Implementación de un modelo in vitro de cáncer que permita evaluar simultáneamente el efecto de estrés (hipoxia y pH) y el silenciamiento de genes.

Cáncer es un término que se usa para definir enfermedades en las que las células se dividen anormalmente sin control y pueden invadir otros tejidos (Courtnay et al., 2015). Actualmente el cáncer sigue siendo una de las principales causas de muerte, a pesar de los avances tecnológicos y los nuevos abordajes para identificar nuevos enfoques terapéuticos (Huber et al., 2010). Estudios recientes demuestran que tanto los factores genéticos (mutaciones en oncogenes y genes supresores de tumor) y factores del microambiente (hipoxia y acidosis) pueden regular el metabolismo glicolítico de las células cancerosas (Justus, Sanderlin, & Yang, 2015). Las células cancerosas presentan una capacidad mejorada de la glucólisis en ausencia o incluso en presencia de oxígeno para apoyar el metabolism

Comparison of Risk Factors for Developing Liver Fibrosis in Subjects With and Without Metabolic Syndrome: A Cohort Study.

Background: Metabolic syndrome (MS), a combination of diabetes, high blood pressure and obesity, is a well-known risk factor for developing non-alcoholic fatty liver disease, condition that can lead to serious liver damage such as liver fibrosis (LF), which is characterized by excessive deposition of connective tissue, progressing to cirrhosis and hepatocellular carcinoma. Nevertheless, subjects without MS may also develop LF. Non-invasive LF predictors based upon anthropometric and biochemical data have been reported. Aim: To compare anthropometric, genetic, and biochemical parameters in subjects with or without MS, and at risk for developing liver fibrosis. Methods: A randomized sample of 200 individuals was taken from the 2015 Nuevo León State Health Survey. Inclusion criteria were age ≥18 and a previously stored blood sample. According to the parameters obtained, subjects were classified as either with or without MS and their NAFLD fibrosis score was calculated considering variables such as age, BMI, glycemia, albumin, platelets, and AST/ALT ratio, to establish a high or low risk of LF. Comparisons of weight, age, BMI, blood glucose, total cholesterol, triglycerides, platelets, albumin, AST/ALT ratio, and HDL were made between groups. DNA was extracted from stored blood samples and genotyped, using q-PCR, according to variants in four genes related to: fatty acid (FA) metabolism (PNPLA3, rs738409), adipocyte differentiation (PLIN2, rs35568725), glucose metabolism (GCKR, rs1260326 and rs780094), and BMI (UCP2, rs659366). Statistical analysis was performed with SPSS v.22. A p value <0.05 was taken as level of significance. Results: A total of 134 subjects were included and divided into four groups (n): With MS+ high risk (35), With MS+ low risk (34), Without MS+ high risk (32), Without MS+ low risk (33). Table 1 shows the main significative findings. Higher age, low platelet count, and increased AST/ALT ratio, were significantly different in high risk subjects, independently of the presence of MS. No association between the polymorphisms and risk for fibrosis was found. In subjects at high risk for LF, statistical significance was found for high cholesterol blood levels (OR= 20.0 (95%CI 2.87;139.38) in carriers of the T allele of GCKR rs780094 polymorphism. Conclusion: Aging, thrombocytopenia, and increased transaminases, the last two indicators of liver disfunction, were found as important risk factors for LF in subjects without metabolic syndrome. None of the genetic variants analyzed resulted associated to risk of LF, although sample size could be a factor. GCKR rs780094 variant was found related with risk for hypercholesterolemia, even though dyslipidemia was not found associated with risk of LF in the present study. &nbsp

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

Evaluación del uso de microarreglos genómicos como herramienta de apoyo para determinar el pronóstico en pacientes con LMA.

Propósito y Método del Estudio: La leucemia mieloide aguda (LMA) es un trastorno clonal causado por la transformación maligna de las células derivadas de la medula ósea. La LMA representa un grupo heterogéneo de desordenes con anormalidades genéticas variables y de respuesta variable al tratamiento.1 Ciertos cambios cromosómicos estructurales (e.g. presencia de las traslocaciones 8;21 y 15;17) y numéricos (presencia de monosomías 5 y 7) se han asociado a un pronóstico favorable o desfavorable, respectivamente. De la misma manera, mutaciones en los genes FLT3,&NPM1& y& C/EBPα& han de mostrado tener una gran importancia pronostica.3,4 No obstante en ausencia de los marcadores comunes de pronóstico desfavorable en pacientes con un fenotipo clínico agresivo, se requiere de la identificación de marcadores alternativos para explicar dicho fenotipo clínico. Actualmente, abordajes genómicos más sensibles con microarreglos de hibridación genómica comparativa (aCGH) son usados para identificar cambios genéticos asociados a la patogénesis del cáncer. Eventualmente, ciertos de estos cambios podrían incluir genes asociados con un pronóstico favorable o desfavorable para los pacientes y permitirá incluir a los aCGH como un método pronóstico en nuestra población. En este estudio se implementó el uso de microarreglos genómicos en 31 pacientes con LMA para determinar el pronóstico en pacientes con LMA. Contribuciones y Conclusiones: Se diseñó, desarrolló e implementó una metodología para la detección de 8 mutaciones de 4 genes diferentes. Se obtuvo la frecuencia de las mutaciones tamizadas; En el análisis de aCGH se encontraron principalmente regiones deletadas en todos los pacientes analizados; en 11 pacientes no se encontró ninguna de las mutaciones tamizadas mientras que en el microarreglo se encontraron regiones deletadas que incluían genes relacionados a cáncer, aberraciones cromosómicas, malignidades hematopoyéticas y LMA; los cromosomas que presentaron más anomalías fueron el 5 y 7. Se determinaron las regiones mínimas comunes que comparten 14 de nuestros pacientes en los cuales se encuentran genes que participan directa o indirectamente en cáncer, vías de señalización como MAPK,JAKXSTAT,p53,proliferación,supervivencia y apoptosis y LMA. Se realizó un análisis personalizado para cada uno de los pacientes con lo que observamos cambios significativos importantes lo que podría explicar la evolución favorable/desfavorable de la LMA

Molecular Characterization of Associated Pathogens in Febrile Patients during Inter-Epidemic Periods of Urban Arboviral Diseases in Tapachula Southern Mexico

Emerging and re-emerging vector-borne infections are a global public health threat. In endemic regions, fever is the main reason for medical attention, and the etiological agent of such fever is not usually identified. In this study, non-specific febrile pathogens were molecularly characterized in serum samples from 253 patients suspected of arbovirus infection. The samples were collected in the southern border region of Mexico from April to June 2015, and February to March 2016. ZIKV, CHIKV, DENV, leptospirosis, and rickettsiosis were detected by qPCR and nested PCR to identify flavivirus and alphavirus genera. The results indicated that 71.93% of the samples were positive for CHIKV, 0.79% for ZIKV, and 0.39% for DENV, with the number positive for CHIKV increasing to 76.67% and those positive for ZIKV increasing to 15.41% under the nested PCR technique. Leptospira Kmetyi was identified for the first time in Mexico, with a prevalence of 3.16%. This is the first report of ZIKV in Mexico, as well the first detection of the virus in early 2015. In conclusion, the etiological agent of fever was determined in 94% of the analyzed samples

Maternal Folic Acid Intake and Methylation Status of Genes Associated with Ventricular Septal Defects in Children: Case–Control Study

Background: DNA methylation is the best epigenetic mechanism for explaining the interactions between nutrients and genes involved in intrauterine growth and development programming. A possible contributor of methylation abnormalities to congenital heart disease is the folate methylation regulatory pathway; however, the mechanisms and methylation patterns of VSD-associated genes are not fully understood. Objective: To determine if maternal dietary intake of folic acid (FA) is related to the methylation status (MS) of VSD-associated genes (AXIN1, MTHFR, TBX1, and TBX20). Methods: Prospective case–control study; 48 mothers and their children were evaluated. The mothers’ dietary variables were collected through a food frequency questionnaire focusing on FA and the consumption of supplements with FA. The MS of promoters of genes was determined in the children. Results: The intake of FA supplements was significantly higher in the control mothers. In terms of maternal folic acid consumption, significant differences were found in the first trimester of pregnancy. Significant differences were observed in the MS of MTHFR and AXIN1 genes in VSD and control children. A correlation between maternal FA supplementation and MS of AXIN1 and TBX20 genes was found in control and VSD children, respectively. Conclusions: A lower MS of AXIN1 genes and a higher MS of TBX20 genes is associated with FA maternal supplementation

Tyrphostin AG17 inhibits adipocyte differentiation in vivo and in vitro

Abstract Background Excessive subcutaneous adiposity in obesity is associated to positive white adipocyte tissue (WAT) differentiation (adipogenesis) and WAT expandability. Here, we hypothesized that supplementation with the insulin inhibitor and mitochondrial uncoupler, Tyrphostin (T-AG17), in vitro and in vivo inhibits adipogenesis and adipocyte hypertrophy. Methods We used a 3T3-L1 proadipocyte cell line to identify the potential effect of T-AG17 on adipocyte differentiation and fat accumulation in vitro. We evaluated the safety of T-AG17 and its effects on physiological and molecular metabolic parameters including hormonal profile, glucose levels, adipogenesis and adipocyte hypertrophy in a diet-induced obesity model using C57BL/6 mice. Results We found that T-AG17 is effective in preventing adipogenesis and lipid synthesis in the 3T3-L1 cell line, as evidenced by a significant decrease in oil red staining (p < 0.05). In obese C57BL/6 mice, oral administration of T-AG17 (0.175 mg/kg for 2 weeks) lead to decreased fat accumulation and WAT hypertrophy. Further, T-AG17 induced adipocyte apoptosis by activating caspase-3. In the hepatocytes of obese mice, T-AG17 promoted an increase in the size of lipid inclusions, which was accompanied by glycogen accumulation. T-AG17 did not alter serum biochemistry, including glucose, insulin, leptin, free fatty acids, creatinine, and aspartate aminotransferase. Conclusion T-AG17 promotes adipocyte apoptosis in vivo and is an effective modulator of adipocyte differentiation and WAT hypertrophy in vitro and in vivo. Therefore, T-AG17 may be useful as a pharmacological obesity treatment