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At least 127 records · Page 7

Free-zone electrophoresis of animal cells. 1: Experiments on cell-cell interactions

The electrophoretically migrating zones wasa monitored. The absence of fluid flows in the direction of migration permits direct measurement of electrophoretic velocities of any material. Sedimentation is orthogonal to electrokinetic motion and the effects of particle-particle interaction on electrophoretic mobility is studied by free zone electrophoresis. Fixed erythrocytes at high concentrations, mixtures of fixed erythrocytes from different animal species, and mixtures of cultured human cells were studied in low ionic strength buffers. The electrophoretic velocity of fixed erythrocytes was not altered by increasing cell concentration or by the mixing of erythrocytes from different species. When zones containing cultured human glial cells and neuroblastoma cells are permitted to interact during electrophoresis, altered migration patterns occur. It is found that cell-cell interactions depends upon cell type.

Todd, P. W.↗

Facilities for Biological Research Aboard the International Space Station

A centrifuge designed as part of an integrated biological facility for installation onboard the International Space Station is presented. The requirements for the 2.5 m diameter centrifuge, which is designed for the support of biological experiments are discussed. The scientific objectives of the facility are to: provide a means of conducting fundamental studies in which gravitational acceleration is a controllable variable; provide a 1g control; determine the threshold acceleration for physiological response, and determine the value of centrifugation as a potential countermeasure for the biomedical problems associated with space flight. The implementation of the facility is reported on, and the following aspects of the facility are described: the host resources systems supply requirements such as power and data control; the habitat holding rack; the life sciences glove box; the centrifuge; the different habitats for cell culture, aquatic studies, plant research and insect research; the egg incubator, and the laboratory support equipment.

Souza, Kenneth A.↗

Constant Applied Force Stimulates Osteoblast Proliferation Via Matrix-Integrin-Signaling Pathways

Reduced weight-bearing caused by immobilization, bed-rest or microgravity leads to atrophy in mechanosensitive tissue such as muscle and bone. We hypothesize that bone tissue requires earth s gravity (1-g) for the maintenance of extracellular matrix, integrin, and kinase-mediated cell growth and survival pathways. We investigate the role of matrix-integrin signaling in bone cells using cell culture centrifugation to provide different levels of hypergravity mechanostimulation. The 10-50-g range we use also mimics physiological intermedullary pressure (1.2 - 5 kPa). 24 hours at 50-g increased primary rat osteoblast proliferation on collagen Type I and fibronectin, but not laminin or uncoated plastic. BrdU incorporation in primary osteoblasts over 24 h showed hypergravity increased the number of cells actively synthesizing DNA from about 60% at 1-g to over 90% at 25-g. Primary rat fibroblasts grown at 50-g (24 h) showed no proliferation increase, suggesting this is a tissue-specific phenomenon. These results suggest that the betal and alpha4 integrins may be involved. To further test this, we used osteocytic-like MLO-Y4 cells that showed increased proliferation at 1-g with stable expression of a betal integrin cytoplasmic tail and transmembrane domain construct. At 50-g, MLO-Y4/betal cells showed greater MAPK activation than MLO-Y4 vector controls, suggesting that betal integrin is involved in transducing mitogenic signals in response to hypergravity. Preliminary results also show that interfering with the alpha4 integrin in primary osteoblasts grown on fibronectin blocked the proliferation response. These results indicate that cells from mechanosensitive bone tissue can respond to gravity-generated forces, and this response involves specific matrix and integrin-dependent signaling pathways.

Vercoutere, W.↗

Cell Culture Unit

The unique capabilities of the Cell Culture Unit and hardware differences between the Science Evaluation Unit and the Flight Unit are briefly presented in this document.

Joe Parrish↗

Stable carbon isotope fractionation by sulfate-reducing bacteria

Biogeochemical transformations occurring in the anoxic zones of stratified sedimentary microbial communities can profoundly influence the isotopic and organic signatures preserved in the fossil record. Accordingly, we have determined carbon isotope discrimination that is associated with both heterotrophic and lithotrophic growth of pure cultures of sulfate-reducing bacteria (SRB). For heterotrophic-growth experiments, substrate consumption was monitored to completion. Sealed vessels containing SRB cultures were harvested at different time intervals, and delta(13)C values were determined for gaseous CO(2), organic substrates, and products such as biomass. For three of the four SRB, carbon isotope effects between the substrates, acetate or lactate and CO(2), and the cell biomass were small, ranging from 0 to 2 per thousand. However, for Desulfotomaculum acetoxidans, the carbon incorporated into biomass was isotopically heavier than the available substrates by 8 to 9 per thousand. SRB grown lithoautotrophically consumed less than 3% of the available CO(2) and exhibited substantial discrimination (calculated as isotope fractionation factors [alpha]), as follows: for Desulfobacterium autotrophicum, alpha values ranged from 1.0100 to 1.0123; for Desulfobacter hydrogenophilus, the alpha value was 0.0138, and for Desulfotomaculum acetoxidans, the alpha value was 1.0310. Mixotrophic growth of Desulfovibrio desulfuricans on acetate and CO(2) resulted in biomass with a delta(13)C composition intermediate to that of the substrates. The extent of fractionation depended on which enzymatic pathways were used, the direction in which the pathways operated, and the growth rate, but fractionation was not dependent on the growth phase. To the extent that environmental conditions affect the availability of organic substrates (e.g., acetate) and reducing power (e.g., H(2)), ecological forces can also influence carbon isotope discrimination by SRB.

Deltaproteobacteria/growth & development/metabolis↗

M-BAND Study of Radiation-Induced Chromosome Aberrations in Human Epithelial Cells: Radiation Quality and Dose Rate Effects

The advantage of the multicolor banding in situ hybridization (mBAND) technique is its ability to identify both inter- (translocation to unpainted chromosomes) and intra- (inversions and deletions within a single painted chromosome) chromosome aberrations simultaneously. To study the detailed rearrangement of low- and high-LET radiation induced chromosome aberrations in human epithelial cells (CH184B5F5/M10) in vitro, we performed a series of experiments with Cs-137 gamma rays of both low and high dose rates, neutrons of low dose rate and 600 MeV/u Fe ions of high dose rate, with chromosome 3 painted with multi-binding colors. We also compared the chromosome aberrations in both 2- and 3-dimensional cell cultures. Results of these experiments revealed the highest chromosome aberration frequencies after low dose rate neutron exposures. However, detailed analysis of the radiation induced inversions revealed that all three radiation types induced a low incidence of simple inversions. Most of the inversions in gamma-ray irradiated samples were accompanied by other types of intra-chromosomal aberrations but few inversions were accompanied by inter-chromosomal aberrations. In contrast, neutrons and Fe ions induced a significant fraction of inversions that involved complex rearrangements of both inter- and intrachromosomal exchanges. The location of the breaks involved in chromosome exchanges was analyzed along the painted chromosome. The breakpoint distribution was found to be randomly localized on chromosome 3 after neutron or Fe ion exposure, whereas non-random distribution with clustering breakpoints was observed after -ray exposure. Our comparison of chromosome aberration yields between 2- and 3-dimensional cell cultures indicated a significant difference for gamma exposures, but not for Fe ion exposures. These experimental results indicated that the track structure of the radiation and the cellular/chromosome structure can both affect radiation-induced chromosome aberrations.

Hada, Megumi↗

Decarbonizing the Building Sector: A Human-Centered Study Focused on Small/Light Commercial Building Energy Equity

Decarbonization of the building sector is no small feat; buildings account for 40% of primary energy consumption, and fossil-fuel combustion in buildings leads to roughly 30% of total greenhouse gas emissions. Energy efficiency, electrification and smart technologies are fundamental strategies to reduce consumption and shift away from fossil-fuel use in buildings. This energy transition carries significant societal risks unless the shift is carried out with equity and justice as a top priority. Low-income, vulnerable and communities of color have higher energy burdens compared to affluent populations. Furthermore, systemic racism and historic exclusionary policies have resulted in increased risks (environmental, climatic, economic, and social) to low-income and communities of color, and underserved communities often do not have financial resources for, or access to, advanced building technologies. The U.S. Department of Energy is funding research to characterize and develop solutions to the challenges of equity and justice that complicate the ability of communities to contribute to goals for decarbonization. Our project has a specific focus on small commercial buildings and the businesses that occupy them. Significantly less is known about the burdens and risks these businesses experience or the challenges they face in pursuing decarbonization, or how those are affected by income and race, in comparison to research on energy equity and justice for diverse households. The project team includes the Pacific Northwest National Laboratory, Arizona State University and Clark Atlanta University. Researchers are conducting semi-structured interviews with small business owners in underserved communities in Phoenix and Atlanta, followed by a survey distributed to the larger community to learn more about the equity and justice issues that communities with different racial, economic, and cultural backgrounds face. Results will help inform an actionable and replicable framework for engaging small commercial building owners/operators to catalyze the reduction of energy burdens and increase equity.

Antonopoulos, Chrissi A.↗

Modernizing the Nuclear Industry and New Ways of Working

Nuclear energy is recognized as the most feasible energy source towards achieving nation-wide net-zero goals (COP 208, MIT). Additionally, the demand for reliable, consistent energy supply is soaring with the construction of energy hungry data centers and AI tools. This means the construction of new nuclear and the continuation of current nuclear are a high national priority. However, this leaves the nuclear industry with a workforce challenge. The rising demand for nuclear power is threatened by an underpopulated workforce. There exist a few contributors to the decreased workforce such as skewed workforce demographics resulting in large-scale workforce retirement. Also, some plants have observed difficulty hiring and retaining skill sets that typically comprise the nuclear power workforce – specifically skilled craftsman. Those who are graduating with desirable skills are seeking employment in sectors that are more modern and culturally more aligned with younger generational values. Lastly, within the nuclear sector, skilled employees will be a competitive commodity as advanced plants come online offering work environments that incorporate modern technologies, skill sets, and opportunities for career advancement. These factors emphasize the need for legacy plants to modernize with a focus on the workplace environment and culture that meets younger generations’ skills, cultural expectations, and desire for advancement opportunities. Nuclear plants are actively engaged in deploying advancements that improve the management of systems, structures, and components as well as process improvements and technologies that can reduce the costs of operation and maintenance. However, the advancements must also be analyzed, reviewed and deployed in a manner that considers the impact on how people within the organization collaborate, communicate, make decisions, and solve problems. Neglecting to consider the cultural impact of modernization risks poor adoption, unrealized opportunities, or insignificant change toward helping attract new employees. The traditional way of working is not necessarily the way new generations want to engage with their employer. For instance, in a survey performed by North American Young Generation in Nuclear the top three reasons for younger nuclear employees to seek other job opportunities was seeking better work-life balance, a lack of advancement opportunities, and work culture and leadership style differences (Smyth et al. 2022). Integrated Operations for Nuclear (ION) is an approach for the nuclear industry to create long-term strategic modernization plans and analyze each advancement for the impact on people and processes and measure how those impacts flow up to support high-level, long-term plant goals and requirements. One principle of ION is to replace the current labor-centric operation style in legacy nuclear plants with data-centric collaborative operation styles. This paper will explain how transitioning operations to leverage centralized skills and responsibility, multi-skilled teams, and collaborative decision making can flatten the hierarchical structure in plants, enable greater individual efficacy while also offering more experiences and advancement opportunities to staff. Adopting the ION way of working shifts the industry mind-set towards more networked, collaborative, and modern way of working that will attract skilled, next-generation workers to operate the legacy nuclear fleet.

99 - GENERAL AND MISCELLANEOUS↗

Coal-dependent Communities in Transition: Identifying Best Practices to Ensure Equitable Outcomes

The U.S. coal industry is experiencing a sharp increase in the numbers of retired and/or decommissioned coal-fired power plants across the U.S. In the years between 2010-2019, around 102 gigawatts (GW) of coal-fired generating capacity has been announced to be decommissioned, representing more than 546 coal-fired power plant units, and an additional 17 GW is planned to be decommissioned by 2025. This change in the energy production landscape presents an impact on the social, environmental, and economic prospects of coal-dependent communities. This report examined the role of communities in the coal power plant decommissioning process and provided community-identified best practices to ensure an equitable process. The experiences of four coal-dependent communities—Wise County, VA, Muskegon, MI, Anderson County, TN, and Becker, MN—are presented as case studies to understand the impacts of the decommissioning process, and associated best practices, from the communities’ perspective. The report results highlight the need to recognize that the decommissioning decision-making process must be community-based to be equitable. Each community’s input is key to the transition away from coal power because there is no one-size-fits-all development plan. In other words, each community’s trajectory through the decommissioning process—from the retirement decision-making stage to the final site redevelopment phase—is unique because each community has distinct needs and wants from the energy transition. What is best for one community may not be suited for another. Ultimately, the framework for site development and community revitalization post-decommissioning cannot be universal because each community’s profile—from a social, cultural, and economic perspective—is different. Community impacts of power plant decommissioning are not limited to job and revenue losses. Communities are likely to be impacted culturally, socially, environmentally, and have long-term health-based impacts that should be acknowledged and addressed in post-retirement plans. Commonly identified decommissioning best practices include: Early and continued engagement throughout, with a number of mediums for communication and feedback (e.g., in-person sessions, virtual meetings, written comment opportunities); Early planning of post-decommissioning projects to replace lost jobs, revenue, and economic activity; Recognition (and mitigation, if possible) of social impacts on the community due to plant closure; Transparency throughout the process, with trusted information being provided about the decommissioning process and timeline; potential impacts on the workforce, economy, and environment; and the feasibility of alternative site uses; Identification of funding sources, technical experts, and/or strategic partnerships to support decommissioning and the affected communities upfront; and, Acknowledgment of communities as stakeholders who have a role in the conversation and right to determine their futures. Three key areas for assisting coal-dependent communities affected by the energy transition: Technical assistance: assessment of site feasibility for alternative uses or to repower with new technologies; Cross-partnership engagement and collaboration: facilitate knowledge-sharing of “lessons learned” about the decommissioning process between communities and provide guidance for decision-making processes; and, Financial assistance: access to grant and/or loan programs to assist with redevelopment survey, bolster community economic security through job creation, and cover environmental clean-up costs. Technical assistance, cross-partnership engagement, and financial aid can be mobilized to help communities throughout various stages of the decommissioning process, including the retirement decision, the site reclamation phase, and eventual revitalization of the site and surrounding community.

01 COAL, LIGNITE, AND PEAT↗

Substrate flexibility regulates growth and apoptosis of normal but not transformed cells

One of the hallmarks of oncogenic transformation is anchorage-independent growth (27). Here we demonstrate that responses to substrate rigidity play a major role in distinguishing the growth behavior of normal cells from that of transformed cells. We cultured normal or H-ras-transformed NIH 3T3 cells on flexible collagen-coated polyacrylamide substrates with similar chemical properties but different rigidity. Compared with cells cultured on stiff substrates, nontransformed cells on flexible substrates showed a decrease in the rate of DNA synthesis and an increase in the rate of apoptosis. These responses on flexible substrates are coupled to decreases in cell spreading area and traction forces. In contrast, transformed cells maintained their growth and apoptotic characteristics regardless of substrate flexibility. The responses in cell spreading area and traction forces to substrate flexibility were similarly diminished. Our results suggest that normal cells are capable of probing substrate rigidity and that proper mechanical feedback is required for regulating cell shape, cell growth, and survival. The loss of this response can explain the unregulated growth of transformed cells.

Non-NASA Center↗

Apollo 12 lunar material - Effects on plant pigments.

Tissue cultures of tobacco grown for 12 weeks in contact with lunar material returned by Apollo 12 contained 21 to 35% more total pigment than control tissues. This difference is due primarily to increased chlorophyll a concentrations per gram fresh weight of tissue in experimental cultures. No differences were noted in the fresh or dry weight of the experimental and control cultures.

Weete, J. D.↗

A spontaneously arising mutation in connexin32 with repeated passage of FRTL-5 cells coincides with increased growth rate and reduced thyroxine release

In this study we examine changes in the cellular properties of FRTL-5 cells as a function of passage number, with particular emphasis on gap junction expression, karyotype, morphology, growth rate and thyroxine (T(4)) release. Early passage FRTL-5 follicular cells transfer dye through gap junctions from injected cell(s) to third-order neighboring cells and beyond within their respective follicles and have immuno-detectable connexin32 (Cx32) type gap junctional plaques in their lateral contacting plasma membranes. By contrast, FRTL-5 cells established as monolayers, or as follicles from cultures passed more than 15 times, did not transfer microinjected Lucifer Yellow dye to contiguous neighboring cells and did not express any immuno-detectable rat thyroid specific connexins (Cx43, Cx32 or Cx26). Western blots confirmed that total, membrane and cytosolic Cx32 protein was present only in early pass follicular cultures. To better understand the passage-dependent loss of Cx32 expression, RT-PCR primers were made to the most unique sequences of the rat Cx32 molecule, the cytoplasmic and carboxyl-terminal regions. These primers were used to screen FRTL-5 RNA from cultures of various passage numbers. The results revealed that later passage cultures had a single base deletion in the middle of the Cx32 cytoplasmic loop region at nucleotide position 378. This base deletion was in the middle position of the codon for amino acid 116, which is normally a CAC (histidine) but read with the frame shift was a CCC (proline). The four amino acids that followed this deletion were also altered with the fourth one becoming UAA, the ochre translation stop codon. This premature stopping of translation resulted in a truncation of 60% of the protein, which included the remaining cytoplasmic loop, third and fourth transmembrane regions and the carboxyl-terminus. The later passage cultures did not produce a carboxyl-terminal RT-PCR product, indicating that the mRNA was also truncated. These regions of the Cx32 molecule contain the sequences and epitopes to which probes and antibodies are directed, and as such alterations of these regions with repeated passage explains reports by others that FRTL-5 cells do not express Cx32, and implies that cultures used for these assessments were passed more than 15 times. To determine if genetic or epigenetic abnormalities existed in FRTL-5 cells we performed chromosome spreads from various passage cultures. FRTL-5 cells have been reported to be diploid and more recently non-diploid; however, we found them to be fully tetraploid. This tetraploidy appears to be unstable in that later passes are tetraploid plus two or three extra chromosomes. There were no obvious translocations, breaks or large-scale interstitial deletions of any chromosomes in the FRTL-5 cultures tested. As FRTL-5 cells were repeatedly passed their morphology changed. Monolayer areas spread from beneath the follicles, and the follicles became flattened in appearance. These physical changes were coincident with dramatically increased growth rates. Early cultures (passed 3-12 times) divided on average every 49+/-1 h, whereas later passes (passes 20-25) divided every 28+/-3 h. To correlate these changes with a measure of thyroid function we assayed T(4) output. Early passage follicular cultures incubated for 6 h with sodium iodide, released on average 5.27+/- 0.33 ng/ml of T(4)/100 follicles. Later passes, or early passes treated with heptanol to down-regulate Cx32, released an average of 3.84+/-0.50 ng/ml of T(4)/100 follicles. There was a 27% difference in T(4) release between early follicular cultures, that were coupled by Cx32, and late or down-regulated early follicular cultures, that were uncoupled (P<0.0001). Collectively, the physical changes documented in this study were coincident with the loss of functional Cx32. This implies a relationship between the loss of intercellular communication and changes in morphogenic appearance, growth rate and reduced thyroid function and supports the previously postulated, tumor-suppressor role for Cx32. FRTL-5 cultures from low passage numbers are an excellent model of primary thyroid cells. However, many reports in the literature ascribe features to FRTL-5 cells that are mutually inconsistent. These differences may be resolved in the future by addressing the passage number and the conditional differences of the cultures being studied.

NASA Discipline Radiation Health↗

Preclinical tumor organoid models in personalized cancer therapy: Not everyone fits the mold

Highlights: • In personalized cancer medicine, each patient receives a specific treatment. • The tumor microenvironment and interpersonal differences can affect treatment. • Organoid culture systems fully recapitulate the tumor microenvironments. In contrast to conventional cancer treatment, in personalized cancer medicine each patient receives a specific treatment. The response to therapy, clinical outcomes, and tumor behavior such as metastases, tumor progression, carcinogenesis can be significantly affected by the heterogeneous tumor microenvironment (TME) and interpersonal differences. Therefore, using native tumor microenvironment mimicking models is necessary to improving personalized cancer therapy. Both in vitro 2D cell culture and in vivo animal models poorly recapitulate the heterogeneous tumor (immune) microenvironments of native tumors. The development of 3D culture models, native tumor microenvironment mimicking models, made it possible to evaluate the chemoresistance of tumor tissue and the functionality of drugs in the presence of cell-extracellular matrix and cell-cell interactions in a 3D construction. Various personalized tumor models have been designed to preserving the native tumor microenvironment, including patient-derived tumor xenografts and organoid culture strategies. In this review, we will discuss the patient-derived organoids as a native tumor microenvironment mimicking model in personalized cancer therapy. In addition, we will also review the potential and the limitations of organoid culture systems for predicting patient outcomes and preclinical drug screening. Finally, we will discuss immunotherapy drug screening in tumor organoids by using microfluidic technology.

60 APPLIED LIFE SCIENCES↗

Fall 2015 NASA Internship, and Space Radiation Health Project

This fall, I was fortunate enough to have been able to participate in an internship at NASA's Lyndon B. Johnson Space Center. I was placed into the Human Health & Performance Directorate, where I was specifically tasked to work with Dr. Zarana Patel, researching the impacts of cosmic level radiation on human cells. Using different laboratory techniques, we were able to examine the cells to see if any damage had been done due to radiation exposure, and if so, how much damage was done. Cell culture samples were exposed at different doses, and fixed at different time points so that we could accumulate a large pool of quantifiable data. After examining quantifiable results relative to the impacts of space radiation on the human body at the cellular and chromosomal level, researchers can defer to different areas of the space program that have to do with astronaut safety, and research and development (extravehicular mobility unit construction, vehicle design and construction, etc.). This experience has been very eye-opening, and I was able to learn quite a bit. I learned some new laboratory techniques, and I did my best to try and learn new ways to balance such a hectic work and school schedule. I also learned some very intimate thing about working at NASA; I learned that far more people want to watch you succeed, rather than watch you fail, and I also learned that this is a place that is alive with innovators and explorers - people who have a sole purpose of exploring space for the betterment of humanity, and not for any other reason. It's truly inspiring. All of these experiences during my internship have impacted me in a really profound way, so much that my educational and career goals are completely different than when I started. I started out as a biotechnology major, and I discovered recently toward the end of the internship, that I don't want to work in a lab, nor was I as enthralled by biological life sciences as a believed myself to be. Taking that all into consideration, I've actually changed my major to mechanical engineering. I discovered that I enjoy building things, and I enjoy learning about materials and interactions between different things. And I quickly became obsessed with rocket and aerospace engineering, so I've decided that after a mechanical engineering degree, I will be pursuing an advanced degree in aerospace engineering. One final way that I was effected by this internship, is that I discovered that I don't want to have a career at NASA. I love this agency with all of my heart, but I refuse to allow my innovation to be bound by a scientifically illiterate congress. As such, I have decided to pursue commercial aerospace companies, such as Space, XCOR, Masten Space Systems, Orbital ATK, and many, many, more. Maybe one day I'll end up back here. I believe in what this agency is doing with my whole heart, and it's unfortunate to see them curtailed in some capacities as a result of budgetary constraints, brought on by people who don't fully understand the effort behind putting people in to space. All in all, this experience has been the best experience of my life - literally a childhood dream came true during this experience - and I cannot adequately explain how grateful I am to have been here for the past sixteen weeks.

Patience, Luke↗

Methods of Securing Chemical and Pharmaceutical Knowledge and Recommendations for International Institutions to Enhance Research Integrity

Here, this paper examines strategies for securing chemical and pharmaceutical expertise in a globalized research environment, focusing on safeguarding intellectual property and preventing the misuse of sensitive and potentially dual-use information. The product of collective efforts between Pacific Northwest National Laboratory, Carol Davila University of Medicine and Pharmacy, and New Bulgarian University, highlights the challenges and opportunities posed by cross-border research collaborations, particularly in the context of differing regulatory frameworks and research cultures. It explores current mechanisms to prevent data loss and unauthorized access to sensitive information while assessing the effectiveness of existing security measures, frameworks, and international export control regimes. The approach examines the differing methodologies for promoting transparency, trust-building, and mutual accountability in joint research projects to cultivate secure data-sharing practices and intellectual property. It provides recommendations for international institutions to implement security guidelines in framing research priorities, encourages continual training and education programs, and the integration of processes for monitoring research compliance. This partnership aims to advance scientific innovation while maintaining global stability, ensuring compliance with international norms, and safeguarding valuable intellectual property as measures in chemical and pharmaceutical research security practices continue to expand due to international collaboration and knowledge exchange.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Nutrient Limitation Predisposes A Cultivate of Burkholderia Contaminans From the ISS Water Processor Assembly to Biofilm Formation Under Simulated Microgravity.

The International Space Station (ISS) Water Processor Assembly (WPA) experiences intermittent dormancy between water recycling events thus promoting biofilm formation within the system. In this work we aimed to gain a deeper understanding of the impact of nutrient limitation on bacterial growth and biofilm formation under microgravity in support of biofilm mitigation efforts in exploration water recovery systems. A representative species of bacteria that is commonly cultured from the ISS WPA was cultured in an WPA influent water ersatz formulation tailored for microbiology studies. Burkholderia contaminans was cultured under a simulated microgravity (SµG) treatment in a vertically rotating high-aspect rotating vessel (HARV), with a rotating control (R) in the horizontal plane at the determined optimal rpm of 15 along with a stationary (S) control. At different time points, the bacterial culture and ersatz were harvested for bacterial counts, transcriptomic and nutrient content analyses. Under the test conditions, the culture under SµG treatment consumed the essential nutrients faster than the R and S control cultures in the early stage of growth, thus approaching a nutrient limited growth condition earlier than the controls. The rapid uptake and subsequent depletion of essential nutrients was further illustrated in the transcriptomic response of the SµG culture when compared to the transcriptomic response of the R and S control conditions. The observed starvation response may serve as one element to explain a moderate enhancement of biofilm formation in the SµG treatment. One implication of this investigation is that biofilm mitigation in the ISS environment could be supported by ensuring a steady flow of water as a vehicle for essential nutrients within the WPA to avoid complete consumption which occurs in times of no flow lending to undesired biofilm formation.

Angie Diaz↗

Does Magnetic Field Affect Malaria Parasite Replication in Human Red Blood Cells?

Digestion of red blood cell (RBC) hemoglobin by the malaria parasite results in the formation of paramagnetic hemazoin crystals inside the parasite body. A number of reports suggest that magnetic field interaction with hamazoin crystals significantly reduces the number of infected cells in culture, and thus magnetic field can be used to combat malaria. We studies the effects of magnetic filed on the Plasmodium falciparum asexual life cycle inside RBCs under various experimental conditions. No effect was found during prolonged exposure of infected RBCs to constant magnetic fields up to 6000 Gauss. Infected RBCs were also exposed, under temperature-controlled conditions, to oscillating magnetic fields with frequencies in the range of 500-20000 kHz, and field strength 30-600 Gauss. This exposure often changed the proportion of different parasite stages in treated culture compared to controls. However, no significant effect on parasitemia was observed in treated cultures. This result indicates that the magnetic field effect on Plasmodium falciparum is negligible, or that hypothetical negative and positive effects on different stages within one 48-hour compensate each other.

Chanturiya, Alexandr N.↗

Impact of Simulated Microgravity on Mesenchymal Stem Cell Proliferation

Background Long term exposure to microgravity has adverse effects on human body such as bone loss. Due to the vital role of mesenchymal stem cell (MSC) in bone regeneration, studying MSC under microgravity conditions has been of interest during the past decade. However, high cost and limited access to space flight can dampen enthusiasm of researchers. Microgravity simulators provide a way to investigate the effects of microgravity in human physiology. Methods In this study, we compared MSC cultures under simulated microgravity using random positioning machine (RPM) and Gravite 3D simulators. Two culture durations, 1 week and 2 weeks, were selected for MSC cultures without media change. To evaluate impact of simulated microgravity on cell proliferation, cell counting was performed using trypan blue exclusion assay. Additionally, RNA samples and conditioned media were collected for further analysis. Results After 1 week and 2 weeks, no cell morphology change was observed from RPM and Gravite cultures compared to control. Notably, distinct lower cell confluence at the edge of the flasks on the RPM was observed, which could be caused by fluid sheer forces. MSCs cultured on both microgravity simulators showed decreased cell proliferation after 1-week culture. After 2-week culture, cells grown on the RPM showed no significant difference in cell proliferation compared with control while cells cultured on the Gravite bioreactor showed significantly enhanced MSC proliferation. Conclusion Type of microgravity simulator and duration of culture significantly affect MSC proliferation. Further investigation into how simulated microgravity influences MSC gene expression and cytokine secretion related to bone loss is currently on going in our lab.

Cuiping Zhang↗