Methods in molecular biology
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The growth and development of plants during spaceflight have important implications for both basic and applied research supported by NASA and other international space agencies. While there have been many reviews of plant space biology, the present chapter attempts to fill a gap in the literature on the actual process and methods of performing plant research in the spaceflight environment. The author has been a principal investigator on six spaceflight projects and has another two space experiments in development. ⋯ Additional issues considered are working at NASA centers, hardware development, safety concerns, and the engineering versus science culture in space agencies. The difficulties of publishing the results from spaceflight research based on such factors as the lack of controls, limited sample size, and the indirect effects of the spaceflight environment also are summarized. Finally, lessons learned from these spaceflight experiences are discussed in the context of improvements for future space-based research projects with plants.
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Clinical epidemiology is the science of human disease investigation with a focus on diagnosis, prognosis, and treatment. The generation of a reasonable question requires definition of patients, interventions, controls, and outcomes. ⋯ The hierarchy of evidence for clinical decision-making places randomized controlled trials (RCT) or systematic review of good quality RCTs at the top of the evidence pyramid. Prognostic and etiologic questions are best addressed with longitudinal cohort studies.
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Simulated microgravity and partial gravity research on Earth is highly convenient for every space biology researcher due to limitations of access to spaceflight. However, the use of ground-based facilities for microgravity simulation is far from simple. ⋯ Furthermore, ground-based facilities are also capable of generating hypergravity or fractional gravity conditions, which are worth being tested and compared with the results of microgravity exposure. Multiple technologies (2D clinorotation, random positioning machines, magnetic levitators or centrifuges), experimental hardware (proper use of containers and substrates for the seedlings or cell cultures), and experimental requirements (some life support/environmental parameters are more difficult to provide in certain facilities) should be collectively considered in defining the optimal experimental design that will allow us to anticipate, modify, or redefine the findings provided by the scarce spaceflight opportunities that have been (and will be) available.
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Recent studies of our group have shown that suPAR may complement APACHE II score for risk assessment in sepsis. suPAR may be measured in serum of patients by an enzyme immunosorbent assay developed by Virogates (suPARnostic™). Production of suPAR from circulating neutrophils and monocytes may be assessed after isolation of neutrophils and monocytes and ex vivo culture. This is followed by measurement of suPAR in culture supernatants.
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Reprogramming of somatic cells, such as skin fibroblasts, to pluripotency was first achieved by forced expression of four transcription factors using integrating retroviral or lentiviral vectors, which result in integration of exogenous DNA into cellular genome and present a formidable barrier to therapeutic application of induced pluripotent stem cells (iPSCs). To facilitate the translation of iPSC technology to clinical practice, mRNA reprogramming method that generates transgene-free iPSCs is a safe and efficient method, eliminating bio-containment concerns associated with viral vectors, as well as the need for weeks of screening of cells to confirm that viral material has been completely eliminated during cell passaging.