Neonatal exposure to xenobiotic estrogen alters the adult immune response and exacerbates endometriosis in mice [abstract]

Faculty Mentor: Dr. Susan C. Nagel, Obstetrics/Gynecology, and Women's HealthAbstract only availableEndometriosis is a common medical condition affecting 5-10% of women worldwide, and results in severe cramps, pelvic pain, and infertility. The cause of the disease is still unknown. Endometriosis occurs when endometrial tissue, which escapes into the peritoneal cavity via retrograde menstruation, adheres to other tissues in the cavity and causes irritated, inflamed lesions. Studies have suggested that the risk of developing endometriosis increases in women who have been exposed to xenobiotic (foreign to the body) estrogens during developmental stages of life. Thus, it is our hypothesis that programming of the immune system by xenoestrogens during development could potentially exacerbate endometriosis. This could occur by altering the peritoneal environment and/or the invading endometrial tissue. Therefore, it is our goal to study the effects of neonatal xenoestrogen exposure on the immune system; and ultimately, on the establishment of endometriosis in adulthood. In order to study this response, we dosed two strains of mice (CD1 and C57) with xenobiotic estrogens on postnatal days 2-14. In experiment A, CD1 mice were dosed with vehicle control (corn oil), 20 µg/kg/day, or 200 µg/kg/day bisphenol A. In experiment B, C57 mice were dosed with a vehicle control (corn oil) or 0.1 µg/kg/day diethylstilbestrol. At 8 weeks of age, endometriosis was induced in each strain via both a surgical induction and an injection technique. At 12 weeks, the endometriotic implants were counted and weighed to determine which mice had a greater susceptibility to the condition. Our next objective will be to analyze peritoneal fluid from the treated mice to identify key immune functions (for example, the release of certain cytokines) that may have been programmed by developmental xenoestrogen exposure.Endometriosis is a common medical condition affecting 5-10% of women worldwide, and results in severe cramps, pelvic pain, and infertility.  The cause of the disease is still unknown.  Endometriosis occurs when endometrial tissue, which escapes into the peritoneal cavity via retrograde menstruation, adheres to other tissues in the cavity and causes irritated, inflamed lesions.  Studies have suggested that the risk of developing endometriosis increases in women who have been exposed to xenobiotic (foreign to the body) estrogens during developmental stages of life.  Thus, it is our hypothesis that programming of the immune system by xenoestrogens during development could potentially exacerbate endometriosis.  This could occur by altering the peritoneal environment and/or the invading endometrial tissue.  Therefore, it is our goal to study the effects of neonatal xenoestrogen exposure on the immune system; and ultimately, on the establishment of endometriosis in adulthood.  In order to study this response, we dosed two strains of mice (CD1 and C57) with xenobiotic estrogens on postnatal days 2-14.  In experiment A, CD1 mice were dosed with vehicle control (corn oil), 20 µg/kg/day, or 200 µg/kg/day bisphenol A.  In experiment B, C57 mice were dosed with a vehicle control (corn oil) or 0.1 µg/kg/day diethylstilbestrol.  At 8 weeks of age, endometriosis was induced in each strain via both a surgical induction and an injection technique.  At 12 weeks, the endometriotic implants were counted and weighed to determine which mice had a greater susceptibility to the condition.  Our next objective will be to analyze peritoneal fluid from the treated mice to identify key immune functions (for example, the release of certain cytokines) that may have been programmed by developmental xenoestrogen exposure

Neonatal exposure to xenobiotic estrogen may alter the adult immune response and exacerbate endometriosis in mice [abstract]

Abstract only availableEndometriosis is a common medical condition affecting 5-10% of women worldwide and often results in severe cramps, pelvic pain, and infertility. The condition occurs when endometrial tissue, which escapes into the peritoneal cavity via retrograde menstruation, adheres to peritoneal cavity tissues and causes irritated, inflamed lesions. Studies have suggested that the risk of developing endometriosis increases in women who have been exposed to xenobiotic (foreign to the body) estrogens during development. This could be due to developmental programming of the peritoneal environment, and specifically, an altered immune function within this environment. Therefore, it is our hypothesis that developmental programming by xenoestrogens alters the immune response to shed endometrial tissue and exacerbates endometriosis. To better understand the role of xenoestrogens in immune programming, we are conducting our studies using a mouse model of surgically induced endometriosis. In particular, we are concentrating on two major aspects of immunity: 1) the presence of immune cells and 2) the function of those cells. Our study of the former is being largely performed using methods of immunohistochemistry (IHC). IHC allows us to quantify the macrophages present in the peritoneal fluid of experimental mice (exposed to diethylstilbestrol) versus control mice (no xenoestrogen exposure). In order to study our second focus, immune cell function, we are using a cytokine antibody array to determine the relative cytokine concentrations in the peritoneal fluid samples. By identifying the degree to which certain cytokine concentrations differ, we hope to better understand the effect of xenoestrogen exposure on immune cell function.Life Sciences Undergraduate Research Opportunity Progra

DULIP: A dual luminescence-based co-immunoprecipitation assay for interactome mapping in mammalian cells

Mapping of protein-protein interactions (PPIs) is critical for understanding protein function and complex biological processes. Here, we present DULIP, a dual luminescence-based co-immunoprecipitation assay, for systematic PPI mapping in mammalian cells. DULIP is a second-generation luminescence-based PPI screening method for the systematic and quantitative analysis of co-immunoprecipitations using two different luciferase tags. Benchmarking studies with positive and negative PPI reference sets revealed that DULIP allows the detection of interactions with high sensitivity and specificity. Furthermore, the analysis of a PPI reference set with known binding affinities demonstrated that both low- and high-affinity interactions can be detected with DULIP assays. Finally, using the well-characterized interaction between Syntaxin-1 and Munc18, we found that DULIP is capable of detecting the effects of point mutations on interaction strength. Taken together, our studies demonstrate that DULIP is a sensitive and reliable method of great utility for systematic interactome research. It can be applied for interaction screening as well as for the validation of PPIs in mammalian cells. Moreover, DULIP permits the specific analysis of mutation-dependent binding patterns

The Effects of Port Water Injection on Spark Ignition Engine Performance and Emissions Fueled by Pure Gasoline, E5 and E10

It has been proven that vehicle emissions such as oxides of nitrogen (NOx) are negatively affecting the health of human beings as well as the environment. In addition, it was recently highlighted that air pollution may result in people being more vulnerable to the deadly COVID-19 virus. The use of biofuels such as E5 and E10 as alternatives of gasoline fuel have been recommended by different researchers. In this paper, the impacts of port injection of water to a spark ignition engine fueled by gasoline, E5 and E10 on its performance and NOx production have been investigated. The experimental work was undertaken using a KIA Cerato engine and the results were used to validate an AVL BOOST model. To develop the numerical analysis, design of experiment (DOE) method was employed. The results showed that by increasing the ethanol fraction in gasoline/ethanol blend, the brake specific fuel consumption (BSFC) improved between 2.3% and 4.5%. However, the level of NOx increased between 22% to 48%. With port injection of water up to 8%, there was up to 1% increase in engine power whereas NOx and BSFC were reduced by 8% and 1%, respectively. The impacts of simultaneous changing of the start of combustion (SOC) and water injection rate on engine power and NOx production was also investigated. It was found that the NOx concentration is very sensitive to SOC variation

Alertness Training Increases Visual Processing Speed in Healthy Older Adults

In this study, we investigated whether alertness training in healthy older adults increases visual processing speed (VPS) and whether functional connectivity in the cingulo-opercular network predicts training gain. Using the theory of visual attention, we derived quantitative estimates of VPS before and after training. In Study 1, 75 healthy older adults participated in alertness training, active-control training, or no training (n = 25 each). A significant Group × Session interaction indicated an increase in VPS in the alertness-training group but not in the control group, despite VPS not differing significantly between groups before training. In Study 2, 29 healthy older adults underwent resting-state functional MRI and then participated in alertness training. Pretraining functional connectivity in the cingulo-opercular network correlated with the individual training-induced change in VPS. In conclusion, results indicate that alertness training improves visual processing in older adults and that functional connectivity in the cingulo-opercular network provides a neural marker for predicting individual training gain

OmoMYC blunts promoter invasion by oncogenic MYC to inhibit gene expression characteristic of MYC-dependent tumors.

MYC genes have both essential roles during normal development and exert oncogenic functions during tumorigenesis. Expression of a dominant-negative allele of MYC, termed OmoMYC, can induce rapid tumor regression in mouse models with little toxicity for normal tissues. How OmoMYC discriminates between physiological and oncogenic functions of MYC is unclear. We have solved the crystal structure of OmoMYC and show that it forms a stable homodimer and as such recognizes DNA in the same manner as the MYC/MAX heterodimer. OmoMYC attenuates both MYC-dependent activation and repression by competing with MYC/MAX for binding to chromatin, effectively lowering MYC/MAX occupancy at its cognate binding sites. OmoMYC causes the largest decreases in promoter occupancy and changes in expression on genes that are invaded by oncogenic MYC levels. A signature of OmoMYC-regulated genes defines subgroups with high MYC levels in multiple tumor entities and identifies novel targets for the eradication of MYC-driven tumors

Efectos sobre la salud humana de los campos magnéticos y eléctricos de muy baja frecuencia (ELF)

El presente documento, promovido por la Dirección General de Seguridad y Salud Laboral de la Consejería de Empleo de la Junta de Andalucía, dentro de la Acción 77 del Plan General de Prevención de Riesgos Laborales 2003-2008, pretende establecer el marco teórico actual de conocimiento sobre los campos ELF; explicar los conceptos fundamentales de los mismos y las fuentes que los generan; establecer los principales efectos biológicos que se producen en los seres humanos a causa de su exposición, y las posibles enfermedades que son motivo de estudio. Asimismo, se establece una comparación entre las dos guías comentadas anteriormente sobre niveles de exposición, y que tienen un reconocimiento importante a nivel internacional. También son objeto de análisis los principales tipos de estudios (epidemiológicos, de laboratorio, etc…); los niveles de exposición poblacionales, tanto para el público en general como para el ocupacional; los métodos de medida e instrumentación empleados; las posibles medidas de protección y prevención; y la normativa relacionada con los campos magnéticos ELF. Se citan además las líneas de investigación recomendadas por la Organización Mundial de la Salud para completar las lagunas existentes de la base científica desarrollada. Finalmente, cabe decir que se ha pretendido citar los principales documentos de recopilaciones bibliográficas, cuya consulta pueda proporcionar información más detallada cuando ésta se requiera.Consejería de Empleo. Junta de Andalucía. Españ

Fructose Intake: Metabolism and Role in Diseases

Fructose consumption has dramatically increased worldwide over the past decades. There are numerous clinical, experimental, and epidemiological studies evidenced that increased consumption of fructose negatively impacts carbohydrate metabolism and lactate formed from fructose can also affect whole-body energy balance. Excessive fructose intake stimulates endogenous glucose production and lipid synthesis in the liver. Currently fructose is believed to be a major contributing factor to chronic metabolic diseases, including obesity, insulin resistance, hypertriglyceridemia, and non-alcoholic fatty liver disease, hyperglycemia, type 2 diabetes, and cancer. These new findings bring challenges to researchers today because of what is still to be discovered, and how to apply what has been discovered to modern health. Further investigation should seek to analyze and understand specific mechanistic effects of fructose in metabolic pathways, and how to apply this knowledge to our daily lives. Conducting this monosaccharide research is important to improve the diet of the general population and to attenuate the epidemics of metabolic disease and associated diseases. Here, we focus on the mechanism and role of fructose in diseases as well as its potential as a dietary interventional target

Cardiac-Specific Expression of the Tetracycline Transactivator Confers Increased Heart Function and Survival Following Ischemia Reperfusion Injury

Mice expressing the tetracycline transactivator (tTA) transcription factor driven by the rat α-myosin heavy chain promoter (α-MHC-tTA) are widely used to dissect the molecular mechanisms involved in cardiac development and disease. However, these α-MHC-tTA mice exhibit a gain-of-function phenotype consisting of robust protection against ischemia/reperfusion injury in both in vitro and in vivo models in the absence of associated cardiac hypertrophy or remodeling. Cardiac function, as assessed by echocardiography, did not differ between α-MHC-tTA and control animals, and there were no noticeable differences observed between the two groups in HW/TL ratio or LV end-diastolic and end-systolic dimensions. Protection against ischemia/reperfusion injury was assessed using isolated perfused hearts where α-MHC-tTA mice had robust protection against ischemia/reperfusion injury which was not blocked by pharmacological inhibition of PI3Ks with LY294002. Furthermore, α-MHC-tTA mice subjected to coronary artery ligation exhibited significantly reduced infarct size compared to control animals. Our findings reveal that α-MHC-tTA transgenic mice exhibit a gain-of-function phenotype consisting of robust protection against ischemia/reperfusion injury similar to cardiac pre- and post-conditioning effects. However, in contrast to classical pre- and post-conditioning, the α-MHC-tTA phenotype is not inhibited by the classic preconditioning inhibitor LY294002 suggesting involvement of a non-PI3K-AKT signaling pathway in this phenotype. Thus, further study of the α-MHC-tTA model may reveal novel molecular targets for therapeutic intervention during ischemic injury