Biochemical Links between Hormonal Mediators of Psychological Stress and the Expression of MUC1 in Prostate Cancer Cells

A growing body of scientific evidence has demonstrated that chronic psychological stress can not only increase the growth and metastasis of tumors through a number of mechanisms, such as by an increase of VEGF and Bcl-2, but also can decrease the survival of cancer patients. However, no studies have reported the effect of psychological stress with respect to the tumor marker MUC1. Therefore, building upon previous research, the purpose of the present study was to investigate the effect of the stress hormones cortisol and norepinephrine on the tumor marker MUC1, which is highly associated with tumor cell metastasis and is aberrantly glycosylated in most human epithelial carcinomas. Thus, it has been widely used in clinics as an important prognostic marker of disease progression and response to treatment. Overexpression of MUC1 in prostate cancer has been associated with more aggressive disease and an increased risk of recurrence. Using the DU-145 prostate cancer cell line as an experimental model, we sought to determine whether the glucocorticoid cortisol and the catecholamine norepinephrine enhanced the expression of MUC1 at the transcriptional and protein levels, and whether increased MUC1 altered the invasive potential of DU-145 cells. The levels of MUC1 protein expression were assayed by ELISA, flow cytometry, and the colorimetric bradford assay. The mRNA levels of MUC1 were measured by RT-PCR. In addition, cell invasiveness and migration were assayed by the matrigel migration assay. The results indicate that physiologically relevant concentrations of cortisol found in tumor microenvironment (10[superscript -7] M) enhanced the expression of MUC1 by approximately 2-fold after 6 or 10 days of treatment as assayed by ELISA. In addition, flow cytometric analyses revealed that DU-145 cells treated for 3 or 6 days with cortisol up-regulated the cell-surface expression of MUC1 by approximately 2-fold, whereas a 10 day exposure up-regulated the expression by 7-fold. Norepinephrine alone did not alter the expression of MUC1 at any time point in any of the experiments. In addition to these elevated levels of MUC1 protein, the mRNA levels of MUC1 were increased by 6-fold when cells were treated with cortisol for 6 days and by 4-fold when cells were treated for 10 days, while norepinephrine had no effect on mRNA levels. In addition, the matrigel migration assay indicated that cells treated with cortisol for 6-10 days migrated faster through the membrane as compared to untreated cells. Together, data generated from this thesis provide novel evidence of a biochemical link between the glucocorticoid cortisol, a hormonal mediator of psychological stress, and increased levels of the tumor marker MUC 1. Findings arising from this thesis raise the possibility that in prostate cancer the interaction of MUC1 with stress hormones, such as cortisol may increase the expression of MUC1 resulting in the observed increase in disease in psychologically stressed individuals. These novel findings highlight the necessity for future studies designed to investigate further the relationship between hormonal mediators of psychological stress and increased levels of MUC1

High Altitude Science Experiments aboard NASA’s WB-57 Airborne Research Platform

Purpose: Exposure to space radiation may place astronauts at significant health risks. This is an under-investigated area of research and therefore more knowledge is needed to better plan long-term space missions. The purpose of this study was to assess the effect of radiation on murine naïve and activated T lymphocytes (T cells) and to test the effectiveness of thermal, radiation and flight tracking technology in biological scientific payloads. We cultured cells in specific cytokines known to increase their viability and exposed them to either flight or had them as ground controls. Flight cells were kept under proper environmental conditions by using an active thermal system, whereas the levels of radiation were measured by NASA’s Timepix radiation sensor during ascent, cruise at 60,000 feet, and descent. In addition, an Automatic Dependent Surveillance Broadcast (ADS-B) device was utilized to track the state vector of the aircraft during flight

Assessment of Scientific Payload Carrying Spirulina Onboard Blue Origin’s New Shepard Vehicle

The research team at ERAU and UTHSCSA analyzed the effects of suborbital flight stressors and various light conditions (red, white, no light) on the Arthrospira platensis (Spirulina), onboard Blue Origin’s New Shepard vehicle. Commercially available cyanobacterium species were cultivated and closely monitored in mother colonies several months before the suborbital flight mission. The aim of this study was to estimate the biomass production and growth as a potential dietary alternative for prospective human spaceflight\u27s life support system. Spirulina samples were flown in a NanoLab with adjacent avionics supporting the light conditions and sensors to monitor the temperature, relative humidity, and accelerations. The various flight parameters measured in the NanoLab were validated with the flight data gathered by Nanoracks, the flight integrator. Thus, we also assessed the effect of microgravity and different light conditions on the gene expression. Our data indicates that the Spirulina samples onboard the rocket had significant (p \u3c 0.01-0.0001) downregulation of majority of the gene expression

[23327774 - Gravitational and Space Research] Investigation of Murine T-Cells and Cancer Cells under Thermal Stressors and 2D Slow Rotating System Effects as a Testbed for Suborbital Flights.pdf

Research indicates that exposure to microgravity leads to immune system dysregulation. However, there is a lack of clear evidence on the specific reasons and precise mechanisms accounting for these immune system changes. Past studies investigating space travel-induced alterations in immunological parameters report many conflicting results, explained by the role of certain confounders, such as cosmic radiation, individual body environment, or differences in experimental design. To minimize the variability in results and to eliminate some technical challenges, we advocate conducting thorough feasibility studies prior to actual suborbital or orbital space experiments. We show how exposure to suborbital flight stressors and the use of a two-dimensional slow rotating device affect T-cells and cancer cells survivability. To enhance T-cell activation and viability, we primed them alone or in combination with IL-2 and IL-12 cytokines. Viability of T-cells was assessed before, during the experiment, and at the end of the experiment for which T-cells were counted every day for the last 4 days to allow the cells to form clear structures and do not disturb their evolution into various geometries. The slow rotating device could be considered a good system to perform T-cell activation studies and develop cell aggregates for various types of cells that react differently to thermal stressors

Examination of Molecular Mechanisms on Vascular Formation and Stress Response in Zebrafish by Different Microgravity Environments

It has been proved that the presence of humans in space requires meticulous mission design and critical understanding of physiological parameters. Space is a hostile environment that has caused numerous health hazards in astronauts, including alterations in vascularization systemand high rates of muscle atrophy. Therefore, understanding the molecular pathways mediating space-induced alterations on human physiology is a necessity in making future missions a success. The goal of this study is to use zebrafish (Danio rerio) embryos as a unique model, which has genomic similarities to humans, to study molecular mechanisms of simulated and real microgravity effect on vascularization system and stress response. To simulate microgravity, we used a two-dimensional clinorotation device to expose zebrafish embryos at 1-day postembryonic fertilization and lasting for four days. Changes of 38 genes expression was measured by q-PCR. Thus, we used a unique zebrafish strain labelled with fluorescent protein allowing to image vascularization system using state of the art confocal microscopy. Our preliminary results indicate that only a small proportion of genes are affected by early simulation of microgravity. Our next goal is to confirm our findings by exposing zebrafish embryos to microgravity during suborbital flight. Our project entitled Muscular characterization in Microgravity Universal Spacelab (McXIMUS) is a joint research collaboration between the Embry- Riddle Aeronautical University (ERAU) and the University of Texas Health Science Center in San Antonio (UTHSCSA) to fly a suborbital payload aboard Blue Origin’s New Sheppard vehicle this Spring, 2019. To ensure the safe environment for zebrafish embryos during the suborbital flight, we designed a NanoLab to guarantee stable thermal conditions inside the payload. Our team has established proper procedures and validation checks to maximize the outcome of this novel scientific experiment. To the best of our knowledge, this is a first time when Danio rerio will be flown on suborbital flight mission to assess microgravity induced alterations on vascularization system and stress. Data obtained from this experiment will give insights into molecular pathways mediating vascular system and stress response and will assist in mapping out the strategies aimed to minimize the antagonizing effect of space travel. A comparative analysis of the pool of genes affected by different types of microgravity platforms and flight stressors (temperature, accelerations, and vibrations) across flight, ground and lab controls will be provided. Thus, this study has a potential to lay foundation for orbital experiments

T3-F: Educational Experiences and Lessons Learned in the Multidisciplinary Design, Fabrication, Integration and Flight Testing of Embry-Riddle High Altitude Science Engineering Rig (ERHASER) Payload aboard NASA’s WB-57 Aircraft

This paper describes the unique educational experiences and highlights the lessons learned during the multidisciplinary design, fabrication, integration and flight testing preparation of our prospective payload as part of the NASA’s Student Opportunities in Airborne Research (SOAR) pilot program aboard the WB57 aircraft. Our payload was comprised of several modular experiments referred as the Embry-Riddle High Altitude Science Engineering Rig (ERHASER), which was tested at about 60,000 feet during an analog suborbital trajectory over the Gulf of Mexico. One of the ERHASER’s experiments was dedicated to fly an ADS-B technology kit that can enhance students’ knowledge in Science Technology Engineering and Mathematics (STEM) with emphasis in aviation, and understand some of the challenges the Federal Administration Aviation (FAA) is facing with integrating new emerging era of suborbital space vehicles into the National Air Space. Understanding suborbital requirements, procedures and ADS-B performance are critical to better assess prospective point-to-point suborbital flights. This is a great opportunity for Embry-Riddle students to use the WB-57 research platform as a high-altitude performance aircraft in testing the functionality of ADS-B technologies during these analog suborbital trajectories. ERHASER’s second experiment consisted of radiation environmental measurements to test its effects on in-vitro biological alterations. Our biological system was composed of murine T-cells primed with different cytokines and cells treated with medicinal plant supercritical extracts. The goal of this study is to investigate the radiation induced cellular damage on these murine immune cells during the WB-57 flight, and to determine the role of supercritical extracts in reversing the epigenetic changes potentially induced by exposure to radiation. These unique experiences provide guidelines that helped faculty to work with students from different disciplines to design, fabricate, integrate and conduct flight tests successfully

Investigation of Zebrafish Larvae Behavior as Precursor for Suborbital Flights: Feasibility Study

Suborbital spaceflights, carrying scientific payloads, allow scientists not only to test the feasibility of their payloads, but they also provide the basis for refining scientific hypotheses to be later tested on the International Space Station (ISS). Therefore, it is essential to establish robust pre-flight procedures in order to take advantage of this unique research platform to facilitate payload delivery. In the present study, we assessed zebrafish larvae behavior as a precursor for the future suborbital spaceflight involving research on the musculoskeletal system. Zebrafish larvae were exposed to the same physiological stressors they would encounter during suborbital spaceflight: alterations in light, thermal, and centrifugation conditions. Their behavioral responses were analyzed using the DanioVision (Noldus) behavioral tracking system. Our results showed that zebrafish were most active when kept in a dark environment as measured by swim distance. Also, thermal alterations revealed that zebrafish larvae adapted well to the different temperatures ranging from 25°C to 32°C with the highest levels of locomotor activity observed at 32°C. Finally, the centrifugation tests demonstrated that although zebrafish were exhausted initially, their recovery process was short, lasting for approximately five minutes. Taken together, our findings support the hypothesis that using zebrafish larvae is a feasible model for future suborbital flights. Thus, the lessons learned allow us to propel this research with more refined and realistic procedures as a precursor for orbital flights to the ISS and to cis-lunar space

Operations and Testing of a Suborbital Research Payload

Embry-Riddle Aeronautical University (ERAU) has secured the opportunity to send a 2U (10cm x 10cm x 20cm) size payload onboard Blue Origin’s New Shepard capsule in the fall of 2017 to conduct a science research experiment. ERAU’s students will be working to design and test a NanoLab payload capable of withstanding the forces of a suborbital space launch. This NanoLab will be recording data through various phases of the 11-minutes suborbital flight. This is a research collaboration between two research teams at ERAU, as well as students and faculty from the University of Texas Health Science Center at San Antonio (UTHSCSA) and the Medical University of South Carolina (MUSC). The two research teams from Embry-Riddle consist of an operations and testing team and an engineering team who will work together to develop and safe operations of the suborbital payload

Educational Experiences and Lessons Learned in the Multidisciplinary Design, Fabrication, Integration and Pre-Flight Testing of Embry-Riddle High Altitude Science Engineering Rig (ERHASER) Payload Aboard NASA\u27s WB-57 Aircraft

This paper describes the unique educational experiences and highlights the lessons learned during the multidisciplinary design, fabrication, integration and flight testing preparation of our prospective payload as part of the NASA’s Student Opportunities in Airborne Research (SOAR) pilot program aboard the WB57 aircraft. Our payload was comprised of several modular experiments referred as the Embry-Riddle High Altitude Science Engineering Rig (ERHASER), which was tested at about 60,000 feet during an analog suborbital trajectory over the Gulf of Mexico. One of the ERHASER’s experiments was dedicated to fly an ADS-B technology kit that can enhance students’ knowledge in Science Technology Engineering and Mathematics (STEM) with emphasis in aviation, and understand some of the challenges the Federal Administration Aviation (FAA) is facing with integrating new emerging era of suborbital space vehicles into the National Air Space. Understanding suborbital requirements, procedures and ADS-B performance are critical to better assess prospective point-to-point suborbital flights. This is a great opportunity for Embry-Riddle students to use the WB-57 research platform as a high-altitude performance aircraft in testing the functionality of ADS-B technologies during these analog suborbital trajectories. ERHASER’s second experiment consisted of radiation environmental measurements to test its effects on in-vitro biological alterations. Our biological system was composed of murine T-cells primed with different cytokines and cells treated with medicinal plant supercritical extracts. The goal of this study is to investigate the radiation induced cellular damage on these murine immune cells during the WB-57 flight, and to determine the role of supercritical extracts in reversing the epigenetic changes potentially induced by exposure to radiation. These unique experiences provide guidelines that helped faculty to work with students from different disciplines to design, fabricate, integrate and conduct flight tests successfully