Vascular adaptation to deconditioning and the effect of an exercise countermeasure: results of the Berlin Bed Rest study.

Deconditioning is a risk factor for cardiovascular disease. The physiology of vascular adaptation to deconditioning has not been elucidated. The purpose of the present study was to assess the effects of bed rest deconditioning on vascular dimension and function of leg conduit arteries. In addition, the effectiveness of resistive vibration exercise as a countermeasure for vascular deconditioning during bed rest was evaluated. Sixteen healthy men were randomly assigned to bed rest (BR-Ctrl) or to bed rest with resistive vibration exercise (BR-RVE). Before and after 25 and 52 days of strict horizontal bed rest, arterial diameter, blood flow, flow-mediated dilatation (FMD), and nitroglycerin-mediated dilatation were measured by echo Doppler ultrasound. In the BR-Ctrl group, the diameter of the common femoral artery decreased by 13 +/- 3% after 25 and 17 +/- 1% after 52 days of bed rest (P < 0.001). In the BR-RVE group this decrease in diameter was significantly attenuated (5 +/- 2% after 25 days and 6 +/- 2% after 52 days, P < 0.01 vs. BR-Ctrl). Baseline blood flow did not change after bed rest in either group. After 52 days of bed rest, FMD and nitroglycerin-mediated dilatation of the superficial femoral artery were increased in both groups, possibly by increased nitric oxide sensitivity. In conclusion, bed rest deconditioning is accompanied by a reduction in the diameter of the conduit arteries and by an increased reactivity to nitric oxide. Resistive vibration exercise effectively attenuates the diameter decrease of leg conduit arteries after bed rest.     

Mobility of the early equine conceptus

Movement of the conceptus within the uterine lumen of barren mares was studied by daily ultrasound examinations on days 11-20 and by rectal palpation on days 15-48 (Experiment 1) and by ultrasound examinations 3 or 4 times per day at 2-4 hour intervals on days 11-16 (Experiment 2). In addition, broodfarm records were analyzed to compare side of ovulation with side of embryo attachment (Experiment 3). The vesicle was found in opposite uterine horns for 43% of the successive, daily, ultrasound examinations on days 11 and 12, 12 and 13, 13 and 14, and 14 and 15; 24% of the successive examinations on days 15 and 16; and 8% on days 16 and 17. No movement was detected after day 17. The vesicle was found in opposite horns during 41% of the successive examinations at 2-4 hour intervals on days 11, 12, 13, 14, and 15, but no movement was detected on day 16. In addition, no transuterine migration was found by rectal palpation between the day of first detection of an embryonal enlargement (mean, day 17) and day 48. During ultrasound examination on days 11-15, the vesicle was found significantly more frequently in the left horn (66% of the observations) than in the right horn (34%); however, final attachment occurred more frequently in the right horn (63% of the mares). In analyses of brood-farm records, ovulation occurred with equal frequency in left and right ovaries in barren and lactating mares, but with significantly greater frequency in the left ovary (63%) in maiden mares. Regardless of the side of ovulation, final attachment of the conceptus occurred significantly more frequently in the right horn (66%) in barren and maiden mares, but not in lactating mares.     

Anatomic transformation of the lymphatic bed of the small intestine following immunization]

The experimental investigation of the lymphatic bed of the small intestine after immunization was performed in 15 rabbits. Ten rabbits were taken as control. The rabbits were immunized by administration of kidney antigen with the complete Freund adjuvant. The animals were killed on the 7-8th days after two cycles of immunization, three injections in each cycle. The interval between the injections was three days, between the cycles - one month. Polychrome injection of arteries and veins was made. The lymphatic bed was filled with Gerota's mass. The chylus sinuses, lymph capillaries and vessels of experimental animals were found to be dilated 2-6 times as compared with normal. There appear many blind processes on the walls of lymph capillaries and vessels. Intraorganic lymphatic pathways were more dilated as compared with extraorganic. The ileum was found to be more reactive in immunization than other two parts of the small intestine.     

Effect of ultrasound on the formation of arterial and lymphatic collaterals in the rat pelvic extremities]

Pronounced morphological alterations combined with destruction of muscular tissue make their appearance in intact arterial and lymphatic bed of pelvic extremities in rats after treatment with ultrasound of high intensity. Ultrasound of low intensity failed to cause morphological changes either in the arterial or the lymphatic bed of extremities. A course of exposure of injured arterial and lymphatic bed of rat's pelvic extremities to ultrasound of small intensity considerably accelerated processes of formation of the arterial and lymphatic collaterals.     

Prolonged bed rest impairs response of the small arteries to cold pressor test

Cold pressor test (CPT) has been known to increase vascular resistance through adrenergic stimulation. To assess the effect of prolonged bed rest (BR) on sympathetic neural control, blood flow velocity of the proper palmar digital artery (resistance vessel) was measured in 10 healthy volunteers during a 90 seconds CPT by pulsed Doppler ultrasound using a 7.5 MHz linear transducer before and after 20 days BR. Blood flow velocity in the proper palmar digital artery decreased during the first half of the CPT both before and after BR, but decreases in peak flow velocity became less prominent (p<0.05) after BR (-48+/-20%) than before bed rest (-61+/-25%). The only subject whose lower body negative pressure tolerance time was prolonged after BR showed prominent decreases in peak flow velocity during the first half of the CPT after BR. Peak flow velocity returned to the pre-test value during the latter half of the CPT in 7 subjects before BR, but in only 2 subjects after BR (p<0.05). Thus, 20 days BR impairs the response of the small digital arteries to CPT, suggesting that a decrease in a adrenergic stimulation occurs after prolonged BR.     

Gestational stage sensitivity to ultrasound effect on postnatal growth and development of mice

BACKGROUND: An experiment was conducted to find out whether ultrasound exposure leads to changes in postnatal growth and development in the mouse. METHODS: A total of 15 pregnant Swiss albino mice were exposed to diagnostic levels of ultrasound (3.5 MHz, 65 mW/cm2, I(SPTP) = 1 mW/cm2 Intensity(Spatial Peak-Temporal Peak), I(SATA) = 240 mW/cm2 Intensity(Spatial Average-Temporal Average)) for 30 min for a single day between days 10 and 18 of gestation (GD 10-18). Virgin female mice were placed with same age group males for mating in the ratio 2 females : 1 male and examined the next morning for the presence of vaginal plug, a sign of successful copulation. The females with vaginal plugs were separated and labeled as 0-day pregnant. Maternal vaginal temperature was also measured. A sham exposed control group of 15 pregnant mice was maintained for comparison. All exposed as well as control animals were left to complete gestation and parturition. Their offspring were used in our further studies. They were monitored during early postnatal life for standard developmental markers, postnatal mortality, body weight, body length, head length, and head width, and growth restriction was recorded up to 6 weeks of age. RESULTS: An exposure to ultrasound induced nonsignificant deviations in the maternal vaginal temperature or developmental markers. Significant low birth weight was observed in the present study, after exposure at GD 11, 12, 14, and 16. However, 14 and 16 days postcoitus during the fetal period appears to be the most sensitive to the ultrasound effect, in view of the number of different effects as well as severity of most of the observed effects when exposed on these gestation days. CONCLUSIONS: The results indicate that diagnostic ultrasound can induce harmful effects on mouse growth and development when given at certain critical periods of gestation.     

Effect of bed rest on the adenine nucleotides concentration in human blood platelets.

The effect of immobilization in bed on metabolism and function of human blood platelet was studied. Blood platelets taken from patients with bone fractures after long term bed rest (14 days and 28 days) demonstrated significantly reduced concentration of total adenine nucleotides (after 28 days reduction about 30%). This decrease of total platelet adenine nucleotides after immobilization in bed is probably caused by stimulation of platelet secretory process. Thrombin which released from control platelets 58.2% +/- 1.5% of total adenine nucleotides liberated decreased amounts (only 23.1% +/- 3.3% of total) of nucleotides from patient platelets isolated after 28 days of immobilization in bed. Loss of nucleotides from platelets was accompanied by slightly increased extent of platelet aggregation. It is concluded that during bed rest the reactivity of blood platelets (aggregation and release reaction) is stimulated.     

Seven days of bed rest decrease insulin action on glucose uptake in leg and whole body

Impaired glucose tolerance develops in normal humans after short-term bed rest. To elucidate the mechanism, insulin action on whole body glucose uptake rate (WBGUR) and leg glucose uptake rate (LGUR) was measured by sequential euglycemic clamp technique combined with femoral arterial and venous cannulation at insulin concentrations of 10 +/- 1, 18 +/- 1, 37 +/- 2, and 360 +/- 15 microU/ml. Studies were performed before (C) and after (BR) 7 days of strict bed rest. WBGUR was significantly lower after bed rest than before (5.5 +/- 0.4 and 7.2 +/- 0.8 mg.min-1.kg-1, respectively) when insulin was 37 microU/ml. LGUR was even more markedly depressed by bed rest, being 0.6 +/- 0.1, 0.9 +/- 0.2, and 2.8 +/- 0.4 mg.min-1.kg leg-1 (BR) compared with 0.9 +/- 0.1, 1.7 +/- 0.4, and 5.9 +/- 0.5 mg.min-1.kg leg-1 (C) (P less than 0.05) at the three lower insulin concentrations. At these insulin concentrations also, lactate release and glucose oxidation and glycogen storage estimated by indirect calorimetry were lower in the leg after bed rest. At the highest insulin dose WBGUR was similar on BR and C days, while LGUR was lower after bed rest. In conclusion, 7 days of bed rest decrease whole body insulin action, a fact that is explained by decreased insulin action in inactive muscle.     

Dual-frequency ultrasound enhances functional neuron differentiation from neural stem cells by modulating Ca2+ dynamics and the ERK1/2 signaling pathway

Our previous study demonstrated that ultrasound is able to promote differentiation on neural stem cells (NSCs), and dual-frequency ultrasound promotes this effect due to enhanced acoustic cavitation compared with single-frequency ultrasound. However, the underlying biological reasons have not been well disclosed. The purpose of this study was to investigate the underlying bioeffects, mechanisms and signaling pathways of dual-frequency ultrasound on NSC differentiation. The morphology, neurite outgrowth, and differentiation percentages were investigated under various dual-frequency simulation parameters with exposure periods varying from 5 to 15 min. Morphological observations identified that dual-frequency ultrasound stimulation promoted ultrasound dose-dependent neurite outgrowth. In particular, cells exposed for 10 min/2 days showed optimal neurite outgrowth and neuron differentiation percentages. In addition, live cell calcium images showed that dual-frequency ultrasound enhanced the internal calcium content of the cells, and calcium ions entering cells from the extracellular environment could be observed. Dual frequency ultrasound exposure enhanced extracellular calcium influx and upregulated extracellular signal-regulated kinases 1/2 (ERK1/2) expression. Observations from immunostaining and protein expression examinations also identified that dual-frequency ultrasound promoted brain-derived neurotrophic factor (BDNF) secretion from astrocytes derived from NSCs. In summary, evidence supports that dual-frequency ultrasound effectively enhances functional neuron differentiation via calcium channel regulation via the downstream ERK1/2 pathway and promotes BDNF secretion to serve as feedback to cascade neuron differentiation. The results may provide an alternative for cell-based therapy in brain injury.     

Growth and regression of ovarian follicles during the follicular phase of the oestrous cycle in heifers undergoing spontaneous and PGF-2 alpha-induced luteolysis

Ultrasonography was used to monitor the growth, ovulation and regression of individual ovarian follicles greater than or equal to 5 mm during the late luteal and follicular phases of the oestrous cycle in heifers treated with injections of PGF-2 alpha to induce luteolysis and in heifers undergoing spontaneous luteolysis. Six heifers were given a single injection of PGF-2 alpha between Day 12 and 15 of the oestrous cycle and their ovaries were examined daily by transrectal ultrasonography until ovulation occurred. Another group of 5 heifers was examined daily by ultrasound from Day 14 or 15 of the cycle through spontaneous luteolysis and ovulation. Blood samples were taken twice daily from this group and analysed for progesterone to determine when luteolysis occurred. All heifers were checked for oestrous behaviour twice daily. Mean diameters of ovulatory follicles on each of the 3 days before oestrus were not different between PGF-2 alpha-treated and untreated heifers. In both groups there was large variation among heifers in the sizes and growth rates of the ovulatory follicles. At 3 days before oestrus the diameters of ovulatory follicles were between 7.5 and 11 mm in PGF-2 alpha-treated heifers and between 6 and 11.5 mm in untreated heifers. Non-ovulatory follicles decreased in size during the 3 days before oestrus and the number of non-ovulatory follicles within the size ranges of ovulatory follicles decreased. The ovulatory follicle was not consistently the largest follicle on the ovaries until the day of oestrus but was always one of the 2 largest follicles during the 3 days before oestrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of 20 days bed rest on muscle morphology

Using ultrasound, muscle thickness and fascicle angles from aponeurosis were evaluated before, during and after 20 days bed rest (BR). Subjects were healthy adults (4 women and 4 men). Measurements were carried out before and after BR and after 10 weeks of recovery, respectively. Muscle measurements were taken at nine sites in trunk and upper and lower extremities, respectively. For the m. triceps brachii, m. vastus lateralis, and m. gastrocnemius medialis, fascicle angles from the aponeurosis as well as muscle thickness were measured. There was a high statistical significant correlation between muscle thickness and cross-sectional area for quadriceps muscles, suggesting applicability of muscle thickness for evaluation of muscle size. Muscle thickness decreased in muscles of the lower extremity by 2.1-4.4 % after bed rest. In triceps brachii and vastus lateralis muscles, there were no prominent changes in muscle thickness and fascicle angles. It was concluded that muscle morphology deteriorates with changes in muscle architecture by bed rest but the response is small and muscle-specific. It was also suggested that bed rest affects not only muscle mass but muscle tone as well.     

Regulation of muscle sympathetic nerve activity after bed rest deconditioning

Cardiovascular deconditioning reduces orthostatic tolerance. To determine whether changes in autonomic function might produce this effect, we developed stimulus-response curves relating limb vascular resistance, muscle sympathetic nerve activity (MSNA), and pulmonary capillary wedge pressure (PCWP) with seven subjects before and after 18 days of -6 degrees head-down bed rest. Both lower body negative pressure (LBNP; -15 and -30 mmHg) and rapid saline infusion (15 and 30 ml/kg body wt) were used to produce a wide variation in PCWP. Orthostatic tolerance was assessed with graded LBNP to presyncope. Bed rest reduced LBNP tolerance from 23.9 +/- 2.1 to 21.2 +/- 1.5 min, respectively (means +/- SE, P = 0.02). The MSNA-PCWP relationship was unchanged after bed rest, though at any stage of the LBNP protocol PCWP was lower, and MSNA was greater. Thus bed rest deconditioning produced hypovolemia, causing a shift in operating point on the stimulus-response curve. The relationship between limb vascular resistance and MSNA was not significantly altered after bed rest. We conclude that bed rest deconditioning does not alter reflex control of MSNA, but may produce orthostatic intolerance through a combination of hypovolemia and cardiac atrophy.     

Photooptical and ultrastructural changes in the microcirculatory bed of the kidney after functional vasoconstriction

In experiments on rats it was found that at the early stages (5 to 15 minutes) after vasoconstriction of the kidney caused by adrenaline solution there occurred sharp narrowing of the intraorganic arterial bed lumen, particularly that of the afferent arterioles. Ultrastructural changes in the glomerular renal capillary components observed were morphological expression of the effect of angiospasm and circulatory hypoxia. Residual phenomena of constriction of the renal microcirculatory bed still persisted at later stages--in 3, 7 days. These changes characterized the stage of peculiar mobility and contractile properties of their endothelial cells, caused by spasmogenic disturbances of microcirculation.     

Migration of myogenic cells in the rat extensor digitorum longus muscle studied with a split autograft model

Summary The ability of myogenic cells to migrate perpendicular to the long axis of freely autografted muscles was examined. Rat extensor digitorum longus muscles were divided, and one half was devitalized by repeated freezing in liquid nitrogen while the other half was kept viable in physiologic saline. The halves were reunited with sutures and grafted back into the original muscle bed. At intervals between 5 and 25 days the grafts were removed and examined histologically for the presence of myotubes within the devitalized region. Myotubes were first seen in the devitalized half 10 days postgrafting with the maximum number of myotubes observed after 12 to 15 days. These results indicate that myogenic cells are capable of migration perpendicular to the long axis of the muscle fibers in an autograft.     

State of the rat thyroid gland in coarctation of the abdominal aorta]

In order to study connections between blood vessels and follicular thyrocytes, the method of modulation (a purposeful change of state in one element with registration of states in other elements of the system) was used. In rats chronic increase of blood stream was produced in the thyroid gland; in 15 days it was 54% as high as in the control. The volume of the vascular bed increased by 28% and that of follicles by 26%. Volumetric ratio between the thyroid epithelium and colloid did not changed. Follicular thyrocytes grew high and the nuclear volumes of these cells increased. Thyrocytes greately varied in their height. The number of mast cells in the thyroid gland remained the same. Iodine absorption by the thyroid gland increased as it is dependent on the volume of the vascular bed of the organism (+0.82). The data obtained demonstrate a significant connection existing between the follicular thyrocytes and blood vessels.     

Ultrasound induced damages and time bound recovery in mouse liver

Therapeutic ultrasound at 875 kHz at 10 and 15 W/cm2 intensity induced extensive damages in the liver of mouse. Total exposure of 5 min was spread over 5 days. Aqueous medium was avoided by coupling the transducer directly to the skin surface. Mild to extensive damages were noted. Complete distortion of hepatocellular architecture was noted in 15 W irradiated mice. However, there was almost complete recovery by 10th day following the last exposure.     

Bed rest reduces metabolic protein content and abolishes exercise-induced mRNA responses in human skeletal muscle

The aim was to test the hypothesis that 7 days of bed rest reduces mitochondrial number and expression and activity of oxidative proteins in human skeletal muscle but that exercise-induced intracellular signaling as well as mRNA and microRNA (miR) responses are maintained after bed rest. Twelve young, healthy male subjects completed 7 days of bed rest with vastus lateralis muscle biopsies taken before and after bed rest. In addition, muscle biopsies were obtained from six of the subjects prior to, immediately after, and 3 h after 45 min of one-legged knee extensor exercise performed before and after bed rest. Maximal oxygen uptake decreased by 4%, and exercise endurance decreased nonsignificantly, by 11%, by bed rest. Bed rest reduced skeletal muscle mitochondrial DNA/nuclear DNA content 15%, hexokinase II and sirtuin 1 protein content ～45%, 3-hydroxyacyl-CoA dehydrogenase and citrate synthase activity ～8%, and miR-1 and miR-133a content ～10%. However, cytochrome c and vascular endothelial growth factor (VEGF) protein content as well as capillarization did not change significantly with bed rest. Acute exercise increased AMP-activated protein kinase phosphorylation, peroxisome proliferator activated receptor-γ coactivator-1α, and VEGF mRNA content in skeletal muscle before bed rest, but the responses were abolished after bed rest. The present findings indicate that only 7 days of physical inactivity reduces skeletal muscle metabolic capacity as well as abolishes exercise-induced adaptive gene responses, likely reflecting an interference with the ability of skeletal muscle to adapt to exercise.     

Metabolic Effects of 2 Days of Strict Bed Rest

ObjectiveTo examine the possibility of whether 2 days of strict hospitalized bed rest would alter the metabolic profile (including insulin resistance as calculated by the quantitative insulin sensitivity check index or QUIC- KI) in both normal subjects and patients with type 2 diabetes in comparison with 2 days of normal activity.MethodsThe design of this pilot study was a randomized, crossover protocol that evaluated the effects of strict bed rest versus normal activity in 5 healthy normal subjects and 5 healthy patients with type 2 diabetes. All study participants completed a screening visit for assessment of baseline health.ResultsAll 10 study subjects completed the protocol without adverse events. Fasting plasma glucose, insulin, and C-peptide levels as well as several known risk factors for atherosclerosis were unchanged in both the subjects without diabetes and the patients with type 2 diabetes after 2 days of hospitalized bed rest. Insulin resistance demonstrated no significant change during the 48 hours of bed rest when compared with the mean value at baseline.ConclusionThis study demonstrates that 48 hours of bed rest has no significant effect on insulin resistance or standard metabolic variables in normal subjects and patients with type 2 diabetes. Therefore, achieving good glucose control in patients hospitalized for a period of 2 days or less does not necessitate early ambulation to prevent an increase in insulin resistance. (Endocr Pract. 2008;14:564-569)     

Branched-chain amino acid supplementation during bed rest: effect on recovery.

Bed rest is associated with a loss of protein from the weight-bearing muscle. The objectives of this study are to determine whether increasing dietary branched-chain amino acids (BCAAs) during bed rest improves the anabolic response after bed rest. The study consisted of a 1-day ambulatory period, 14 days of bed rest, and a 4-day recovery period. During bed rest, dietary intake was supplemented with either 30 mmol/day each of glycine, serine, and alanine (group 1) or with 30 mmol/day each of the three BCAAs (group 2). Whole body protein synthesis was determined with U-(15)N-labeled amino acids, muscle, and selected plasma protein synthesis with l-[(2)H(5)]phenylalanine. Total glucose production and gluconeogenesis from alanine were determined with l-[U-(13)C(3)]alanine and [6,6-(2)H(2)]glucose. During bed rest, nitrogen (N) retention was greater with BCAA feeding (56 +/- 6 vs. 26 +/- 12 mg N. kg(-1). day(-1), P < 0.05). There was no effect of BCAA supplementation on either whole body, muscle, or plasma protein synthesis or the rate of 3-MeH excretion. Muscle tissue free amino acid concentrations were increased during bed rest with BCAA (0.214 +/- 0.066 vs. 0.088 +/- 0.12 nmol/mg protein, P < 0.05). Total glucose production and gluconeogenesis from alanine were unchanged with bed rest but were significantly reduced (P < 0.05) with the BCAA group in the recovery phase. In conclusion, the improved N retention during bed rest is due, at least in part, to accretion of amino acids in the tissue free amino acid pools. The amount accreted is not enough to impact protein kinetics in the recovery phase but does improve N retention by providing additional essential amino acids in the early recovery phase.     

Effects of chronic physical activity and of ultrasound treatment on bone consolidation

The objective of this study was to investigate the effects of ultrasound treatment and physical exercise on the velocity of bone consolidation and resistance to deformation. We performed osteotomy in the upper third of the right tibia of rats. Physical training consisted of swimming 1 h per day with a load of 5% b.w. for 30 days. Therapy with medium-intensity ultrasound was applied daily on the damaged area. Wistar rats were divided into the following groups: osteotomized sedentary animals with no ultrasound treatment (1.OSnUS), osteotomized trained animals with no ultrasound treatment (2.OTnUS), osteotomized sedentary animals with ultrasound treatment (3.OSwUS), and osteotomized trained animals with ultrasound treatment (4.OTwUS). The animals were sacrificed for the following analyses: muscle glycogen, serum alkaline phosphatase at the 5th, 10th, 20th, and 30th days, test of maximum resistance to flexion, rupture flexion and mean tibial rigidity at the 30th day. Muscle glycogen was increased at the 20th day; alkaline phosphatase was elevated at the 5th and 20th days in groups 3.OSwUS and 4.OTwUS, and decreased at the 10th day. Groups 1.OSnUS and 2.OTnUS did not show significant variations. In the mechanical resistance tests, we noted that ultrasound therapy and the association of physical activity used in the present study showed significant differences in bone resistance and bone rigidity after 30 days of treatment. These facts suggest that ultrasound or physical activity, or their combination may accelerate the process of bone tissue repair.