Etiology of Microcephaly and Central Nervous System Defects during the Zika Epidemic in Colombia.

OBJECTIVE: To estimate the prevalence of microcephaly and central nervous system (CNS) defects during the Zika virus (ZIKV) epidemic in Colombia and proportion attributable to congenital ZIKV infection. STUDY DESIGN: Clinical and laboratory data for cases of microcephaly and/or CNS defects reported to national surveillance between 2015 and 2017 were reviewed and classified by a panel of clinical subject matter experts. Maternal and fetal/infant biologic specimens were tested for congenital infection and chromosomal abnormalities. Infants/fetuses with microcephaly and/or CNS defects (cases) were classified into broad etiologic categories (teratogenic, genetic, multifactorial, and unknown). Cases classified as potentially attributable to congenital ZIKV infection were stratified by strength of evidence for ZIKV etiology (strong, moderate, or limited) using a novel strategy considering birth defects unique or specific to ZIKV or other infections and laboratory evidence. RESULTS: Among 858 reported cases with sufficient information supporting a diagnosis of microcephaly or CNS defects, 503 were classified as potentially attributable to congenital ZIKV infection. Of these, the strength of evidence was considered strong in 124 (24.7%) cases; moderate in 232 (46.1%) cases; and limited in 147 (29.2%). Of the remaining, 355 (41.4%) were attributed to etiologies other than ZIKV infection (syphilis, toxoplasmosis, rubella, cytomegalovirus, herpes 1 and herpes 2 viruses only, n = 32 [3.7%]; genetic, n = 16 [1.9%]; multifactorial, n = 42 [4.9%]; unknown, n = 265 [30.9%]). CONCLUSIONS: Fifty-eight percent of cases of microcephaly and/or CNS defects were potentially attributable to congenital ZIKV infection; however, the strength of evidence varied considerably. This surveillance protocol might serve as a model approach for investigation and etiologic classification of complex congenital conditions.

Análisis de hemoglobinopatías en regiones afrocolombianas usando muestras de sangre seca de cordón umbilical / Analysis of hemoglobinopathies in afro-colombian regions using dried blood samples of umbilical cord

Introducción: las hemoglobinopatías constituyen el grupo de trastornos genéticos más frecuentes a nivel mundial, con mayor prevalencia en población negra. En Colombia hay regiones de alta densidad poblacional de afrodescendientes, lo cual plantea la necesidad de establecer estrategias para la identificación temprana de los portadores y enfermos con propósitos de prevención. Objetivo: realizar la identificación temprana de hemoglobinopatías en muestras de sangre seca de cordón umbilical. Material y métodos: para las muestras de sangre seca se utilizaron tarjetas de tamizaje neonatal rutinario de hipotiroidismo congénito. La sangre es eluida por medio de soluciones buffer acuosas y luego se aplican para separación por electroforesis en geles de isoelectroenfoque y en columna de electroforesis capilar. Resultados: de 5141 muestras recibidas de sangre seca de cordón umbilical, se descartaron 1294 por problemas de calidad, se procesaron 3847 (75%) y de éstas 3244 (84%) fueron útiles para el estudio. Se encontraron 288 (7%) con alguna variante de hemoglobina, FC (1,88%), FS(4,65%), SC(0.06%), SS(0,12%), Bart (0,18%), FC/otro (0,12%). Conclusiones: las muestras de sangre seca de cordón umbilical permiten realizar el estudio por electroforesis capilar y por isoelectroenfoque de las diferentes variantes de hemoglobinas anormales, que puede ser aplicable al diagnóstico temprano, siempre que se cumplan requisitos de calidad, en la aplicación de la muestra y en el tiempo de entrega para análisis. (Acta Med Colomb 2012: 37: 118-124).

Introduction: the hemoglobinopathies represent the most common group of genetic disorders worldwide, with higher prevalence in black population. In Colombia there are regions with high density population of African descent, which raises the need to develop strategies for early identification of carriers and patients with preventive purposes. Objective: realize early identification of hemoglobinopathies in dried blood samples from the umbilical cord. Material and methods: for the dried blood samples we used routine neonatal screening cards for congenital hypothyroidism. The blood is eluted by means of aqueous buffer solutions and then applied for separation by electrophoresis in isoelectric focusing gels and in capillary electrophoresis column. Results: of the 5141cord dried blood samples received, 1294 were discarded due to quality problems; 3847 (75%) were processed and of these, 3244 (84%) were appropriate for the study. We found 288 (7%) with some hemoglobin variant, FC (1.88%), FS (4.65%), SC (0.06%), SS (0.12%), Bart (0.18%) , FC / other (0.12%). Conclusions: dried blood samples of umbilical cord allow the study of the different variants of abnormal hemoglobins by capillary electrophoresis and isoelectric focusing, which may be applicable to an early diagnosis in the neonatal screening, provided that quality requirements in the sample application and in the delivery time for analysis are accomplished. (Acta Med Colomb 2012: 37: 118-124).

Uso del bajo peso al nacer como criterio seleccionador para la vigilancia rutinaria de anomalías congénitas de origen infeccioso / Use of low birth weight as selecting criterion for the routine surveillance of congenital anomalies of infectious origin

Introducción: La vigilancia de las anomalías congénitas cobra importancia en el contexto mundial de erradicar el síndrome de rubéola congénita. Objetivo: Identificar las anomalías congénitas y considerar el bajo peso como criterios para hacer pruebas rutinarias de IgM para el complejo TORCH. Metodología: Vigilancia de las anomalías congénitas, específicas y no específicas del síndrome de rubéola congénita (SRC) y bajo peso al nacer en diez centros hospitalarios. Se consideró como caso, todo recién nacido con alguna anomalía congénita o con bajo peso para la edad gestacional. Se practicaron pruebas en suero para rubéola, toxoplasmosis, citomegalovirus, herpes y parvovirus. Para los casos negativos se practicó cariotipo. Resultados: Se captaron 840 casos, 669 por peso bajo para la edad gestacional, 52 por anomalía no relacionada con SRC, 105 por anomalías que se podrían relacionar con el SRC. Las mßs frecuentes fueron cardiopatías, 5.1%; hepatoesplenomegalias, 3.9%; y microcefalias, 1.2%. Para sífilis congénita la tasa de positividad fue 3.7%; rubéola, 0.5%; toxoplasmosis, 1.4%; citomegalovirus, 1.5%; parvovirus, 1.2%; y herpes, 0.5%. El riesgo relativo para bajo peso con IgM rubéola positivo fue RR = 2.83 (IC: 1.26:6.36-0.95). Discusión y conclusiones: La vigilancia del SRC se hizo sobre todo por medio de la vigilancia de febriles en el primer a±o y por la presencia de algunas anomalías congénitas. La sensibilidad se mejoró por medio de la vigilancia rutinaria de anomalías congénitas, con la inclusión de bajo peso al nacer, como criterio seleccionador para estudiar los agentes infecciosos.

Introduction: Surveillance of congenital anomalies receives importance in the world-wide context of eradicating the congenital rubella syndrome. Objective: To identify the congenital anomalies and to consider the low birth weightá a criterion to test IgM for the complex TORCH. Methodology: Surveillance of the congenital specific and non specific anomalies of the congenital rubella syndrome(CRS) and low birth weight in ten hospitals. It was considered as case, everything new born with some congenital anomaly or low weight for the gestational age. Serum tests for rubella, toxoplasmosis, citomegalovirus, herpes and parvovirus were practiced. For the negative cases cariotype was performed. Results: A total of 840 cases were selected, 669 by low weight for the gestational age, 52 by anomalies not related to CRS, 52; by anomalies that could be related to the CRS, 105. The most frequent anomalies were congenital heart diseases,á 5.1%; hepatosplenomegalies, 3.9%; and microcephalies, 1.2%. There were confirmatory IgM titles for rubella in 0.5% of cases; toxoplasmosis 1.4%; citomegalovirus 1.5%; parvovirus 1.2%; herpes 0.5%; and positive test for congenital syphilis, 3.7%. In total there were 8.8% positive results for any congenital infectious disease. The relative risk for low birth weight having IgM positive rubella was RR = 2.83 (IC: 1.26:6.36-0.95). Discussion and conclusions: The surveillance for CRS, through the monitoring of febrile in the first year and by the presence of some specific congenital anomalies, could be improved in sensitivity by means of the routine monitoring of congenital anomalies, with the inclusion of low birth weight, like a selecting criterion to study the infectious agents.

Estudio inmunocitoquímico y molecular de cultivo primario de tejido molar / Immunocytochemical and molecular studies with primary cultures of molar tissue

Introducción. La enfermedad trofoblástica gestacional comprende un conjunto de patologías caracterizadas por crecimiento e invasión anómalos del trofoblasto. Las bases moleculares de esta patología son desconocidas, en parte por la dificultad para disponer de modelos biológicos adecuados. Se plantea que el sistema de factores de crecimiento similares a la insulina puede tener un papel fundamental en el desarrollo de la enfermedad. Objetivo. Caracterizar cultivos primarios de placentas de primer trimestre provenientes de pacientes con mola hidatidiforme completa y aborto espontáneo no molar mediante morfología, inmunocitoquímica y expresión diferencial de algunos genes del sistema de factores de crecimiento similares a la insulina. Materiales y métodos. Se empleó inmunocitoquímica para determinar células trofoblásticas y detección por transcripción reversa y reacción en cadena de la polimerasa de genes del sistema de factores de crecimiento similares a la insulina asociados al tipo celular. Resultados. La morfología evidenció heterogeneidad de los cultivos, incluidas células mesenquimales, trofoblásticas y de decidua. El contenido de células de trofoblasto con citoqueratina-7 (marcador específico) estuvo entre 16 y 37 por ciento. La expresión de genes corroboró la presencia de trofoblasto por medio del ARNm del factor II de crecimiento similar a la insulina, en tanto que los transcritos de la hormona de crecimiento variante evidenciaron la presencia de sincitiotrofoblasto. El factor I de crecimiento similar a la insulina y la proteína de unión tipo 1 se relacionaron con células mesenquimales y de decidua. Se observó una mayor expresión del factor II de crecimiento similar a la insulina en tejidos molares en comparación con aborto no molar. Conclusiones. Los resultados mostraron la utilidad de combinar tres metodologías, morfología, inmunocitoquímica y expresión de genes, como herramientas para la caracterización y seguimiento de cultivos placentarios a partir....

Introduction. Gestational trophoblastic disease includes a group of pathologies characterized by abnormal trophoblast growth and invasion. The molecular bases of the disease are largely unknown, due in part to the lack of appropriate biological models. The insulin-like growth factor (IGF) system plays a fundamental role in the growth and development of many tissues and is involved in the progression of several diseases. Objectives. Primary cell cultures derived from first trimester placenta were characterized from patients with complete hydatidiform mole and spontaneous non molar abortion by immunocytochemical and molecular methods. Materials and Methods. The immunocytochemical method used specific markers for trophoblastic cells, whereas RT-PCR was used to identify insulin-like growth factor gene expression. Results. Histochemical staining with hematoxilin-eosin revealed that the cultures contained heterogeneous cell types, including trophoblast and endometrial decidual cells. The ratio of trophoblast cells in the cultures varied between 16% and 37%, as detected by cytokeratine-7 as the specific trophoblast marker. Gene expression analysis corroborated the presence of trophoblasts by detecting insulin-like growth factor II mRNA, whereas GH-V transcripts were correlated with the presence of syncitiotrophoblasts. Insulin-like growth factor I and insulin-like growth factor binding protein 1 mRNAs were related to mesenchyimal and decidual cells, respectively. Higher insulin-like growth factor II expression levels were found in molar tissues in comparison with non-molar abortions. Conclusion. By combining three methodologies—morphology, immunocytochemistry and gene expression, characterization and follow-up of placenta cultures from abnormal tissues is found to facilitate diagnosis.

[Immunocytochemical and molecular studies with primary cultures of molar tissue]. / Estudio inmunocitoquímico y molecular de cultivo primario de tejido molar.

INTRODUCTION: Gestational trophoblastic disease includes a group of pathologies characterized by abnormal trophoblast growth and invasion. The molecular bases of the disease are largely unknown, due in part to the lack of appropriate biological models. The insulin-like growth factor (IGF) system plays a fundamental role in the growth and development of many tissues and is involved in the progression of several diseases. OBJECTIVES: Primary cell cultures derived from first trimester placenta were characterized from patients with complete hydatidiform mole and spontaneous non molar abortion by immunocytochemical and molecular methods. MATERIALS AND METHODS: The immunocytochemical method used specific markers for trophoblastic cells, whereas RT-PCR was used to identify insulin-like growth factor gene expression. RESULTS: Histochemical staining with hematoxilin-eosin revealed that the cultures contained heterogeneous cell types, including trophoblast and endometrial decidual cells. The ratio of trophoblast cells in the cultures varied between 16% and 37%, as detected by cytokeratine-7 as the specific trophoblast marker. Gene expression analysis corroborated the presence of trophoblasts by detecting insulin-like growth factor II mRNA, whereas GH-V transcripts were correlated with the presence of syncitiotrophoblasts. Insulin-like growth factor I and insulin-like growth factor binding protein 1 mRNAs were related to mesenchyimal and decidual cells, respectively. Higher insulin-like growth factor II expression levels were found in molar tissues in comparison with non-molar abortions. CONCLUSION: By combining three methodologies-morphology, immunocytochemistry and gene expression, characterization and follow-up of placenta cultures from abnormal tissues is found to facilitate diagnosis.