Variantes genéticas en la región 3’UTR de los genes BRCA1 y BRCA2 y su reconocimiento por miARNs en cáncer de mama y ovario hereditario

El síndrome de cáncer de mama y ovario hereditario (HBOC) es una patología causada principalmente por mutaciones en los genes BRCA1 y BRCA2. La región 3'UTR de los genes permite la unión de la mayoría de los microARNs involucrados en la regulación genética fina. El objetivo de esta tesis doctoral fue identificar variantes alélicas en los sitios de unión de miARN en las regiones 3'UTR en los genes BRCA1 y BRCA2 en pacientes diagnosticados con HBOC. La estrategia experimental consistió en colectar muestras de sangre de 50 pacientes con HBOC y de 50 controles de una población del Noreste de México y obtener el ADN genómico. Las regiones 3'UTR de BRCA1 y BRCA2 se amplificaron por PCR punto final y se secuenciaron para identificar variantes genéticas utilizando herramientas de bioinformática. Los resultados mostraron nueve polimorfismos (SNPs): cuatro en BRCA1 (rs3092995 (C/G), rs8176318 (C/T), rs111791349 (G/A) y rs12516 (C/T)) y cinco en BRCA2 (rs15869 (A/C), rs7334543 (A/G), rs1157836 (A/G) y rs75353978 (TT/del TT). También se identificó una nueva variante en la posición c.* 457 (A / C) en 3'UTR de BRCA2. Las variantes rs111791349-A, rs15869-C y c.*457-C aumentaron el riesgo de HBOC en la población de estudio: (odds ratio (OR) rango 3.7- 15.4; p <0.05). Las variantes genéticas en el 3'UTR de BRCA1 y BRCA2 aumentaron el riesgo de HBOC entre 3.7-15.4 veces en la población con esta condición. La presencia/ausencia de estos SNPs puede influir en la pérdida / creación de sitios de unión de miARN, como hsa-miR-1248 en BRCA1 3'UTR o el sitio de unión de la familia hsamiR-548 en BRCA2. Estos resultados agregan nuevas pruebas de participación de miARN en la patogénesis de HBOC. En conclusión la presencia los SNPs rs8176318 (G/T), rs15869 (A/C) y la variante c.*457 (A/C) fueron significativas y están relacionadas a un aumento en el riesgo en el grupo de HBOC. La nueva variante descrita en la región 3’UTR de BRCA2, tanto en pacientes HBOC como en los controles, ubicada en sitio c.*457 posiblemente sea específica para población de la región del Noreste de México. Finalmente, el análisis in silico se postula la pérdida/creación de sitios de unión de miARN, como hsa-miR-1248 en BRCA1 3'UTR o el sitio de unión de la familia hsa-miR548 en BRCA2 que sugiere la participación de miARNs en la patogénesis del HBOC. Abstract Hereditary breast and ovarian cancer (HBOC) syndrome are mainly caused by mutations in the BRCA1 and BRCA2 genes. The 3’UTR region allows the binding of microRNAs, which are involved in genetic tune regulation. We aimed to identify allelic variants on 3’UTR miRNA-binding sites in the BRCA1 and BRCA2 genes in HBOC patients. Blood samples were obtained from 50 patients with HBOC and from 50 controls. The 3’UTR regions of BRCA1 and BRCA2 were amplified by PCR and sequenced to identify genetic variants using bioinformatics tools. We detected nine polymorphisms in 3’UTR, namely: four in BRCA1 (rs3092995 (C/G), rs8176318 (C/T), rs111791349 (G/A), and rs12516 (C/T)) and five in BRCA2 (rs15869 (A/C), rs7334543 (A/G), rs1157836 (A/G), and rs75353978 (TT/del TT)). A new variant in position c.*457 (A/C) on 3’UTR of BRCA2 was also identified. The following three variants increased the risk of HBOC in the study population: rs111791349-A, rs15869-C, and c.*457-C (odds ratio (OR) range 3.7–15.4; p < 0.05). Genetic variants into the 3’UTR of BRCA1 and BRCA2 increased the risk of HBOC between 3.7–15.4 times in the study population. The presence/absence of these polymorphisms may influence the loss/creation of miRNA binding sites, such as hsa-miR1248 in BRCA1 3′UTR or the hsa-miR-548 family binding site in BRCA2. Our results add new evidence of miRNA participation in the pathogenesis of HBOC

Aspergillus in liquid-based cervicovaginal cytology in a postmenopausal patient: A case report

Abstract. Aspergillus is an opportunistic fungus present in humid environments, whose natural environment is in soil, hay and compost. It is a frequent contaminant in the clinical laboratory. Because of this, the fungus is often inhaled, affecting those with an underlying pulmonary disease or immune deficiency. Fungal genitourinary tract infections are relatively common. A rare Aspergillus spp cervical infection diagnosed via liquid-based cytology is presented in the current study. The 57-year-old woman attended her annual check-up without any relevant medical history. The result of a gynecological examination by Papanicolaou smear was normal and routine liquid-based cytology was performed. The specimen exhibited fungal organisms characterized by septate hyphae branching at acute angles, most consistent with the Aspergillus species. Subsequent cytology demonstrated the same results. Antifungal treatment was initiated and a second post-treatment smear only exhibited atrophy. The cytomorphological features of Aspergillus spp. are discussed in the current study and a brief review of the few reported cases of a primary cervical infection in the literature is provided. In addition, the liquid-based cytology was established as a tool to diagnose the rare Aspergillus infection

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

Genetic Variants in the 3’UTR of BRCA1 and BRCA2 Genes and Their Putative Effects on the microRNA Mechanism in Hereditary Breast and Ovarian Cancer

Hereditary breast and ovarian cancer (HBOC) syndrome is mainly caused by mutations in the BRCA1 and BRCA2 genes. The 3&rsquo;UTR region allows for the binding of microRNAs, which are involved in genetic tune regulation. We aimed to identify allelic variants on 3&rsquo;UTR miRNA-binding sites in the BRCA1 and BRCA2 genes in HBOC patients. Blood samples were obtained from 50 patients with HBOC and from 50 controls. The 3&rsquo;UTR regions of BRCA1 and BRCA2 were amplified by PCR and sequenced to identify genetic variants using bioinformatics tools. We detected nine polymorphisms in 3&rsquo;UTR, namely: four in BRCA1 (rs3092995 (C/G), rs8176318 (C/T), rs111791349 (G/A), and rs12516 (C/T)) and five in BRCA2 (rs15869 (A/C), rs7334543 (A/G), rs1157836 (A/G), and rs75353978 (TT/del TT)). A new variant in position c.*457 (A/C) on 3&rsquo;UTR of BRCA2 was also identified. The following three variants increased the risk of HBOC in the study population: rs111791349-A, rs15869-C, and c.*457-C (odds ratio (OR) range 3.7&ndash;15.4; p &lt; 0.05). Genetic variants into the 3&rsquo;UTR of BRCA1 and BRCA2 increased the risk of HBOC between 3.7&ndash;15.4 times in the study population. The presence/absence of these polymorphisms may influence the loss/creation of miRNA binding sites, such as hsa-miR-1248 in BRCA1 3&prime;UTR or the hsa-miR-548 family binding site in BRCA2. Our results add new evidence of miRNA participation in the pathogenesis of HBOC

Molecular detection of Helicobacter pylori based on the presence of cagA and vacA virulence genes in dental plaque from patients with periodontitis

Background/purpose: Helicobacter pylori (H. pylori) infection is the most common in the world and is associated with various gastrointestinal pathologies, including chronic gastritis, peptic ulcers, and gastric cancer. The prevalence is associated with socioeconomic conditions, with this infection being more common in developing countries than in developed countries. The presence and permanence of H. pylori in the oral cavity has been reported, but its role is controversial. The aim of this study was to determine the prevalence of H. pylori in dental plaque of patients with periodontitis. Materials and methods: A cross-sectional study was carried out and Periodontal Screening and Recording (PSR) index was determined. 38 dental plaque samples were taken and total DNA was extracted and qPCR was performed. Results: 60.5% of the samples (n = 23) were positive for the presence of H. pylori by the amplification of the 16S rRNA and vacA genes. In addition, cagA gene was detected in 21.7% (n = 5) of H. pylori-positive. A significant relationship between periodontal status and H. pylori oral infection was found (P ≤ 0.05); patients with initial and moderate periodontitis were the most affected with 39.1% and 30.4%, respectively. Conclusion: Our results suggest that the prevalence of H. pylori in the oral cavity could be related to the progression of periodontal disease. Therefore, oral hygiene and treatment for the elimination of oral H. pylori could stop the progression of periodontal disease

International Nosocomial Infection Control Consortiu (INICC) report, data summary of 43 countries for 2007-2012. Device-associated module

We report the results of an International Nosocomial Infection Control Consortium (INICC) surveillance study from January 2007-December 2012 in 503 intensive care units (ICUs) in Latin America, Asia, Africa, and Europe. During the 6-year study using the Centers for Disease Control and Prevention's (CDC) U.S. National Healthcare Safety Network (NHSN) definitions for device-associated health care–associated infection (DA-HAI), we collected prospective data from 605,310 patients hospitalized in the INICC's ICUs for an aggregate of 3,338,396 days. Although device utilization in the INICC's ICUs was similar to that reported from ICUs in the U.S. in the CDC's NHSN, rates of device-associated nosocomial infection were higher in the ICUs of the INICC hospitals: the pooled rate of central line–associated bloodstream infection in the INICC's ICUs, 4.9 per 1,000 central line days, is nearly 5-fold higher than the 0.9 per 1,000 central line days reported from comparable U.S. ICUs. The overall rate of ventilator-associated pneumonia was also higher (16.8 vs 1.1 per 1,000 ventilator days) as was the rate of catheter-associated urinary tract infection (5.5 vs 1.3 per 1,000 catheter days). Frequencies of resistance of Pseudomonas isolates to amikacin (42.8% vs 10%) and imipenem (42.4% vs 26.1%) and Klebsiella pneumoniae isolates to ceftazidime (71.2% vs 28.8%) and imipenem (19.6% vs 12.8%) were also higher in the INICC's ICUs compared with the ICUs of the CDC's NHSN