Job Stability and Gender Perspectives: Application of a Logistic Regression Model

Several changes have been taking place in the labour market since the 1970s that have created the right climate to spur organisations and workers to demand greater flexibility in employment. In this context, temporary employment has been the focus of many research papers and temporary contracts have been used as a tool to achieve labour flexibility. In order to understand the situation in Andalusia (Spain), this paper aims to identify the decisive factors in permanent employment. To this end, starting hypotheses will be defined about the decisive factors in permanent employment and the positive or negative significance of their influence; the starting hypotheses will then be tested empirically using a logistic regression model on a sample population of wage earners in Andalusia. In the second stage, given that the ratio of temporary contracts is much higher among women, the variable ‘gender’ is likely to be decisive in the construction of the regression model, therefore the decisive factors for permanent employment in Andalusia will be evaluated separately for men and women, in order to calibrate the impact of gender on job stability. Finally, based on the estimated probabilities of having a permanent job depending on gender, the degree of labour discrimination faced by women in the Andalusian labour market will be analysed

YKT6 expression, exosome release, and survival in non-small cell lung cancer

BACKGROUND: Cancer-derived exosomes are involved in metastasis. YKT6 is a SNARE protein that participates in the regulation of exosome production and release, but its role in non-small cell lung cancer (NSCLC) has not been examined. MATERIALS AND METHODS: Ultracentrifugation-purified exosomes from the A549 cell line were studied by CRYO-TEM, nanoparticle tracking analysis and western blot (TSG101 marker). YKT6 was inhibited using a DsiRNA and selected pre-microRNAs. MicroRNAs targeting YKT6 were validated by Renilla/Luciferase assay and western blot. YKT6 expression and its prognostic impact were analyzed in 98 tissue specimens from resected NSCLC patients. RESULTS: Membranous nanosized vesicles (mode size: 128nm) with TSG101 protein were purified from A549 cells. YKT6 inhibition reduced exosome release by 80.9%. We validated miR-134 and miR-135b as miRNAs targeting YKT6, and transfection with the pre-miRNAs also produced a significant reduction in exosome release. The analysis of YKT6 in tumor samples showed that patients with high levels had shorter disease-free and overall survival. CONCLUSIONS: YKT6 is a key molecule in the regulation of exosome release in lung cancer cells and is in turn precisely regulated by miR-134 and miR-135b. Moreover, YKT6 levels impact prognosis of resected NSCLC patients

miR-141 and miR-200c as markers of overall survival in early stage non-small cell lung cancer adenocarcinoma

Several treatments in non-small cell lung cancer (NSCLC) are histology-dependent, and the need for histology-related markers is increasing. MicroRNAs (miRNAs) are promising molecular markers in multiple cancers and show differences in expression depending on histological subtype. The miRNA family miR-200 has been associated with the regulation of epithelial-mesenchymal (EMT)/mesenchymal-epithelial transition (MET). EMT involves profound phenotypic changes that include the loss of cell-cell adhesion, the loss of cell polarity, and the acquisition of migratory and invasive properties that facilitates metastasis. A dual role for the miR-200 family in the prognosis of several tumors has been related to tumor cell origin. However, the prognostic role and function of miR-200 family in early-stage NSCLC adenocarcinoma and squamous cell carcinoma (SCC) have not been well established. Methods: miRNA expression was determined using TaqMan assays in 155 tumors from resected NSCLC patients. Functional studies were conducted in three NSCLC cell lines: H23, A-549 and HCC-44. Results: High miR-200c expression was associated with shorter overall survival (OS) in the entire cohort (p = 0.024). High miR-200c (p = 0.0004) and miR-141 (p = 0.009) expression correlated with shorter OS in adenocarcinoma - but not in SCC. In the multivariate analysis, a risk score based on miR-141 and miR-200c expression emerged as an independent prognostic factor for OS in the entire cohort (OR, 2.787; p = 0.033) and in adenocarcinoma patients (OR, 10.649; p = 0.002). Functional analyses showed that miR-200c, was related to mesenchymal-epithelial transition (MET) and affected cell migration and E-cadherin levels, while overexpression of miR-141 reduced KLF6 protein levels and produced an increase of secretion of VEGFA in vitro (H23, p = 0.04; A-549, p = 0.03; HCC-44, p = 0.02) and was associated with higher blood microvessel density in patient tumor samples (p<0.001). Conclusion: High miR-141 and miR-200c expression are associated with shorter OS in NSCLC patients with adenocarcinoma through MET and angiogenesis

The significance of PIWI family expression in human lung embryogenesis and non-small cell lung cancer

The expression of Piwi-interacting RNAs, small RNAs that bind to PIWI proteins, was until recently believed to be limited to germinal stem cells. We have studied the expression of PIWI genes during human lung embryogenesis and in paired tumor and normal tissue prospectively collected from 71 resected non-small-cell lung cancer patients. The mRNA expression analysis showed that PIWIL1 was highly expressed in 7-week embryos and downregulated during the subsequent weeks of development. PIWIL1 was expressed in 11 of the tumor samples but in none of the normal tissue samples. These results were validated by immunohistochemistry, showing faint cytoplasmic reactivity in the PIWIL1-positive samples. Interestingly, the patients expressing PIWIL1 had a shorter time to relapse (TTR) (p = 0.006) and overall survival (OS) (p = 0.0076) than those without PIWIL1 expression. PIWIL2 and 4 were downregulated in tumor tissue in comparison to the normal tissue (p < 0.001) and the patients with lower levels of PIWIL4 had shorter TTR (p = 0.048) and OS (p = 0.033). In the multivariate analysis, PIWIL1 expression emerged as an independent prognostic marker. Using 5-Aza-dC treatment and bisulfite sequencing, we observed that PIWIL1 expression could be regulated in part by methylation. Finally, an in silico study identified a stem-cell expression signature associated with PIWIL1 expression

Desarrollo de biomarcadores serológicos para la evaluación de la eficacia terapéutica del tratamiento con benznidazole de pacientes con enfermedad de Chagas

La enfermedad de Chagas, causada por Trypanosoma cruzi, afecta aproximadamente a 7 millones de personas en el mundo, existiendo actualmente mas de 50.000 casos en Espa~na. Esta enfermedad parasitaria presenta dos fases claramente diferenciadas, aguda y cronica. La mayora de los pacientes permanecen en una fase cronica asintomatica denominada fase indeterminada, que deriva (en un 30-40% de los pacientes) a una fase sintomatica caracterizada, principalmente, por alteraciones cardacas, digestivas, neurologicas o mixtas. En la fase cronica indeterminada Benznidazol y Nifurtimox son los farmacos de eleccion. Si bien se recomienda su administracion, los efectos secundarios son frecuentes. Ademas, el impacto del tratamiento solo puede ser determinado a traves de la seroconversion medida con test serologicos convencionales, lo cual tarda decadas en establecerse. Es por ello, que un reto para el control de la enfermedad de Chagas es el establecimiento de biomarcadores (BMKs) que permitan determinar la e#12;cacia del tratamiento en un corto periodo de tiempo. En los ultimos a~nos se han llevado a cabo numerosos estudios para evaluar diferentes moleculas candidatas como BMKs de progresion y de e#12;cacia terapeutica en la enfermedad de Chagas. En nuestro laboratorio identi#12;camos los antgenos KMP11, PFR2, HSP70 y 3973d de T. cruzi los cuales son reconocidos con una alta especi#12;cidad y sensibilidad por el suero de pacientes en fase cronica de la enfermedad, produci endose una cada de la reactividad del suero frente a ellos tras un corto periodo de tiempo tras el tratamiento con benznidazol (6 a 9 meses). En este contexto y, con el #12;n de evaluar la utilidad de estos BMKs como herramienta para determinar la e#12;cacia terapeutica, se ha analizado la reactividad del suero de 66 pacientes con Chagas cronico en fase indeterminada, frente al mencionado set de antgenos, antes (T0) y tras el tratamiento con benznidazol (a T9, T24 y T48 meses). Se establecio un algoritmo de medida sobre la base del criterio de e#12;cacia terapeutica asociado al hecho de detectar una cada continua en la reactividad frente a los 4 BMKs, as como que la disminucion en la reactividad a los 24/48 meses post-tratamiento debe de ser, al menos, para dos de los biomarcadores del 40% (para los antgenos KMP11, PFR2 y 3973d) o del 30% (para HSP70), respecto a la reactividad observada previamente al tratamiento. Los resultados obtenidos muestran que un 42.4% de los pacientes cumplen el criterio de e#12;cacia terapeutica a 24 meses (con valor predictivo del 85 %) y un 68.8% de pacientes a los 48 meses. Ninguno de los pacientes que cumplen el criterio de e#12;cacia terapeutica presenta una PCR positiva tras el tratamiento con benznidazol. En resumen, estimamos que el mencionado set de BMKs constituye una herramienta util para monitorizar, en un corto periodo de tiempo, el impacto del tratamiento en pacientes con Chagas cronico

YKT6 expression, exosome release, and survival in non-small cell lung cancer

BACKGROUND: Cancer-derived exosomes are involved in metastasis. YKT6 is a SNARE protein that participates in the regulation of exosome production and release, but its role in non-small cell lung cancer (NSCLC) has not been examined. MATERIALS AND METHODS: Ultracentrifugation-purified exosomes from the A549 cell line were studied by CRYO-TEM, nanoparticle tracking analysis and western blot (TSG101 marker). YKT6 was inhibited using a DsiRNA and selected pre-microRNAs. MicroRNAs targeting YKT6 were validated by Renilla/Luciferase assay and western blot. YKT6 expression and its prognostic impact were analyzed in 98 tissue specimens from resected NSCLC patients. RESULTS: Membranous nanosized vesicles (mode size: 128nm) with TSG101 protein were purified from A549 cells. YKT6 inhibition reduced exosome release by 80.9%. We validated miR-134 and miR-135b as miRNAs targeting YKT6, and transfection with the pre-miRNAs also produced a significant reduction in exosome release. The analysis of YKT6 in tumor samples showed that patients with high levels had shorter disease-free and overall survival. CONCLUSIONS: YKT6 is a key molecule in the regulation of exosome release in lung cancer cells and is in turn precisely regulated by miR-134 and miR-135b. Moreover, YKT6 levels impact prognosis of resected NSCLC patients

miR-141 and miR-200c as markers of overall survival in early stage non-small cell lung cancer adenocarcinoma

Several treatments in non-small cell lung cancer (NSCLC) are histology-dependent, and the need for histology-related markers is increasing. MicroRNAs (miRNAs) are promising molecular markers in multiple cancers and show differences in expression depending on histological subtype. The miRNA family miR-200 has been associated with the regulation of epithelial-mesenchymal (EMT)/mesenchymal-epithelial transition (MET). EMT involves profound phenotypic changes that include the loss of cell-cell adhesion, the loss of cell polarity, and the acquisition of migratory and invasive properties that facilitates metastasis. A dual role for the miR-200 family in the prognosis of several tumors has been related to tumor cell origin. However, the prognostic role and function of miR-200 family in early-stage NSCLC adenocarcinoma and squamous cell carcinoma (SCC) have not been well established. Methods: miRNA expression was determined using TaqMan assays in 155 tumors from resected NSCLC patients. Functional studies were conducted in three NSCLC cell lines: H23, A-549 and HCC-44. Results: High miR-200c expression was associated with shorter overall survival (OS) in the entire cohort (p = 0.024). High miR-200c (p = 0.0004) and miR-141 (p = 0.009) expression correlated with shorter OS in adenocarcinoma - but not in SCC. In the multivariate analysis, a risk score based on miR-141 and miR-200c expression emerged as an independent prognostic factor for OS in the entire cohort (OR, 2.787; p = 0.033) and in adenocarcinoma patients (OR, 10.649; p = 0.002). Functional analyses showed that miR-200c, was related to mesenchymal-epithelial transition (MET) and affected cell migration and E-cadherin levels, while overexpression of miR-141 reduced KLF6 protein levels and produced an increase of secretion of VEGFA in vitro (H23, p = 0.04; A-549, p = 0.03; HCC-44, p = 0.02) and was associated with higher blood microvessel density in patient tumor samples (p<0.001). Conclusion: High miR-141 and miR-200c expression are associated with shorter OS in NSCLC patients with adenocarcinoma through MET and angiogenesis

miR-141 and miR-200c as markers of overall survival in early stage non-small cell lung cancer adenocarcinoma

Several treatments in non-small cell lung cancer (NSCLC) are histology-dependent, and the need for histology-related markers is increasing. MicroRNAs (miRNAs) are promising molecular markers in multiple cancers and show differences in expression depending on histological subtype. The miRNA family miR-200 has been associated with the regulation of epithelial-mesenchymal (EMT)/mesenchymal-epithelial transition (MET). EMT involves profound phenotypic changes that include the loss of cell-cell adhesion, the loss of cell polarity, and the acquisition of migratory and invasive properties that facilitates metastasis. A dual role for the miR-200 family in the prognosis of several tumors has been related to tumor cell origin. However, the prognostic role and function of miR-200 family in early-stage NSCLC adenocarcinoma and squamous cell carcinoma (SCC) have not been well established. Methods: miRNA expression was determined using TaqMan assays in 155 tumors from resected NSCLC patients. Functional studies were conducted in three NSCLC cell lines: H23, A-549 and HCC-44. Results: High miR-200c expression was associated with shorter overall survival (OS) in the entire cohort (p = 0.024). High miR-200c (p = 0.0004) and miR-141 (p = 0.009) expression correlated with shorter OS in adenocarcinoma - but not in SCC. In the multivariate analysis, a risk score based on miR-141 and miR-200c expression emerged as an independent prognostic factor for OS in the entire cohort (OR, 2.787; p = 0.033) and in adenocarcinoma patients (OR, 10.649; p = 0.002). Functional analyses showed that miR-200c, was related to mesenchymal-epithelial transition (MET) and affected cell migration and E-cadherin levels, while overexpression of miR-141 reduced KLF6 protein levels and produced an increase of secretion of VEGFA in vitro (H23, p = 0.04; A-549, p = 0.03; HCC-44, p = 0.02) and was associated with higher blood microvessel density in patient tumor samples (p<0.001). Conclusion: High miR-141 and miR-200c expression are associated with shorter OS in NSCLC patients with adenocarcinoma through MET and angiogenesis