Research Containing: Molecular biology

Microflow1, a sheathless fiber-optic flow cytometry biomedical platform: demonstration onboard the international space station

by cfynanon 9 June 2015in Technology Development & Demonstration No comment

A fiber-optic based flow cytometry platform was designed to build a portable and robust instrument for space applications. At the core of the Microflow1 is a unique fiber-optic flow cell fitted to a fluidic system and fiber coupled to the source and detection channels. A Microflow1 engineering unit was first tested and benchmarked against a commercial flow cytometer as a reference in a standard laboratory environment. Testing in parabolic flight campaigns was performed to establish Microflow1's performance in weightlessness, before operating the new platform on the International Space Station. Microflow1 had comparable performances to commercial systems, and operated remarkably and robustly in weightlessness (microgravity). Microflow1 supported immunophenotyping as well as microbead-based multiplexed cytokine assays in the space environment and independently of gravity levels. Results presented here provide evidence that this fiber-optic cytometer technology is inherently compatible with the space environment with negligible compromise to analytical performance.

A Field-Based Cleaning Protocol for Sampling Devices Used in Life-Detection Studies

by cfynanon 9 June 2015in Technology Development & Demonstration No comment

Abstract Analytical approaches to extant and extinct life detection involve molecular detection often at trace levels. Thus, removal of biological materials and other organic molecules from the surfaces of devices used for sampling is essential for ascertaining meaningful results. Organic decontamination to levels consistent with null values on life-detection instruments is particularly challenging at remote field locations where Mars analog field investigations are carried out. Here, we present a seven-step, multi-reagent decontamination method that can be applied to sampling devices while in the field. In situ lipopolysaccharide detection via low-level endotoxin assays and molecular detection via gas chromatography?mass spectrometry were used to test the effectiveness of the decontamination protocol for sampling of glacial ice with a coring device and for sampling of sediments with a rover scoop during deployment at Arctic Mars-analog sites in Svalbard, Norway. Our results indicate that the protocols and detection technique sufficiently remove and detect low levels of molecular constituents necessary for life-detection tests.

Related URLs:
http://dx.doi.org/10.1089/ast.2008.0275

New molecular technologies against infectious diseases during space flight

by cfynanon 9 June 2015in Technology Development & Demonstration No comment

Latent virus reactivation, reduction in the number of immune cells, decreased cell activation and increased sensitivity of astronauts to infections following their return on Earth demonstrate that the immune system is less efficient during space flight. This dysfunction during long-term missions could result in the appearance of opportunistic infections or a decrease in the immuno-surveillance mechanisms that eradicate cancer cells. On the other hand, monitoring of the microbial environment is essential to prevent infectious diseases in space. Therefore, both aspects will have to be monitored continuously during long-term missions in space, using miniature and semi-automated diagnostic systems. In the short term, such equipment will allow the study of the causes of space-related immunodeficiency, developing countermeasures to maintain an optimal immune function and improving our capacity to prevent infectious diseases during space missions. In order to achieve these objectives, a new diagnostic system has been designed to perform a set of biological and immunological assays on board spacecrafts. Through flow cytometric assays and molecular biology analyses, this diagnostic system will improve medical surveillance of astronauts and will be used to test countermeasures aimed at preventing immune deficiency during space missions.

Comparison of three rapid and easy bacterial DNA extraction methods for use with quantitative real-time PCR

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The development of fast and easy on-site molecular detection and quantification methods for hazardous microbes on solid surfaces is desirable for several applications where specialised laboratory facilities are absent. The quantification of bacterial contamination necessitates the assessment of the efficiency of the used methodology as a whole, including the preceding steps of sampling and sample processing. We used quantitative real-time polymerase chain reaction (qrtPCR) for Escherichia coli and Staphylococcus aureus to measure the recovery of DNA from defined numbers of bacterial cells that were subjected to three different DNA extraction methods: the QIAamp DNA Mini Kit, Reischl et al.'s method and FTA Elute. FTA Elute significantly showed the highest median DNA extraction efficiency of 76.9% for E. coli and 108.9% for S. aureus. The Reischl et al. method and QIAamp DNA Mini Kit inhibited the E. coli qrtPCR assay with a 10-fold decrease of detectable DNA. None of the methods inhibited the S. aureus qrtPCR assay. The FTA Elute applicability was demonstrated with swab samples taken from the International Space Station (ISS) interior. Overall, the FTA Elute method was found to be the most suitable to selected criteria in terms of rapidity, easiness of use, DNA extraction efficiency, toxicity, and transport and storage conditions.

Physiological effects of microgravity on osteoblast morphology and cell biology

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Biochemical and Molecular Biological Analyses of space-flown nematodes in Japan, the First International Caenorhabditis elegans Experiment (ICE-First)

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The first International Caenorhabditis elegans Experiment (ICE-First) was carried out using a Russian Soyuz spacecraft from April 19-30, 2004. This experiment was a part of the program of the DELTA (Dutch Expedition for Life science Technology and Atmospheric research) mission, and the space agencies that participate in the International Space Station (ISS) program formed international research teams. A Japanese research team that conducted by Japan aerospace Exploration Agency (JAXA) investigated the following aspects of the organism: (1) whether meiotic chromosomal dynamics and apoptosis in the germ cells were normal under microgravity conditions, (2) the effect of the space flight on muscle cell development, and (3) the effect of the space flight on protein aggregation. In this article, we summarize the results of these biochemical and molecular biological analyses.

Cytogenetic biodosimetry using the blood lymphocytes of astronauts

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Cytogenetic analysis of peripheral blood lymphocytes is the most sensitive and reliable method currently available for in vivo assessment of the biological effects of exposure to radiation and provides the most informative measurement of radiation induced health risks. Data indicates that space missions of a few months or more can induce measureable increases in the yield of chromosome damage in the blood lymphocytes of astronauts that can be used to estimate an organ dose equivalent, and biodosimetry estimates lie within the range expected from physical dosimetry. Space biodosimetry poses some unique challenges compared to terrestrial biological assessments of radiation exposures, but data provides a direct measurement of space radiation damage, which takes into account individual radiosensitivity in the presence of confounding factors such as microgravity and other stress conditions. Moreover if chromosome damage persists in the blood for many years, results can be used for retrospective dose reconstruction. In contrast to physical measurements, which are external to body and require multiple devices to detect all radiation types all of which have poor sensitivity to neutrons, biodosimetry is internal and includes the effects of shielding provided by the body itself plus chromosome damage shows excellent sensitivity to protons, heavy ions, and neutrons. In addition, chromosome damage is reflective of cancer risk and biodosimetry values can therefore be used to validate and develop risk assessment models that can be used to characterize health risk incurred by crewmembers. The current paper presents a review of astronaut biodosimetry data, along with recently derived data on the relative cancer risk estimated using the quantitative approach derived from the European Study Group on Cytogenetic Biomarkers and Health database.

New molecular technologies against infectious diseases during space flight

by cfynanon 9 June 2015in Biology & Biotechnology No comment

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Research Containing: Molecular biology

Microflow1, a sheathless fiber-optic flow cytometry biomedical platform: demonstration onboard the international space station

A Field-Based Cleaning Protocol for Sampling Devices Used in Life-Detection Studies

New molecular technologies against infectious diseases during space flight

Comparison of three rapid and easy bacterial DNA extraction methods for use with quantitative real-time PCR

Physiological effects of microgravity on osteoblast morphology and cell biology

Biochemical and Molecular Biological Analyses of space-flown nematodes in Japan, the First International Caenorhabditis elegans Experiment (ICE-First)

Cytogenetic biodosimetry using the blood lymphocytes of astronauts

New molecular technologies against infectious diseases during space flight