Regulación y función de microRNAs en las alteraciones del testículo de ratón inducidas por exposición a mezclas de ftalatos y alquifenoles.

Los seres humanos y la vida animal están diariamente expuesta a mezclas de compuestos conocidos como perturbadores endocrinos (PEs), entre ellos destacan los ftalatos: [bis 2-etil-hexil ftalato (DEHP), dipentil ftalato (DBP), bencil-butil ftalato (BBP)] y los alquifenoles: [4-Nonilfenol (NP) y 4-terc-Octilfenol (OP)], por formar parte de la manufactura de productos de uso cotidiano, como plásticos y utensilios de limpieza, y liberarse de sus contenedores contaminando el medioambiente. Estos compuestos pueden alterar la homeostasis del sistema endocrino principalmente a nivel reproductivo, afectando la biosíntesis de testosterona o estrógenos. Dado que la producción de ambas hormonas controla la gametogénesis y la espermatogénesis la exposición a estos compuestos desde etapas muy tempranas del desarrollo gestacional, ha sido asociado a patologías reproductivas e infertilidad en los machos adultos. Los mecanismos por los cuales ejercen su acción aun son pocos conocidos. Sin embargo, involucran cambios en la expresión de transcritos que codifican proteínas importantes para el desarrollo y mantenimiento de la función testicular. En este aspecto toma relevancia el estudio de los microRNAs, que son RNAs pequeños no codificantes, que actúan como reguladores negativos de la expresión génica a nivel post-transcripcional. Sin embargo, se desconoce la participación de los microRNAs en las alteraciones testiculares inducidas por la exposición a mezclas de ftalatos y alquifenoles. Por ello, en esta tesis doctoral se planteó la hipótesis de “La exposición crónica a una mezcla definida de ftalatos y alquifenoles desregula los niveles de microRNAs, implicados en el balance testosterona/estrógenos lo cual afecta la fertilidad en ratones machos”. Para responder esta hipótesis, diseñamos en ratones un modelo de exposición crónica a una mezcla de los ftalatos DEHP, DBP y BBP en dosis de 0,3 mg/Kg/día cada uno y los alquifenoles NP y OP en dosis de 0,05 mg/Kg/día cada uno, estas dosis son equivalentes a los niveles de exposición en humanos. Esta mezcla fue administrada en el agua de ingesta de hembras preñadas desde el día 0,5 post-coitum hasta la adultez de las crías machos (60/75 días). Como grupo control a unos animales se les administro en el agua de bebida, únicamente los compuestos en que se diluyeron los ftalatos y alquifenoles. Nuestros principales hallazgos mostraron que, los testículos de ratones expuestos a la mezcla de ftalatos y alquifenoles, presentan una disminución en los niveles intratesticulares de estradiol vía cambios en los niveles de mensajero y proteínas de enzimas de la vía esteroidogénica (principalmente una disminución de aromatasa), alteraciones en la espermatogénesis y apoptosis de células germinales vía cambios en la expresión de proteínas implicadas en este proceso. Estas alteraciones fueron asociadas con un incremento en la expresión de transcritos que participan en biogénesis (Drosha), edición (Adar-1S) y estabilidad de miRNAs (Tut-4). Además, en los testículos de los animales expuestos detectamos por secuenciación masiva (NGS) 2 microRNAs sobre-expresados y 8 miRNAs disminuidos, estos datos fueron validados por qPCR, Junto con ello , se detecto por NGS 36 variantes de miRNAs (isomiRs) que presentaron cambios en sus niveles de expresión, siendo algunos de estos isomiRs, variantes de los miRNA canónicos alterados. Analizando las interacciones entre microRNAs/mRNAs se encontró que, la pérdida del control pos-transcripcional de miR-18a-5p hacia LxR-β estaría implicado en la sobreexpresión de Star, Cyp17a1 y en la disminución de Cyp19a1, lo cual serelaciona directamente con la disminución en la síntesis de estradiol que observamos en los animales expuestos.Peer reviewe

Auditoría tributaria a la Compañía OMECIV Constructora Cía. Ltda., de la provincia de Morona Santiago, cantón Taisha, del periodo 2015

La Auditoría Tributaria a la Compañía Omeciv Constructora Cía. Ltda., de la Provincia de Morona Santiago, Cantón Taisha, del Periodo 2015, con el propósito de evaluar el cumplimiento tributario de la empresa de acuerdo a los requerimientos de la Ley de Régimen Tributario Interno y su Reglamento interno. En la investigación preliminar se analizó la información en función de aspectos importantes tales como: identificación, base legal, misión, visión. Se analizó la documentación existente para verificar y analizar los datos además con la investigación de campo, mediante encuestas a los empleados y la entrevista al gerente de la compañía se determinó el entorno actual de la institución. En la planificación se elaboró un programa de Auditoría específica relacionado con el Impuesto al Valor Agregado y el Impuesto a la Renta, en la ejecución, se desarrolló el programa de Auditoría. Se pudo determinar que los principales retrasos se producen por las personas que laboran en el departamento de contabilidad, además se han encontrado inconsistencias en los formularios 104, formulario 103, analizado los estados financieros y los datos que se encuentran registrados en las declaraciones del Impuesto a la Renta anual. Se recomienda al gerente aplicar políticas tributarias en la compañía OMECIV y cumpla con las disposiciones legales de la normativa vigente y sus obligaciones con el propósito de evitar de parte de la administración tributaria multas e intereses.The tax audit to the Omeciv Constructora Company Cia. Ltda., from Morona Santiago Province, Canton Taisha, during the period 2015 was development with the purpose of evaluating the tax fulfillment of the company according to the requirements of the law of internal tax regime and its internal regulation. In the preliminary investigation, the information was analyzed in function of important aspects such as identification, legal basis, mission, vision. The existing documentation was analyzed to verify and analyze the data. Moreover, the current environment of the institution was identified with the field investigation supported on the employee surveys and the interview to the manager of the company. In the planning, it was elaborated a specific auditing program related to the Value-Added Tax and the Income Tax. In the execution, the audit program was developed. It was possible to determine that the people working in the accounting department produced the main delays. Besides, there were inconsistencies in the 104 and 103 forms once analyzed the financial statements and the data registered in the annual Income Tax return. It is recommended to the manager applying tax policies in the OMECIV Company and fulfill with the legal provisions of the current regulations and their obligations with the purpose of avoiding fines and interest by the tax administration

Proyecto para el incremento del servicio de transporte turístico a nivel internacional en la compañía Interangeles Tours Cía. Ltda. Riobamba - Ecuador

En el presente trabajo investigativo se propone un Proyecto para el incremento del Servicio de Transporte Turístico a nivel Internacional en la Compañía INTERANGELES TOURS CIA LTDA ubicada en la ciudad de Riobamba Ecuador. Para el levantamiento de la información se aplicó una encuesta al mercado objetivo, la cual posibilitó recolectar datos primarios necesarios para estructurar la oferta de paquetes. Una vez identificadas las fortalezas y debilidades a nivel interno e identificada claramente la demanda insatisfecha se procedió a diseñar un conjunto de paquetes turísticos con destinos en Colombia, Perú y Venezuela tendientes a captar la creciente demanda de este servicio; por otro lado se desarrolló el estudio técnico que entre otras cosas plantea la incorporación de nuevas unidades de transporte para la Compañía que le permitan brindar un servicio de calidad; y, finalmente se detalla el estudio financiero que consolida en términos monetarios los resultados de los estudios anteriores para posteriormente evaluar la propuesta y determinar la factibilidad de ejecutar el proyecto desde el punto de vista financiero. La principal conclusión gira en torno a la factibilidad de ejecutar la propuesta puesto que consideramos se ajusta perfectamente a las expectativas de crecimiento de la empresa, y sobre todo los resultados de la evaluación en términos financieros comprueban que es viable ejecutarla ya que arroja indicadores atractivos para sus inversionistas sobre cualquier otra alternativa de inversión en el mercado financiero. Finalmente se recomienda a la empresa tomar la decisión de ejecutar el proyecto, aprovechando la experiencia y acogida que ha venido ganando durante los últimos tiempos en el sector del transporte turístico, la cual combinada con una adecuada campaña de marketing de seguro le permitirá obtener y sobrepasar las proyecciones realizadas en la presente propuesta sin mayor dificultad.In this research a project for increasing Tourist Transport Service at INTERANGELES TOURS CIA LTA ternational Company located in Riobamba-Ecuador is proposed. data uprising a target market survey was applied, which enabled primary data collection needed to structure packages offering Once internal strengths and weaknesses and unmet demand were identified, a set of touristic packages as designed with destinations to Colombia, Peru and Venezuela, the purpose was to capture this service gowing demand; a technical study was also conducted which suggests new transportation units ncorporation for the Company to offer a quality service; and finally the financial study is detailed to patsolidate in monetary terms the results of previous studies to further evaluate the proposal and Beterminate the feasibility of implementing the project from the f the feasibility of implementing the project from the financial standpoint. The main conclusion deals with the feasibility to implement the proposal because it is considered to be perfectly adjusted to the company growth expectations, and the evaluation results in financial terms prove that it is feasible due to the attractive indicators for its investors over any other investment in the financial market. Finally it is recommended for the company to implement the project, drawing on the experience and acceptance that it has gained in recent years in the tourist transport sector, which combined with a proper marketing campaign will get and exceed projections made in this proposal without much difficulty

Chronic low-dose exposure to a mixture of environmental endocrine disruptors induces microRNAs/isomiRs deregulation in mouse concomitant with intratesticular estradiol reduction

16 p.-7 fig.-3 tab.Humans are environmentally exposed not only to single endocrine-disrupting chemicals (EDCs) but to mixtures that affect their reproductive health. In reproductive tissues, microRNAs (miRNAs) are emerging as key targets of EDCs. Here, we analysed changes in the testis “miRNome” (and their biogenesis mechanism) in chronically exposed adult mice to a cocktail of five EDCs containing 0.3 mg/kg-body weight (BW)/day of each phthalate (DEHP, DBP, BBP) and 0.05 mg/kg-BW/day of each alkylphenol (NP, OP), from conception to adulthood. The testis “miRNome” was characterised using next-generation sequencing (NGS). Expression levels of genes involved in miRNA biogenesis were measured by RT-qPCR, as well as several physiological and cytological parameters. We found two upregulated,and eight down-regulated miRNAs and thirty-six differentially expressed isomiRs along with an over-expression of Drosha, Adar and Zcchc11. A significant decrease of intratesticular estradiol but not testosterone was detected. Functional analysis showed altered spermatogenesis, germ cell apoptosis and negative correlation of miR-18a-5p with Nr1h2 involved in the deregulation of the steroidogenesis pathway. Here, we present the first association between miRNA/isomiRs deregulation, their mechanisms of biogenesis and histopathological and hormonal alterations in testes of adult mice exposed to a mixture of low-dose EDCs, which can play a role in male infertility.This work was supported by grants from FONDECYT (1150352) and CONICYT (21120505), Chile, and MINECO (BFU2013-42164-R), Spain.Peer reviewe

Frecuencia de leucemias y linfomas diagnosticados en el hospital de la sociedad de lucha contra el cáncer (SOLCA), de la ciudad de Cuenca durante el período 1996 a 2005

Se realizó un estudio descriptivo, a partir de la recolección de datos de las fichas médicas del Hospital de la Sociedad de Lucha contra el Cáncer (SOLCA), y se realizo una base de datos mediante el software Epi Info versión 3.5.3. y Microsoft Excel 2007. Resultados: Durante el período de estudio se diagnosticaron 1026 personas con Leucemias y Linfomas distribuidas en 453 Leucemias y 573 Linfomas, en relación a las Leucemias se encontró que el tipo más frecuente es Leucemia Linfoblástica Aguda con un 51.43% del total de casos diagnosticados de leucemias, la edad de mayor incidencia fue de 20 44 años con el 28.5%, el genero masculino afectado en 51.66% frente al femenino en 48.34%, procedentes de Azuay en el 55.41%, seguido por El Oro con el 22.96%. En relación a los Linfomas se encontró que el tipo más frecuente es el Linfoma no Hodgkin de células grandes (difuso) con un 38.57%, la edad de mayor incidencia fue de 61- 80 años 20.77% durante cada década, el genero masculino afectado en 50.3% frente al femenino en 49.7%, procedentes de Azuay en el 58.46%, seguido por El Oro con el 15.18%.auOBJECTIVE: To determine the frequency of leukemia’s and lymphomas diagnosed at the “Hospital de la Sociedad de Lucha Contra el Cancer (SOLCA) in the city of Cuenca during the period 1996 - 2005”. METHODS AND MATERIALS: A descriptive study from the collection of data from medical records of the Hospital of the Society against Cancer (SOLCA), and we made a database using Epi Info software version 3.5.3 and Microsoft Excel 2007. RESULTS: During the study period, 1026 people were diagnosed with leukemia and lymphoma leukemia distributed in 453 and 573 lymphomas in relation to leukemia was found that the most common type is acute lymphoblastic leukemia with a 51.43% of all diagnosed cases of leukemia The highest incidence age was 20 – 44 years with 28.5%, the male gender affected 51.66% versus 48.34% female, from Azuay in 55.41%, followed by the gold with 22.96%. In relation to the lymphoma was found that the most common type is the non-Hodgkin’s lymphoma large cell (diffuse) with 38.57% the highest incidence age was 65 years or more to 40.1%, the affected male gender 50.3 % vs. female 49.7%, from 58.46% in Azuay, followed by gold with 15.18%.MédicoCuenc

Effects of single vs mixed EDCs in mouse testes

11 p.-5 fig.-2 tab.Endocrine-disrupting chemicals (EDCs) generate reproductive dysfunctions affecting the biosynthesis of steroid hormones and genes of the steroidogenic pathway. EDCs effects are mainly reported as a result of exposure to single compounds. However, humans are environmentally exposed to a mixture of EDCs. Herein, we assess chronic exposure to single alkylphenols and phthalates versus a mixture in mouse testes histology and steroidogenesis. Pregnant mice were exposed through drinking water to: 0.3 mg/Kg-body weight (BW)/day of each phthalate (DEHP, DBP, BBP), 0.05 mg/Kg-BW/day of each alkylphenol (NP, OP), or their mixture, covering from 0.5 post-coital day to weaning, continuing in the male offspring each exposure until adulthood (60 days old). Body and relative testis weight were increased in mixture-exposed mice along with histological alterations. Intratesticular testosterone (T) changed only in mice exposed to DBP, whereas estradiol (E2) levels were altered in all groups (except BBP). mRNA levels of genes encoding hormones of the steroid pathway (Cyp11a1, Hsd3b1, Cyp17a1 and Cyp19a1), cholesterol transporters (Star), and transcriptional factors (Sp1) showed that mice exposed to single or mixed compounds had alterations in at least two transcripts. However, none of the different types of exposure induced changes in all transcripts. In addition, changes at the mRNA or protein levels with single compounds were not always the same as those with a mixture. In conclusion, the effects of a chronic exposure to a mixture of EDCs on the expression of genes and proteins of the steroidogenic pathway and hormonal status were different from those exposed to single EDC.This work was supported by grants from FONDECYT (1150352) and CONICYT (21120505),Chile, and MINECO (BFU2013-42164-R), Spain.Peer reviewe

Combined proteomic and miRNome analyses of mouse testis exposed to an endocrine disruptors chemicals mixture reveals altered toxicological pathways involved in male infertility

50 p.-6 fig.-1 tab.The increase in male idiopathic infertility has been associated with daily exposure to endocrine disruptors chemicals (EDCs). Nevertheless, the mechanisms of action in relation to dysregulating proteins and regulatory microRNAs are unknown.We combined proteomic and miRNome analyses of mouse testis chronically exposed to low doses of a define mixture of EDCs [phthalates: bis (2-ethylhexyl), dibutyl and benzyl-butyl; 4-nonylphenol and 4-tert-octylphenol], administered in the drinking water from conception until adulthood (post-natal day 60/75) and compared them with no-exposed control mice.We analysed fertility parameters and global changes in the patterns of mice testis proteome by 2D-electrophoresis/mass spectrometry, along with bioinformatic analyses of dysregulated microRNAs, and their association with published data in human infertile patients.We detected a decrease in the potential fertility of exposed mice associated with changes in the expression of 18 proteins (10 up-regulated, 8 down-regulated). Functional analysis showed that 89% were involved in cell death. Furthermore, we found a group of 23 microRNAs/isomiRs (down-regulated) correlated with six of the up-regulated target proteins (DIABLO, PGAM1, RTRAF, EIF4E, IVD and CNDP2). Regarding this, PGAM1 up-regulation was validated by Western blot and mainly detected in Sertoli cells. Some of these microRNA/protein dysregulations were reported in human testis with spermatogenic failure.Overall, a chronic exposure to EDCs mixture in human males could potentially lead to spermatogenic failure through changes in microRNA expression, which could post-transcriptionally dysregulate mRNA targets that encode proteins participating in cell death in testicular cells. Finally, these microRNA/protein dysregulations need to be validated with other EDCs mixtures and concentrations.This work was supported by grants from FONDECYT 1150352 to R.D.M., FONDECYT 3160273 to P.U-M and CONICYT 21120505 to J.B., Chile, and MINECO BFU2013-42164-R and BFU2017-87095-R to J. d. M., SpainPeer reviewe

Selective treatments of prostate tumor cells with a cold atmospheric plasma jet

International audienceBackground: Low temperature plasma is an ionised gas generated at atmospheric pressure with the temperature of heavy particles (ions, molecules, atoms) close to room temperature. Recent progress highlighted the efficiency of plasma to induce cell death in many cancer cell lines. In the current context of the emergence of the use of cold plasma for medical purposes, this article aims to study the effect of a cold plasma of helium on ex-vivo prostate cells.Methods: A cold atmospheric plasma torch was developed to treat different tumor cells. Parameters like electric voltage, gas flow, cell/torch distance, and exposure time were studied to improve the effects of plasma onto two human prostate tumor cells lines, with different metastatic potential, LNCaP and PC3 and then, compared to human prostate epithelial cells P69. The plasma torch operated by a voltage up to 10 kV, without any risk of arcing, exhibited a visible plasma plume up to about 70 mm, with a temperature close to room temperature. The induced temperature rise due to the plasma treatment is measured to ensure that observed results are not thermal effects. After plasma treatment, microscopy and MTT analyses are achieved to measure cells viability. An ordinary one-way ANOVA test were used between two groups with p-value less than 0.05.Results: An optimized protocol leads to the tumor cells death with regard to cell viability. Cold plasma treatment exhibits differential effect, regarding tumor status of cells without any significant influence of thermal effect (temperature increases of only few degrees Celcius above room temperature). The results show that non-tumor cells P69 are less sensitive to cold plasma exposure with a cell viability of 67% after 120 s of treatment, compared to tumor cell lines LNCaP and PC3, respectively at cell viability of 29% and 23% for the same treatment time.Conclusion: The cold helium plasma treatment showed a selective effect on prostate cells, with an enhanced apoptosis way for tumor cells, whereas the healthy cells are more resistant to the treatment. Moreover, during the treatment, the surface temperature rises only few degrees Celcius. This result is very promising for future in-vivo treatments

Selective treatments of prostate tumor cells with a cold atmospheric plasma jet

International audienc

Selective treatments of prostate tumor cells with a cold atmospheric plasma jet

International audienceBackground: Low temperature plasma is an ionised gas generated at atmospheric pressure with the temperature of heavy particles (ions, molecules, atoms) close to room temperature. Recent progress highlighted the efficiency of plasma to induce cell death in many cancer cell lines. In the current context of the emergence of the use of cold plasma for medical purposes, this article aims to study the effect of a cold plasma of helium on ex-vivo prostate cells.Methods: A cold atmospheric plasma torch was developed to treat different tumor cells. Parameters like electric voltage, gas flow, cell/torch distance, and exposure time were studied to improve the effects of plasma onto two human prostate tumor cells lines, with different metastatic potential, LNCaP and PC3 and then, compared to human prostate epithelial cells P69. The plasma torch operated by a voltage up to 10 kV, without any risk of arcing, exhibited a visible plasma plume up to about 70 mm, with a temperature close to room temperature. The induced temperature rise due to the plasma treatment is measured to ensure that observed results are not thermal effects. After plasma treatment, microscopy and MTT analyses are achieved to measure cells viability. An ordinary one-way ANOVA test were used between two groups with p-value less than 0.05.Results: An optimized protocol leads to the tumor cells death with regard to cell viability. Cold plasma treatment exhibits differential effect, regarding tumor status of cells without any significant influence of thermal effect (temperature increases of only few degrees Celcius above room temperature). The results show that non-tumor cells P69 are less sensitive to cold plasma exposure with a cell viability of 67% after 120 s of treatment, compared to tumor cell lines LNCaP and PC3, respectively at cell viability of 29% and 23% for the same treatment time.Conclusion: The cold helium plasma treatment showed a selective effect on prostate cells, with an enhanced apoptosis way for tumor cells, whereas the healthy cells are more resistant to the treatment. Moreover, during the treatment, the surface temperature rises only few degrees Celcius. This result is very promising for future in-vivo treatments