Estudio molecular de la familia GH

Tesis (Maestría en Ciencias con Especialidad en Biología Molecular e Ingeniería Genética) UANLUANLhttp://www.uanl.mx

Estudio de la etiología de meningitis aséptica y meningoencefalitis en el Hospital Universitario Dr. José Eleuterio González, mediante estrategias de virología molecular

Antecedentes: La meningitis aséptica y la meningoencefalitis constituyen problemas de gran importancia clínica. Sus agentes etiológicos más frecuentes son los enterovirus (ECHOvirus, Coxsackievirus, Enterovirus no-polio), Togavirus, Flavivirus, Bunyavirus y otros grupos antes conocidos como ARBOvirus (Arthopod-borne viruses) y el virus del herpes humano. Objetivo: En este trabajo nos propusimos identificar, mediante la reacción en cadena de la polimerasa, los genomas virales de los virus del herpes simple (VHS tipos 1 y 2), del enterovirus y del virus del oeste del Nilo (VON) como causantes de meningitis aséptica y de meningoencefalitis. Material y métodos: Se estudiaron 30 pacientes que ingresaron al Hospital Universitario Dr. José Eleuterio González durante el período de febrero de 2005 a junio de 2008. Se analizaron muestras de líquido cefalorraquídeo (2 mL) para buscar, mediante la reacción en cadena de la polimerasa, el genoma del enterovirus, del VHS tipos 1 y 2, y del virus del oeste del Nilo, siguiendo protocolos estandarizados. Resultados: Ninguno de los pacientes estudiados fue positivo para los virus buscados mediante amplificación por la reacción en cadena de la polimerasa. Entre los datos clínicos relevantes encontramos que la presencia de estupor o el coma se relacionó significativamente con la muerte (p = 0.005). Encontramos, además, que tres muertes ocurrieron en el grupo de pacientes con meningoencefalitis (p = 0.021). Conclusiones: No encontramos el genoma de los virus analizados en el líquido cefalorraquídeo de los pacientes estudiados, probablemente debido al tamaño limitado de la muestra, a su calidad, o a diversos factores epidemiológicos en nuestra región. La información clínica y del laboratorio de los pacientes estudiados es similar a la que está publicada en estudios anteriores. Es necesario realizar estudios más amplios, incluyendo a un mayor número de pacientes que nos permitan conocer la etiología de este tipo de enfermedades en nuestra comunidad. Esto hará posible mejorar la metodología para el diagnóstico en los pacientes con patología del sistema nervioso central

Presencia del virus del oeste del Nilo en el noreste de México

Objetivo. Detectar la presencia del virus del oeste del Nilo (VON) en aves, equinos y seres humanos en el noreste de México. Material y métodos. Se buscó en diferentes localidades del noreste de México la presencia de anticuerpos antivirus del oeste del Nilo (anti-VON) en suero de 33 aves, 24 caballos y 237 personas mediante pruebas de ELISA durante el periodo de julio de 2003 a julio de 2006. En los sueros humanos se buscó también el RNA-VON mediante RT-PCR. Resultados. Se encontraron tres aves seropositivas y 15 equinos. En el hombre, 40% de los sueros fue positivo para anticuerpos IgG y ninguno para anticuerpos IgM. Conclusiones. El VON se encuentra activo en México y se suma a otras enfermedades emergentes transmitidas por vectores que representan un reto a la investigación y a los programas de prevención

Biobanks: Experience of the School of Medicine and the “Dr. José Eleuterio González” University Hospital of the Universidad Autónoma de Nuevo León

Medical research has greatly beneited from molecular biology and increasingly relies on tools from the “omics” disciplines (mainly genomics, transcriptomics, proteomics and metabolomics). The availability of biological samples preserved with high quality standards is a sine qua non condition for such studies and their repositories are referred to as biobanks. Biobanks support the transportation, storage, preservation, and initial pathological and analytical examinations of biospecimens, as well as the protection of relevant information and the comparison of clinical and laboratory findings. A biobank facility is one of the most valuable tools the academic medicine organizations can offer to their researchers to improve the competitiveness of their current and future medical research. it acts as an essential bridge and an effective catalyst for research synergies between basic and clinical sciences, and it can be potentiated with efforts to raise funds for acquiring and maintaining cutting-edge analytical infrastructure to better serve its clinical, pharmaceutical and biotech clients

Identification of a Novel Pathogenic Rearrangement Variant of the APC Gene Associated with a Variable Spectrum of Familial Cancer

Familial adenomatous polyposis (FAP) is an autosomal-dominant condition characterized by the presence of multiple colorectal adenomas, caused by germline variants in the adenomatous polyposis coli (APC) gene. More than 300 germline variants have been characterized. The detection of novel variants is important to understand the mechanisms of pathophysiology. We identified a novel pathogenic germline variant using next-generation sequencing (NGS) in a proband patient. The variant is a complex rearrangement (c.422+1123_532-577 del ins 423-1933_423-1687 inv) that generates a complete deletion of exon 5 of the APC gene. To study the variant in other family members, we designed an endpoint PCR method followed by Sanger sequencing. The variant was identified in the proband patient’s mother, one daughter, her brother, two cousins, a niece, and a second nephew. In patients where the variant was identified, we found atypical clinical symptoms, including mandibular, ovarian, breast, pancreatic, and gastric cancer. Genetic counseling and cancer prevention strategies were provided for the family. According to the American College of Medical Genetics (ACMG) guidelines, this novel variant is considered a PVS1 variant (very strong evidence of pathogenicity), and it can be useful in association with clinical data for early surveillance and suitable treatment. View Full-Tex

Molecular cloning of the myo-inositol oxygenase gene from the kidney of baboons

Abstract. The enzyme myo-Inositol oxygenase (MIOX) is also termed ALDRL6. It is a kidney‑specific member of the aldo‑keto reductase family. MIOX catalyzes the first reaction involved in the myo‑inositol metabolism signaling pathway and is fully expressed in mammalian tissues. MIOX catalyzes the oxidative cleavage of myo‑Inositol and its epimer, D-chiro-Inositol to D-glucuronate. The dioxygen-dependent cleavage of the C6 and C1 bond in myo‑Inositol is achieved by utilizing the Fe2+/Fe3+ binuclear iron center of MIOX. This enzyme has also been implicated in the complications of diabetes, including diabetic nephropathy. The MIOX gene was amplified with reverse transcription‑polymerase chain reaction from baboon tissue samples, and the product was cloned and sequenced. MIOX expression in the baboon kidney is described in the present study. The percentages of nucleotide and amino acid similarities between baboons and humans were 95 and 96%, respectively. The MIOX protein of the baboon may be structurally identical to that of humans. Furthermore, the evolutionary changes, which have affected these sequences, have resulted from purifying forces. Key words: animal models, gene expression, kidney, myo-inositol oxygenase, Old World monke

Epidemiological Algorithm and Early Molecular Testing to Prevent COVID-19 Outbreaks in a Mexican Oncologic Center

Introduction: Prevention strategies and detection of latent COVID-19 infections in oncology staff and oncologic patients are essential to prevent outbreaks in a cancer center. In this study, we used two statistical predictive models in oncology staff and patients from the radiotherapy area to prevent outbreaks and detect COVID-19 cases. Methods: Staff and patients answered a questionnaire (electronic and paper surveys, respectively) with clinical and epidemiological information. The data was collected through two online survey tools: Real-Time Tracking (R-Track) and Summary of Factors (S-Facts). According to the algorithm\u27s models, cut-off values were established. SARS-CoV-2 qRT-PCR tests confirmed the algorithm\u27s positive individuals. Results: Oncology staff members (n=142) were tested, and 14% (n=20) were positives for the R-Track algorithm; 75% (n=15) were qRT-PCR positive. The S-Facts algorithm identified 7.75% (n=11) positive oncology staff members, and 81.82% (n=9) were qRT-PCR positive. Oncology patients (n=369) were evaluated, and 1.36% (n=5) were positive for the algorithms. The 5 patients (100%) were confirmed by qRT-PCR at a very early stage. Conclusions: The proposed algorithms could prove to become an essential prevention tool in countries where qRT-PCR tests and vaccines are insufficient for the population

Risk Association of TOX3 and MMP7 Gene Polymorphisms with Sporadic Breast Cancer in Mexican Women

Breast cancer (BC) has one of the highest incidences and mortality worldwide. Single nucleotide polymorphisms (SNPs) in TOX3 rs3803662 and MMP7 rs1943779 have been associated with susceptibility to BC. In this case-control study, we evaluated the association of rs3803662 (TOX3)/rs1943779 (MMP7) SNPs with clinical features, immunohistochemical reactivity, and risk association with BC in women from northeastern Mexico. We compared 212 BC cases and 212 controls. DNA was isolated from peripheral blood to perform the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. We calculated genotype frequencies, odds ratios, and 95% confidence intervals. We found that CT (Cytocine-Thymine) and TT (Thymine -Thymine) genotypes, and T alleles of TOX3 rs3803662, were associated with BC risk (p = 0.034, p = 0.011, respectively). SNP TOX3 rs3803662 was associated with positive progesterone receptors (PR) and triple-negative BC (TNBC) but not with estrogen receptor (ER) or HER2 reactivity. CT and TT genotypes (p = 0.006) and T alleles (p = 0.002) of SNP MMP7 rs1943779 were associated with risk of BC. We found that T alleles of TOX3 rs3803662 and MMP7 rs1943779 SNPs are associated with BC risk. These findings contribute to personalized medicine in Mexican women

Olfactomedin‑like 2 A and B (OLFML2A and OLFML2B) expression profile in primates (human and baboon)

Background: The olfactomedin‑like domain (OLFML) is present in at least four families of proteins, including OLFML2A and OLFML2B, which are expressed in adult rat retina cells. However, no expression of their orthologous has ever been reported in human and baboon. Objective: The aim of this study was to investigate the expression of OLFML2A and OLFML2B in ocular tissues of baboons (Papio hamadryas) and humans, as a key to elucidate OLFML function in eye physiology. Methods: OLFML2A and OLFML2B cDNA detection in ocular tissues of these species was performed by RT‑PCR. The amplicons were cloned and sequenced, phylogenetically analyzed and their proteins products were confirmed by immunofluorescence assays. Results: OLFML2A and OLFML2B transcripts were found in human cornea, lens and retina and in baboon cornea, lens, iris and retina. The baboon OLFML2A and OLFML2B ORF sequences have 96% similarity with their human’s orthologous. OLFML2A and OLFML2B evolution fits the hypothesis of purifying selection. Phylogenetic analysis shows clear orthology in OLFML2A genes, while OLFML2B orthology is not clear. Conclusions: Expression of OLFML2A and OLFML2B in human and baboon ocular tissues, including their high simi‑ larity, make the baboon a powerful model to deduce the physiological and/or metabolic function of these proteins in the eye