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At least 343 records · Page 19

Resistance exercise as a countermeasure to disuse-induced bone loss

During spaceflight, skeletal unloading results in loss of bone mineral density (BMD). This occurs primarily in the spine and lower body regions. This loss of skeletal mass could prove hazardous to astronauts on flights of long duration. In this study, intense resistance exercise was used to test whether a training regimen would prevent the loss of BMD that accompanies disuse. Nine subjects (5 men, 4 women) participated in a supine maximal resistance exercise training program during 17 wk of horizontal bed rest. These subjects were compared with 18 control subjects (13 men, 5 women) who followed the same bed rest protocol without exercise. Determination of treatment effect was based on measures of BMD, bone metabolism markers, and calcium balance obtained before, during, and after bed rest. Exercisers and controls had significantly (P < 0.05) different means, represented by the respective following percent changes: lumbar spine BMD, +3% vs. -1%; total hip BMD, +1% vs. -3%; calcaneus BMD, +1% vs. -9%; pelvis BMD, -0.5% vs. -3%; total body BMD, 0% vs. -1%; bone-specific alkaline phosphatase, +64% vs. 0%; alkaline phosphatase, +31% vs. +5%; osteocalcin, +43% vs. +10%; 1,25 dihydroxyvitamin D, +12% vs. -15%; parathyroid hormone intact molecule, +18% vs. -25%; and serum and ionized calcium, -1% vs. +1%. The difference in net calcium balance was also significant (+21 mg/day vs. -199 mg/day, exercise vs. control). The gastrocnemius and soleus muscle volumes decreased significantly in the exercise group, but the loss was significantly less than observed in the control group. The results indicate that resistance exercise had a positive treatment effect and thus might be useful as a countermeasure to prevent the deleterious skeletal changes associated with long-duration spaceflight.

Clinical Trial↗

Exercise Thermoregulation Following 13 Days of Bed Rest

This investigation examined two potential mechanisms, altered skin blood flow (SBF) and sweating rate (SR) responses, that may be responsible for an elevated core temperature during exercise after bed rest (BR) and space flight. Seven healthy men (29 +/- 5 yr, 179.6 +/- 7.1 cm, 77.2 +/- 17.0kg; mean +/- SD) underwent 13 days of 6 deg head-down BR. Pre- and post-BR, subjects completed supine submaximal cycle ergometry (20 min at 40% and 20 min at 65% of pre-BR supine VO2pk) in a thermoneutral room (23.4 +/- 0.5 C, 56 +/- 8 %RH) during, heat production (VO2 ; indirect calorimetry), intestinal temperature (T(sub in) ; ingestible pill), SBF (laser Doppler velocimetry), local SR (dew point hygrometry), and total sweat loss (TSL; Delta body weight) were measured. Pre- and post- BR plasma volume (PV) was measured using I-125 dilution. After BR, T(sub in) was elevated at rest (36.99 +/- 0.14 vs. 37.30 +/- 0.06 C; p<_0.05) and at the end of exercise (37.57 +/- 0.13 vs. 37.90 +/- 0.09 C; P less than or equal to 0.05). However, the increase in T(sub in) from rest to the end of exercise was not different after BR (0.59 +/- 0.07 vs. 0.60 +/- 0.07 C). There was no difference in VO2 pre- to post-BR during rest (0.28 +/- 0.04 vs. 0.24 +/- 0.03 1 multiplied by min(exp -1) ) or 40% VO2pk (0.95 +/- 0.08 vs. 0.96 +/- 0.05 1 multiplied by min(exp -1)), but VO2 was significantly less at the end of the 65% VO2pk stage (1.53 +/- 0.09 vs. 1.42 +/- 0.11 1 multiplies by min(exp - 1); p less than or equal to 0.05). The percent change in SBF from rest to end of exercise was less after BR (211 +/- 53 vs. 96 +/- 31%; p less than or equal to 0.05), the threshold for the onset of SBF was greater (37.17 +/- 0.18vs. 37.51 +/- 0.17 C; p less than or equal to 0.05), and the slope of the response tended to be reduced (536 +/- 184 vs. 201 +/- 46 %A PC; p=0.08). TSL was not different after BR (0.42+0.06 vs. 0.44 +/- 0.08 kg), but the T in threshold at the onset of sweating was delayed significantly (37.06 +/- 0.1 1 vs. 37.34 +/- 0.06 C; p less than or equal to 0.05). However, the slope of SR was not changed after BR (3.45 +/- 1.22 vs. 2.58 +/- 0.71 mg multiplied by min(exp -1) multiplied by cm sq). PV was decreased by 11% after BR (3,259 +/- 177 vs. 2,894 +/- 138 ml; p less than or equal to 0.05). These results suggest that exercise SBF and SR responses after BR are altered, and heat production is unchanged or reduced, consistent with observations following space flight. The higher resting T(sub in) with a proportional increase in T(sub in) during exercise and delayed onset of SBF and SR suggest a centrally-mediated elevation in the thermoregulatory set point during microgravity exposure.

Lee, Stuart M. C.↗

ECASTAR: Energy conservation. An assessment of systems, technologies and requirements

A methodology was presented for a systems approach to energy conservation actions and their potentials and impacts in the United States. Constraints affecting the approach were ranked, and the most important ones are the present economic and technical conditions. The following unresolved issues were identified: consumptive lifestyles vs. conservation ethic, environmental standards vs. energy conservation, capital availability, decentralization and vertical integration vs. centralization, fuel rich regions vs. fuel poor regions, supply vs. end use conservation, life cycle costing vs. initial cost, mandatory savings vs. voluntary savings, labor intensive vs. capital intensive, price control vs. free market. The following recommendations were made: provide action/impact assessment, establish regional energy centers, improve technology articulation with government, design total energy systems, utilize existing systems approach expertise.

Source record↗

Temperature Regulation in Crewmembers After a 115-Day Space Flight

Impaired thermoregulation, which has been observed during exercise following bed rest, may significantly impact crewmembers during space flight operations by decreasing exercise capacity and orthostatic tolerance. Impaired temperature regulation would cause higher levels of core temperature, due to an attenuated cutaneous vasodilatory reflex and sweating response, for a given oxygen consumption. Two mate crewmembers of the Mir 18 mission performed supine cycle exercise se (20 min @ 40% and 20 min @ 65% preflight VO2pk) 145 days preflight and 5 days postflight. Core temperature (Tcore) was measured by an ingestible telemetry pill, skin blood flow (SBF) by laser Doppler velocimetry, and sweat rate (SR) by dew point hygrometry. Tcore at the time of test termination was similar (37.8 C) for both subjects before and after flight despite a shorter test duration (40 vs 28-29 minutes) postflight. The slopes of the SBF/Tcore relationship (Subj 1: 396 vs 214; Subj 2: 704 vs 143 Perfusion Unit/degC) and SR/Tcore relationship (Subj 1: 4.5 vs 2.1; Subj 2: 11.0 vs 3.6mg/min/sq cm/degC) were reduced postflight. Tcore thresholds for both SR (Subj 1: 37.4 vs 37.6; Subj 2: 37.6 vs 37.6 C) and SBF (Subj 1: 37.3 vs 37.5; Subj 2: 37.6 vs 37.7 C) were similar pre- to postflight. For these 2 crewmembers, it appeared that thermoregulation during exercise was impaired as evidenced by compromised heat loss responses after long-duration space flight.

Lee, S. M. C.↗

Impaired bone formation in male idiopathic osteoporosis: further reduction in the presence of concomitant hypercalciuria

We present iliac bone histomorphometric data and related biochemical data from 16 nonalcoholic men (50 +/- 11 (SD) years) referred for evaluation of spontaneous skeletal and/or appendicular fractures and reduced spinal bone density. All men were eugonadal and had no known underlying disorder associated with osteopenia. For the group, mean serum chemistry values were within normal limits including immunoreactive parathyroid hormone, osteocalcin and serum 1,25-dihydroxyvitamin D [1,25(OH)2D]. Nine men demonstrated hypercalciuria (greater than or equal to 0.1 mmol/kg per day) while on a constant metabolic diet of 20 mmol/day Ca. Their 24-hour urinary calcium was significantly greater than that for the remaining 7 men (7.4 +/- 1.6 vs. 5.0 +/- 0.8 mmol/day, p = 0.003), as was their calciuric response to a 1 g oral calcium load (0.23 +/- 0.06 vs. 0.15 +/- 0.05 Ca/creatinine, p = 0.042). Serum parameters (including parathyroid hormone and 1,25(OH)2D) of hypercalciuric and normocalciuric men were not significantly different. Histomorphometric indices for cancellous bone demonstrated significant differences between the entire group of osteoporotic men and age-adjusted normal values for bone volume (11.4 +/- 4.0% vs. 23.2 +/- 4.4%), osteoid surface (5.6 +/- 3.9% vs. 12.1 +/- 4.6%), osteoblastic surface (2.0 +/- 2.3% vs. 3.9 +/- 1.9%), and mineralizing surface (1.9 +/- 2.4% vs. 5.1 +/- 2.7%); there were also significant differences in bone formation rate (total surface referent) (0.004 +/- 0.001 vs. 0.011 +/- 0.006 mm3/mm2 per year). Compared with the normocalciuric group the 9 hypercalciuric men had significantly lower osteoblastic surfaces (1.6 +/- 1.9% vs. 2.5 +/- 2.6%) and mineralizing surfaces (1.4 +/- 1.5% vs. 2.7 +/- 3.2%).(ABSTRACT TRUNCATED AT 250 WORDS).

NASA Discipline Regulatory Physiology↗

Relationship Between the Sub-Micron Fraction (SMF) and Fine-Mode Fraction (FMF) in the Context of AERONET Retrievals

The sub-micron (SM) aerosol optical depth (AOD) is an optical separation based on the fraction of particles below a specified cutoff radius of the particle size distribution (PSD) at a given particle radius. It is fundamentally different from spectrally separated FM (fine-mode) AOD. We present a simple (AOD-normalized) SM fraction versus FM fraction (SMF vs. FMF) linear equation that explains the well-recognized empirical result of SMF generally being greater than the FMF. The AERONET inversion (AERinv) products (combined inputs of spectral AOD and sky radiance) and the spectral deconvolution algorithm (SDA) products (input of AOD spectra) enable, respectively, an empirical SMF vs. FMF comparison at similar (columnar) remote sensing scales across a variety of aerosol types. SMF (AERinv-derived) vs. FMF (SDA-derived) behavior is primarily dependent on the relative truncated portion (ε c ) of the coarse-mode (CM) AOD associated with the cutoff portion of the CM PSD and, to a second order, the cutoff FM PSD and FM AOD (ε f ). The SMF vs. FMF equation largely explains the SMF vs. FMF behavior of the AERinv vs. SDA products as a function of PSD cutoff radius (“inflection point”) across an ensemble of AERONET sites and aerosol types (urban-industrial, biomass burning, dust, maritime and a mixed class of Arctic aerosols). The overarching dynamic was that the linear SMF vs. FMF relation pivots clockwise about the approximate (SMF, FMF) singularity of (1, 1) in a “linearly inverse” fashion (slope and intercept of approximately 1−ε c and ε c ) with increasing cutoff radius. SMF vs. FMF slopes and intercepts derived from AERinv and SDA retrievals confirmed the general domination of ε c over ε f in controlling that dynamic. A more general conclusion is the apparent confirmation that the optical impact of truncating modal (whole) PSD features can be detected by an SMF vs. FMF analysis.

Norman T. O'Neill↗

Body Composition of Rats on NASA Rodent Foodbar Versus Purina Lab Chow

The objective of this study is to test the nutritional adequacy of the NASA Rodent Foodbar for long term use for rats. The Foodbar was tested on two rat strains, Sprague Dawley and Fischer 344. Rats were fed either Foodbar or the control diet, Purina Chow #5012. Body composition analysis was performed on a selected number of samples (n=6 for each gender and treatment group) as part of extensive testing of the Foodbar. Water, fat, ash, and protein (and their percentages) for both treatment groups were compared. Sprague Dawley Foodbar fed rats had a lower % protein than Chow fed rats (25.8% vs. 30.0%). In addition, Sprague Dawley females fed Foodbar had higher total fat (42.1g vs. 24.6g) and % fat (16.4% vs. 10.5%), but lower % water (51.3% vs. 54.8%) than Sprague Dawley females fed Chow. Fischer 344 Foodbar fed rats had a lower % water (47.4% vs. 49,3%) and total water (93.2g vs. 99.6g). In addition, Fischer 344 Foodbar males had higher % fat (17.0% vs. 13.8%) and total fat (43.0g vs. 34.7g). The data suggests that body composition of Foodbar fed rats tends to have lower water content but a higher fat content compared to controls, but not all results are the same for different strains and even different genders within the same strains. The data obtained here, in addition to other data, helps provide a better understanding of the nutritional adequacy of the Foodbar and whether the formula may need to be modified.

Yu, Diane Sau Mun↗

The neuropathic postural tachycardia syndrome

BACKGROUND: The postural tachycardia syndrome is a common disorder that is characterized by chronic orthostatic symptoms and a dramatic increase in heart rate on standing, but that does not involve orthostatic hypotension. Several lines of evidence indicate that this disorder may result from sympathetic denervation of the legs. METHODS: We measured norepinephrine spillover (the rate of entry of norepinephrine into the venous circulation) in the arms and legs both before and in response to exposure to three stimuli (the cold pressor test, sodium nitroprusside infusion, and tyramine infusion) in 10 patients with the postural tachycardia syndrome and in 8 age- and sex-matched normal subjects. RESULTS: At base line, the mean (+/-SD) plasma norepinephrine concentration in the femoral vein was lower in the patients with the postural tachycardia syndrome than in the normal subjects (135+/-30 vs. 215+/-55 pg per milliliter [0.80+/-0.18 vs. 1.27+/-0.32 nmol per liter], P=0.001). Norepinephrine spillover in the arms increased to a similar extent in the two groups in response to each of the three stimuli, but the increases in the legs were smaller in the patients with the postural tachycardia syndrome than in the normal subjects (0.001+/-0.09 vs. 0.12+/-0.12 ng per minute per deciliter of tissue [0.006+/-0.53 vs. 0.71+/-0.71 nmol per minute per deciliter] with the cold pressor test, P=0.02; 0.02+/-0.07 vs. 0.23+/-0.17 ng per minute per deciliter [0.12+/-0.41 vs. 1.36+/-1.00 nmol per minute per deciliter] with nitroprusside infusion, P=0.01; and 0.008+/-0.09 vs. 0.19+/-0.25 ng per minute per deciliter [0.05+/-0.53 vs. 1.12+/-1.47 nmol per minute per deciliter] with tyramine infusion, P=0.04). CONCLUSIONS: The neuropathic postural tachycardia syndrome results from partial sympathetic denervation, especially in the legs.

NASA Discipline Regulatory Physiology↗

Endocrine relationships during human spaceflight

Human spaceflight is associated with a chronic loss of protein from muscle. The objective of this study was to determine whether changes in urinary hormone excretion could identify a hormonal role for this loss. Urine samples were collected from the crews of two Life Sciences Space Shuttle missions before and during spaceflight. Data are means +/- SE with the number of subjects in parentheses. The first value is the mean preflight measurement, and the second value is the mean inflight measurement. Adrenocorticotropic hormone (ACTH) [27.7 +/- 4.4 (9) vs. 25.1 +/- 3.4 (9) ng/day], growth hormone [724 +/- 251 (9) vs. 710 +/- 206 (9) ng/day], insulin-like growth factor I [6.81 +/- 0.62 vs. 6.04 +/- 0.51 (8) nM/day], and C-peptide [44.9 +/- 8.3 (9) vs. 50.7 +/- 10.3 (9) micrograms/day] were unchanged with spaceflight. In contrast, free 3,5,3'-triiodothyronine [791 +/- 159 (9) vs. 371 +/- 41 (9) pg/day, P < 0.05], prostaglandin E2 (PGE2) [1, 064 +/- 391 (8) vs. 465 +/- 146 (8) ng/day, P < 0.05], and its metabolite PGE-M [1,015 +/- 98 (9) vs. 678 +/- 105 (9) ng/day, P < 0. 05] were decreased inflight. The urinary excretion of most hormones returned to their preflight levels during the postflight period, with the exception of ACTH [47.5 +/- 10.3 (9) ng/day], PGE2 [1,433 +/- 327 (8) ng/day], PGF2alpha, [2,786 +/- 313 (8) ng/day], and its metabolite PGF-M [4,814 +/- 402 (9) ng/day], which were all increased compared with the preflight measurement (P < 0.05). There was a trend for urinary cortisol to be elevated inflight [55.3 +/- 5. 9 (9) vs. 72.5 +/- 11.1 micrograms/day, P = 0.27] and postflight [82.7 +/- 8.6 (8) micrograms/day, P = 0.13]. The inflight human data support ground-based in vitro work showing that prostaglandins have a major role in modulating the changes in muscle protein content in response to tension or the lack thereof.

Flight Experiment↗

The variety of variables in mathematical expressions

Various distinctions are discussed which can be made regarding evaluation of mathematical expressions: regular evaluation vs. infinite evaluation vs. finite evaluation, regular variables vs. mathematical variables vs. shadow variables vs. labels, simplification vs. evaluation vs. solution of equations. The unsatisfactory state of evaluation strategies in symbolic systems is due to insufficient use of such distinctions in the past.

Moses, J.↗

Outcome of patients with hypertrophic obstructive cardiomyopathy after percutaneous transluminal septal myocardial ablation and septal myectomy surgery

OBJECTIVES: This study was conducted to evaluate follow-up results in patients with hypertrophic obstructive cardiomyopathy (HOCM) who underwent either percutaneous transluminal septal myocardial ablation (PTSMA) or septal myectomy. BACKGROUND: Controversy exists with regard to these two forms of treatment for patients with HOCM. METHODS: Of 51 patients with HOCM treated, 25 were treated by PTSMA and 26 patients via myectomy. Two-dimensional echocardiograms were performed before both procedures, immediately afterwards and at a three-month follow-up. The New York Heart Association (NYHA) functional class was obtained before the procedures and at follow-up. RESULTS: Interventricular septal thickness was significantly reduced at follow-up in both groups (2.3 +/- 0.4 cm vs. 1.9 +/- 0.4 cm for septal ablation and 2.4 +/- 0.6 cm vs. 1.7 +/- 0.2 cm for myectomy, both p < 0.001). Estimated by continuous-wave Doppler, the resting pressure gradient (PG) across the left ventricular outflow tract (LVOT) significantly decreased immediately after the procedures in both groups (64 +/- 39 mm Hg vs. 28 +/- 29 mm Hg for PTSMA, 62 +/- 43 mm Hg vs. 7 +/- 7 mm Hg for myectomy, both p < 0.0001). At three-month follow-up, the resting PG remained lower in the PTSMA and myectomy groups (24 +/- 19 mm Hg and 11 +/- 6 mm Hg, respectively, vs. those before procedures, both p < 0.0001). The NYHA functional class was also significantly improved in both groups (3.5 +/- 0.5 vs. 1.9 +/- 0.7 for PTSMA, 3.3 +/- 0.5 vs. 1.5 +/- 0.7 for myectomy, both p < 0.0001). CONCLUSIONS: Both myectomy and PTSMA reduce LVOT obstruction and significantly improve NYHA functional class in patients with HOCM. However, there are benefits and drawbacks for each therapeutic method that must be counterbalanced when deciding on treatment for LVOT obstruction.

Non-NASA Center↗

Haloanaerobium kushneri sp. nov., an obligately halophilic, anaerobic bacterium from an oil brine

Three strains, designated VS-751T, VS-511 and VS-732, of a strictly anaerobic, moderately halophilic, Gram-negative, rod-shaped bacterium were isolated from a highly saline (15-20%) brine from an oil reservoir in central Oklahoma, USA. The optimal concentration of NaCl for growth of these three strains was 2 M (12%), and the strains also grew in the presence of an additional 1 M MgCl2. The strains were mesophilic and grew at a pH range of 6-8. Carbohydrates used by all three strains included glucose, fructose, arabinose, galactose, maltose, mannose, cellobiose, sucrose and inulin. Glucose fermentation products included ethanol, acetate, H2 and CO2, with formate produced by two of the three strains. Differences were noted among strains in the optimal temperature and pH for growth, the maximum and minimum NaCl concentration that supported growth, substrate utilization and cellular fatty acid composition. Despite the phenotypic differences among the three strains, analysis of the 16S rRNA gene sequences and DNA-DNA hybridizations showed that these three strains were members of the same genospecies which belonged to the genus Haloanaerobium. The phenotypic and genotypic characteristics of strains VS-751T, VS-511 and VS-732 are different from those of previously described species of Haloanaerobium. It is proposed that strain VS-751T (ATCC 700103T) be established as the type strain of a new species, Haloanaerobium kushneri.

NASA Discipline Exobiology↗

Comparison of Knee and Ankle Dynamometry between NASA's X1 Exoskeleton and Biodex System 4

Pre- and post-flight dynamometry is performed on International Space Station crewmembers to characterize microgravity-induced strength changes. Strength is not assessed in flight due to hardware limitations and there is poor understanding of the time course of in-flight changes. PURPOSE: To assess the reliability of a prototype dynamometer, the X1 Exoskeleton (EXO) and its agreement with a Biodex System 4 (BIO). METHODS: Eight subjects (4 M/4 F) completed 2 counterbalanced testing sessions of knee extension/flexion (KE/KF), 1 with BIO and 1 with EXO, with repeated measures within each session in normal gravity. Test-retest reliability (test 1 and 2) and device agreement (BIO vs. EXO) were evaluated. Later, to assess device agreement for ankle plantarflexion (PF), 10 subjects (4 M/6 F) completed 3 test conditions (BIO, EXO, and BIOEXO); BIOEXO was a hybrid condition comprised of the Biodex dynamometer motor and the X1 footplate and ankle frame. Ankle comparisons were: BIO vs. BIOEXO (footplate differences), BIOEXO vs. EXO (motor differences), and BIO vs. EXO (all differences). Reliability for KE/KF was determined by intraclass correlation (ICC). Device agreement was assessed with: 1) repeated measures ANOVA, 2) a measure of concordance (rho), and 3) average difference. RESULTS: ICCs for KE/KF were 0.99 for BIO and 0.96 to 0.99 for EXO. Agreement was high for KE (concordance: 0.86 to 0.95; average differences: -7 to +9 Nm) and low to moderate for KF (concordance: 0.64 to 0.78; average differences: -4 to -29 Nm, P<0.05). BIO vs. BIOEXO PF concordance ranged from 0.89 to 0.92 and mean differences ranged from -9 to +3 Nm (BIO < BIOEXO). BIOEXO vs. EXO PF concordance ranged from 0.73 to 0.80 while mean differences were -18 to -36 Nm (BIOEXO < EXO, P<0.05). PF concordance for BIO vs. EXO was slightly lower (0.61 to 0.84) and mean differences were greater (-27 to -33 Nm; BIO < EXO, P<0.05). CONCLUSION: BIO and EXO were similarly reliable for KE and KF. KE measures produced high agreement between devices; KF did not. For ankle PF, torque differences due to the two footplates were small. However, the X1 motor reports greater torques than the Biodex motor during PF. This first prototype provides proof of concept for a reliable, robotic-based exoskeleton to perform portable dynamometry for large muscle groups of the lower body.

English, K. L.↗

Female Astronauts Exhibit Greater Reductions in Aerobic Peak Power Following Long-Duration Space Flight than Males

BACKGROUND: Long-duration exposure to microgravity during space flight causes cardiovascular deconditioning, ultimately reducing aerobic fitness. While sex-based differences play an integral role in health on Earth, there is a paucity of data to inform space flight-induced health and performance implications for female astronauts, which may be critical to ensure health, safety, and mission success. Therefore, this retrospective investigation characterized aerobic capacity in female and male astronauts to inform whether sex differences exist from pre to postflight. METHODS: A NASA historical database (n=47 crew; 11 female, 36 male) of International Space Station (ISS) crewmembers was used to characterize sex differences in aerobic capacity changes from pre to postflight to update the NASA Aerobic and Muscle Risk Evidence Report. Aerobic capacity (VO2pk; ml/kg/min) and peak power (W) were assessed using a graded cycling exercise protocol that included a 3-min warmup at 50 W, with stepwise increases in resistance (25 W/min) until volitional fatigue. Group differences in baseline demographics and the percent change in aerobic capacity from preflight (~90 days before launch) to postflight (~3 days after return) were compared using independent t-tests. Significance was set at P<0.05 (Mean ±SD). RESULTS: At preflight, females were younger (44.5±6.6 vs 48.6±5.6 yrs; p=0.041), weighed less (68.8±10.0 vs 81.0±8.8 kg; p< 0.001), with lower VO2pk (32.8±5.9 vs 37.8±6.0 ml/kg/min; p=0.021) and aerobic peak power (229±42 vs 299±52 W; p< 0.001) than males, but had similar heart rate at VO2pk (176.0±7.3 vs 173.0±10.8 beats/min; p=0.406). ISS flight durations (F: 205.0±58.3 vs M: 176.0±40.5 days; p=0.061) and reductions in VO2pk from pre to postflight were comparable between sexes (F: -11.8±10.7%; M: -8.8%±7.9%; p=0.32). However, females had a greater reduction in aerobic peak power than males (−13.3%±8.9% vs −4.9%±8.8%; p < 0.01). CONCLUSIONS: These data indicate that some indicators of fitness (i.e., aerobic peak power) are more impacted in female astronauts compared to males after long-duration space flight missions. Future investigations with larger sample sizes are necessary to identify the physiological basis of sex-based differences and develop effective countermeasure approaches for both sexes.

Nicole C Strock↗

Does Collection Time Bias the Ecology of Cleanroom Air Samples?

Microbial monitoring of astromaterials collections has taken on increased importance with the return of biologically sensitive samples from the asteroids Ryugu and Bennu and the initiation of the Mars Sample Return Program. Terrestrial bacteria and fungi can alter the mineralogy and organic composition of our collections causing irreversible contamination of pristine samples and increasing the risk of false positives for life detection measurements. NASA has conducted routine microbial monitoring of its existing collections since 20181. Initial monitoring focused on surface samples collected with foam swabs. Although, airborne microbiology is often decoupled from surface microbiology in the built environment2 culture-based air sampling techniques like impactors were not compliant with existing contamination control requirements. Bringing organic rich media, gelatin or liquids into curation cleanrooms presents an unacceptable risk to pristine samples. In 2022 NASA purchased a materials complaint air sampler and began collecting air samples from the cleanrooms in addition to surface samples3. The new instrument uses an electret filter to collect samples that are suitable for cultivating organisms or for direct DNA sequencing. Preliminary DNA sequencing results appeared to indicate that longer sampling times biased the microbial community in favor of hearty, spore-forming bacteria3. We present the results of a study comparing overnight sampling (17 hours) to short (1 hour) sampling of unoccupied curation cleanrooms. The results will help us optimize our monitoring protocols and develop a more detailed inventory of the ecology of astromaterials curation cleanrooms. Methods: We analyzed 72 paired air samples from six different cleanrooms including the meteorite processing lab (ISO 7 equivalent, 16 samples), the lunar lab (ISO 6 equivalent, 10 samples), the stardust lab (ISO 5 equivalent 14 samples), the OSIRIS-REx lab (ISO 5 equivalent, 12 samples), the Hayabusa2 lab (ISO 5 equivalent, 14 samples), and the Genesis lab (ISO 4 equivalent, 6 samples). All the samples were collected with an InnovaPrep Bobcat air sampler operating at a sampling rate of 200 L/min. The sampler operates for 5 minutes out of every 20 minute period. Half of the samples were collected by filtering 3,000L (15 min. of active sampling) of air across an electret filter for one hour. The rest of the samples were collected by filtering approximately 51,000 L air across the filter overnight (~17 hours, 255 min. of active sampling). Cells were eluted from the filter using 6-7 ml of pressurized 0.15% tween 20 in PBS (phosphate buffered saline). This liquid was used to cultivate bacteria according to previously published methods1,4,5 and for DNA extraction and next generation sequencing. DNA was extracted with a Qiagen MagAttract PowerMicrobiome kit6. To identify bacteria and archaea, the 16S rRNA gene was amplified using Earth Microbiome primers for the V4 region 7. The amplified DNA was sequenced on an Illumina MiSeq using a V3 reagent kit. The resulting sequences were processed using DADA2 and QIIME2 as implemented on the EDGE bioinformatics platform8–10. Results: Only two of the 72 samples had no amplifiable DNA. Amplified DNA concentrations ranged from 2.67 – 0.272 ng/µl. The median concentration of amplified DNA for the 1 hour samples was 0.770 ± 0.368 ng/µl. The median concentration of amplified DNA for the overnight samples was 0.877 ± 0.434 ng/µl. On average the overnight samples had slightly more sequences (58,960 vs. 59,456) and ASV’s (amplicon sequence variants) (60 vs 64.5) than the one hour samples, but these differences are not statistically significant. The most abundant ASV in every sample mapped to the genus Cupravidus. ASV’s mapping to the genuses Bacillus, Schlegelella, Thermus, and Staphylococcus were also common. Discussion and Future Work: Alpha diversity statistics like Shannon Entropy and Faith Phylogenetic Diversity are used to describe the diversity of organisms in a single sample. If a longer sampling time was biasing the data, we would expect to see a change in these diversity statistics vs. sample time. However, we did not observe this in our data. The median Shannon entropy was slightly higher for the overnight samples (3.773 vs 3.611) as was the Faith Phylogenetic Diversity (4.042 vs 3.596), but both values were within a standard deviation of each other for the two sampling times (Fig. 1). It is unlikely, that the longer sampling time is introducing bias into our data. We do observe a significant decrease in diversity when comparing the air samples by lab. The Genesis lab (ISO 4 equivalent) has a lower median number of ASV’s (45.5) than the other labs (62). Median values for Shannon Entropy (3.717 vs. 3.430) and Faith Phylogenetic Diversity (3.796 vs. 3.548) are also lower for Genesis, but those values are with one standard deviation of each other for the different sampling times. This is consistent with previous culture-based results suggesting that the environment in cleanrooms tends to select for a core group of organisms capable of surviving under dry, low nutrient, conditions. The presence of the ASV’s mapping to Cupravidus and Thermus in our sequencing blanks and controls suggests that several of the most common organisms in our samples represent contaminants from the reagents used to perform the DNA extractions and sequencing. Further work is needed to identify these contaminants, remove them from our data and recalculate the diversity statistics. This is a systematic error. Therefore, we do not expect removing the sequencing contaminants to change our conclusions. Longer air sample collection times appear to result in slightly higher diversity and do not bias the results towards “hardy” bacteria like spore-formers. Based on these preliminary results we conclude that sampling at least 3,000 liters of air is sufficient to capture the microbial diversity of cleanrooms, and that air samples can also be collected overnight without negatively impacting diversity. These results allow us to be flexible when designing microbial monitoring plans so that they do not interfere with routine lab activity. References: 1. Regberg, A. B. et al. 49th Lunar and Planetary Science Conference (2018). 2. The United States Pharmacopeial Convention. USP General Chapter <1116> (2013). 3. Regberg, A. B., et al. 54th Lunar and Planetary Science Conference (2023). 4. Regberg, A. B. et al. 53rd Lunar and Planetary Science Conference ( 2022). 5. Davis, R. E.,et al. 50th Lunar and Planetary Science Conference (2019). 6. Qiagen. MagAttract® PowerMicrobiome® DNA/RNA EP Kit Handbook. (2018). 7. Walters, W. et al. mSystems 1, (2015). 8. Callahan, B. J. et al. Nat. Methods 13, 581–583 (2016). 9. Hall, M. & Beiko, R. G. Microbiome Analysis: Methods and Protocols113–129 (Springer, 2018). 10. Philipson, C. et al. Bio-Protoc. 7, e2622 (2017).

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