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La propagación de los Agaves spp. en México es muy extensa y el país cuenta con 151 especies endémicas. Sin embargo, prácticas como la construcción, el desarrollo de fibras, la elaboración de alimentos y bebidas alcohólicas, así como la elaboración de productos ornamentales han ocasionado una considerable disminución entre las poblaciones silvestres. En este sentido, el presente trabajo tiene como objetivo realizar la caracterización morfométrica por el método de I.S.T.A de las semillas de Agave spp. colectadas en los diferentes municipios del estado de Guerrero (Agave cupreata Trel. & A. Berger y Agave angustifolia Haw.). Los resultados obtenidos se sometieron a análisis de varianza y pruebas de medias (Tukey, p ≤ 0.05), tomándose el largo, ancho, grosor de las semillas y su volumen. El sitio de muestreo que presentó el mayor número de semillas por kilogramo (233,590) fue Lamazintla. Por otra parte, las semillas de mayor longitud, anchura y grosor se encontraron en el municipio de Alpoyeca (8.6; 6.4; 0.62 mm, respectivamente). Además el análisis de pureza demostró que la muestra obtenida en Amojileca cuenta con el mayor número de semillas maduras con un promedio del 57%.

Introducción. Los efectos adversos de los químicos en la reproducción masculina es un área de preocupación creciente, actualmente el Tl es utilizado principalmente en la fabricación de dispositivos electrónicos, interruptores y cierres; así como en plantas generadoras de energía, fábricas de cemento y fundiciones. Objetivo. Evaluar la calidad espermática y el daño genético de los espermatozoides en ratones expuestos oralmente a Tl. Materiales y métodos. Se utilizaron ratones macho y dosis de Tl de 5, 15 y 25 mg/kg/5d, se evaluaron los parámetros de calidad espermática y el daño al ADN por la técnica de SCSA. Resultados. La exposición a Tl afecto la motilidad, la viabilidad y la morfología espermática. Se observaron alteraciones en la estructura del ADN (%DFI y %HSD) de los espermatozoides. El Tl se podría considerar reprotóxico, ya que altera capacidad reproductiva.

Leptin is a cytokine-like hormone that functions as a link between obesity and breast cancer (BC). Leptin treatment induces Epithelial to Mesenchymal Transition (EMT) in BC cell lines. In non-tumoral breast epithelial MCF10A cells, acute leptin treatment induces partial EMT. However, the effect of chronic leptin treatment on EMT in non-tumorigenic breast cells has not been fully explored. This study aimed to evaluate the effect of chronic leptin treatment on the induction of EMT in MCF10A cells. We found that chronic leptin treatment induces a switch from an epithelial to a mesenchymal morphology, partial loss of E-cadherin and gain of vimentin expression. Immunolocalization experiments showed a partial loss of E-cadherin at cell junctions and increased cytoplasmic localization of vimentin in leptin-treated cells. Moreover, chronic leptin treatment increased collective cell migration and invasion. Furthermore, when cultured in non-adherent conditions leptin treated cells exhibited reduced cell aggregation, increased survival, and decreased apoptosis, which correlates with increased FAK and AKT phosphorylation. Finally, bioinformatic analysis in two publicly available RNAseq datasets from normal breast tissue shows that high levels of leptin mRNA correlate positively with the expression of mesenchymal markers, and negatively with epithelial markers. Thus, our results demonstrate that chronic leptin treatment induces EMT in non-tumorigenic MCF10A cells and suggest that high leptin expression in normal breast tissue may induce EMT and contribute to increased risk of breast cancer.

Benzopyrene is a high molecular weight polycyclic aromatic hydrocarbon with highly recalcitrant and develops carcinogenic effects. CsrA is a conserved regulatory protein, which controls the translation and stability of its target transcripts, having negative or positive effects depending on the target mRNAs. It is known that Bacillus licheniformis M2-7 has the ability to grow and survive in concentrations of hydrocarbons as benzopyrene, which prompted in part by CsrA, as it occurs in the presence of gasoline. However, there are a few studies that reveal the genes involved in that process. In order to know the involved genes in the Bacillus licheniformis M2-7 degradation pathway, the plasmid pCAT-sp containing a mutation in the catEgen was constructed and used to transform B. licheniformis M2-7 and generate the CAT1 strain. We determined the capacity of the mutant B. licheniformis(CAT1) to grow in the presence of glucose and benzopyrene as a carbon source. We observed that CAT1 strain presented an increase in growth in the presence of glucose, but a statistically considerable decrease in the presence of benzopyrene with respect to the wild strain M2-7. Also we demonstrated that the Csr system regulates its expression positively since it was observed that the expression of the gene in the mutant strain LYA12 (M2-7 csrA:: Sp, SpR) decreased considerably with respect to the wild strain. This allowed to characterize the metabolic pathway that Bacillus licheniformis M2-7 implements in the presence of benzopyrene.

ZO-1α+ and ZO-1α− proteins are expressed in hermetic and leaky tight junctions, respectively. Two cis-acting distant exonic elements partly activate the 240 nucleotide-long α exon producing the ZO-1α+ isoform. However, the elements within and around the α exon and their respective factors involved in its splicing are unknown. To study the dynamic interaction between SRSF3 and its bioinformatically predicted target sites around the 3’ss upstream of the α exon during its activation, we performed EMSA, crosslinking, and in vivo splicing assays by ZO-1 minigene expression and siRNA-mediated silencing in transfected cells. Using V1 RNase, we probed the possible formation of a hairpin RNA structure between the intronic and proximal exonic SRSF3 binding sites. The hairpin sufficed for complex formations in the EMSA. The interaction of SRSF3 with the intronic site promoted the cooperative binding of SRSF3 to the exonic site. Finally, SRSF3 restored α exon activation in SRSF3 knockdown transfectants. Altogether, our results show that SRSF3–hairpin RNA interaction is crucial in the early recognition of 3’ss for α exon activation. It remains to be explored whether SRSF3 recruits or stabilizes the binding of other factors or brings separate splice sites into proximity.

Cancer is a public health problem worldwide, and one of the crucial steps within tumor progression is the invasion and metastasis of cancer cells, which are directly related to cancer-associated deaths in patients. Recognizing the molecular markers involved in invasion and metastasis is essential to find targeted therapies in cancer. Interestingly, about 50% of the discovered drugs used in chemotherapy have been obtained from natural sources such as plants, including isoflavonoids. Until now, most drugs are used in chemotherapy targeting proliferation and apoptosis-related molecules. Here, we review recent studies about the effect of isoflavonoids on molecular targets and signaling pathways related to invasion and metastasis in cancer cell cultures, in vivo assays, and clinical trials. This review also reports that glycitein, daidzein, and genistein are the isoflavonoids most studied in preclinical and clinical trials and displayed the most anticancer activity targeting invasion-related proteins such as MMP-2 and MMP-9 and also EMT-associated proteins. Therefore, the diversity of isoflavonoids is promising molecules to be used as chemotherapeutic in invasive cancer. In the future, more clinical trials are needed to validate the effectiveness of the various natural isoflavonoids in the treatment of invasive cancer.

BackgroundSome species of theFicusgenus show pharmacological activity, including antiproliferative activity, in cell lines of several cancer Types. ficus crocatais distributed in Mexico and used in traditional medicine, as it is believed to possess anti-inflammatory, analgesic, and antioxidant properties. However, as of yet, there are no scientific reports on its biological activity. This study aims to evaluate the phytochemical profile ofF. crocataleaf extracts and their effects on breast cancer MDA-MB-231 cells proliferation. Moreover, the study aims to unearth possible mechanisms involved in the decrease of cell proliferation.MethodsThe extracts were obtained by the maceration of leaves with the solvents hexane, dichloromethane, and acetone. The phytochemical profile of the extracts was determined using gas chromatography coupled with mass analysis. Cell proliferation, apoptosis, and cell cycle analysis in MDA-MB-231 cells were determined using a Crystal violet assay, MTT assay, and Annexin-V/PI assay using flow cytometry. The data were analyzed using ANOVA and Dunnett’s test.ResultsThe hexane (Hex-EFc), dichloromethane (Dic-EFc), and acetone (Ace-EFc) extracts ofF. crocatadecreased the proliferation of MDA-MB-231 cells, with Dic-EFc having the strongest effect. Dic-EFc was fractioned and its antiproliferative activity was potentiated, which enhanced its ability to induce apoptosis in MDA-MB-231 cells, as well as increased p53, procaspase-8, and procaspase-3 expression.ConclusionsThis study provides information on the biological activity ofF. crocataextracts and suggests their potential use against triple-negative breast cancer.

Breast cancer is the most common invasive neoplasia, and the second leading cause of the cancer deaths in women worldwide. Mammary tumorigenesis is severely linked to obesity, one potential connection is leptin. Leptin is a hormone secreted by adipocytes, which contributes to the progression of breast cancer. Cell migration, metalloproteases secretion, and invasion are cellular processes associated with various stages of metastasis. These processes are regulated by the kinases FAK and Src. In this study, we utilized the breast cancer cell lines MCF7 and MDA-MB-231 to determine the effect of leptin on FAK and Src kinases activation, cell migration, metalloprotease secretion, and invasion. We found that leptin activates FAK and Src and induces the localization of FAK to the focal adhesions. Interestingly, leptin promotes the activation of FAK through a Src- and STAT3-dependent canonical pathway. Specific inhibitors of FAK, Src and STAT3 showed that the effect exerted by leptin in cell migration in breast cancer cells is dependent on these proteins. Moreover, we established that leptin promotes the secretion of the extracellular matrix remodelers, MMP-2 and MMP-9 and invasion in a FAK and Src-dependent manner. Our findings strongly suggest that leptin promotes the development of a more aggressive invasive phenotype in mammary cancer cells.

Background/aims Ouabain, a well-known plant-derived toxin, is also a hormone found in mammals at nanomolar levels that binds to a site located in the a-subunit of Na⁺,K⁺-ATPase. Our main goal was to understand the physiological roles of ouabain. Previously, we found that ouabain increases the degree of tight junction sealing, GAP junction-mediated communication and ciliogenesis. Considering our previous results, we investigated the effect of ouabain on adherens junctions. Methods We used immunofluorescence and immunoblot methods to measure the effect of 10 nM ouabain on the cellular and nuclear content of E-cadherin, β-catenin and γ-catenin in cultured monolayers of Marin Darby canine renal cells (MDCK). We also studied the effect of ouabain on adherens junction biogenesis through sequential Ca²⁺ removal and replenishment. Then, we investigated whether c-Src and ERK1/2 kinases are involved in these responses. Results Ouabain enhanced the cellular content of the adherens junction proteins E-cadherin, β-catenin and γ-catenin and displaced β-catenin and γ-catenin from the plasma membrane into the nucleus. Ouabain also increased the expression levels of E-cadherin and β-catenin in the plasma membrane after Ca²⁺ replenishment. These effects on adherens junctions were sensitive to PP2 and PD98059, suggesting that they depend on c-Src and ERK1/2 signaling. The translocation of β-catenin and γ-catenin into the nucleus was specific because ouabain did not change the localization of the tight junction proteins ZO-1 and ZO-2. Moreover, in ouabain-resistant MDCK cells, which express a Na⁺,K⁺-ATPase α1-subunit with low affinity for ouabain, this hormone was unable to regulate adherens junctions, indicating that the ouabain receptor that regulates adherens junctions is Na⁺,K⁺-ATPase. Conclusion Ouabain (10 nM) upregulated adherens junctions. This novel result supports the proposition that one of the physiological roles of this hormone is the modulation of cell contacts.

Background To improve the efficiency of early diagnosis systems for cervical cancer, the use of cellular and viral markers for identifying precancerous lesions with a greater probability to progress to cancer has been proposed. Several cellular proteins and markers of oxidative DNA damage have been suggested as possible biomarkers of cervical carcinogenesis; however, they have not been evaluated together. In this study, we analyzed the expression of the cellular markers p16INK4a, Ki-67, CyclinE1, TOP2A/MCM2, and telomerase, as well as the DNA oxidative damage markers ROS and 8-OHdG. The analyses were performed in liquid-based cervical cytology samples or biopsies with premalignant lesions or cervical cancer diagnosis, with the purpose of selecting a panel of biomarkers that allow the identification of precursor lesions with greater risk of progression to cervical cancer. Methods We analyzed 1485 liquid-based cytology samples, including 239 non-squamous intraepithelial lesions (NSIL), 901 low-grade squamous intraepithelial lesions (LSIL), 54 high-grade squamous intraepithelial lesions (HSIL), and 291 cervical cancers (CC). The biomarkers were analyzed by immunocytochemistry and Human Papilloma Virus (HPV) genotyping with the INNO-LiPA genotyping Extra kit. Results We found that all tested cellular biomarkers were overexpressed in samples with high risk-HPV infection, and the expression levels increased with the severity of the lesion. TOP2A/MCM2 was the best biomarker for discriminating between LSIL and HSIL, followed by p16INK4a and cyclinE1. Statistical analysis showed that TOP2A/MCM2 provided the largest explanation of HSIL and CC cases (93.8%), followed by p16INK4a (91%), cyclin E1 (91%), Ki-67 (89.3%), and telomerase (88.9%). Conclusions We propose that the detection of TOP2A/MCM2, p16INK4a and cyclin E1 expression levels is useful as a panel of biomarkers that allow identification of cervical lesions with a higher risk for progression to CC with high sensitivity and precision; this can be done inexpensively, in a single and non-invasive liquid-based cytology sample.

Foodborne illnesses, such as infections or food poisoning, can be caused by bacterial biofilms present in food matrices or machinery. The production of biofilms by several strains of Bacillus cereus on different materials under different culture conditions was determined, as well as the relationship of biofilms with motility, in addition to the enterotoxigenic profile and candidate genes that participate in the production of biofilms. Biofilm production of B. cereus strains was determined on five materials: glass, polystyrene, polyethylene, polyvinylchloride (PVC), PVC/glass, in three culture media: Phenol red broth, tryptic soy broth, and brain heart infusion broth, in two different temperatures (37 &deg, C and 25 &deg, C), and in two different oxygen conditions (oxygen and CO2 tension). Furthermore, the strains were molecularly characterized by end-point polymerase chain reaction. Motility was determined on semi-solid agar. The B. cereus strains in this study were mainly characterized as enterotoxigenic strains, statistically significant differences were found in the PVC material and biofilm production. Motility was positively associated with the production of biofilm in glass/PVC. The sipW and tasA genes were found in two strains. The results of this study are important in the food industry because the strains carry at least one enterotoxin gene and produce biofilms on different materials

The aim of this work was to isolate pulp (PH) and seed hydrocolloids (SH) of the fruit of Hymenaea corbaril L. and determinate chemical, functional and rheological properties, as a proposal of a new attractive product for functional applications in food systems. Both hydrocolloids have a high content of carbohydrates, minerals and fiber. The solubility of PH was higher 94.1%. The emulsifying capacity of SH, PH and GG increases with the hydrocolloid concentration. The emulsions showing a good stabilization stability with drop sizes of 5–20 μm. The emulsions PH and SH at 1% were higher stability than guar gum (GG), due to slower down the cremation index after 3 days of storage at 35 and 40°C. The hydrocolloids exhibited a pseudoplastic behavior at concentrations of 0.5–2%. The elastic behavior suggests its application to edible films, while the viscous behavior suggests its incorporation in food systems. Los hidrocoloides son utilizados como aditivos alimentarios, obtenidos de endospermos de semillas de dicotiledóneas. Una interesante alternativa para su uso como fuente biológica del estado de Guerrero es la extracción de hidrocoloides de pulpa (PH) y semilla (SH) del fruto de Hymenaea corbaril L. El objetivo de este trabajo fue aislar hidrocoloides de pulpa y semilla y determinar sus propiedades químicas, funcionales y reológicas para proponerlos como un nuevo producto atractivo con aplicación a los sistemas alimenticios. Los resultados fisicoquímicos mostraron que ambos hidrocoloides poseen un alto contenido en carbohidratos, minerales y fibra. La solubilidad del PH es mayor con 94.1%. Se observó buena estabilización de emulsiones con tamaños de gota de 5- 20 μm. Las emulsiones de PH y SH al 1% mostraron mayor estabilidad que GG debido a que mostraron un descenso más lento en el índice de cremado durante los tres días de almacenamiento a 35 y 40°C. Ambos hidrocoloides muestran comportamientos de tipo pseudoplástico n

Epithelial–mesenchymal transition (EMT) is a reversible cellular process, characterized by changes in gene expression and activation of proteins, favoring the trans-differentiation of the epithelial phenotype to a mesenchymal phenotype. This process increases cell migration and invasion of tumor cells, progression of the cell cycle, and resistance to apoptosis and chemotherapy, all of which support tumor progression. One of the signaling pathways involved in tumor progression is the MAPK pathway. Within this family, the ERK subfamily of proteins is known for its contributions to EMT. The ERK subfamily is divided into typical (ERK 1/2/5), and atypical (ERK 3/4/7/8) members. These kinases are overexpressed and hyperactive in various types of cancer. They regulate diverse cellular processes such as proliferation, migration, metastasis, resistance to chemotherapy, and EMT. In this context, in vitro and in vivo assays, as well as studies in human patients, have shown that ERK favors the expression, function, and subcellular relocalization of various proteins that regulate EMT, thus promoting tumor progression. In this review, we discuss the mechanistic roles of the ERK subfamily members in EMT and tumor progression in diverse biological systems.

Breast cancer is the most common invasive neoplasia, and the second leading cause of death associated with cancer in women worldwide. Mammary tumorigenesis is severely linked to obesity, the potential connection is leptin. Leptin is a hormone secreted by adipocytes, which contributes to the progression of breast cancer. Cell migration, metalloproteases secretion, and invasion are cellular processes associated with various stages of metastasis. These processes are regulated by the kinases FAK and Src. In this study, we utilized the breast cancer cell lines MCF7 and MDA-MB-231 to determine the effect of leptin on FAK and Src kinases activation, cell migration, metalloproteases secretion, and invasion. By Western blot we found that leptin activates FAK and Src, and induces the localization of FAK to the focal adhesions. Specific inhibitors of FAK and Src showed that the effect exerted by leptin in cell migration, and invasion in breast cancer cells is dependent on these kinases. Moreover, by gelatin zymmography we established that leptin promotes the secretion of the extracellular matrix remodelers, MMP-2 and MMP-9, in a FAK and Src dependent manner. Our findings strongly suggest that leptin promotes the development of a more aggressive invasive phenotype in mammary cancer cells.

The use of monoclonal antibodies for the detection of cellular biomarkers during carcinogenesis provides new strategies for cancer diagnosis or prognosis in patients. Loss of the Restrictive Element 1-Silencing Transcription (REST) factor has been observed in previous molecular and immunological approaches in aggressive breast cancer, small cell lung cancer, liver carcinoma, and colo-rectal cancer; however, for clinic diagnosis, monoclonal antibodies for REST recognition are unavailable. The goal of this work was to design, produce and characterize monoclonal antibodies against the REST DNA binding damain (DBD) that would be suitable for immunoassays. We searched for conserved domains, and immunogenic and antigenic sites in the REST structure via in silico analysis. For mice immunization, we used a recombinant REST DBD purified by affinity chromatography, and then Hybridomas were generated by mouse spleen fusion with myeloma cells. Finally, for monoclonal antibody characterization, we performed enzyme-linked immunosorbent (ELISA), western blot, dot blot, immunocytochemistry (ICC) and immunoprecipitation assays. Results showed that the DBD is conserved in REST isoforms and contains immunogenic and antigenic sites. We generated three clones producing monoclonal antibodies against REST DBD, one of them specifically recognized native REST and was suitable for ICC in samples from patients.

Leptin is a hormone secreted mainly by adipocytes; physiologically, it participates in the control of appetite and energy expenditure. However, it has also been linked to tumor progression in different epithelial cancers. In this review, we describe the effect of leptin on epithelial–mesenchymal transition (EMT) markers in different study models, including in vitro, in vivo, and patient studies and in various types of cancer, including breast, prostate, lung, and ovarian cancer. The different studies report that leptin promotes the expression of mesenchymal markers and a decrease in epithelial markers, in addition to promoting EMT-related processes such as cell migration and invasion and poor prognosis in patients with cancer. Finally, we report that leptin has the greatest biological relevance in EMT and tumor progression in breast, lung, prostate, esophageal, and ovarian cancer. This relationship could be due to the key role played by the enriched tumor microenvironment in adipose tissue. Together, these findings demonstrate that leptin is a key biomolecule that drives EMT and metastasis in cancer.

Background The repressor element 1-silencing transcription factor (REST) is a regulator of gene expression, and the Ras association domain family member 1 A (RASSF1A) is an important tumor suppressor gene involved in cancer development. Although extensive characterization of the roles of REST and RASSF1A in cancer development have been reported in cellular models, the link between them and their possible role in the development of squamous intraepithelial lesions (SIL) and squamous cell carcinoma (SCC) of the cervix have not been explored. The aim of this study was to evaluate the expression of REST and RASSF1A in cervical cytological and histological samples from patients diagnosed with SIL or SCC and in CC-derived cell lines. Materials and methods We analyzed the expression of REST and RASSF1A by immunocyto/histochemistry in cervical samples from patients (n = 271) and in cancer cell lines. Data analyses were performed using the Kruskal–Wallis test and generalized linear models. Results We identified binding sites for REST in RASSF1A and observed a significant reduction in REST and RASSF1A nuclear expression in samples from patients with high-grade SIL (HSIL) and SCC. For REST, we observed an average decrease of 334 and 423 r.u.d. for HSIL (n = 21) and SCC (n = 18) compared with non-LSIL (n = 72), whereas for RASSF1A, this decrease was 126 and 217 r.u.d., respectively (p Conclusion Our results provide evidence of the altered expression of REST and RASSF1A in SIL and SCC, with a significant decrease in HSIL, SCC, and SCC-derived cell lines; findings that can contribute to the diagnosis, prognosis, and post-treatment follow-up of patients diagnosed with SIL or SCC.

Background Cervical cancer (CC) is the fourth cause of mortality by neoplasia in women worldwide. The use of immunomarkers is an alternative tool to complement currently used algorithms for detection of cancer, and to improve selection of therapeutic schemes. Aberrant expression of Ezrin and E-cadherin play an important role in tumor invasion. In this study we analyzed Ezrin and E-cadherin expression in liquid-based cervical cytology samples, and evaluated their potential use as prognostic immunomarkers. Methods Immunocytochemical staining of Ezrin and E-cadherin was performed in cervical samples of 125 patients. The cytological or histological diagnostic was performed by Papanicolaou staining or H&E staining, respectively. HPV genotyping was determined using INNO-LIPA Genotyping Extra kit and the HPV physical status by in situ hybridization. Ezrin expression in HaCaT, HeLa and SiHa cell lines was determined by immunocytochemistry, immunofluorescence and Western blot. Results High Ezrin expression was observed in cervical cancer samples (70%), samples with multiple infection by HR-HPV (43%), and samples with integrated viral genome (47%). High Ezrin expression was associated with degree of SIL, viral genotype and physical status. In contrast, low E-cadherin expression was found in cervical cancer samples (95%), samples with multiple infection by HR-HPV/LR-HPV (87%) and integrated viral genome (72%). Low E-cadherin expression was associated with degree of SIL and viral genotype. Interestingly, Ezrin nuclear staining was associated with degree of SIL and viral genotype. High Ezrin expression, high percent of nuclear Ezrin and low E-cadherin expression behaved as risk factors for progression to HSIL and cervical cancer. Conclusions Ezrin and E-cadherin expression profile in cervical cytology samples could be a potential prognostic marker, useful for identifying cervical lesions with a high-risk of progression to cervical cancer. Electronic supplementary material The online version of this article (10.1186/s12885-018-4243-7) contains supplementary material, which is available to authorized users.

Variantes genéticas se han relacionado con enfermedades de rasgos complejos y factores de riesgo asociados a éstas. El propósito de este estudio fue evaluar la asociación entre diversos polimorfismos de un solo nucleótido (SNP, por sus siglas en inglés) con el síndrome metabólico (SM) y con el índice aterogénico de plasma (IAP). Se estudiaron mujeres guerrerenses, a quienes se les realizaron mediciones somatométricas, biomarcadores sanguíneos y se identificaron SNP en siete genes por reacción en cadena de la polimerasa (PCR, por sus siglas en inglés) en tiempo real. Se observó disminución de adiponectina en mujeres con SM y con el mayor riesgo aterogénico, y se encontraron asociaciones entre apoproteina E (APOE) con niveles elevados de lipoproteínas de baja densidad (LDL, por sus siglas en inglés) y triglicéridos, del SNP rs1501299 con el IAP, de los genotipos AA/rs708272 y GG/rs1884051, con los niveles altos de LDL, y del genotipo AA/rs854560 con LDL y con la actividad paraoxonasa. Los resultados indican que biomarcadores sanguíneos y genéticos pudiesen estar implicados en alteraciones metabólicas y, por tanto, incrementar el riesgo individual del desarrollo de enfermedades.

Introduction: Gardnerella vaginalis is the main etiological agent of bacterial vaginosis (BV), this bacteria has been classified in eight biotypes based on the production of beta-galactosidase, hydrolysis of hippurate and lipase, additionally produces several virulence factors, among them vaginolysin (VLY) is the most important. VLY is a pore-forming cytolysin that damages cells structurally and functionally through the binding with the human receptor CD59 and cholesterol. To date there are no studies linking the production of VLY with the biotypes of these bacteria, which was the main objective of this study. Methodology 90 strains of G. vaginalis were analysed, of which 60 were associated with normal flora and 30 with BV, biotyping was performed according to the scheme proposed by Piot et al. 1984. Dot Blot and haemagglutination evaluated VLY production and lytic capacity. The Dot Blot results were analysed densitometrically and classified as low, moderate and abundant. The lytic capacity was expressed in percentage. Results We identified in the analysed population biotypes 1, 2, 5 and 6 of G. vaginalis . In the group with normal flora: biotype 1 was identified in 22%, biotype 2 in 12%, biotype 5 in 32% and biotype 6 in 35%, while in the group associated with BV biotype 1 was identified in 33%, biotype 2 in 10%, biotype 5 in 10% and biotype 6 in 47%. Low production of VLY was observed in 48%, moderate in 37% and abundant in 15% of the cases in the group with normal flora. While in the group with BV low production was observed in 30%, moderate in 43% and abundant in 27% of the cases. Additionally, we observed that the biotype 6 (normal flora) and 2 (BV) have the highest lytic capacity, without finding significant differences between both groups. Conclusion We isolated the biotypes 1, 2, 5, and 6 in both study groups. The biotypes associated with VB showed higher production of vaginolysin and a greater lytic capacity. These results suggest that VLY produced by biotypes of G. vaginalis could be a key factor in the establishment and maintenance of BV.

Introduction: Apolipoprotein E (ApoE) 4 isoform has been associated with elevated levels of cholesterol, low-density lipoprotein cholesterol (LDL-C), and triglycerides (TGs), meanwhile several polymorphisms in the LDL receptor ( LDLR) gene have been associated with increased levels of total cholesterol and LDL-C. Material and Methods: We studied 400 women from Southwest Mexico. Anthropometric features and biochemical profile were evaluated, and genotyping of single nucleotide polymorphisms rs429358 and rs7412 in the APOE gene and rs688 in the LDLR gene was determined by TaqMan assays. Results: We found significant association between LDL-C (odds ratio [OR] = 3.3, 95% confidence interval [CI]: 1.9-5.7) and marginal association with TG (OR = 1.7, 95% CI: 1.0-2.9) of atherogenic risk in women carriers of the ApoE4 isoform compared to ApoE3. The TT genotype of rs688 in the LDLR gene was not found to be associated with elevated levels of total cholesterol or LDL-C. Conclusion: Our results show that carrier women of the ApoE4 isoform are more likely to have elevated levels of LDL-C and therefore increased risk of developing atherosclerosis.

The splicing of the N exon in the pre-mRNA coding for the RE1-silencing transcription factor (REST) results in a truncated protein that modifies the expression pattern of some of its target genes. A weak 3'ss, three alternative 5'ss (N4-, N50-, and N62-5'ss) and a variety of putative target sites for splicing regulatory proteins are found around the N exon; two GGGG codes (G2-G3) and a poly-Uridine tract (N-PU) are found in front of the N50-5'ss. In this work we analyzed some of the regulatory factors and elements involved in the preferred selection of the N50-5'ss (N50 activation) in the small cell lung cancer cell line H69. Wild type and mutant N exon/β-globin minigenes recapitulated N50 exon splicing in H69 cells, and showed that the N-PU and the G2-G3 elements are required for N50 exon splicing. Biochemical and knockdown experiments identified these elements as U2AF65 and hnRNP H targets, respectively, and that they are also required for N50 exon activation. Compared to normal MRC5 cells, and in keeping with N50 exon activation, U2AF65, hnRNP H and other splicing factors were highly expressed in H69 cells. CLIP experiments revealed that hnRNP H RNA-binding occurs first and is a prerequisite for U2AF65 RNA binding, and EMSA and CLIP experiments suggest that U2AF65-RNA recognition displaces hnRNP H and helps to recruit other splicing factors (at least U1 70K) to the N50-5'ss. Our results evidenced novel hnRNP H and U2AF65 functions: respectively, U2AF65-recruiting to a 5'ss in humans and the hnRNP H-displacing function from two juxtaposed GGGG codes.