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Rats were the first mammals to be domesticated for scientific research, and abundant physiological data are available on them. Rats are expected to continue to play an important role as experimental animals, especially with advancements such as CRISPR/Cas9 technology. Environmental enrichment aims to promote species-specific behaviors and psychological well-being. In the present study, we designed a double-decker (DD) cage, which utilizes two stacked plastic cages for rat enrichment, and investigated the influence of housing in the DD cage on rat mating behavior. The results indicated that mount frequency, total mount counts, and total ejaculation latency were significantly lower in the DD cages than in the single-decker (SD) cages. Notably, in the DD cages, the body weight loss of male rats after mating behavior was lower than that observed in the SD cage. Water consumption per day during mating behavior was also significantly lower in the DD cages, although no significant differences were observed in daily food intake during mating behavior. In addition, reproductive performance, including pregnancy rate and birth rate, did not change in the DD cages. In summary, our study demonstrated that DD cages reduce mount frequency and ejaculation latency during rat mating, resulting in decreased water consumption and weight loss in male rats. Therefore, housing in DD cages may serve as a beneficial enrichment for rats.

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An in vitro assay system using patient-derived tumor models represents a promising preclinical cancer model that replicates the disease better than traditional cell culture models. Patient-derived tumor organoid (PDO) and patient-derived tumor xenograft (PDX) models have been previously established from different types of human tumors to recapitulate accurately and efficiently their tissue architecture and function. However, these models have low throughput and are challenging to construct. Thus, the present study aimed to establish a simple in vitro high-throughput assay system using PDO and PDX models. Furthermore, the current study aimed to evaluate different classes of anticancer drugs, including chemotherapeutic, molecular targeted and antibody drugs, using PDO and PDX models. First, an in vitro high-throughput assay system was constructed using PDO and PDX established from solid and hematopoietic tumors cultured in 384-well plates to evaluate anticancer agents. In addition, an in vitro evaluation system of the immune response was developed using PDO and PDX. Novel cancer immunotherapeutic agents with marked efficacy have been used against various types of tumor. Thus, there is an urgent need for in vitro functional potency assays that can simulate the complex interaction of immune cells with tumor cells and can rapidly test the efficacy of different immunotherapies or antibody drugs. An evaluation system for the antibody-dependent cellular cytotoxic activity of anti-epidermal growth factor receptor antibody and the cytotoxic activity of activated lymphocytes, such as cytotoxic T lymphocytes and natural killer cells, was constructed. Moreover, immune response assay systems with bispecific T-cell engagers were developed using effector cells. The present results demonstrated that in vitro assay systems using PDO and PDX may be suitable for evaluating anticancer agents and immunotherapy potency with high reproducibility and simplicity.

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BACKGROUND/AIM: Clear cell sarcoma (CCS) of soft tissue is exceedingly rare and frequently exhibits aggressive behavior. Toward the goals of improving the aggressive course and poor prognosis of CCS, and establish new therapeutic methods, molecular genetic and biological characterizations of CCS are required. MATERIALS AND METHODS: A new human CCS cell line (designated RSAR001) was established from the pleural effusion of a 44-year-old man with multiple lung metastases and pleural dissemination. The cell line and its xenograft were characterized including their morphology, immunohistochemistry, cytogenetic analysis, reverse transcription-polymerase chain reaction, direct sequencing analysis, and fluorescence in situ hybridization analysis. RESULTS: The cell line has been maintained for over 12 months with more than 50 passages. RSAR001 cells exhibited a fascicular or diffuse growth pattern of short spindle- or oval-shaped cells with clear cytoplasm in heterotransplanted tumor, that was similar to the primary tumor. Immunophenotypically, RSAR001 cells in vitro and in vivo exhibited almost the same characteristics as the primary tumor. Cytogenetic analyses revealed a translocation, t(12;22)(q13;q12). Reverse transcription-polymerase chain reaction and direct sequencing analysis detected transcripts of the Ewing sarcoma breakpoint region 1-activating transcription factor 1 (EWSR1-ATF1) type 1 fusion gene. Fluorescence in situ hybridization using a break-apart probe for the EWSR1 gene on 22q12 showed a rearrangement. CONCLUSION: These findings indicate that the RSAR001 cell line harbors EWSR1-ATF1 type 1 chimeric fusion gene, which is specific to CCS. RSAR001 cells might be useful for investigating biological behaviors and developing new treatments such as molecular-targeting antitumor drugs or immunological drugs for CCS.

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Patient-derived tumor xenograft models represent a promising preclinical cancer model that better replicates disease, compared with traditional cell culture; however, their use is low-throughput and costly. To overcome this limitation, patient-derived tumor organoids (PDOs) were established from human lung, ovarian and uterine tumor tissues, among others, to accurately and efficiently recapitulate the tissue architecture and function. PDOs were able to be cultured for >6 months, and formed cell clusters with similar morphologies to their source tumors. Comparative histological and comprehensive gene expression analyses proved that the characteristics of PDOs were similar to those of their source tumors, even following long-term expansion in culture. At present, 53 PDOs have been established by the Fukushima Translational Research Project, and were designated as Fukushima PDOs (F­PDOs). In addition, the in vivo tumorigenesis of certain F­PDOs was confirmed using a xenograft model. The present study represents a detailed analysis of three F­PDOs (termed REME9, 11 and 16) established from endometrial cancer tissues. These were used for cell growth inhibition experiments using anticancer agents. A suitable high-throughput assay system, with 96- or 384­well plates, was designed for each F­PDO, and the efficacy of the anticancer agents was subsequently evaluated. REME9 and 11 exhibited distinct responses and increased resistance to the drugs, as compared with conventional cancer cell lines (AN3 CA and RL95-2). REME9 and 11, which were established from tumors that originated in patients who did not respond to paclitaxel and carboplatin (the standard chemotherapy for endometrial cancer), exhibited high resistance (half-maximal inhibitory concentration >10 µM) to the two agents. Therefore, assay systems using F­PDOs may be utilized to evaluate anticancer agents using conditions that better reflect clinical conditions, compared with conventional methods using cancer cell lines, and to discover markers that identify the pharmacological effects of anticancer agents.

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Isoflurane is a widely used anesthetic, but its effects with increase in inspired concentration on cardiovascular function have not yet been clarified in rodents. Additionally, there are only a few studies comparing isoflurane-induced cardiorespiratory effects between rat strains. Thus, we investigated the differences in cardiorespiratory responsiveness to increasing concentration of inspired isoflurane in SHR/Izm, WKY/Izm and Crl:CD (SD) rats, by increasing the setting values of vaporizer's dial indicator. The rats were anesthetized with 1.5% isoflurane, and electrocardiograms, blood pressure, and respiratory rate were recorded simultaneously. Thereafter, the inspired concentration was increased stepwise to 2%, 3%, 4%, and 5%, and cardiorespiratory parameters were obtained at each concentration. Under anesthesia at more than 4%, although prolongation of the RR and PR intervals was observed in all strains, shortening of the QTC interval was found only in SHR/Izm rats. From frequency domain analysis of heart rate variability, an increase in LF/HF ratio and a decrease of HF components were observed in SHR/Izm and WKY/Izm rats, respectively, with 5% isoflurane anesthesia. Blood pressure and heart rate were remarkably reduced in SHR/Izm rats at higher concentrations, whereas the reduction was smallest in WKY/Izm rats among the three strains examined. Respiratory rate was inspired concentration-dependently decreased in all strains. These results suggested that SHR/Izm rats are more sensitive to suppressive effects of isoflurane anesthesia on cardiovascular function among these rat strains.

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We investigated effect of microgravity environment during spaceflight on postnatal development of the rheological properties of the aorta in rats. The neonate rats were randomly divided at 7 days of age into the spaceflight, asynchronous ground control, and vivarium control groups (8 pups for one dam). The spaceflight group rats at 9 days of age were exposed to microgravity environment for 16 days. A longitudinal wall strip of the proximal descending thoracic aorta was subjected to stress-strain and stress-relaxation tests. Wall tensile force was significantly smaller in the spaceflight group than in the two control groups, whereas there were no significant differences in wall stress or incremental elastic modulus at each strain among the three groups. Wall thickness and number of smooth muscle fibers were significantly smaller in the spaceflight group than in the two control groups, but there were no significant differences in amounts of either the elastin or collagen fibers among the three groups. The decreased thickness was mainly caused by the decreased number of smooth muscle cells. Plastic deformation was observed only in the spaceflight group in the stress-strain test. A microgravity environment during spaceflight could affect postnatal development of the morphological and rheological properties of the aorta.

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A magnitude 9.0 great earthquake, the 2011 off the Pacific coast of Tohoku Earthquake, occurred on March 11, 2011, and subsequent Fukushima Daiichi Nuclear Power Station (Fukushima NPS) accidents stirred up natural radiation around the campus of Fukushima Medical University (FMU). FMU is located in Fukushima City, and is 57 km to the northwest of Fukushima NPS. Due to temporary failure of the steam boilers, the air conditioning system for the animal rooms, all autoclaves, and a cage washer could not be used at the Laboratory Animal Research Center (LARC) of FMU. The outside air temperature dropped to zero overnight, and the temperature inside the animal rooms fell to 10°C for several hours. We placed sterilized nesting materials inside all cages to encourage rodents to create nests. The main water supply was cut off for 8 days in all, while supply of steam and hot water remained unavailable for 12 days. It took 20 days to restore the air conditioning system to normal operation at the facility. We measured radiation levels in the animal rooms to confirm the safety of care staff and researchers. On April 21, May 9, and June 17, the average radiation levels at a central work table in the animal rooms with HEPA filters were 46.5, 44.4, and 43.4 cpm, respectively, which is equal to the background level of the equipment. We sincerely hope our experiences will be a useful reference regarding crisis management for many institutes having laboratory animals.

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Measles virus (MeV) vaccine strain, AIK-C, is temperature sensitive (ts), which is thought to be associated with attenuation of virus pathogenicity. In this study, replication and antibody response were examined in cotton rats using viruses carrying different forms of the P gene, which is responsible for the ts phenotype of strain AIK-C and its parental Edmonston strain. When cotton rats were inoculated intranasally, ts viruses neither replicated in lungs, nor reproducibly generated an antibody response. When inoculated intramusculary (i.m.), however, ts strains raised an antibody titer in all animals. This response was not observed when ultraviolet-inactivated virus was used. ts virus, inoculated i.m., was recovered from cotton rat drainage lymph nodes. These results suggest that ts virus, inoculated i.m., could replicate in the cotton rat, presumably at the superficial lymph node, and induce an antibody response. Therefore, cotton rats can serve as a small-animal model for investigating immune responses to safer ts vaccine, as well as recombinant vaccine using AIK-C as a vector for protection against other infectious agents.

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Although rats often show an upright standing behavior the cardiovascular response during the behavior has not yet been fully clarified. In this study we quantified the activity of upright standing behavior in rats using infrared beam detectors and measured cardiovascular variables during the behavior. Rats demonstrated a high level of upright standing activity as they showed the upright posture more than 500 times per day at 10 weeks of age. The average upright standing duration time was less than 10s. Arterial pressure slightly decreased while heart rate increased in response to the behavior and these responses were not affected by sino-aortic denervation. Our results indicate that other mechanisms such as the vestibulo-cardiovascular reflex may completely compensate the lack of the baroreceptor reflex to maintain cardiovascular homeostasis in response to acute positional changes in rats. Moreover rats demonstrate complex integrative mechanisms maintaining cardiovascular homeostasis against the upright standing behavior which frequently occurs in rats.

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To investigate whether the rate of change in blood pressure affects the sensitivity of the aortic baroreceptor afferent response, the change in aortic nerve activity (ANA) to two different rates of ramp increase in mean blood pressure (MBP), elicited by phenylephrine administration, was determined in the rat under urethane (1.5 g kg(-1)) anesthesia. The sensitivity of the increase in ANA following a rapid (average ramp rate, 9.14+/-0.60 mm Hg s(-1), n=11) or gradual (1.78+/-0.24 mm Hg s(-1), n=11) increase in MBP was 2.03+/-0.14% and 1.81+/-0.20% of baseline mm Hg(-1), respectively. These values were not significantly different from each other (P=0.16). Furthermore, we found no correlation between the rate of ramp increase in MBP and the sensitivity of the increase in ANA (r=0.24, P=0.29, n=22). These results suggest that, at least within the normal physiological range of MBP, the rate of the ramp change in blood pressure does not affect aortic baroreceptor afferent sensitivity in the anesthetized rat.

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Deep-sea water is rich in minerals, e.g., Mg, Ca, and K which have been considered to be associated with prevention of cardiovascular disease. We investigated the effect of deep-sea water on cardiovascular hemodynamics in Kurosawa and Kusanagi-Hypercholesterolemic (KHC) rabbits. Deep-sea water was pumped in the offing of Cape Muroto in Japan and the mineral constituents were refined to a degree of hardness of 1,000. Twenty four 4-month-old KHC rabbits were given refined deep-sea water (n=12) and tap water (n=12) for 6 months. Pressure and flow waves at the ascending aorta were recorded under pentobarbital anesthesia. Systolic, diastolic, pulse and mean arterial pressures and total peripheral resistance were significantly lower in the deep-sea water group than in the control group. There were no significant differences in changes in serum lipid levels, plasma renin and angiotensin converting enzyme activities and electrolyte levels except for Mg(2+) after the feeding of the water between the two groups. A slight increase in serum Mg(2+) level in the deep-sea water group may not account for the inhibition of mild hypertension. From our results, we conclude that deep-sea water could improve cardiovascular hemodynamics, even though the factors which affect the blood pressure are still unknown.

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To investigate postnatal developmental changes in functional characteristics of the afferent pathway of the aortic baroreceptor reflex, the responses of aortic nerve activity (ANA) to blood pressure (BP) changes elicited by phenylephrine and sodium nitroprusside administration were tested in 3-, 8- and 20-week-old male rats under chloralose (60 mg kg(-1) i.p.) and urethane (600 mg kg(-1) i.p.) anaesthesia. The function curve of ANA in response to BP changes showed a sigmoid shape that shifted to the right from 3 to 8 weeks of age. The maximal activity and maximal gain of the aortic nerve, which were calculated by a logistic function analysis, were significantly higher in 20-week-old rats (maximal activity, 532 +/- 47% of baseline; maximal gain, 7.9 +/- 0.8% of baseline mmHg(-1); n = 9) than in 3-week-old rats (maximal ANA, 268 +/- 25% of baseline, P < 0.001; maximal gain, 4.9 +/- 0.5% of baseline mmHg(-1), P < 0.01, n = 9) and 8-week-old rats (maximal ANA, 309 +/- 18% of baseline, P < 0.001; maximal gain, 4.9 +/- 0.3% of baseline mmHg(-1), P < 0.01, n = 11). These results suggest that the operating point of aortic baroreceptor afferents is reset to the higher pressure level during development from 3 to 8 weeks of age and, thereafter, the afferent gain increases from 8 to 20 weeks of age. This functional change may be an important factor to prevent an excess increase of BP, which would result in pathophysiological problems.

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Junctional adhesion molecule-1 (JAM-1) forms part of the tight junction between adjacent endothelial cells. Using microarray technology, we showed previously that JAM-1 was differentially expressed in the brain stem of spontaneously hypertensive rats compared with normotensive Wistar-Kyoto (WKY) rats. In this study, we quantified the expression of JAM-1 in the brain stem of spontaneously hypertensive rats and WKY rats and established whether any differential expression was confined to this region of the brain or was ubiquitous throughout the central nervous system and, indeed, the whole body. Because the nucleus tractus solitarii plays a pivotal role in arterial pressure regulation, we assessed whether JAM-1 in this region affects the chronic regulation of arterial pressure. Real time RT-PCR revealed that JAM-1 mRNA was upregulated in multiple regions of the brain and all of the peripheral vascular beds studied. In the nucleus tractus solitarii, the level of JAM-1 mRNA was significantly higher in both young (3-week-old, prehypertensive) and adult male spontaneously hypertensive rats (15 to 18 weeks old) than that of age-matched WKY rats (fold differences; prehypertensives: 1.01+/-0.06 versus 1.59+/-0.13; n=10; P<0.01; adult: 1.08+/-0.14 versus 2.86+/-0.57; n=10; P<0.01). After adenoviral-mediated expression of JAM-1 in the nucleus tractus solitarii of adult WKY rats (15 weeks old; n=6), systolic pressure was increased from 120+/-4 to 132+/-4 mm Hg (P<0.01). Our data suggest that JAM-1 expression in the spontaneously hypertensive rat is upregulated throughout the body compared with the WKY rat and that this is not secondary to the hypertension. When JAM-1 is expressed in the nucleus tractus solitarii, it raises arterial pressure, suggesting a novel prohypertensive role for this protein within the brain stem.

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At present, there is no single software package that provides a comprehensive power spectral analysis of pulse interval (PI) and arterial blood pressure (BP), spontaneous cardiac baroreceptor reflex gain (sBRG) and respiratory rate. Furthermore, scientific validation of the software that is currently commercially available and employed has not been published. We introduce 'Hey-Presto' software, which fully evaluates cardiovascular autonomic function from the BP signal obtained from rats. The program performs power spectral analysis of HR and BP variability, respiratory rate and, based on a time-series method, spontaneous cardiac baroreceptor (sBRG). We have validated Hey-Presto with conventional pharmacological agents to block cardiac vagal and cardiac sympathetic transmission in conscious rats fitted with a radio-telemetery BP transducer. Following administration of atropine (1 mg kg(-1), I.V.), high-frequency (HF) power of the PI decreased (P < 0.01) and was associated with the expected increase in HR. Subsequent cardiac sympathetic blockade (atenolol, 1 mg kg(-1), I.V.) reduced the low frequency (LF) to HF ratio (LF:HF) of the PI (P < 0.01), which was consistent with the observed reduction in HR. We also found that alterations in sBRG after blockade of cardiac autonomic transmission were highly comparable to values computed manually using vasoactive drugs administered intravenously. The software also detected circadian rhythms in sBRG, HF component of the PI, LF:HF of the PI and LF component of the BP as well as BP and HR during continuous 24 h recording. By demonstrating its application to humans, we found appropriate changes in the power of PI and the LF power of the BP during postural changes. These results demonstrate that Hey-Presto allows a fully automated, reliable, fast and comprehensive evaluation of cardiovascular autonomic function based on chronic measurements of BP in rats. Moreover, we have confirmed its versatility by demonstrating its application to man.

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The effects of microgravity on the histological characteristics of the aortic depressor nerve, which is the afferent of the aortic baroreflex arc, were determined in 10 female adult rats. The rats were assigned for nursing neonates in the Space Shuttle Columbia or in the animal facility on the ground (NASA Neurolab, STS-90), and were housed for 16 days under microgravity in space (microg, n=5) or under one force of gravity on Earth (one-g, n=5). In the Schwann cell unit in which the axons of unmyelinated fibers are surrounded by one Schwann cell, the average number of axons per unit in the microg group was 2.1 +/- 1.6 (mean +/- SD, n=312) and significantly less than that in the one-g group (3.0 +/- 2.9, n=397, p<0.05). The proportion of unmyelinated fibers in the aortic depressor nerve in the microg group was 64.5 +/- 4.4% and significantly less than that in the one-g group (74.0 +/- 7.3%, p<0.05). These results show that there is a decrease in the number of high-threshold unmyelinated fibers in the aortic depressor nerve in adult rats flown on the Shuttle Orbiter, suggesting that the aortic baroreflex is depressed under microgravity during space flight.

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International efforts to standardize regulations and study designs and to promote the principles of Reduction, Replacement, and Refinement (the 3 Rs) have reduced and refined animal use. In NASA ARC and KSC, researchers are responsible only for activities related directly to the conduct of their animal experiments. The IACUC plays an important role in conformity with NIH policies. Even if researchers design protocols of the space life science in Japan, the animal experiments should be carried out under the global harmonized conditions in accordance with NIH/NASA policies, guides and rules. It is important that researchers himself must look forward the ethical animal experiment.

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The aim of this study is to examine the structural development in kidney and lung macroscopically which relate with cardiovascular system in rats raised in space. Twenty three nine-day old rats and six fifteen-day old rats, which were launched at these ages and nursed by their dams in the Space Shuttle Colombia for 16 days (STS-90; Neurolab). Seventeen animals of the nine-day old rats were defined as the nine-day group, and the rest was defined as the re-adaptation group, which were reared on the ground for 30 more days after landing. The organs were weighed and the ratio of the organ weight to the body weight (body weight ratio) was calculated. Both of lung and kidney in flight rats were significantly heavier than ground controls in the body weight ratio. We found that the kidney in the nine-day and the fifteen-day group tended to extend of dorsal-ventral length in macroscopic observations. However, this difference was not observed in the re-adaptation group. These results suggest that space environment may affect in kidney development. On the other hand, the lung had no differences in macroscopic structure among flight and control groups.

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Abdominal arterial pressure during parabolic flight was measured using a telemetry system to clarify the acute effect of microgravity on hemodynamics in conscious rats. The microgravity condition was elicited by three different levels of entry gravity, i.e. 2 G, 1.5 G and 1 G. On exposure to 2 G, mean aortic pressure (MBP) increased up to 118.7 mm Hg +/- 7.3 compared with the value at 1 G (107.0 +/- 6.3 mm Hg, n=6). The value at microgravity preceded by 2 G was 118.0 mmHg +/- 5.2 mm HG and it was still higher than at 1 G. When 1.5 G was elicited before microgravity exposure, MBP also increased (1.5 G: 114.9 +/- 5.3 vs 1 G: 105.8+/-5.0 mm Hg) and the value at microgravity was 117.3 + /- 5.3 mmHg. During pre-microgravity maneuver with 1 G, no changes were observed compared with the control level at 1 G (pre-microgravity: 105.0 +/- 5.0 vs 1G: 104.8 +/- 5.1 mm Hg ), whereas the MBP increased up to 117.0 +/- 6.5 mm Hg on exposure to microgravity. From these results, we found that in conscious rat MBP increase during acute microgravity exposure with either 1 G or hyper-G entry.

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In humans, it has been reported that dynamic exercise attenuates spontaneous baroreceptor reflex sensitivity (sBRS), which is an index of the gain of the baroreceptor-cardiac reflex. We demonstrated previously that endogenously produced NO from endothelial nitric oxide synthase (eNOS) within the nucleus tractus solitarii (NTS), the central terminal site of baroreceptor afferents, depressed sBRS. In this study, we investigated whether eNOS activity within the NTS plays any role in down-modulating the sBRS during dynamic exercise. In conscious Wistar rats arterial pressure and heart rate (HR) were monitored continuously and chronically using radiotelemetry before and during wheel cage running at 6 m min(-1) for 10 min. sBRS was determined by a time-series method. During dynamic exercise systolic blood pressure (SBP) and HR were significantly increased (SBP: 138 +/- 2 vs. 125 +/- 2 mmHg, P < 0.001; HR: 447 +/- 6 vs. 362 +/- 8 beats min(-1), P < 0.001) while sBRS was significantly decreased (0.53 +/- 0.03 vs. 1.08 +/- 0.08 ms mmHg(-1), P < 0.001). In sino-aortic denervated rats the change in SBP in response to dynamic exercise was significantly larger than that in baroreceptor-intact rats (denervated: 21.6 +/- 2.5 mmHg; intact: 12.0 +/- 2.8 mmHg, P < 0.05). In contrast, denervation made no difference to the change in HR. Although disabling eNOS activity in the NTS by adenoviral-directed expression of a dominant negative mutant form of eNOS increased resting sBRS (1.48 +/- 0.20 vs. 1.09 +/- 0.15 ms mmHg(-1), P < 0.05), the absolute level reached during dynamic exercise was identical to control. These results demonstrate that during dynamic exercise (i) the sBRS decreases around the operating point of the baroreceptor-cardiac reflex function curve in normotensive rats, (ii) the baroreceptor reflex operates to limit the rise in arterial pressure, and (iii) the attenuation of sBRS is not mediated by changes in eNOS activity within the NTS.

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Abdominal aortic pressure (AAP), heart rate (HR), and aortic nerve activity (ANA) during parabolic flight were measured by using a telemetry system to clarify the acute effect of microgravity (microG) on hemodynamics in rats. While the animals were conscious, AAP increased up to 119 +/- 3 mmHg on exposure to microG compared with the value at 1 G (95 +/- 3 mmHg; P < 0.001), whereas AAP decreased immediately on exposure to microG under urethane anesthesia (microG: 72 +/- 9 mmHg vs. 1 G: 78 +/- 8 mmHg; P < 0.05). HR also increased during microG in conscious animals (microG: 349 +/- 12 beats/min vs. 1 G: 324+9 beats/min; P < 0.01), although no change was observed under anesthesia. ANA, which was measured under anesthesia, decreased in response to acute microG exposure (microG: 33 +/- 7 counts/s vs. 1 G: 49 +/- 5 counts/s; P < 0.01). These results suggest that microG essentially induces a decrease of arterial pressure; however, emotional stress and body movements affect the responses of arterial pressure and HR during exposure to acute microG.

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