The discovery and comparative expression analysis of three distinct type I interferons in the perciform fish, meagre (Argyrosomus regius).

Type I interferons (IFN) play an important role in anti-viral responses. In teleost fish multiple genes exist, that are classified by group/subgroup. That multiple subgroups are present in Acanthopterygian fish has only become apparent recently, and 3 subgroups are now known to be expressed, including a new subgroup termed IFNh. However, the potential to express multiple IFN subgroups and their interplay is not well defined. Hence this study aims to clarify the situation and undertook the first in-depth analysis into the nature and expression of IFNc, IFNd and IFNh in the perciform fish, meagre. Constitutive expression was analysed initially during larval development and in adult tissues (gills, mid-gut, head kidney, spleen). During early ontogeny IFNc was the highest expressed IFN, and this was also the case in adult tissues with the exception of gills where IFNd was highest. However, comparison between tissues for individual isoforms showed that spleen had high transcript levels of all three IFNs, IFNd/IFNh were also highly expressed in gills. The expression of each sub-group was increased significantly in the four tissues following injection of poly I:C, however, this increase was only seen in the mid-gut for IFNh. Following in vitro stimulation with poly I:C again all three isoforms were upregulated, although with differences in kinetics and the cell source used. For example, early induction was seen for IFNc/IFNh in gill cells, IFNd/IFNh in splenocytes and all three isoforms in head kidney cells. Induction was sustained in splenocytes and head kidney cells, but in gut cells only a late induction was seen. These results demonstrate a complex pattern of regulation between the different IFN isoforms present in meagre and highlights potential sub-functionalisation of these IFN subgroups during perciform anti-viral responses.

Two types of TNFα in meagre (Argyrosomus regius): Discovery, distribution and expression modulation.

TNFα is a key cytokine involved in systemic inflammation and regulation of immune cells and is important during development. In the present study, 2 isoforms of TNFα were discovered in meagre, an emerging species in aquaculture. Phylogenetic analysis suggests these isoforms represent the type I and type II TNFα classes previously described in other teleost species. This study is the first to compare how these 2 types of TNFα behave in meagre and aims to provide insights into their expression in teleost fish by interrogating expression in whole tissues and isolated cell populations in four immunologically important sites (gills, intestine, head kidney and spleen) following PAMP stimulation, as well as monitoring gene expression during meagre development. Differential expression was seen in head kidney and gills, where TNFα1 was more highly expressed. Both isoforms increased in head kidney of meagre following injection with LPS, but this was not seen in other tissues or after injection with other PAMPs. However, in vitro studies hinted at a possible mucosal bias for TNFα1, which was more highly induced in gill and intestinal cell suspensions by PAMPs. In contrast TNFα2 was more highly induced in cells from systemic tissues. Through early development expression of both types of TNFα decreased as the meagre matured, with the exception of a transient increase shortly after the move to a dry feed diet. However, during the later stages of development expression of both isoforms increased in the gills. This data demonstrates a degree of differential expression of TNFα1 and TNFα2 in meagre with regard to expression regulation, and highlights the importance of TNFα during early development of teleost fish.

Prevalencia de genotipos del papiloma virus en mujeres de Cuenca / Prevalence of genotypes of papilloma virus in women in Cuenca

Objetivos: Determinar la prevalencia de los genotipos del papiloma virus de alto y bajo grado oncogénico; relacionar con los grupos de edad, con los genotipos que evitan las dos vacunas existentes y con los fatores de riesgo para cáncer cervical uterino. Metodología: Estudio epidemiológico, observacional, transversal. La muestra aleatorizada fue de 500 mujeres de la zona urbana de la ciudad de Cuenca. Se efecturaon dos tomas del cuello uterino. Se utilizó el método de reacción de cadena de polimerasa y el estudio citopatológico Papanicolaou. Resultados: La prevalencia de los genotipos del papiloma virus de alto grado oncogénico fue del 35.9% y para bajo grado el 14.3%. La prevalencia de las alteraciones citológicas, Papanicolaou, fue del 16%. Estas prevalencias fueron más frecuentes en los grupos de 30 a 39 años y 40 a 50 años. Por inferencia, la cobertura de la vacuna Cervarix sería del 8% y para el Gardasil del 13%. Fueron factores de riesgo los relacionados con el número y frecuencia de relaciones sexuales, número de compañeros sexuales. Conclusiones. La prevalencia de los genotipos fue alta y la cobertura de las vacunas baja.

OBJECTIVES. To determine the prevalence of papilloma virus genotypes of high and low oncogenic grade, to relate to age groups, with genotypes that avoid both existing vaccines and the risk factors for uterine cervical cancer. METHODS. Epidemiological, observational, cross-sectional study. The random sample was 500 women from the urban area of Cuenca city. Two samples of the cervix were taken. Polimeraza chain reaction method and cytopathology Papanicolaou study were used. RESULTS. The prevalence of papilloma virus genotypes of high oncogenic grade was 35,9% and 14,3% low oncogenic grade. The prevalence of cytologic abnormalities, Papanicolaou, was 16%. This prevalence was more frequent in the group of 30-39 years, and 40-50 years. By inference, the coverage of the Cervarix vaccine would be 8% and 13% for Gardasil. Risk factors were related to the number and frequency of intercourse and number of sexual partners. CONCLUSION. The prevalence of genotypes was high and the vaccine coverage was low.

Antígenos HLA de clase ii en pacientes con artritis reumatoide ecuatorianos y asociación con el factor reumatoide, anticuerpos anti-citrulinados y nódulos reumatoides / HLA antigens class ii in patients with ecuadorian rheumatoid arthritis and association with the rheumatoid factor, anti-citrulinated antibodies and rheumatoid nodules

Objetivo Determinar los antígenos de Clase II en 52 pacientes con artritis reumatoide (AR) y 52 controles, y establecer la asociación de los antígenos de Clase II DRB1* con el factor reumatoide (FR), anticuerpos anti-péptido cíclico citrulinados (anti-CCP) y nódulos reumatoides. Material y Métodos Estudio de casos y controles, realizado en pacientes del Hospital Vicente Corral Moscoso y José Carrasco Arteaga de la ciudad de Cuenca-Ecuador; se incluyeron 52 pacientes con AR que cumplen los criterios del Colegio Americano de Reumatología (ACR) y 52 controles. Se identificaron a través de la Reacción en Cadena de la Polimerasa (PCR), los diferentes tipos de HLA que tienen cada uno de los 52 pacientes con AR y controles. Se buscó asociación de los HLA-DR1* con la AR y las pruebas de laboratorio: FR, anticuerpos anti-CCP y con nódulos reumatoides. El análisis estadístico se realizó con el cálculo de OR (IC 95%) y 2 de Pearson con corrección de Fisher y la información se procesó en SPSS v15. Resultados Los alelos más frecuentes fueron HLA-DRB1*09 (17,3%) OR 3,42 (IC95% 1,21 ­ 10,14) y HLADRB1*04 (32,7%) OR 1,81 (IC95% 0,93 ­ 3,55). Tanto el factor reumatoide como los anticuerpos anti-péptido cíclico citrulinados estuvieron presentes en el 82,7% de pacientes con artritis reumatoide y los nódulos reumatoides se detectaron en el 19,2% de pacientes con AR. Se encontró asociación entre los nódulos reumatoides con valores altos de FR y anti-CCP y estos a su vez con los alelos HLA-DRB1*14 y HLADRB1*04. La mayoría de pacientes con alelos HLA-DRB1*09 con predominio heterocigoto tuvieron anticuerpos anti-CCP y FR positivo. Conclusión La presencia de los alelos HLA-DRB1*04 y HLADRB1*09 está relacionada con la susceptibilidad de presentar AR en pacientes ecuatorianos, además se relaciona con valores elevados de factor reumatoide y anticuerpos anti-péptido cíclico citrulinados.

Objective To determine Class II antigens in 52 patients with rheumatoid arthritis (RA) and 52 controls and establish the association of Class II antigens DRB1* with rheumatoid anti- cyclic citrullinated peptide (anti-CCP) rheumatoid factor (RF), and antibodies rheumatoid nodules. Methods and Materials Case and control study in patients from the Hospital Vicente Corral Moscoso and José Carrasco Arteaga of Cuenca-Ecuador; 52 patients with rheumatoid arthritis who satisfy the criteria of ACR-American College of Rheumatology and 52 healthy patients. The different types of HLA from each 52 patients with rheumatoid arthritis and controls were identified through PCR - Polymerase Chain Reaction. Association of HLA-DR1* with rheumatoid arthritis and rheumatoid factor, anti-citrullinated and rheumatoid nodules. The statistical analysis was done by the OR (IC 95%) and 2 of Pearson with Fisher correction and the information was processed in SPSS v15. Results The most frequency alleles were HLA-DRB1*09 (17,3%) OR 3,42 (IC95% 1,21 ­ 10,14) and HLADRB1*04 (32,7%) OR 1,81 (IC95% 0,93 ­ 3,55). The rheumatoid factor and anti-cyclic citrullinated peptide antibodies were presented in 82,7% of patients with rheumatoid arthritis and rheumatoid nodules was detected in the 19.2% of patients with rheumatoid arthritis . An association between rheumatoid nodules with high values of rheumatoid factor and anti-ccp and these in turn with HLA-DRB1*14 y HLA-DRB1*04 alleles were found. The majority of patients with HLA-DRB1*09 alleles with heterozygote predomination had positive anti-ccp and rheumatoid factor. Conclusion The presence of HLA-DRB1*04 and HLA-DRB1*09 alleles is related with the susceptibility of present rheumatoid arthritis in Ecuadorian patients, and it is related with higher values of rheumatoid factor and anti-cyclic citrullinated peptide.

Correlación del recuento de cuerpos laminares con el índice de lecitina/esfingomielina y/o la presencia de fosfadilglicerol para la determinación de madurez pulmonar fetal / Correlation of the lamellar bodies count with lecithin/sphingomyelin ratio and/or the presence of phosphadylglicerol for determination of fetal lung maturity

El objetivo del estudio fue determinar si el recuento de Cuerpos Laminares mayor a 30.000 por uL se correlaciona con pruebas "Gold Standard" cuando estas indican que existe madurez pulmonar fetal (38, 39, 41). Se correlacionó el recuento obtenido en 22 muestras de líquido amniótico de amniocentesis realizadas en el primer semestre del año 2.000, con 2 pruebas de referencia, como son el índice de leticina/Esfingomielina > 2 (19, 22) y/o la presencia de fosfalidilglicerol (20, 26), El recuento de cuerpos laminares se efectuó en un contador de plaquetas Celldyn 1400 de Abbott y las muestras debieron cumplir con los requisitos de nuestro protocolo, los cuales fueron: embarazos con edad gestacional entre 32 a 36 semanas que requirieron amniocentesis por alguna indicación médica y embarazadas con diabetes insulino requirientes de 32 a 40 semanas a quienes se las realizó amniocentesis antes de su parto; además que las muestras no estuvieran contaminadas con sangre ni meconio. Los resultados demostraron que un recuento mayor a 30.000 cuerpos laminares por uL, (12 pacientes), se correlacionaron en un 100 por ciento con una o ambas pruebas de referencia cuando estas indican madurez pulmonar fetal. El recuento de cuerpos laminares es de muy bajo costo (33, 34, 41) y técnicamente accesible en los servicios públicos ya que solo se requiere de un equipo para el recuento de plaquetas, pudiendo ser utilizada para determinar madurez pulmonar fetal cuando otras pruebas clásicas como el Test de Clements dan resultado negativo (porcentaje de falsos inmaduros elevado)