Biobancos y COVID-19: implicaciones éticas / Biobanks and COVID-19: Ethical implications / Biobanks e COVID-19: Implicações éticas

Propósito/Contexto: De acuerdo con los antecedentes de epidemias anteriores, y en el contexto de la emergencia actual por COVID19, se ha reconocido como prioritario el desarrollo de investigaciones y la creación de biobancos o biorepositorios, que permitan el acceso a muestras e información para comprender la enfermedad, formular intervenciones, evaluar la seguridad y eficacia de exámenes, pruebas de diagnóstico, tratamientos, vacunas y estrategias de manejo. Aunque la investigación durante eventos de importancia en salud pública es fundamental, los estudios con seres humanos durante las emergencias deben contar con mayores garantías éticas que en las situaciones ordinarias. Metodología/Enfoque: para esta revisión se realizó la búsqueda de literatura relacionada con normatividad y legislación, biobancos, colecciones de muestras, y su relevancia en el contexto de la pandemia. Discusión/Conclusiones/Contribuciones: En esta revisión incluimos algunas reflexiones sobre el establecimiento y uso de los biobancos en el contexto de la pandemia que aqueja al mundo actualmente.

Purpose/Context. According to the history of previous epidemics, and in the context of the current emergency caused by COVID-19, it is a priority to carry out research and create biobanks or biorepositories. These allow accessing samples and information to understand the disease, formulate interventions, and evaluate the safety and efficacy of examinations, diagnostic tests, treatments, vaccines, and management strategies. Although research during significant public health emergencies is essential, studies on human beings in these cases must have higher ethical guarantees than ordinary situations. Method/Approach. This review includes literature related to regulations and legislation, biobanks, sample collections, and their relevance to the context of the pandemic. Results/Findings. Accelerated sample collection and the establishment of biobanks have been typical during this pandemic, which induces the adoption of sample management guidelines with major ethical, legal, and social differences among countries. Discussion/Conclusions/Contributions. This review comprises reflections on the establishment and use of biobanks during the pandemic that is currently afflicting the world.

Objetivo/Contexto. De acordo com os antecedentes de epidemias anteriores e no contexto da atual emergência causada pelo COVID-19, tem-se reconhecido como prioritário o desenvolvimento de pesquisas e a criação de biobancos ou bio-repositórios, que permitam o acesso a amostras e informações para entender a doença, formular intervenções, avaliar a segurança e eficácia dos exames, dos testes de diagnóstico, dos tratamentos, as vacinas e as estratégias de manejo. Embora a pesquisa durante eventos importantes em saúde pública seja essencial, os estudos com seres humanos em situações de emergência devem ter maiores garantias éticas do que em situações comuns. Metodologia/Abordagem. Para esta revisão, se efetuo a busca de literatura relacionada com o regulamento e legislação, biobancos, coletas de amostras e sua relevância no contexto da pandemia. Resultados/Descobertas. Achamos que, no contexto da pandemia, é comum a coleta acelerada de amostras e o estabelecimento de biobancos, o que induz à adoção de diretrizes para o tratamento dessas amostras, com diferenças importantes em aspectos éticos, legais e sociais entre os diferentes países. Discussão/Conclusões/Contribuições. Nesta revisão, incluímos algumas reflexões sobre o estabelecimento e o uso de biobancos no contexto da pandemia que atualmente afeta o mundo.

Complete Genome Sequence of a Colombian Zika Virus Strain Obtained from BALB/c Mouse Brain after Intraperitoneal Inoculation.

A Zika virus (ZIKV) strain was isolated from an acute febrile patient during the Zika epidemics in Colombia. The strain was intraperitoneally inoculated into BALB/c mice, and 7 days postinoculation, neurological manifestations and ZIKV infection in the brain were demonstrated. The reported genome sequence is highly related to strains circulating in the Americas.

Congenital cerebral toxoplasmosis, Zika and chikungunya virus infections: a case report

We report a case of intrauterine infection by Toxoplasma gondii, Chikungunya and Zika viruses in a Colombian woman from the southern part of the country. The patient attended prenatal care in the second trimester of her pregnancy and she informed that in the first trimester she had presented with clinical symptoms compatible with Zika virus infection. Amniotic fluid PCR assays showed infection by T. gondii, chikungunya and Zika viruses. Diagnostic imaging showed fetal malformation of the central nervous system. At 29 weeks of gestation, pregnancy was terminated medically.

Severe Neurologic Disorders in 2 Fetuses with Zika Virus Infection, Colombia.

We report the results of pathologic examinations of 2 fetuses from women in Colombia with Zika virus infection during pregnancy that revealed severe central nervous system defects and potential associated abnormalities of the eye, spleen, and placenta. Amniotic fluid and tissues from multiple fetal organs tested positive for Zika virus.

Preliminary Report of Microcephaly Potentially Associated with Zika Virus Infection During Pregnancy - Colombia, January-November 2016.

In Colombia, approximately 105,000 suspected cases of Zika virus disease (diagnosed based on clinical symptoms, regardless of laboratory confirmation) were reported during August 9, 2015-November 12, 2016, including nearly 20,000 in pregnant women (1,2). Zika virus infection during pregnancy is a known cause of microcephaly and serious congenital brain abnormalities and has been associated with other birth defects related to central nervous system damage (3). Colombia's Instituto Nacional de Salud (INS) maintains national surveillance for birth defects, including microcephaly and other central nervous system defects. This report provides preliminary information on cases of congenital microcephaly identified in Colombia during epidemiologic weeks 5-45 (January 31-November 12) in 2016. During this period, 476 cases of microcephaly were reported, compared with 110 cases reported during the same period in 2015. The temporal association between reported Zika virus infections and the occurrence of microcephaly, with the peak number of reported microcephaly cases occurring approximately 24 weeks after the peak of the Zika virus disease outbreak, provides evidence suggesting that the period of highest risk is during the first trimester of pregnancy and early in the second trimester of pregnancy. Microcephaly prevalence increased more than fourfold overall during the study period, from 2.1 per 10,000 live births in 2015 to 9.6 in 2016. Ongoing population-based birth defects surveillance is essential for monitoring the impact of Zika virus infection during pregnancy on birth defects prevalence and measuring the success in preventing Zika virus infection and its consequences, including microcephaly.

[Identification and differential expression of the microphthalmia-associated transcription factor in heart and isolated cardiomyocytes from Guinea pigs: possible role in hypertrophy and viability]. / Identificación y expresión diferencial del factor de transcripción asociado a microftalmia en corazón y cardiomiocitos aislados de cobayo: su posible papel en la hipertrofia y la viabilidad.

INTRODUCTION: The microphthalmia -associated transcription factor ( MITF ) regulates the expression of specific genes and its cardiac expression and function is not known. OBJECTIVES: To identify the expression of MITF in hearts and isolated cardiomyocytes from Guinea pigs, to describe morphological changes associated with mRNA interference of MITF and to evaluate their relative changes in expression in isolated cardiomyocytes under ischemic preconditioning. MATERIALS AND METHODS: The cardiac specific isoform, MITF-H, and relative expression level analysis, was determined by semi-quantitative real time PCR, sequencing and Western blotting. Reduction of mRNA-MITF-H was induced by transduction of specific-MITF-shRNAi interference. The cardiac morphological changes, diameter and number of cardiac fibers were evaluated by direct observation and light microscopy. RESULTS: A cDNA fragment of 281 bp was amplified from heart and isolated ventricular cardiac myocytes. Sequence analysis confirmed the identity of the isoform MITF-H, exon 1. The MITF silencing was associated with an increase in cardiac index (heart weight/body weight vs . 5.46 x 10 -3 vs 4.6 x 10 -3 ) and higher diameter of cardiac fibers (50.2±16 µ m vs 38,7±14,7 µ m p<0.05, n=150). In isolated cardiac myocytes under ischemic preconditioning we observed a higher relative expression compared with that measured in myocytes exposed to normoxia and simulated ischemia (eighty and one hundred times, p <0.05, n = 5). Conclusion. The results suggest that MITF-H isoform may be involved in Guinea pig cardiac hypertrophy, response to stress by ischemia and cardiomyocytes survival.

[Gene polymorphisms in the dihydrofolate reductase ( dhfr ) and dihydropteroate synthase ( dhps ) genes and structural modelling of the dhps gene in Colombian isolates of Toxoplasma gondii]. / Polimorfismos en los genes de dihidrofolato-reductasa ( dhfr ) y dihidropteroato-sintasa ( dhps ) y modelado estructural del gen dhps en aislamientos colombianos de Toxoplasma gondii.

INTRODUCTION: There are no reports describing polymorphisms in target genes of anti- Toxoplasma drugs in South American isolates. OBJECTIVE: This study sought to perform cloning and sequencing of the dihydrofolate reductase ( dhfr ) and dihydropteroate-synthase ( dhps ) genes of the reference Rh strain and two Colombian isolates of Toxoplasma gondii . MATERIALS AND METHODS: Two isolates were obtained from the cerebrospinal fluid of HIV-infected patients with cerebral toxoplasmosis. A DNA extraction technique and PCR assay for the dhfr and dhps genes were standardized, and the products of amplification were cloned into Escherichia coli and sequenced. RESULTS: One polymorphism (A « G) was found at position 235 of exon 2 in the dhps gene. In addition, two polymorphisms (G « C) at positions 259 and 260 and one polymorphism (T « G) at position 371 within exon 4 of the dhps gene were detected. In this last exon, a bioinformatic analysis revealed a non-synonymous polymorphism in the coding region that could lead to the substitution of Glu (CAA or CAG) for His (encoded by codons AAU or AAC). A structural model of the T. gondii DHPS protein was calculated, and the results revealed modifications in secondary structure due to mutations. CONCLUSIONS: The methods described in this study can be used as a tool to search for polymorphisms in samples from patients with different clinical manifestations of toxoplasmosis and to examine their relationship with the therapeutic response.

Polimorfismos en los genes de dihidrofolato-reductasa ( dhfr ) y dihidropteroato-sintasa ( dhps ) y modelado estructural del gen dhps en aislamientos colombianos de Toxoplasma gondii / Gene polymorphisms in the dihydrofolate reductase ( dhfr ) and dihydropteroate synthase ( dhps ) genes and structural modelling of the dhps gene in Colombian isolates of Toxoplasma gondii

Introducción. No existen reportes sobre las variaciones en la secuencia de los genes blanco de los medicamentos anti- Toxoplasma en aislamientos provenientes de Suramérica. Objetivo. Clonar y secuenciar los genes de la dihidrofolato-reductasa ( dhfr ) y la dihidropteroato-sintetasa ( dhps ) de la cepa de referencia RH y de dos aislamientos colombianos de Toxoplasma gondii. Materiales y métodos. Se obtuvieron dos aislamientos de T. gondii en líquido céfalorraquídeo de pacientes colombianos positivos para HIV con toxoplasmosis cerebral. Se extrajo el ADN de los genes dhfr y dhps y se amplificaron mediante reacción en cadena de la polimerasa (PCR). Los productos fueron clonados en el vector pGEM-T y secuenciados. Resultados. Se encontró un cambio de adenina por guanina (A « G) en la posición 235 del exón 2 del gen dhps , dos cambios de guanina por citocina (G « C) en las posiciones 259 y 260 y un cambio de timina por guanina (T « G) en la posición 371 del exón 4 del gen dhps. Por análisis bioinformático, en este último exón se identificó un polimorfismo no sinónimo en la región codificante, que podría llevar al cambio de una Glu (CAA o CAG) por una His (codificada por los codones AAU o AAC). Se calculó el modelo estructural de la enzima dihidropteroato-sintetasa (DHPS) de T. gondii y se identificaron las modificaciones en la estructura secundaria ocasionadas por las mutaciones. Conclusiones. La metodología estandarizada puede servir como base para la búsqueda de polimorfismos en muestras de pacientes con diferentes manifestaciones clínicas de toxoplasmosis y para establecer su posible relación con los cambios en la sensibilidad a los antifolatos y la reacción al tratamiento.

Introduction: There are no reports describing polymorphisms in target genes of anti- Toxoplasma drugs in South American isolates. Objective: This study sought to perform cloning and sequencing of the dihydrofolate reductase ( dhfr ) and dihydropteroate-synthase ( dhps ) genes of the reference Rh strain and two Colombian isolates of Toxoplasma gondii . Materials and methods: Two isolates were obtained from the cerebrospinal fluid of HIV-infected patients with cerebral toxoplasmosis. A DNA extraction technique and PCR assay for the dhfr and dhps genes were standardized, and the products of amplification were cloned into Escherichia coli and sequenced. Results: One polymorphism (A « G) was found at position 235 of exon 2 in the dhps gene. In addition, two polymorphisms (G « C) at positions 259 and 260 and one polymorphism (T « G) at position 371 within exon 4 of the dhps gene were detected. In this last exon, a bioinformatic analysis revealed a non-synonymous polymorphism in the coding region that could lead to the substitution of Glu (CAA or CAG) for His (encoded by codons AAU or AAC). A structural model of the T. gondii DHPS protein was calculated, and the results revealed modifications in secondary structure due to mutations. Conclusions: The methods described in this study can be used as a tool to search for polymorphisms in samples from patients with different clinical manifestations of toxoplasmosis and to examine their relationship with the therapeutic response.

Identificación y expresión diferencial del factor de transcripción asociado a microftalmia en corazón y cardiomiocitos aislados de cobayo: su posible papel en la hipertrofia y la viabilidad / Identification and differential expression of the microphthalmia-associated transcription factor in heart and isolated cardiomyocytes from Guinea pigs: Possible role in hypertrophy and viability

Introducción. El factor de transcripción asociado a la microftalmia ( Microphtalmia-Associated Transcription Factor , MITF) regula la expresión de genes específicos, pero no se conoce su expresión y su función a nivel cardiaco. Objetivos. Identificar la expresión del MITF en corazón y en cardiomiocitos aislados de cobayo, describir los cambios morfológicos asociados con su disminución y evaluar los niveles relativos de su expresión en cardiomiocitos aislados en condiciones de preacondicionamiento isquémico. Materiales y métodos. El análisis de la expresión relativa de la isoforma específica de tejido cardiaco ( heart-type MITF, MITF-H), se determinó mediante reacción en cadena de la polimerasa (PCR) en tiempo real semicuantitativa, secuenciación y Western blot . La disminución del ARNm del MITF se indujo con un ARN pequeño de interferencia ( short hairpin RNA interference , shRNAi) específico. El tamaño, el diámetro y el número de fibras musculares se evaluaron por observación directa con microscopía de luz. Resultados. Se amplificó un fragmento de 281 pb de ADNc; el análisis de la secuencia confirmó la identidad del exón 1 y la isoforma H del MITF. La interferencia del ARNm del MITF se asoció con un mayor índice cardiaco (peso corazón/peso corporal: 5,46 x 10 -3 Vs. 4,6 x 10 -3 ) y un incremento del diámetro de las fibras cardiacas (50,2±16 µm Vs. 38,7±14,7 µm; p<0,05, n=150). En los cardiomiocitos aislados en condiciones de preacondicionamiento isquémico, se observó una expresión relativa del MITF-H mayor que en los miocitos en normoxia y expuestos a lesión por isquemia simulada (80 y 100 veces más, n=5, p<0,05, n=3). Conclusión. Los resultados sugieren que el MITF-H podría estar involucrado en la hipertrofia, la respuesta al estrés por isquemia y la supervivencia de cardiomiocitos de cobayo.

Introduction: The microphthalmia -associated transcription factor ( MITF ) regulates the expression of specific genes and its cardiac expression and function is not known. Objectives: To identify the expression of MITF in hearts and isolated cardiomyocytes from Guinea pigs, to describe morphological changes associated with mRNA interference of MITF and to evaluate their relative changes in expression in isolated cardiomyocytes under ischemic preconditioning. Materials and methods: The cardiac specific isoform, MITF-H, and relative expression level analysis, was determined by semi-quantitative real time PCR, sequencing and Western blotting. Reduction of mRNA-MITF-H was induced by transduction of specific-MITF-shRNAi interference. The cardiac morphological changes, diameter and number of cardiac fibers were evaluated by direct observation and light microscopy. Results: A cDNA fragment of 281 bp was amplified from heart and isolated ventricular cardiac myocytes. Sequence analysis confirmed the identity of the isoform MITF-H, exon 1. The MITF silencing was associated with an increase in cardiac index (heart weight/body weight vs . 5.46 x 10 -3 vs 4.6 x 10 -3 ) and higher diameter of cardiac fibers (50.2±16 µ m vs 38,7±14,7 µ m p<0.05, n=150). In isolated cardiac myocytes under ischemic preconditioning we observed a higher relative expression compared with that measured in myocytes exposed to normoxia and simulated ischemia (eighty and one hundred times, p <0.05, n = 5). Conclusion. The results suggest that MITF-H isoform may be involved in Guinea pig cardiac hypertrophy, response to stress by ischemia and cardiomyocytes survival.

Presentación diferencial de ARN mensajeros e identificación del gen selenocisteína liasa en células de carcinoma hepatocelular con expresión transitoria de la proteína core del virus de la hepatitis C / Differential display of messenger RNA and identification of selenocysteine luase gene in hepatocellular carcinoma cells transiently expressing hepatitis C virus Core protein

Introducción. El virus de la hepatitis C se asocia a diversas hepatopatías como hepatitis aguda, hepatitis crónica, esteatosis, cirrosis y carcinoma hepatocelular. Numerosos estudios han explorado mecanismos virales implicados en el establecimiento de la infección persistente y en las propiedades oncogénicas e inmunomoduladoras de la proteína core del virus de la hepatitis C. Las investigaciones orientadas a evaluar los cambios en la expresión de genes celulares endógenos inducidos por la proteína core son importantes para identificar genes candidatos responsables de los mecanismos de patogenicidad del virus de la hepatitis C. Objetivos. Comparar perfiles de expresión e identificar genes celulares endógenos en la línea celular derivada de carcinoma hepatocelular humano, HepG2, con expresión transitoria de la proteína core del virus de la hepatitis C. Materiales y métodos. Se utilizó la técnica de presentación diferencial de ARN mensajero por RT-PCR en células HepG2 con y sin expresión transitoria de la proteína core del virus de la hepatitis C o de la proteína verde fluorescente, obtenidas previamente con el sistema de expresión del Semliki Forest Virus, mediante transducción de partículas recombinantes rSFV-Core o rSFV-GFP. Resultados. Se observaron diferencias en las intensidades de las bandas de ARNm expresadas en células HepG2 transducidas con rSFV-Core comparadas con células sin transducir y trasducidas con rSFV-GFP. Un ARNm de 258 pb expresado diferencialmente en células HepG2 transducidas con rSFV-Core fue clonado e identificado como selenocisteína liasa. Conclusión. Los resultados confirman que la expresión de la proteína core del virus de la hepatitis C se asocia con cambios en la expresión de ARN mensajeros específicos, incluido al gen selenocisteina liasa, el cual puede estar involucrado en la fisiopatología del carcinoma hepatocelular