Research Containing: Humans

Development of a Novel Three-Dimensional, Automatable and Integrated Bioprocess for the Differentiation of Embryonic Stem Cells into Pulmonary Alveolar Cells in a Rotating Vessel Bioreactor System

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Application of stem cells for cell therapy of respiratory diseases is a developing field. We have previously established several protocols for the differentiation of embryonic stem cells (ESC) into alveolar epithelial cells, which require a high degree of operator interference and result in a low yield of target cells. Herein, we have shown that, by provision of a medium conditioned using A549 cells and by integration of classic steps of ESC differentiation into a single step through encapsulation in hydrogels (three-dimensional) and culture in a rotary bioreactor, murine ESC (mESC) could be directed to differentiate into distal respiratory epithelial cells. Type I and II pneumocytes (with a yield of 50% for type II) and Clara cells were demonstrated by the expression of aquaporin 5, surfactant protein C, and Clara cell secretory protein, respectively. We identified target cells as early as day 5 of culture and stably maintained our differentiated cells in vitro for 100 days. Electron microscopy demonstrated microvilli and intracellular lamellar bodies (LB), and fluorescent staining confirmed the active process of exocytosis of these LB in differentiated type II cells. When these cells were decapsulated and cultured in static conditions in flask cultures (two-dimensional), they retained their characteristic type II phenotype and morphology. In conclusion, our protocol offers integrated bioprocessing, shorter time of differentiation, lower cost, no use of growth factors, high reproducibility, and high phenotypic and functional stability, as well as being amenable to automation and being scalable, which would move this field closer to future clinical applications.

Related URLs:
<Go to ISI>://WOS:000302136300003

Space shuttle flight crew spatial orientation survey results

by cfynanon 9 June 2015in Biology & Biotechnology No comment

BACKGROUND & METHOD: A survey was distributed to 77 Space Shuttle flight crew members; 40 responded covering 71 missions. The goal was to capture historical information before Shuttle retirement and to better understand subjective experiences of illusory sensations due to the transition from 1-G to microgravity and back. RESULTS: We analyzed the response data to answer four questions: 1) Do older astronauts suffer more from illusory sensations than younger astronauts? We conclude that they do not because younger flight crew had about twice the rate of illusory sensations as older flight crew. 2) Do trial head motions during re-entry in an effort to hasten readaptation to 1-G really help? Apparently not because those who made trial head motions had a 38% rate of illusory sensations whereas those who did not make trial head motions had a 15% rate of illusory sensations. 3) Do symptoms decrease as flight experience increases? Yes, as reported in other publications, although there are individual exceptions. 4) Do longer duration missions lead to more illusory sensations and readaptation difficulties than shorter duration missions? Yes, the rate of illusory sensations for longer missions was 38%, whereas it was 24% for shorter missions. DISCUSSION: Based upon our results, long-duration missions may induce orientation problems that could have significant mission impacts.

Long-duration space flight and bed rest effects on testosterone and other steroids

by cfynanon 9 June 2015in Biology & Biotechnology No comment

CONTEXT: Limited data suggest that testosterone is decreased during space flight, which could contribute to bone and muscle loss. OBJECTIVE: The main objective was to assess testosterone and hormone status in long- and short-duration space flight and bed rest environments and to determine relationships with other physiological systems, including bone and muscle. DESIGN: Blood and urine samples were collected before, during, and after long-duration space flight. Samples were also collected before and after 12- to 14-d missions and from participants in 30- to 90-d bed rest studies. SETTING: Space flight studies were conducted on the International Space Station and before and after Space Shuttle missions. Bed rest studies were conducted in a clinical research center setting. Data from Skylab missions are also presented. PARTICIPANTS: All of the participants were male, and they included 15 long-duration and nine short-duration mission crew members and 30 bed rest subjects. MAIN OUTCOME MEASURES: Serum total, free, and bioavailable testosterone were measured along with serum and urinary cortisol, serum dehydroepiandrosterone, dehydroepiandrosterone sulfate, and SHBG. RESULTS: Total, free, and bioavailable testosterone was not changed during long-duration space flight but were decreased (P < 0.01) on landing day after these flights and after short-duration space flight. There were no changes in other hormones measured. Testosterone concentrations dropped before and soon after bed rest, but bed rest itself had no effect on testosterone. CONCLUSIONS: There was no evidence for decrements in testosterone during long-duration space flight or bed rest.

Benefits for bone from resistance exercise and nutrition in long-duration spaceflight: Evidence from biochemistry and densitometry

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Exercise has shown little success in mitigating bone loss from long-duration spaceflight. The first crews of the International Space Station (ISS) used the "interim resistive exercise device" (iRED), which allowed loads of up to 297 lb(f) (or 1337 N) but provided little protection of bone or no greater protection than aerobic exercise. In 2008, the Advanced Resistive Exercise Device (ARED), which allowed absolute loads of up to 600 lb(f) (1675 N), was launched to the ISS. We report dietary intake, bone densitometry, and biochemical markers in 13 crewmembers on ISS missions from 2006 to 2009. Of these 13, 8 had access to the iRED and 5 had access to the ARED. In both groups, bone-specific alkaline phosphatase tended to increase during flight toward the end of the mission (p = 0.06) and increased 30 days after landing (p < 0.001). Most markers of bone resorption were also increased in both groups during flight and 30 days after landing (p < 0.05). Bone densitometry revealed significant interactions (time and exercise device) for pelvis bone mineral density (BMD) and bone mineral content (p < 0.01), hip femoral neck BMD (p < 0.05), trochanter BMD (p < 0.05), and total hip BMD (p < 0.05). These variables were unchanged from preflight only for ARED crewmembers, who also returned from flight with higher percent lean mass and lower percent fat mass. Body mass was unchanged after flight in both groups. All crewmembers had nominal vitamin D status (75 +/- 17 nmol/L) before and during flight. These data document that resistance exercise, coupled with adequate energy intake (shown by maintenance of body mass determined by dual-energy X-ray absorptiometry [DXA]) and vitamin D, can maintain bone in most regions during 4- to 6-month missions in microgravity. This is the first evidence that improving nutrition and resistance exercise during spaceflight can attenuate the expected BMD deficits previously observed after prolonged missions.

The nutritional status of astronauts is altered after long-term space flight aboard the International Space Station

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Defining optimal nutrient requirements is critical for ensuring crew health during long-duration space exploration missions. Data pertaining to such nutrient requirements are extremely limited. The primary goal of this study was to better understand nutritional changes that occur during long-duration space flight. We examined body composition, bone metabolism, hematology, general blood chemistry, and blood levels of selected vitamins and minerals in 11 astronauts before and after long-duration (128-195 d) space flight aboard the International Space Station. Dietary intake and limited biochemical measures were assessed during flight. Crew members consumed a mean of 80% of their recommended energy intake, and on landing day their body weight was less (P = 0.051) than before flight. Hematocrit, serum iron, ferritin saturation, and transferrin were decreased and serum ferritin was increased after flight (P < 0.05). The finding that other acute-phase proteins were unchanged after flight suggests that the changes in iron metabolism are not likely to be solely a result of an inflammatory response. Urinary 8-hydroxy-2'-deoxyguanosine concentration was greater and RBC superoxide dismutase was less after flight (P < 0.05), indicating increased oxidative damage. Despite vitamin D supplement use during flight, serum 25-hydroxycholecalciferol was decreased after flight (P < 0.01). Bone resorption was increased after flight, as indicated by several markers. Bone formation, assessed by several markers, did not consistently rise 1 d after landing. These data provide evidence that bone loss, compromised vitamin D status, and oxidative damage are among critical nutritional concerns for long-duration space travelers.

Men and women in space: bone loss and kidney stone risk after long-duration spaceflight

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Bone loss, a key concern for long-duration space travelers, is typically considered a female issue. The number of women who have flown long-duration space missions is now great enough to allow a quantitative comparison of changes in bone and renal stone risk by sex. Participants were 42 astronauts (33 men and 9 women) on long-duration missions to the International Space Station. Bone mineral density (by dual-energy X-ray absorptiometry) and biochemical markers of bone metabolism (from blood and urine samples) were evaluated before and after flight. Data were analyzed in two groups, based on available resistance exercise equipment. Missions were 49 to 215 days in duration, flown between 2000 and 2012. The bone density response to spaceflight was the same for men and women in both exercise groups. The bone mineral density response to flight was the same for men and women, and the typical decrease in bone mineral density (whole body and/or regional) after flight was not observed for either sex for those using an advanced resistive exercise device. Biochemical markers of bone formation and resorption responded similarly in male and female astronauts. The response of urinary supersaturation risk to spaceflight was not significantly different between men and women, although risks were typically increased after flight in both groups, and risks were greater in men than in women before and after flight. The responses of men and women to spaceflight with respect to these measures of bone health were not different.

Thyroid status of Space Shuttle crewmembers: effects of iodine removal

by cfynanon 9 June 2015in Biology & Biotechnology No comment

INTRODUCTION: Iodine is often used for water purification and has been used throughout the U.S. space program. Because of concern about potential effects on crewmembers' thyroid function, in 1997 a system was implemented on board the Space Shuttles to remove iodine from water before it was consumed. We report here thyroid hormone data from crews flying before and after this system was implemented. METHODS: Blood samples were collected and analyzed for thyroid hormone content during routine medical exams before and after Space Shuttle missions. Data are reported for 224 male and 49 female astronauts (about two-thirds of them before implementation of iodine removal). RESULTS: Serum concentrations of total thyroxine (T4) and the free T4 index were elevated in men after flight and triiodothyronine (T3) was lower after flight, regardless of iodine removal status. T4 was higher, even before flight, in the group of men who flew after iodine removal was implemented. Conversely, T3 was lower in men who flew during that period. Before iodine removal was implemented, thyroid stimulating hormone (TSH) was elevated in male and tended to be elevated in female astronauts, with average increases of 27% and 19% after flight, respectively. After iodine removal was implemented, postflight TSH was not significantly different from preflight values. DISCUSSION: These data provide evidence that crewmembers' increase in serum TSH on landing day after early Shuttle flights resulted from their consumption of iodinated water during spaceflight, because the same increase was not observed after implementation of the iodine removal system.

Simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood were carried out using bioreactors. The co-culture of umbilical cord blood (UCB)-derived hematopoietic stem cells (HSCs) and mesenchymal stem cells (MSCs) was performed within spinner flasks and a rotating wall vessel (RWV) bioreactor using glass-coated styrene copolymer (GCSC) microcarriers. The medium used was composed of serum-free IMDM containing a cocktail of SCF 15 ng center dot mL(-1), FL 5 ng center dot mL(-1), TPO 6 ng center dot mL(-1), IL-3 15 ng center dot mL(-1), G-CSF 1 ng center dot mL(-1) and GM-CSF 5 ng center dot mL(-1). Accessory stromal cells derived from normal allogeneic adipose tissue were encapsulated in alginate-chitosan (AC) beads and used as feeding cells. The quality of the harvested UCB-HSCs and MSCs was assessed by immunophenotype analysis, methylcellulose colony and multi-lineage differentiation assays. After 12 days of culture, the fold-expansion of total cell numbers, colony-forming units (CFU-C), CD34(+)/CD45(+)/CD105(-) (HSCs) cells and CD34(-)/CD45(-)/CD105(+) (MSCs) cells using the RWV bioreactor were (3.7 +/- A 0.3)- , (5.1 +/- A 1.2)- , (5.2 +/- A 0.4)- , and (13.9 +/- A 1.2)-fold respectively, significantly better than those obtained using spinner flasks. Moreover, UCB-HSCs and UCB-MSCs could be easily separated by gravity sedimentation after the co-culture period as only UCB-MSCs adhered on to the microcarriers. Simultaneously, we found that the fibroblast-like cells growing on the surface of the GCSC microcarriers could be induced and differentiated towards the osteoblastic, chondrocytic and adipocytic lineages. Phenotypically, these cells were very similarly to the MSCs derived from bone marrow positively expressing the MSCs-related markers CD13, CD44, CD73 and CD105, while negatively expressing the HSCs-related markers CD34, CD45 and HLA-DR. It was thus demonstrated that the simultaneous expansion and harvest of UCB-HSCs and UCB-MSCs is possible to be accomplished using a feasible bioreactor culture system such as the RWV bioreactor with the support of GCSC microcarriers.

Related URLs:
<Go to ISI>://WOS:000284600400014

Comparison of antibiotic resistance, biofilm formation and conjugative transfer of Staphylococcus and Enterococcus isolates from International Space Station and Antarctic Research Station Concordia

by cfynanon 9 June 2015in Biology & Biotechnology No comment

The International Space Station (ISS) and the Antarctic Research Station Concordia are confined and isolated habitats in extreme and hostile environments. The human and habitat microflora can alter due to the special environmental conditions resulting in microbial contamination and health risk for the crew. In this study, 29 isolates from the ISS and 55 from the Antarctic Research Station Concordia belonging to the genera Staphylococcus and Enterococcus were investigated. Resistance to one or more antibiotics was detected in 75.8 % of the ISS and in 43.6 % of the Concordia strains. The corresponding resistance genes were identified by polymerase chain reaction in 86 % of the resistant ISS strains and in 18.2 % of the resistant Concordia strains. Plasmids are present in 86.2 % of the ISS and in 78.2 % of the Concordia strains. Eight Enterococcus faecalis strains (ISS) harbor plasmids of about 130 kb. Relaxase and/or transfer genes encoded on plasmids from gram-positive bacteria like pIP501, pRE25, pSK41, pGO1 and pT181 were detected in 86.2 % of the ISS and in 52.7 % of the Concordia strains. Most pSK41-homologous transfer genes were detected in ISS isolates belonging to coagulase-negative staphylococci. We demonstrated through mating experiments that Staphylococcus haemolyticus F2 (ISS) and the Concordia strain Staphylococcus hominis subsp. hominis G2 can transfer resistance genes to E. faecalis and Staphylococcus aureus, respectively. Biofilm formation was observed in 83 % of the ISS and in 92.7 % of the Concordia strains. In conclusion, the ISS isolates were shown to encode more resistance genes and possess a higher gene transfer capacity due to the presence of three vir signature genes, virB1, virB4 and virD4 than the Concordia isolates.

[Reproductive function of the male rat after a flight on the Kosmos-1129 biosatellite]

by cfynanon 9 June 2015in Biology & Biotechnology No comment

Male rats that were flown for 18.5 days on Cosmos-1129 were mated postflight with intact females. The mating 5 days postflight when the ejaculate consisted of spermatozoids that were exposed to zero-g effects in the mature stage yielded the litter which lagged behind the controls with respect to the growth and development during the first postnatal month. The mating 2.5-3 months postflight when the ejaculate consisted of spermatozoids that were exposed to zero-g effects at the stem cell stage yielded the litter which did not differ from the control.

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Research Containing: Humans

Development of a Novel Three-Dimensional, Automatable and Integrated Bioprocess for the Differentiation of Embryonic Stem Cells into Pulmonary Alveolar Cells in a Rotating Vessel Bioreactor System

Space shuttle flight crew spatial orientation survey results

Long-duration space flight and bed rest effects on testosterone and other steroids

Benefits for bone from resistance exercise and nutrition in long-duration spaceflight: Evidence from biochemistry and densitometry

The nutritional status of astronauts is altered after long-term space flight aboard the International Space Station

Men and women in space: bone loss and kidney stone risk after long-duration spaceflight

Thyroid status of Space Shuttle crewmembers: effects of iodine removal

Simultaneous expansion and harvest of hematopoietic stem cells and mesenchymal stem cells derived from umbilical cord blood

Comparison of antibiotic resistance, biofilm formation and conjugative transfer of Staphylococcus and Enterococcus isolates from International Space Station and Antarctic Research Station Concordia

[Reproductive function of the male rat after a flight on the Kosmos-1129 biosatellite]