. In this article, we aim to identify and summarize key papers that would be helpful for faculty members interested in exploring program evaluation. In November of 2016, the 2015-2016 Academic life in emergency medicine (ALiEM) Faculty Incubator program highlighted key papers in a discussion of program evaluation. This list of papers was augmented with suggestions by guest experts and by an open call on Twitter. This resulted in a list of 30 papers on program evaluation. Our authorship group then engaged in a process akin to a Delphi study to build consensus on the most important papers about program evaluation for medical education faculty. We present our group's top five most highly rated papers on program evaluation. We also summarize these papers with respect to their relevance to junior medical education faculty members and faculty developers. Program evaluation is challenging. The described papers will be informative for junior faculty members as they aim to design literature-informed evaluations for their educational programs.

Academic Primer Series: Eight Key Papers about Education Theory.

INTRODUCTION: Many teachers adopt instructional methods based on assumptions of best practices without attention to or knowledge of supporting education theory. Familiarity with a variety of theories informs education that is efficient, strategic, and evidence-based. As part of the Academic Life in Emergency Medicine Faculty Incubator Program, a list of key education theories for junior faculty was developed. METHODS: A list of key papers on theories relevant to medical education was generated using an expert panel, a virtual community of practice synthetic discussion, and a social media call for resources. A three-round, Delphi-informed voting methodology including novice and expert educators produced a rank order of the top papers. RESULTS: These educators identified 34 unique papers. Eleven papers described the general use of education theory, while 23 papers focused on a specific theory. The top three papers on general education theories and top five papers on specific education theory were selected and summarized. The relevance of each paper for junior faculty and faculty developers is also presented. CONCLUSION: This paper presents a reading list of key papers for junior faculty in medical education roles. Three papers about general education theories and five papers about specific educational theories are identified and annotated. These papers may help provide foundational knowledge in education theory to inform junior faculty teaching practice.

Características clínicas de los pacientes menores de 18 años con atrofia muscular espinal en Medellín, 2008 - 2013 / Clinical characteristics of patients under 18 years-old with spinal muscular atrophy in Medellín, 2008 - 2013

Introducción: La atrofia muscular espinal (AME) es una enfermedad degenerativa que afecta las neuronas motoras del asta anterior de la médula espinal, se manifiesta por debilidad muscular progresiva de predominio proximal, hipotonía y arreflexia osteotendinosa, la etiología es una mutación en el gen de supervivencia neuronal SMN. Objetivo: determinar las características clínicas de los pacientes menores de 18 años con atrofia muscular espinal en de Medellín, durante el período 2008-2013. Materiales y métodos: se realizó un estudio descriptivo retrospectivo de los pacientes con AME que consultaron en el Hospital Universitario San Vicente Fundación y un consultorio privado de neuropediatría en Medellín durante el período 2008-2013, en total se recopilaron datos de 29 pacientes, se revisaron las características clínicas, las ayudas diagnósticas practicadas y los tratamientos realizados. Resultados: la AME tipo II resultó la forma clínica más frecuente (62%) seguida por la AME tipo I (24.13%), las principales manifestaciones fueron la hipotonía (100%) debilidad muscular (93.1%) y la arreflexia osteotendinosa (82.8%). Las fasciculaciones en la lengua se presentaron en el 48.3% de los pacientes. La prueba molecular fue realizada en 6 pacientes y en todos se encontró deleción del exón 7 del gen SMN1. Conclusión: la atrofia muscular espinal es una enfermedad degenerativa y de progresión variable de acuerdo a su clasificación. Clínicamente, se debe sospechar cuando exista síndrome motoneuronal y fasciculaciones linguales. El diagnóstico molecular es el método más acertado para confirmar la enfermedad.

Introduction: Spinal muscular atrophy (SMA) is a degenerative disease that affects motor neurons in the anterior horn of the spinal cord, it is manifested by progressive muscle weakness predominantly proximal, hypotonia andosteotendinous arreflexia, the cause is a mutation in neuronal survival gene SMN1 Objective: Establish the clinical, electromyographics and genetics characteristics of patients younger than 18 years with spinal muscular atrophy in the Medellín city, during the period 2008-2013. Materials and methods: A retrospective study of patients with SMA seen in the Hospital Universitario San Vicente Foundation and private center of Neuro- pediatric of Medellín during the period 2008-2013 was performed. Data from 29 patientes were available, were reviewed clinical feature, diagnostic aids and treatments practiced. Results: SMA type II resulted the most frequent clinical presentation (62%) followed by SMA type I (24.13%), the main manifestations were hypotonia (100 %), muscle weakness (93.1%) and osteotendinous arreflexia (82.8%), tongue fasciculations occurred in 48.3% of patients. The molecular test was performed in 6 patients and in all the deletion of exon 7 of the SMN1 gene was found. Conclusion: spinal muscular atrophy is a degenerative and progressive disease according to their clinical classification. It should be suspected when there are motoneuronal syndrome and lingual twitches. Molecular diagnosis is the most accurate to confirm the disease.

Novel cerebrotendinous xanthomatosis mutation causes familial early dementia in Colombia.

INTRODUCTION: Cerebrotendinous xanthomatosis is an infrequent cause of dementia. It is an autosomal recessive disorder with clinical and molecular heterogeneity. OBJECTIVE: To identify the presence of a possible mutation in a Colombian family with several affected siblings and clinical characteristics compatible with cerebrotendinous xanthomatosis associated to early dementia. MATERIALS AND METHODS: We studied a series of cases with longitudinal follow-up and genetic analysis. RESULTS: These individuals had xanthomas, mental retardation, psychiatric disorders, behavioral changes, and multiple domains cognitive impairment with dysexecutive dominance that progressed to early dementia. CYP27A1 gene coding region sequencing revealed a novel mutation (c.1183_1184insT). CONCLUSION: The mutation found in this family is responsible for the described dementia features. Early identification of familial history with mental retardation, xanthomas and cognitive impairment might prevent the progression to this treatable type of dementia. Even though this mutation lies in the most frequently mutated codon of CYP27A1 gene, it has not been reported previously.

Síndromes de Bartter y Gitelman: revisión de los aspectos genéticos, fisiopatológicos y clínicos / Bartter and Gitelman syndromes: a review of genetic, physiopathological and clinical aspects

Los síndromes de Bartter y Gitelman son trastornos hereditarios caracterizados por una reducción marcada del transporte de sales por el asa ascendente gruesa de Henle. Los pacientes con el síndrome de Bartter presentan grandes pérdidas renales de agua, hipotensión arterial, alcalosis metabólica, hipokalemia e hipercalciuria y tienen un riesgo alto de sufrir nefrolitiasis. Estudios recientes han permitido subdividir el síndrome de Bartter en cinco tipos de acuerdo con el defecto genético y el fenotipo. El tipo 1 es causado por una mutación en el gen que codifica para el cotransportador cloro-sodio-potasio; el tipo 2 se debe a una mutación en el gen que codifica para el canal de potasio ROMK. Estas dos variantes genéticas se denominan conjuntamente síndrome de Bartter neonatal por el comienzo temprano de los síntomas, con polihidramnios materno, prematuridad, poliuria grave y nivel elevado de prostaglandina E2. El tipo 3, conocido como síndrome de Bartter clásico, es causado por mutaciones en el gen que codifica para el canal de cloro CLC-Kb; se detecta desde la niñez con retardo del crecimiento, poliuria, polidipsia y anorexia. El tipo 4 se asocia a sordera neurosensorial y se caracteriza por una mutación en el gen que codifica para la proteína Barttin la cual hace parte de los canales de cloro CLC-Kb y CLC-Ka; el tipo 5 se debe a una mutación en el gen que codifica para un receptor de calcio ubicado en la membrana basolateral del asa ascendente de Henle; estos pacientes, además de los síntomas comunes a los de otros tipos, presentan déficit de paratohormona. El síndrome de Gitelman tiene un fenotipo más leve y una presentación más tardía que el de Bartter; se diferencia de este porque los pacientes tienen hipomagnesemia e hipocalciuria; pueden ser asintomáticos o presentar debilidad muscular transitoria, parestesias, parálisis e incluso alteraciones del ritmo cardíaco. Los avances en genética molecular han permitido la clasificación adecuada de estos síndromes y han abierto puertas para diferentes opciones terapéuticas. Esta revisión incluye aspectos genéticos, fisiopatológicos y clínicos de estos síndromes.

Bartter and Gitelman syndromes are hereditary disorders characterized by a remarkable reduction of salt transportation by the thick ascending limb of the Henle‘s loop. Consequently, patients suffering from Bartter syndrome present with renal salt wasting, low blood pressure, hypokalemic metabolic alkalosis and hypercalciuria, and are at risk of developing renal stones. Based on recent studies, the Bartter syndrome has been subdivided into five types according to the genetic defect involved and the phenotype: type 1 is caused by a mutation in the gene coding for the chloride, sodium, and potassium co-transporter; type 2 is due to a mutation in the gene coding for the ROMK potassium channel. These two genetic variations are jointly denominated neonatal Bartter syndrome, because of their early clinical presentation, with maternal polyhydramnios, prematurity, severe polyuria and high levels of E2 prostaglandine. Type 3, also known as classic Bartter syndrome, is produced by mutations in the gen that codes for the chloride CLC-Kb channel; this type is detected since childhood with growth delay, polyuria, polydipsia and anorexia. Type 4, which is associated with neurosensorial deafness, is characterized by a mutation in the gen that codes for the Barttin protein which is a part of the CLC-Kb and CLC-Ka chloride channels. Type 5 appears because of a mutation in the gene that codes for a calcium receptor located at the basolateral membrane of the ascending limb of Henle‘s loop; patients with this type develop parathormone deficit, as well as the symptoms that are common to all types of the syndrome. The phenotype of Gitelman syndrome is less severe and its clinical presentation is delayed; it differs from the Bartter syndrome in that patients have hypomagnesemia and hipocalciuria. They may be asymptomatic or show transitory muscular weakness, paresthesias, paralysis and even cardiac rhythm alterations. Recent advances in molecular genetics have made it possible to distinguish between the different clinical types and are the basis for several therapeutic options. This review includes genetic, physiopathological and clinical aspects of the Bartter and Gitelman syndromes.

Análisis de ligamento genético de la diabetes mellitus tipo 1, a marcadores de los cromosomas 2 y 11 en familias antioqueñas / Genetic linkage analysis of type 1 diabetes mellitus to markers on chromosomes 2 and 11 in families from Antioquia, Colombia

La Diabetes Mellitus (DM) comprende un grupo heterogéneo de desordenes hiperglucémicos clasificados en subgrupos de acuerdo a su fisiopatología y etiología, entre los cuales se destacan la Diabetes Mellitus tipo1 (DM1) y la diabetes mellitus tipo 2 (DM2). La DM1 es de aparición temprana y una absoluta escasez de insulina hace que los pacientes sean insulino dependientes desde el inicio de los síntomas y la (DM2) que se manifiesta en la edad adulta y no todos los pacientes que la sufren son insulino dependientes

Diabetes mellitus (DM) comprises e heterogeneous group of hypoglycemic disorders, that are grouped according to their physiopathology and etiology; the most notorious ones are type 1 DM (DM1) and type 2 DM (DM2); DM1 is characterized by early onset and absolute lack of insulin; therefore, patients suffering from it depend on insulin since the beginning of their symptoms; in contrast, DM2 manifests during adult life and not all patients depend on insulin

Análisis molecular en 29 pacientes con diagnóstico presuntivo de mitocondriopatía: encefalopatía mitocondrial con acidosis láctica y aplopejía (MELAS) / Molecular analysis in mitochondrial encephalophathy with lactic acidosis and stroke (MELAS)

El objetivo de la investigación fue identificar en pacientes con diagnóstico presuntivo de MELAS la mutación A3243G, tradicionalmente asociada con ella. Se secuenciaron los fragmentos 533 pb del genoma mitocondrial correspondiente al tRNA-Leu y parte del gen NDI (posiciones 3162 a 3695) en 29 pacientes con diagnóstico presuntivo de MELAS y en cuatro personas sanas. Todos los pacientes fueron negativos para la mutación A3243G, tampoco se hallaron otras mutaciones descritas en MELAS. Se halló una transición (A3547G) en el gen NDI, hasta ahora no informada en la literatura, en 14 de las muestras secuenciadas. Esta transición, identificada en pacientes y controles, cambia el codón ATC(i) por GTC(V). Al realizar el genotipo del mtDNA de la PCR, se encontró que de las 14 muestras con G en la posición 3547 de NDI, 13 expresaban el haplotipo B. Teniendo en cuenta que la transición detectada se presentó tanto en pacientes como en controles y que I y V son aminoácidos de estructura química semejante, hidrofóbicos, que se localizan en un motivo altamente hidrofóbico de la proteína (LALTIALLL), es posible concluír que este cambio consiste en un polimorfismo. La presencia del Haplotipo B, propia de la población amerindia, indica la alta frecuencia de mitocondria de origen antioqueño. La transición 3547G podría ser un marcador de la población indoamericana con la cual se mezclaron los españoles para constituír la población antioqueña