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Research Containing: Base Sequence

Space Environmental Factor Impacts upon Murine Colon Microbiota and Mucosal Homeostasis

by cfynanon 22 August 2016in Biology & Biotechnology No comment

Astronaut intestinal health may be impacted by microgravity, radiation, and diet. The aim of this study was to characterize how high and low linear energy transfer (LET) radiation, microgravity, and elevated dietary iron affect colon microbiota (determined by 16S rDNA pyrosequencing) and colon function. Three independent experiments were conducted to achieve these goals: 1) fractionated low LET gamma radiation (137Cs, 3 Gy, RAD), high Fe diet (IRON) (650 mg/kg diet), and a combination of low LET gamma radiation and high Fe diet (IRON+RAD) in male Sprague-Dawley rats; 2) high LET 38Si particle exposure (0.050 Gy), 1/6 G partial weight bearing (PWB), and a combination of high LET38Si particle exposure and PWB in female BalbC/ByJ mice; and 3) 13 d spaceflight in female C57BL/6 mice. Low LET radiation, IRON and spaceflight increased Bacteroidetes and decreased Firmicutes. RAD and IRON+RAD increased Lactobacillales and lowered Clostridiales compared to the control (CON) and IRON treatments. Low LET radiation, IRON, and spaceflight did not significantly affect diversity or richness, or elevate pathogenic genera. Spaceflight increased Clostridiales and decreased Lactobacillales, and similar trends were observed in the experiment using a ground-based model of microgravity, suggesting altered gravity may affect colonic microbiota. Although we noted no differences in colon epithelial injury or inflammation, spaceflight elevated TGFbeta gene expression. Microbiota and mucosal characterization in these models is a first step in understanding the impact of the space environment on intestinal health.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/26083373

MIZ1, an essential protein for root hydrotropism, is associated with the cytoplasmic face of the endoplasmic reticulum membrane in Arabidopsis root cells

by cfynanon 9 June 2015in Biology & Biotechnology No comment

MIZ1 is encoded by a gene essential for root hydrotropism in Arabidopsis. To characterize the property of MIZ1, we used transgenic plants expressing GFP-tagged MIZ1 (MIZ1-GFP) and mutant MIZ1 (MIZ1(G235E)-GFP) in a miz1-1 mutant. Although both chimeric genes were transcribed, the translational products of MIZ1(G235E)-GFP did not accumulate in roots. Moreover, MIZ1-GFP complemented the mutant phenotype but not MIZ1(G235E)-GFP. The signal corresponding to MIZ1-GFP was detected at high levels in cortical cells and lateral root cap cells and accumulated in compartments in cortical cells. MIZ1-GFP was fractionated into a soluble protein fraction and an endoplasmic reticulum (ER) membrane fraction, where it was bound to the surface of the ER membrane at the cytosolic side.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/22285304

Microgravity during spaceflight directly affects in vitro osteoclastogenesis and bone resorption

by cfynanon 9 June 2015in Biology & Biotechnology No comment

During space flight, severe losses of bone mass are observed. Both bone formation and resorption are probably involved, but their relative importance remains unclear. The purpose of this research is to understand the role of osteoclasts and their precursors in microgravity-induced bone loss. Three experiments on isolated osteoclasts (OCs) and on their precursors, OSTEO, OCLAST, and PITS, were launched in the FOTON-M3 mission. The OSTEO experiment was conducted for 10 d in microgravity within bioreactors with a perfusion system, where the differentiation of precursors, cultured on a synthetic 3-dimensional bonelike biomaterial, skelite, toward mature OCs was assessed. In OCLAST and in PITS experiments, differentiated OCs were cultured on devitalized bovine bone slices for 4 d in microgravity. All of the experiments were replicated on ground in the same bioreactors, and OCLAST also had an inflight centrifuge as a control. Gene expression in microgravity, compared with ground controls, demonstrated a severalfold increase in genes involved in osteoclast maturation and activity. Increased bone resorption, proved by an increased amount of collagen telopeptides released VS ground and centrifuge control, was also found. These results indicate for the first time osteoclasts and their precursors as direct targets for microgravity and mechanical forces.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=medl&AN=19329761
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:medline&id=pmid:19329761&id=doi:10.1096%2Ffj.08-127951&issn=0892-6638&isbn=&volume=23&issue=8&spage=2549&pages=2549-54&date=2009&title=FASEB+Journal&atitle=Microgravity+during+spaceflight+directly+affects+in+vitro+osteoclastogenesis+and+bone+resorption.&aulast=Tamma&pid=%3Cauthor%3ETamma+R%3C%2Fauthor%3E&%3CAN%3E19329761%3C%2FAN%3E

Microarray analysis of spaceflown murine thymus tissue reveals changes in gene expression regulating stress and glucocorticoid receptors

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The detrimental effects of spaceflight and simulated microgravity on the immune system have been extensively documented. We report here microarray gene expression analysis, in concert with quantitative RT-PCR, in young adult C57BL/6NTac mice at 8 weeks of age after exposure to spaceflight aboard the space shuttle (STS-118) for a period of 13 days. Upon conclusion of the mission, thymus lobes were extracted from space flown mice (FLT) as well as age- and sex-matched ground control mice similarly housed in animal enclosure modules (AEM). mRNA was extracted and an automated array analysis for gene expression was performed. Examination of the microarray data revealed 970 individual probes that had a 1.5-fold or greater change. When these data were averaged (n = 4), we identified 12 genes that were significantly up- or down-regulated by at least 1.5-fold after spaceflight (P < or = 0.05). The genes that significantly differed from the AEM controls and that were also confirmed via QRT-PCR were as follows: Rbm3 (up-regulated) and Hsph110, Hsp90aa1, Cxcl10, Stip1, Fkbp4 (down-regulated). QRT-PCR confirmed the microarray results and demonstrated additional gene expression alteration in other T cell related genes, including: Ctla-4, IFN-alpha2a (up-regulated) and CD44 (down-regulated). Together, these data demonstrate that spaceflight induces significant changes in the thymic mRNA expression of genes that regulate stress, glucocorticoid receptor metabolism, and T cell signaling activity. These data explain, in part, the reported systemic compromise of the immune system after exposure to the microgravity of space.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/20213684

LIF-free embryonic stem cell culture in simulated microgravity

by cfynanon 9 June 2015in Biology & Biotechnology No comment

BACKGROUND: Leukemia inhibitory factor (LIF) is an indispensable factor for maintaining mouse embryonic stem (ES) cell pluripotency. A feeder layer and serum are also needed to maintain an undifferentiated state, however, such animal derived materials need to be eliminated for clinical applications. Therefore, a more reliable ES cell culture technique is required. METHODOLOGY/PRINCIPAL FINDINGS: We cultured mouse ES cells in simulated microgravity using a 3D-clinostat. We used feeder-free and serum-free media without LIF. CONCLUSIONS/SIGNIFICANCE: Here we show that simulated microgravity allows novel LIF-free and animal derived material-free culture methods for mouse ES cells.

Related URLs:
http://ovidsp.ovid.com/ovidweb.cgi?T=JS&CSC=Y&NEWS=N&PAGE=fulltext&D=emed9&AN=2009387611
http://sfxhosted.exlibrisgroup.com/mayo?sid=OVID:embase&id=pmid:&id=doi:10.1371%2Fjournal.pone.0006343&issn=1932-6203&isbn=&volume=4&issue=7&spage=e6343&pages=&date=2009&title=PLoS+ONE&atitle=LIF-free+embryonic+stem+cell+culture+in+simulated+microgravity&aulast=Kawahara&pid=%3Cauthor%3EKawahara+Y.%3C%2Fauthor%3E&%3CAN%3E2009387611%3C%2FAN%3E

Gravity changes during animal development affect IgM heavy-chain transcription and probably lymphopoiesis

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Our previous research demonstrated that spaceflight conditions affect antibody production in response to an antigenic stimulation in adult amphibians. Here, we investigated whether antibody synthesis is affected when animal development occurs onboard a space station. To answer this question, embryos of the Iberian ribbed newt, Pleurodeles waltl, were sent to the International Space Station (ISS) before the initiation of immunoglobulin heavy-chain expression. Thus, antibody synthesis began in space. On landing, we determined the effects of spaceflight on P. waltl development and IgM heavy-chain transcription. Results were compared with those obtained using embryos that developed on Earth. We find that IgM heavy-chain transcription is doubled at landing and that spaceflight does not affect P. waltl development and does not induce inflammation. We also recreated the environmental modifications encountered by the embryos during their development onboard the ISS. This strategy allowed us to demonstrate that gravity change is the factor responsible for antibody heavy-chain transcription modifications that are associated with NF-kappaB mRNA level variations. Taken together, and given that the larvae were not immunized, these data suggest a modification of lymphopoiesis when gravity changes occur during ontogeny.

Related URLs:
http://www.ncbi.nlm.nih.gov/pubmed/22993194

Scale-Up of Mouse Embryonic Stem Cell Expansion in Stirred Bioreactors

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The aim of this study was to develop a robust, quality controlled and reproducible large-scale culture system using serum-free (SF) medium to obtain vast numbers of embryonic stem (ES) cells as a starting source for potential applications in tissue regeneration, as well as for drug screening studies. Mouse ES (mES) cells were firstly cultured on microcarriers in spinner flasks to investigate the effect of different parameters such as the agitation rate and the feeding regimen. Cells were successfully expanded at agitation rates up to 60 rpm using the SF medium and no significant differences in terms of growth kinetics or metabolic profiles were found between the two feeding regimens evaluated: 50% medium renewal every 24 h or 25% every 12 h. Overall, cells reached maximum concentrations of (4.2 +/- 0.4) and (5.6 +/- 0.8) x 10(6) cells/mL at Day 8 for cells fed once or twice per day; which corresponds to an increase in total cell number of 85 +/- 7 and 108 +/- 16, respectively. To have a more precise control over culture conditions and to yield a higher number of cells, the scale-up of the spinner flask culture system was successfully accomplished by using a fully controlled stirred tank bioreactor. In this case, the concentration of mES cells cultured on microcarriers increased 85 +/- 15-fold over 11 days. Importantly, mES cells expanded under stirred conditions, in both spinner flask and fully controlled stirred tank bioreactor, using SF medium, retained the expression of pluripotency markers such as Oct-4, Nanog, and SSEA-1 and their differentiation potential into cells of the three embryonic germ layers. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 27: 1421-1432, 2011

Related URLs:
<Go to ISI>://WOS:000296061900026

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