Diagnóstico molecular y caracterización genética de patógenos transmitidos por artrópodos vectores, en pacientes con síndrome febril en la región fronteriza del sur de México

Las enfermedades producidas por arbovirus son un problema de salud a nivel mundial, el 17 % de enfermedades infecciosas en el mundo provienen de transmisión por vectores. En regiones endémicas, la fiebre es la razón principal para que las personas busquen atención médica, dada la presentación clínica no específica de diversas infecciones, pocos pacientes reciben un diagnóstico preciso y específico, y el resto sigue siendo desconocido. El objetivo de este trabajo fue explorar la epidemiología y la utilidad de herramientas diagnósticas en pacientes con cuadros febriles, en una región del sur oeste mexicano. Se recolectaron 253 sueros de pacientes con cuadro clínico sugerente de infección por arbovirus. Las manifestaciones clínicas presentadas en los pacientes destacaron fiebre, cefalea y escalofríos; en dos periodos comprendido de abril-junio del 2015 y febrero-marzo del 2016. Las muestras fueron evaluadas por qPCR sonda específico para ZIKV, CHIKV y DENV. Con respecto a las muestras negativas en el primer tamizaje (67 muestras) se les realizo una segunda prueba, PCR-anidada específica para los géneros flavivirus y alfavirus, Las muestras positivas (44 muestras) fueron confirmados por secuenciación. Las muestras negativas restantes (23 muestras) se les determino la presencia de leptospirosis y rickettsiosis por medio de qPCR con SYBR Green, el producto de PCR obtenido de los casos positivos (8 muestras) fueron confirmados por secuenciación. El ensayo por qPCR sonda específica logro detectar el 70.75% casos positivos para CHIKV, 2.37% para ZIKV y 0.39% en DENV. El poder de detección por PCR anidada se vio aumentada para ZIKV y CHIKV, 75.49% y 15.01%, respectivamente. Se encontraron 8 muestras positivas para Leptospira kmetyi en las muestras negativas por qPCR sonda específica y PCR anidada para alfavirus y flavivirus. Se logró diagnóstico en el 94% de los casos totales con la combinación de diferentes técnicas diagnósticas. La introducción de una técnica alternativa aumento el poder de detección en este estudio. Se reporta por primera vez en la región fronteriza de México la presencia de L. kmetyi.

Expression profile of microRNAs in the testes of patients with Klinefelter syndrome

Klinefelter syndrome (KS) is the most common sex chromosome aneuploidy. A distinctive characteristic of KS is oligozoospermia. Despite multiple studies that have described the natural history of the degenerative process of germ cells in patients with KS, the molecular mechanisms that initiate this process are not well characterized. MicroRNA (miRNA)-mediated post-transcriptional control mechanisms have been increasingly recognized as important regulators of spermatogenesis; however, only a few studies have evaluated the role of miRNAs in the gonadal failure of these patients. Here, we describe a differential expression profile for the miRNAs in testicular tissue samples taken from KS patients. We analysed testicular tissue samples from 4 KS patients and 5 control patients (obstructive azoospermia) through next-generation sequencing, which can provide information about the mechanisms involved in the degeneration of germ cells. A distinctive differential expression profile was identified for 166 miRNAs in the KS patients: 66 were upregulated, and 100 were downregulated. An interactome analysis was performed for 7 of the upregulated and the 20 downregulated miRNAs. The results showed that the target genes are involved in the development, proliferation, and differentiation processes of spermatogenesis, which may explain their role in the development of infertility. This is the first report of a miRNA expression profile generated from testicular tissue samples of KS patients

Comparison of specific expression profile in two in�vitro hypoxia models

Abstract. The microenvironment plays a fundamental role in carcinogenesis: Acidity and hypoxia are actively involved in this process. It is important to have in vitro models to study these mechanisms. The models that are most commonly referred to are the hypoxia chamber and the chemical induction [Cobalt (II) chloride]. It is not yet defined if these models are interchangeable if the metabolic effect is the same, and if the results may be compared in these models. In the present study, the response to the effect of stress (hypoxia and acidity) in both models was evaluated. The results indicated that in the chemical model, the effect of hypoxia appeared in an early form at 6 h; whereas in the gas chamber the effect was slow and gradual and at 72 h there was an overexpression of erythropoietin (EPO), vascular endothelial growth factor (VEGF), carbonic anhydrase 9 (CA9) and hypoxia-inducible factor 1α (HIF1α). In addition to the genes analyzed by reverse transcription-quantitative polymerase chain reaction, the global expression analysis between both models revealed the 9 most affected genes in common. The present study additionally identified 3 potential genes (lysyl oxidase, ankyrin repeat domain 37, B-cell lymphoma 2 interacting protein 3 like) previously identified in other studies, which may be considered as universal hypoxia genes along with HIF1α, EPO, VEGF, glucose transporter 1 (GLUT1), CA9, and LDH. To the best of the author's knowledge, this is the first time that both hypoxia models have been compared, and it was demonstrated that the effect of hypoxia induction was time sensitive in each model. These observations must be considered prior to selecting one of these models to identify selective hypoxia genes and their effects in cancer

Gene Copy Number Quantification of SHOX , VAMP7 , and SRY for the Detection of Sex Chromosome Aneuploidies in Neonates

Aims: To explore the feasibility of detecting sex chromosome aneuploidies (SCAs) by means of gene copy number quantification of short stature homeobox (SHOX), vesicle-associated membrane protein 7 (VAMP7), and SRY in newborns. Materials and Methods: Gene doses of SHOX, VAMP7, and SRY were determined by quantitative polymerase chain reaction (qPCR) using DNA obtained from dried blood samples from newborns. Relative quantification values were obtained. An aneuploidy profile was established according to cutoff values. Samples with ≥2 gene doses (out of range) were reanalyzed, and those with aneuploidy profiles were confirmed by karyotyping. Sensitivity, specificity, and positive and negative predictive values were obtained. Results: A total of 10,033 samples were collected (4945 females and 5088 males). Of 244 (2.43%) samples with ≥2 gene doses that were retested, 20 cases were confirmed. The overall incidence of SCAs was 1 in 500 live newborns. There were six cases of Turner syndrome (1/824), 3 cases of XXX (1/1648), 7 cases of Klinefelter syndrome (1/726), and 4 cases of of XYY (1/1272). The sensitivity was 0.952 (95.24%), specificity of 0.975 (97.56%), positive predictive value of 0.909 (90.91%), and negative predictive value of 0.987 (98.77%). Conclusions: Gene copy number analyses of VAMP7, SHOX, and SRY genes by qPCR from blood samples spotted onto filter paper is a highly reliable method for the early detection of male and female SCAs

Rothmund-Thomson Syndrome: A 13-Year Follow-Up

Rothmund-Thomson syndrome (RTS) is a rare autosomal recessive disorder presenting with poikiloderma and other clinical features, affecting the bones and eyes and, in type II RTS, presenting an increased risk for malignancy. With about 300 cases reported so far, we present a 13-year follow-up including clinical images, X-rays and genetic analysis. A 13-month-old female started with a facial rash with blisters on her cheeks and limbs at the age of 3 months along with congenital hypoplastic thumbs, frontal bossing and fine hair, eyebrows and eyelashes. The patient was lost to follow-up and returned 12 years later with palmoplantar hyperkeratotic lesions, short stature, disseminated poikiloderma and sparse scalp hair, with absence of eyelashes and eyebrows. Radiographic analysis showed radial ray defect, absence of the thumb and three wrist carpal bones, and reduced bone density. Gene sequencing for the RECQL4 helicase gene revealed a mutation on each allele. RTS is a rare disease, and in this patient we observed the evolution of her skin lesions and other clinical features, which were important for the classification of type II RTS. The next years will provide even more information on this rare disease