Modulation of the leptin receptors expression in breast cancer cell lines exposed to leptin and tamoxifen

Artículo open accessInvestigación referente a la modulación de la leptina en un modelo de obesidad in vitro y tamoxifeno con líneas celulares de cáncer de mamaSEIA UAE

Electroeluting DNA Fragments

Purified DNA fragments are used for different purposes in Molecular Biology and they can be prepared by several procedures. Most of them require a previous electrophoresis of the DNA fragments in order to separate the band of interest. Then, this band is excised out from an agarose or acrylamide gel and purified by using either: binding and elution from glass or silica particles, DEAE-cellulose membranes, "crush and soak method", electroelution or very often expensive commercial purification kits. Thus, selecting a method will depend mostly of what is available in the laboratory. The electroelution procedure allows one to purify very clean DNA to be used in a large number of applications (sequencing, radiolabeling, enzymatic restriction, enzymatic modification, cloning etc). This procedure consists in placing DNA band-containing agarose or acrylamide slices into sample wells of the electroeluter, then applying current will make the DNA fragment to leave the agarose and thus be trapped in a cushion salt to be recovered later by ethanol precipitation

Expression of AdipoR1 and AdipoR2 Receptors as Leptin-Breast Cancer regulation mechanisms

: La obesidad es considerada como uno de los principales factores de riesgo implicados en el desarrollo del cáncer de mama y se ha documentado el papel que juega el tejido adiposo a través de Leptina y Adiponectina. El estudio de estas proteínas ha determinado su importancia en el desarrollo y progreso del cáncer de mama, pero muy pocos son los que han estudiado a AdipoR1 y AdipoR2 y la influencia de la Leptina sobre ellos. Se planteó el analizó la expresión de AdipoR1 y AdipoR2 modulada por concentraciones diferenciales de Leptina en un modelo de obesidad asociado a cáncer de mama en líneas celulares, utilizando sondas TaqMan®, mediado por concentraciones diferenciales de Leptina en líneas celulares. Seguido se llevó a cabo la extracción de RNA, obtención de cDNA (Retrotranscripción) y PCR en tiempo real. Se caracterizó cada línea celular por medio de Inmunohistoquímica. Concluyendo con base en los resultados que la concentración de normo-peso (10ng/mL) de leptina generó un incremento de la expresión de ambos receptores de adiponectina.The development of breast cancer is influenced by the adipose tissue through the proteins leptin and adiponectin. However, there is little research concerning AdipoR1 and AdipoR2 receptors and the influence of leptin over them. The objective of this work was to analyze the expression of AdipoR1 and AdipoR2, modulated by differential concentrations of leptin in an obesity model (10ng/mL, 100ng/mL, and 1000ng/mL) associated with breast cancer in MCF-7 and HCC1937 cell lines. Each cell was characterized through immunohistochemistry, and the expression of AdipoR1 and AdipoR2 was analyzed by PCR in real time using TaqMan® probes. Leptin induced an increase in cell population of MCF-7 (23.8%, 10ng/m, 48 h) and HCC1937 (17.24%, 1000ng/mL, 72 h). In MCF-7, the expression of AdipoR1 decreased (3.81%, 1000ng/Ml) and the expression of AdipoR2 increased by 13.74 times (10ng/mL) with regard to the control. In HCC1937, the expression of AdipoR1 decreases by 86.28% (10ng/mL), as well as the expression of AdipoR1 and AdipoR2 mRNA

Yield and quality of hybrid tomato grafted and cultivated under shade mesh and greenhouse

The objective was to determine the yield, fruit quality and root development of four grafted tomato hybrids grown under anti-aphid mesh cover and greenhouse. The grafted hybrids were grown under shade mesh and greenhouse from april to november 2014. Variables evaluated were: fruit weight per plant, number of clusters per plant, number of fruits per plant, polar and equatorial fruit diameter, vitamin C and lycopene contents, and root fresh and dry weight. Hybrids grafted and cultivated under anti-aphid mesh had higher quality than the production obtained under greenhouse. However, lycopene and vitamin C contents and accumulated root system dry matter are greater under greenhous

Functional relevance of the switch of VEGF receptors/co-receptors during peritoneal dialysis-induced mesothelial to mesenchymal transition

Vascular endothelial growth factor (VEGF) is up-regulated during mesothelial to mesenchymal transition (MMT) and has been associated with peritoneal membrane dysfunction in peritoneal dialysis (PD) patients. It has been shown that normal and malignant mesothelial cells (MCs) express VEGF receptors (VEGFRs) and co-receptors and that VEGF is an autocrine growth factor for mesothelioma. Hence, we evaluated the expression patterns and the functional relevance of the VEGF/VEGFRs/co-receptors axis during the mesenchymal conversion of MCs induced by peritoneal dialysis. Omentum-derived MCs treated with TGF-β1 plus IL-1β (in vitro MMT) and PD effluent-derived MCs with non-epithelioid phenotype (ex vivo MMT) showed down-regulated expression of the two main receptors Flt-1/VEGFR-1 and KDR/VEGFR-2, whereas the co-receptor neuropilin-1 (Nrp-1) was up-regulated. The expression of the Nrp-1 ligand semaphorin-3A (Sema-3A), a functional VEGF competitor, was repressed throughout the MMT process. These expression pattern changes were accompanied by a reduction of the proliferation capacity and by a parallel induction of the invasive capacity of MCs that had undergone an in vitro or ex vivo MMT. Treatment with neutralizing anti-VEGF or anti-Nrp-1 antibodies showed that these molecules played a relevant role in cellular proliferation only in naïve omentum-derived MCs. Conversely, treatment with these blocking antibodies, as well as with recombinant Sema-3A, indicated that the switched VEGF/VEGFRs/co-receptors axis drove the enhanced invasion capacity of MCs undergoing MMT. In conclusion, the expression patterns of VEGFRs and co-receptors change in MCs during MMT, which in turn would determine their behaviour in terms of proliferation and invasion in response to VEGFThis work was supported by grant SAF2010-21249 from the ‘‘Ministerio de Economía y Competitividad’’ to M.L.C. and by grant S2010/BMD-2321 from ‘‘Comunidad Autónoma de Madrid’’ to M.L.C. and R.S. This work was also partially supported by grants PI 09/0776 from ‘‘Fondo de Investigaciones Sanitarias’’ to A.A., and RETICS 06/0016 (REDinREN, Fondos FEDER, EU) to R.S

Tamoxifen ameliorates peritoneal membrane damage by blocking mesothelial to mesenchymal transition in peritoneal dialysis

Mesothelial-to-mesenchymal transition (MMT) is an auto-regulated physiological process of tissue repair that in uncontrolled conditions such as peritoneal dialysis (PD) can lead to peritoneal fibrosis. The maximum expression of peritoneal fibrosis induced by PD fluids and other peritoneal processes is the encapsulating peritoneal sclerosis (EPS) for which no specific treatment exists. Tamoxifen, a synthetic estrogen, has successfully been used to treat retroperitoneal fibrosis and EPS associated with PD. Hence, we used in vitro and animal model approaches to evaluate the efficacy of Tamoxifen to inhibit the MMT as a trigger of peritoneal fibrosis. In vitro studies were carried out using omentum-derived mesothelial cells (MCs) and effluent-derived MCs. Tamoxifen blocked the MMT induced by transforming growth factor (TGF)-β1, as it preserved the expression of E-cadherin and reduced the expression of mesenchymal-associated molecules such as snail, fibronectin, collagen-I, α-smooth muscle actin, and matrix metalloproteinse-2. Tamoxifen-treatment preserved the fibrinolytic capacity of MCs treated with TGF-β1 and decreased their migration capacity. Tamoxifen did not reverse the MMT of non-epitheliod MCs from effluents, but it reduced the expression of some mesenchymal molecules. In mice PD model, we demonstrated that MMT progressed in parallel with peritoneal membrane thickness. In addition, we observed that Tamoxifen significantly reduced peritoneal thickness, angiogenesis, invasion of the compact zone by mesenchymal MCs and improved peritoneal function. Tamoxifen also reduced the effluent levels of vascular endothelial growth factor and leptin. These results demonstrate that Tamoxifen is a therapeutic option to treat peritoneal fibrosis, and that its protective effect is mediated via modulation of the MMT processThis work was supported by grant SAF2010-21249 from the ‘‘Ministerio de Economia y Competitividad’’ to MLC and by grant S2010/BMD-2321 from ‘‘Comunidad Autónoma de Madrid’’ to MLC and RS. This work was also partially supported by grants PI 09/0776 from ‘‘Fondo de Investigaciones Sanitarias’’ to AA, and RETICS 06/0016 (REDinREN, Fondos FEDER, EU) to R

Evaluation of micronuclei in oral mucosa of individuals exposed to ionizing radiation: a pilot study from Celaya, México

Introduction: Occupational exposure to ionizing radiation can potentially lead to adverse health effects, including cancer and genetic defects. Genetic damage caused by radiation can be detected if micronuclei are observed. The objective of this pilot study was to detect the presence of micronuclei in cells of the oral mucosa in inidividuals occupationally exposed to ionizing radiation.Methods:  We implemented a pilot case-control study in which we compared oral mucosa micronuclei in 30 medical and nursing personnel in radiology centers in Celaya, Mexico, with 30 volunteers not exposed to ionizing radiation recruited from a public University. The oral mucosa was brushed and the amount of micronuclei was quantified. Chi-square test or t-test for two proportions were used to compared ionizing radiation and genetic damage between exposed and non-exposed groups.Results: The exposed group had an average of 5.37 ± 3.49 micronuclei and the non-exposed had 0.37 ± 0.61 (P<0.01). In the exposed group, 90% of participants exhibited genetic damage compared to 6.67% in the unexposed group (P<0.05).Conclusion: In this pilot study, medical and nursing staff from radiology centers presented with higher genetic damage compared to control group. Further studies are needed to identify the prevalence of genetic damage due to occupational radiation exposure in Mexico

Tefroestratigrafía preliminar de erupciones explosivas del volcán Misti (Arequipa, Perú) desde la Autopista y sus implicaciones para los peligros volcánicos

El volcán Misti ha tenido erupciones plinianas y sub-plinianas en el pasado y hay una población aproximada de un millón de habitantes viviendo en sus alrededores. La combinación de la gran población viviendo cerca del volcán y su historia de tales erupciones explosivas hacen al Misti un volcán peligroso. Para entender los peligros volcánicos en tal situación, es necesario entender la cantidad y poder de tales erupciones y las áreas impactadas. Por estudiar la tefroestratigrafía del volcán se puede entender los detalles de sus erupciones. Actualmente el Servicio Geológico de los Estados Unidos e INGEMMET tienen un proyecto colaborativo para estudiar la tefroestratigrafía del volcán Misti desde finales del Pleistoceno. El documento presenta los resultados preliminares del proyecto y sus implicaciones para los peligros volcánicos del Misti

Evaluation of micronuclei in oral mucosa of individuals exposed to ionizing radiation: a pilot study from Celaya, México

Introduction: Occupational exposure to ionizing radiation can potentially lead to adverse health effects, including cancer and genetic defects. Genetic damage caused by radiation can be detected if micronuclei are observed. The objective of this pilot study was to detect the presence of micronuclei in cells of the oral mucosa in inidividuals occupationally exposed to ionizing radiation. Methods:  We implemented a pilot case-control study in which we compared oral mucosa micronuclei in 30 medical and nursing personnel in radiology centers in Celaya, Mexico, with 30 volunteers not exposed to ionizing radiation recruited from a public University. The oral mucosa was brushed and the amount of micronuclei was quantified. Chi-square test or t-test for two proportions were used to compared ionizing radiation and genetic damage between exposed and non-exposed groups. Results: The exposed group had an average of 5.37 ± 3.49 micronuclei and the non-exposed had 0.37 ± 0.61 (P<0.01). In the exposed group, 90% of participants exhibited genetic damage compared to 6.67% in the unexposed group (P<0.05). Conclusion: In this pilot study, medical and nursing staff from radiology centers presented with higher genetic damage compared to control group. Further studies are needed to identify the prevalence of genetic damage due to occupational radiation exposure in Mexico