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At least 19 records

Effect of dietary sodium on fluid/electrolyte regulation during bed rest

BACKGROUND: A negative fluid balance during bed rest (BR) is accompanied by decreased plasma volume (PV) which contributes to cardiovascular deconditioning. HYPOTHESIS: We hypothesized that increasing dietary sodium while controlling fluid intake would increase plasma osmolality (POSM), stimulate fluid conserving hormones, and reduce fluid/electrolyte (F/E) losses during BR; conversely, decreasing dietary sodium would decrease POSM, suppress fluid conserving hormones, and increase F/E losses. METHODS: We controlled fluid intake (30 ml x kg(-1) x d(-1)) in 17 men who consumed either a 4.0 +/- 0.06 g x d(-1) (174 mmol x d(-1)) (CONT; n = 6), 1.0 +/- 0.02 g x d(-1) (43 mmol x d(-1)) (LS; n = 6), or 10.0 +/- 0.04 g x d(-1) (430 mmol x d(-1)) (HS; n = 5) sodium diet before, during, and after 21 d of 6 degrees head-down BR. PV, total body water, urine volume and osmolality, POSM, and F/E controlling hormone concentrations were measured. RESULTS: In HS subjects, plasma renin activity (-92%), plasma/urinary aldosterone (-59%; -64%), and PV (-15.0%; 6.0 ml x kg(-1); p < 0.05) decreased while plasma atrial natriuretic peptide (+34%) and urine antidiuretic hormone (+24%) increased during BR (p < 0.05) compared with CONT. In LS, plasma renin activity (+166%), plasma aldosterone (+167%), plasma antidiuretic hormone (+19%), and urinary aldosterone (+335%) increased with no change in PV compared with CONT (p < 0.05). Total body water did not change in any of the subjects. CONCLUSIONS: Contrary to our hypothesis, increasing dietary sodium while controlling fluid intake during BR resulted in a greater loss of PV compared with the CONT subjects. Reducing dietary sodium while controlling fluid intake did not alter the PV response during BR compared with CONT subjects.

Clinical Trial↗

Dietary Sodium Effects on Bone Loss and Calcium Metabolism During Bed Rest

The acceleration of age-related bone loss is one of the most detrimental effects of space flight. The ability to understand and counteract this loss will be critical for crew health and safety during and after long-duration missions. Studies in healthy ambulatory individuals have linked high salt (sodium) diets, hypercalciuria, and increased renal stone risk. Dietary salt may modulate bone loss through changes in calcium metabolism and the calcium endocrine system. The research proposed here will determine the role of dietary salt in the loss of bone during simulated space flight. Calcium metabolism will be determined through calcium kinetics studies, endocrine and biochemical measurements; and estimates of the mass, distribution and mechanical properties of bone, in subjects fed low (100 mmol sodium/day) or high (250 mmol sodium/day) levels of dietary salt during 28 days of headdown tilt bedrest. This research addresses the role of dietary salt in the loss of bone and calcium in space flight, and integrates the changes in calcium metabolism with those occurring in other physiologic systems. These data will be critical for both countermeasure development, and in determination of nutritional requirements for extended-duration space flight. The potential countermeasures resulting from this research will reduce health risks due to acceleration of age-related osteoporosis and increased risk of renal stone formation..

Smith, Scott M.↗

Orthostatic responses to dietary sodium restriction during heat acclimation

Several studies have shown that individuals consuming low-salt diets and working in the heat have an increased risk or incidence of heat injury, suggestive of inadequate cardiovascular adjustment. Furthermore, others have shown that prolonged work in hot climates can precipitate orthostatic hypotension and syncope. This study was designed to evaluate the effects of moderate-salt (MS) and low-salt (LS) diets on the circulatory responses and incidence of presyncopal symptoms to an orthostatic test (OT) during successive days of heat acclimation (HA). Seventeen unacclimatized male soldiers (mean +/- SE: age 20+/-1 yrs) participated in this two-phase study. The first phase consisted of a seven day dietary stabilization period during which all subjects consumed similar diets of about 4000 kcal/day containing 8g NaCl and lived in a dormitory setting (21 C, 30% RH). The second phase commenced on day eight and consisted of dietary NaCl restriction and 10 days HA (days 8-17). Volunteers were randomly assigned to either the MS diet (n=9) providing 8g NaCl/day or the LS diet (n=8) furnishing just 4g NaCl/day. The acquisition of HA was manifested in both groups by reductions in exercising rectal temperature and heart rate (HR); these characteristics were similar in the MS and LS diets. The OT was performed at 21 C on day seven of the stabilization phase and on days 9, 11, 13, 15, and 17 of the HA phase, before and after 8.5 hr of intermittent treadmill walking in a hot environment. Blood pressure (BP) and HR responses at 1,2, and 4 min and any presyncopal symptoms were recorded after assuming an upright position from recumbency. All subjects completed the OT before and after prolonged exercise in the heat without incidence of either hypotension or presyncopal symptoms irrespective of dietary-salt intake and day of HA. The results indicate that the prolonged work in the heat can be performed without orthostatic hypotension or syncope while consuming 4g NaCl/day with adequate fluid replacement. Furthermore, the circulatory responses to OT showed no improvement with successive days of HA irrespective of dietary-salt intake.

Szlyk, Patricia C.↗

Human cardiovascular response to sympathomimetic agents during head-down bed rest: the effect of dietary sodium

Changes in sympathoadrenal function and cardiovascular deconditioning have long been recognized as a feature of the physiological adaptation to microgravity. The deconditioning process, coupled with altered hydration status, is thought to significantly contribute to orthostatic intolerance upon return to Earth gravity. The cardiovascular response to stimulation by sympathomimetic agents before, during, and after exposure to simulated microgravity was determined in healthy volunteers equilibrated on normal or high sodium diets in order to further the understanding of the deconditioning process.

NASA Discipline Number 18-10↗

Effects of Long Duration Spaceflight on Venous and Arterial Compliance

The visual impairment and intracranial pressure syndrome (VIIP) is a newly described space flight-associated medical condition made up of a constellation of symptoms affecting at least 34% of American astronauts who have flown International Space Station (ISS) missions. VIIP is defined primarily by visual acuity deficits and anatomical changes to eye structures, and is thought to be related to elevated intracranial pressure secondary to space flightinduced cephalad fluid shifts. Loss of visual acuity could be a significant threat to crew health and performance and may be suggestive of other adaptations with implications for years post-flight. Our primary objective is to determine whether vascular compliance is altered by space flight and whether such adaptations are related to the incidence of VIIP. In particular, we will measure ocular parameters and vascular compliance in vessels of the head and neck in astronauts who have no space flight experience, in astronauts before, during, and after space flight, and in bed rest subjects with conditions similar to space flight. Additionally, we will analyze astronaut data from the Lifetime Surveillance of Astronaut Health (LSAH) archive to determine which factors might be predictive of the development of VIIP. The project will be conducted in four separate but related parts. To understand the baseline condition of astronauts without any prior space flight experience, we will study 10 astronauts who have never flown in space by performing a comprehensive evaluation of the vasculature of the head, neck and eyes. Hemodynamic data (stroke volume and blood pressure), ocular (tonometry and ocular ultrasound), venous and arterial parameters will be acquired across a range of tilt angles (20, 10, 0, -10, -20 degrees). Vessels to be studied include the temporal, jugular, and vertebral veins and the cerebral, carotid and vertebral arteries. Ophthalmic data from the annual physical will be obtained through data sharing. To examine the relation between vascular compliance in the head and neck and the development of VIIP after a long duration space flight, we will study 10 astronauts before, during, and after long-duration ISS missions. Pre- and post-flight testing will be identical to that described above. During flight, images of the same vessels of interest will be obtained for later analysis. Ophthalmic data including VIIP scores will be obtained through data sharing from medically-required tests. To investigate the effects of age and elevated sodium intake, two potential contributors to VIIP, we will study 24 men (in two age groups: 25-35 and 45-55) during a 14 day 6deg head-down bed rest, a well-accepted analog of space flight. Standard NASA bed rest conditions will be maintained except for dietary sodium. Sodium intake will be similar to that of ISS astronauts, which is higher than consumed in previous bed rest studies. Pre- and post-bed rest testing procedures will be identical to the testing protocol described above for astronauts. Ophthalmic testing (optical coherence tomography, fundoscopy, and tonometry) will be conducted on the same day that vascular compliance measures are obtained. To identify parameters that may relate to an increase in an astronaut's susceptibility to developing VIIP, we will use data mining techniques to evaluate astronaut data obtained from the LSAH. Medical history, family history, space flight history and its related exposures, and history of high performance jet aircraft exposure will be examined for their potential relationship to ocular data. We hypothesize that the cephalad fluid shift induced by space flight will result in structural and functional adaptations in head and neck vessels leading to decreased vascular compliance and related to the development of VIIP symptoms. Further, although VIIP has not been observed in previous bed rest studies, we hypothesize that an elevated sodium intake will increase the incidence of VIIP symptoms in this space flight analog. Finally, we hypothesize that data mining analyses will reveal relationships between health history, previous exposures (including space flight and high performance aircraft), and the development of VIIP in the astronaut population.

Ribeiro, L. C.↗

Development of Countermeasures and Exercise Protocols to Reduce the Effects of Microgravity

I have helped scientists at NASA-JSC in analyzing data from many projects. Some of the major ones are: (1) cardiovascular responses to lower body negative pressure (LBNP) following bed rest, (2) the effects of dietary sodium, (3) in-flight cycle exercise mitigates reduced oxygen consumption at submaximal heart rates following space flight, (4) exercise thermoregulation after 13 days of head down bed rest, and (5) bed rest induced orthostatic intolerance. Many of the projects have now been completed and some of them are in the process of being published and others have been presented at national meetings. These projects have helped me be a true statistician and given me a real-life perspective of how interesting and complicated data can be. As a by-product of of these involvements I have been able to write and publish some methodological research that have applications in NASA and elsewhere. For instance, while I was at JSC, I happened to meet Dr. Al Feiveson and got into a discussion of the Space Shuttle Reliability. This led us to rethink about the way the data on the accelerated life testing of space shuttle pressure vessels had been analyzed. This has resulted in a major statistical paper and the paper has appeared in one of the top journals in the field of Statistics. A review of the paper by the editor of the journal was published in AmStatNews, a copy is attached with this report. I have presented these findings at the national/international statistics conference and at other places. I have also written another paper on reliability and a paper on calibration techniques that have applications in the engineering and the biomedical branches of NASA. Further, I am currently in the process of writing at least two more papers that have direct applications in NASA related studies.

Kulkarni, Pandurang M.↗

Water and Energy Dietary Requirements and Endocrinology of Human Space Flight

Fluid and energy metabolism and related endocrine changes have been studied nearly from the beginning of human space flight in association with short- and long-duration flights. Fluid and electrolyte nutrition status is affected by many factors including the microgravity environment, stress, changes in body composition, diet, exercise habits, sleep cycles, and ambient temperature and humidity conditions. Space flight exposes astronauts to all these factors and consequently poses significant challenges to establishing dietary water, sodium, potassium, and energy recommendations. The purpose of this article is to review the results of ground-based and space flight research studies that have led to current water, electrolyte, and energy dietary requirements for humans during space flight and to give an overview of related endocrinologic changes that have been observed in humans during short- and long-duration space flight.

short duration↗

Optimized bioregenerative space diet selection with crew choice

Previous studies on optimization of crew diets have not accounted for choice. A diet selection model with crew choice was developed. Scenario analyses were conducted to assess the feasibility and cost of certain crew preferences, such as preferences for numerous-desserts, high-salt, and high-acceptability foods. For comparison purposes, a no-choice and a random-choice scenario were considered. The model was found to be feasible in terms of food variety and overall costs. The numerous-desserts, high-acceptability, and random-choice scenarios all resulted in feasible solutions costing between 13.2 and 17.3 kg ESM/person-day. Only the high-sodium scenario yielded an infeasible solution. This occurred when the foods highest in salt content were selected for the crew-choice portion of the diet. This infeasibility can be avoided by limiting the total sodium content in the crew-choice portion of the diet. Cost savings were found by reducing food variety in scenarios where the preference bias strongly affected nutritional content.

Non-NASA Center↗

Angiotensin converting enzyme inhibition and the kidney

Angiotensin II (Ang II) induces a marked reduction in renal blood flow at doses well below those required to induce a pressor response, and as blood flow falls there is a decline in glomerular filtration rate and sodium excretion. This striking sensitivity of the renal blood supply led many workers to consider the possibility that angiotensin functions as a local renal hormone. As angiotensin converting enzyme (ACE) was found in particular abundance in the lung, it seemed reasonable to suspect that most of the conversion occurred there, and that the function of Ang II would be primarily systemic, rather than intrarenal. In this review, I will explore the evidence that has accumulated on these two possibilities, since they have important implications for our current understanding of normal kidney function and derangements of kidney function in disease.

NASA Program Biomedical Research↗

The extended amygdala and salt appetite

Both chemo- and mechanosensitive receptors are involved in detecting changes in the signals that reflect the status of body fluids and of blood pressure. These receptors are located in the systemic circulatory system and in the sensory circumventricular organs of the brain. Under conditions of body fluid deficit or of marked changes in fluid distribution, multiple inputs derived from these humoral and neural receptors converge on key areas of the brain where the information is integrated. The result of this central processing is the mobilization of homeostatic behaviors (thirst and salt appetite), hormone release, autonomic changes, and cardiovascular adjustments. This review discusses the current understanding of the nature and role of the central and systemic receptors involved in the facilitation and inhibition of thirst and salt appetite and on particular components of the central neural network that receive and process input derived from fluid- and cardiovascular-related sensory systems. Special attention is paid to the structures of the lamina terminalis, the area postrema, the lateral parabrachial nucleus, and their association with the central nucleus of the amygdala and the bed nucleus of the stria terminalis in controlling the behaviors that participate in maintaining body fluid and cardiovascular homeostasis.

NASA Discipline Regulatory Physiology↗

Effects of subfornical organ lesions on acutely induced thirst and salt appetite

We examined the role of the subfornical organ (SFO) in stimulating thirst and salt appetite using two procedures that initiate water and sodium ingestion within 1-2 h of extracellular fluid depletion. The first procedure used injections of a diuretic (furosemide, 10 mg/kg sc) and a vasodilator (minoxidil, 1-3 mg/kg ia) to produce hypotension concurrently with hypovolemia. The resulting water and sodium intakes were inhibited by intravenous administration of ANG II receptor antagonist (sarthran, 8 micrograms . kg(-1). min(-1)) or angiotensin-converting enzyme inhibitor (captopril, 2.5 mg/h). The second procedure used injections of furosemide (10 mg/kg sc) and a low dose of captopril (5 mg/kg sc) to initiate water and sodium ingestion upon formation of ANG II in the brain. Electrolytic lesions of the SFO greatly reduced the water intakes, and nearly abolished the sodium intakes, produced by these relatively acute treatments. These results contrast with earlier findings showing little effect of SFO lesions on sodium ingestion after longer-term extracellular fluid depletion.

NASA Discipline Regulatory Physiology↗

Managing the Risk for Early Onset Osteoporosis in Long-Duration Astronauts Due to Spaceflight

Early Onset Osteoporosis is probably the most recognized but poorly understood long-term health risk due to spaceflight. Osteoporosis management is primarily prophylactic and clinical interventions rely upon the ability to predict fractures which is currently determined by surrogate measures of bone strength. The RMAT for Early Onset Osteoporosis identified some open issues related to the fact that long-duration astronauts compose a unique group of subjects for which clinical approaches for osteoporosis management do not apply. Long-duration astronauts are healthy, young (25 to 55 years of age), predominantly male, and physical fit relative to the typical osteoporosis patient. Moreover, during prolonged space missions (typically 6-month missions) the skeleton not only adapts to weightlessness, but is influenced by numerous risk factors induced by operational constraints, e.g., inability to maintain preflight weight-bearing and aerobic activities, sub-optimal dietary intake (e.g., high sodium content for food stability, lack of fresh fruit and vegetables), suppression of vitamin D metabolism by uv shielding, and remote medicine care. Moreover, adaptation results in novel changes to astronauts bones that cannot be detected by current medically-useful measures. Consequently, a panel of clinicians (recognized leaders and policy-makers in osteoporosis) was convened to review the dataset of bone measures and bone loss risk factors in long-duration astronauts. Driven by the queries in the RMAT, the panel was charged to determine 1) if an intervention is required to prevent this risk, 2) what type and at what time would intervention be optimal, 3) what is the clinical trigger that would require a medical response from flight surgeons and 4) how should research data be used in the clinical care of astronauts. Hence, the RMAT determined that a bone health policy need to be formulated specific for this unique cohort subjected to a novel skeletal condition

Sibonga, Jean D.↗

Studies of acid-base homeostasis during simulated weightlessness: Application of the water immersion model to man

The effects of water immersion on acid-base homeostasis were investigated under carefully controlled conditions. Studies of renal acidification were carried out on seven healthy male subjects, each consuming a diet containing 150 meq sodium and 100 meq potassium. Control and immersion studies were carried out on each subject on the fourth and sixth days, respectively, of dietary equilibration, by which time all subjects had achieved sodium balance. The experimental protocols on study days were similar (except for the amount of water administered).

Epstein, M.↗

Nutritional Status Assessment During the Phase IIA and Phase III Lunar/Mars Life Support Test Project

Nutrition is a critical concern for extended-duration space missions (Smith and Lane, 1999). Loss of body weight is a primary consequence of altered nutrition, and is frequently observed during space flight (Smith and Lane; 1999). Other existing dietary concerns for space flight include excessive intakes of sodium and iron, and insufficient intakes of water and vitamin D (Smith and Lane, 1999). Furthermore, dependence on closed or semi-closed food systems increases the likelihood of inadequate intakes of key nutrients. This is a significant concern for extended-duration space missions. Space nutrition research often necessitates detailed recording of all food consumption. While this yields extremely accurate data, it requires considerable time and effort, and thus is not suitable for routine medical monitoring during space flight. To alleviate this problem, a food frequency questionnaire (FFQ) was designed to provide a quick and easy, yet reasonably accurate, method for crewmembers to provide dietary intake information to the ground. We report here a study which was designed to assess nutritional status before, during, and after the 60-d and 91-d chamber stays. An additional goal of the study was to validate a food frequency questionnaire designed specifically for use with space flight food systems.

Smith, Scott M.↗

Implementation of the NASA AG-Bed Rest Pilot

To examine the efficacy of artificial gravity (AG) as a countermeasure to spaceflight deconditioning, intermittent AG produced by a horizontal short-radius centrifuge (SRC) was utilized on human test subjects deconditioned by bed rest. This poster will present the subject screening, study design, logistics, and implementation of the 41 day pilot study conducted at the University of Texas Medical Branch, Galveston, TX bed rest facility. An extensive screening process was employed to exclude subjects that were dissimilar to the U.S. astronaut population. Candidates underwent a modified U.S. Air Force Class III physical and tests of bone density, cardiovascular fitness, vestibular system function, psychological fitness and centrifuge tolerance. 15 subjects completed the study; 7 control and 8 AG treatment. All provided written consent to volunteer after the nature of the study and its hazards were clearly explained to them. Standard conditions were strictly regulated; Ta = 72 +/- 2 F, humidity = 70 +/- 5%, light/dark cycle 16h:8h. All fluid intake (minimum 28.5 ml/kg body weight/day) and urine output was monitored. Caloric intake was adjusted as necessary to maintain body weight. Carbohydrate, fat and protein were provided in a ratio of 55:30:15. Phosphorus intake was 1400 mg/d, sodium intake was 2 mmol/kg/d, potassium intake was 1.3 mmol/kg/d, and dietary calcium intake was 1000 mg/d. A physician examined each subject daily. During the first 11 days of the study protocol, subjects were ambulatory, but confined to the facility. Subjects participated in multiple baseline tests of bone, muscle, cardiovascular, sensory-motor, immunological, and psychological function. On the 12th day, subjects entered the bed rest phase of the study, during which they were confined to strict 6? head down tilt bed rest for 21 days. Beginning 24 hrs into this period, treatment subjects received 1 hour daily exposures to artificial gravity which was produced by spinning the subjects on a 3.0 m radius SRC. They were oriented radially in the supine position so that the centrifugal force was aligned with their long body axis, and while spinning, they #stood# on a force plate, supporting the centrifugal loading (2.5 g at the feet, 1.0 g at the heart). The subject station allowed free translation over approximately 10 cm to ensure full loading of the lower extremities and to allow for anti-orthostatic muscle contractions. Control subjects were positioned on the centrifuge but did not spin. Following the bed rest phase, subjects were allowed to ambulate again, but remained within the facility for an additional 9 days and participated in multiple follow-up tests of physiological function.

Warren, L. E.↗

Fluid and Electrolyte Nutrition

Studies of fluid and electrolyte homeostasis have been completed since the early human space flight programs, with comprehensive research completed on the Spacelab Life Sciences missions SLS-1 and SLS-2 flights, and more recently on the Mir 18 mission. This work documented the known shifts in fluids, the decrease in total blood volume, and indications of reduced thirst. Data from these flights was used to evaluate the nutritional needs for water, sodium, and potassium. Interpretations of the data are confounded by the inadequate energy intakes routinely observed during space flight. This in turn results in reduced fluid intake, as food provides approximately 70% water intake. Subsequently, body weight, lean body mass, total body water, and total body potassium may decrease. Given these issues, there is evidence to support a minimum required water intake of 2 L per day. Data from previous Shuttle flights indicated that water intake is 2285 +/- 715 ml/day (mean +/- SD, n=26). There are no indications that sodium intake or homeostasis is compromised during space flight. The normal or low aldosterone and urinary sodium levels suggest adequate sodium intake (4047 +/- 902 mg/day, n=26). Because excessive sodium intake is associated with hypercalciuria, the recommended maximum amount of sodium intake during flight is 3500 mg/day (i.e., similar to the Recommended Dietary Allowance, RDA). Potassium metabolism appears to be more complex. Data indicate loss of body potassium related to muscle atrophy and low dietary intake (2407 +/- 548 mg/day, n=26). Although possibly related to measurement error, the elevations in blood potassium suggest alterations in potassium homeostasis. The space RDA for minimum potassium intake is 3500 mg/day. With the documented inadequate intakes, efforts are being made to increase dietary consumption of potassium.

Lane, Helen W.↗

Renal stone risk assessment during Space Shuttle flights

PURPOSE: The metabolic and environmental factors influencing renal stone formation before, during, and after Space Shuttle flights were assessed. We established the contributing roles of dietary factors in relationship to the urinary risk factors associated with renal stone formation. MATERIALS AND METHODS: 24-hr. urine samples were collected prior to, during space flight, and following landing. Urinary and dietary factors associated with renal stone formation were analyzed and the relative urinary supersaturation of calcium oxalate, calcium phosphate (brushite), sodium urate, struvite and uric acid were calculated. RESULTS: Urinary composition changed during flight to favor the crystallization of calcium-forming salts. Factors that contributed to increased potential for stone formation during space flight were significant reductions in urinary pH and increases in urinary calcium. Urinary output and citrate, a potent inhibitor of calcium-containing stones, were slightly reduced during space flight. Dietary intakes were significantly reduced for a number of variables, including fluid, energy, protein, potassium, phosphorus and magnesium. CONCLUSIONS: This is the first in-flight characterization of the renal stone forming potential in astronauts. With the examination of urinary components and nutritional factors, it was possible to determine the factors that contributed to increased risk or protected from risk. In spite of the protective components, the negative contributions to renal stone risk predominated and resulted in a urinary environment that favored the supersaturation of stone-forming salts. Dietary and pharmacologic therapies need to be assessed to minimize the potential for renal stone formation in astronauts during/after space flight.

STS Shuttle Project↗

Zinc, copper, and nitrogen balances during bed rest and fluoride supplementation in healthy adult males

The effects of bed rest and fluoride supplementation on zinc, copper, and nitrogen balances and Zn and Cu serum levels were measured in 15 healthy males. Subjects aged 19-54 y remained on a metabolic research ward for 10 wk. During weeks 1-5, subjects were ambulatory. During wks 6-10 they remained in continuous bed rest. During weeks 3-10 nine subjects received 10 or 20 mg F/d as sodium fluoride. Daily urine and weekly fecal composites were made and biweekly fasting blood samples were taken. Dietary intakes were 1.40 +/- 0.17 mg Cu/d (22.0 +/- 2.7 mumol Cu/d), 10.82 +/- 0.49 mg Zn/d (165.6 +/- 7.6 mumol Zn/d), and 14.27 +/- 0.23 g N/d (1019 +/- 16 mmol N/d). Bed rest increased urinary Zn and N excretions and fecal Zn excretions and decreased Zn balance (p less than 0.05) whereas Cu balance was unchanged. During bed rest, F supplementation increased Zn and N balances compared with untreated control subjects (p less than 0.05). These results are compatible with bone and muscle atrophy during bed rest and increased bone formation with F supplementation.

Clinical Trial↗