Bone tissue changes in rats during prolonged restriction of motor activity

The objective of this investigation was to measure the effect of prolonged restriction of motor activity (hypokinesia) of rats on the mass, density, mineral composition, reconstruction parameters and elemental composition of their bone tissue. The studies were done during 90 days of hypokinesia (HK) on 90 male Wistar rats equally divided into two groups: (1) vivarium control rats (VCR) and (2) hypokinetic rats (HKR). For the simulation of the hypokinetic effect the HKR group was kept for 90 days in small individual cages made of wood that restricted the movements of rats in all directions without hindering food and water intakes. During the prehypokinetic period of 15 days and during the hypokinetic period of 90 days bone mass, bone density, bone calcium and phosphorus concentrations, bone reconstruction parameters and elemental composition of bones were determined. During the same periods food intake and body weight losses were also measured. In the HKR group signs of osteoporosis in the spongy structures of the tubular bones were observed; they also showed significant decrease in rat femur weight, and in cross section of the rat femur, and in mineral concentrations of the femoral head when compared with the VCR group. The HKR group also show a significant decrease in food intake and body weight when compared with the VCR group. The corresponding parameters did not change significantly in the VCR group when compared with the baseline control values. It was concluded that prolonged exposure to HK induced osteoporosis and structural changes in bones. This apparently occurred due to inhibition of bone tissue formation in the HKR group.     

Morphometric examination of glomerulus and juxta glomerural system of rat kidney during prolonged restriction of motor activity

The aim of this study was to examine the structural changes of glomerular and juxta glomerular system (JGS) of the kidney of rats during prolonged restriction of motor activity (hypokinesia). The studies were performed during 90 days of hypokinesia (HK) on 144 male Wistar rats divided into two groups: Group one placed under ordinary vivarium conditions and serving as vivarium control rats (VCR) and Group two subjected to HK and serving as hypokinetic rats (HKR). For the simulation of the hypokinetic effect the HKR group was kept in small individual cages made of wood that restricted the movements of rats in all directions without hindering food and water intake. During a prehypokinetic period of 15 days and the hypokinetic period of 90 days, body weight and food intake were measured and morphometric examinations were done to measure the cortical and juxta medullary glomerulus on kidney sections from the VCR and HKR groups (eight rats from each group). Body weight and food intake decreased significantly in the HKR groups when compared with the VCR group. Kidney weight of rats increased, the superficial volume decreased and that of the juxta medullary glomerules increased, whereas juxta glomerular granularity indexes decreased significantly in the HKR group when compared with the VCR group. The measured parameters did not change significantly in the VCR group when compared with the baseline control values. It was concluded that prolonged exposure to HK induces a significant increase in the kidney weight and a relationship appeared between variations of the volume of cortical and juxta medullary glomerulus and the function of the juxtaglomerular apparatus.     

Muscle electrolyte measurements during and after hypokinesia in determining muscle electrolyte depletion during hypokinesia in the rat

Hypokinesia (diminished movement) induces muscle mineral depletion. However, the mechanism of muscle mineral depletion during hypokinesia (HK) remains unknown. Measuring electrolyte retention and electrolyte values in muscle, plasma, and urine during and after HK, the aim of this study was to discover if HK could depress mineral retention and lead to muscle mineral depletion. Studies were done on 204 13-wk-old male Wistar rats (370–390 g) during 10 d pre-HK period, 98 d HK period, and 15 d post-HK period. Rats were equally divided into two groups: vivarium control rats (VCR) and hypokinetic rats (HKR). All hypokinetic rats were kept for 98 d in small individual cages, which restricted their movements in all directions without hindering food and water intakes. All control rats were housed for 98 d in individual cages under vivarium control conditions. Both groups of rats were pair-fed. During the HK period skeletal muscle sodium (Na), potassium (K), magnesium (Mg), calcium (Ca), and water content and electrolyte retention decreased significantly (p < 0.05), while urinary and plasma electrolyte levels increased significantly (p < 0.05) in HKR compared with their pre-HK values and their respective VCR. During the initial days of the post-HK period, mineral retention increased significantly (p < 0.05), plasma and urinary electrolyte level decreased significantly (p < 0.05), while muscle electrolyte and water content remained significantly (p < 0.05) depressed in HKR compared with VCR. Muscle mineral and water content, electrolyte retention, plasma, and urinary electrolyte values did not change in VCR compared with their pre-HK values. It was concluded that during HK decreased muscle mineral content may suggest muscle mineral depletion, while increased urinary electrolyte loss and muscle mineral depletion may demonstrate reduced mineral retention. Reduced electrolyte excretion and depressed muscle mineral content during post-HK may indicate skeletal muscle mineral depletion during HK. Dissociation between electrolyte retention and muscle mineral depletion may demonstrate the presence of decreased electrolyte retention as the mechanism of muscle electrolyte depletion during prolonged HK.     

Urinary and serum electrolyte changes in athletes during periodic and continuous hypokinetic and ambulatory conditions

Hypokinesia (HK) (diminished movement) induces significant electrolyte changes, but little is known about the effect of periodic hypokinesia (PHK) on minerals. The aim of this study was to measure the effect of PHK and continuous hypokinesia (CHK) on urinary and serum electrolytes. Studies were done during a 30-d period of prehypokinesia (HK) and during 364 d of PHK and CHK periods. Thirty male athletes aged 24.6±7.7 yr were chosen as subjects. They were equally divided into three groups: unrestricted ambulatory control subjects (UACS), continuously hypokinetic subjects (CHKS), and periodically hypokinetic subjects (PHKS). The UACS group experienced no changes in the daily activities and regular training and they were maintained under an average running distance of 11.7 km/d. The CHKS group was limited to an average walking distance of 0.7 km/d; and the PHKS group was limited to an average walking distance of 0.7 and running distance of 11.7 km/d for 5 d and 2 d/wk, respectively, for a period of 364 d. Urinary and serum phosphate (P), calcium (Ca), sodium (Na) and potassium (K), serum intact parathyroid hormone (iPTH), calcitonin (CT), plasma renin activity (PRA) and aldosterone (PA) levels, food and water intakes, and physical characteristics were measured. Urinary P, Ca, Na, and K loss, serum Ca, P, Na, and K, and PRA and PA values increased significantly (p≤0.01), whereas serum iPTH and CT levels decreased significantly (p≤0.01) in the PHKS and CHKS groups when compared with the UACS group. However, significant (p≤0.01) differences were observed between PHKS and CHKS groups regarding urinary and serum electrolytes, serum and plasma hormones. Food and water intakes, body weight, body fat, and peak oxygen uptake decreased significantly (p ≤ 0.01) in the CHKS group when compared with PHKS and UACS groups. Food and fluid intakes, body fat, and body weight increased significantly (p≤0.01), whereas peak oxygen uptake remained significantly (p≤0.01) higher in the PHKS group when compared with the CHKS group. Serum and urinary minerals, serum hormones, food and fluid intakes, and physical characteristics did not change significantly (p>0.01) in the UACS group when compared with their baseline control values. It was shown that both PHK and CHK induce significant serum and urinary electrolyte changes. However, urinary and serum electrolyte changes were significantly (p≤0.01) greater during PHK than CHK. It was concluded that the greater the stability of muscular activity, the smaller the serum and urinary electrolyte changes during prolonged HK.     

Tissue magnesium loss during prolonged hypokinesia in magnesium supplemented and unsupplemented rats

This study aims at showing that during hypokinesia (HK) tissue magnesium (Mg2+) content decreases more with higher Mg2+ intake than with lower Mg2+ intake and that Mg2+ loss increases more with higher than lower tissue Mg2+ depletion due to inability of the body to use Mg2+ during HK. Studies were conducted on male Wistar rats during a pre-HK period and a HK period. Rats were equally divided into four groups: unsupplemented vivarium control rats (UVCR), unsupplemented hypokinetic rats (UHKR), supplemented vivarium control rats (SVCR) and supplemented hypokinetic rats (SHKR). SVCR and SHKR consumed 42 mEq Mg2+ per day. The gastrocnemius muscle and right femur bone Mg2+ content decreased significantly, while plasma Mg2+ level and urine and fecal Mg2+ loss increased significantly in SHKR and UHKR compared with their pre-HK values and their respective vivarium controls (SVCR and UVCR). However, muscle and bone Mg2+ content decreased more significantly and plasma Mg2+ level, and urine and fecal Mg2+ loss increased more significantly in SHKR than in UHKR. The greater tissue Mg2+ loss with higher Mg2+ intake and the lower tissue Mg2+ loss with lower Mg2+ intake shows that the risk of higher tissue Mg2+ depletion is directly related to the magnitude of Mg2+ intake. The higher Mg2+ loss with higher tissue Mg2+ depletion and the lower Mg2+ loss with lower Mg2+ tissue depletion shows that the risk of greater Mg2+ loss is directly related to the magnitude of tissue Mg2+ depletion. It was concluded that tissue Mg2+ depletion increases more when the Mg2+ intake is higher and that Mg2+ loss increases more with higher than lower tissue Mg2+ depletion indicating that during prolonged HK the tissue Mg2+ depletion is not due to the Mg2+ shortage in food but to the inability of the body to use Mg2+.     

Daily magnesium supplementation on serum and urinary magnesium changes in rats during prolonged restriction of motor activity

The objective of this investigation was to determine whether a plentiful magnesium (Mg²⁺) supplementation might be used to normalize or prevent Mg deficiency. This is manifested by increased rather than decreased serum Mg²⁺ concentration as is observed during prolonged hospitalization, which is developed during prolonged hypokinesia (HK) (decreased motor activity). Eighty male Wistar rats with an initial body weight of 370–390 g were used to perform the studies: They were equally divided into four groups:

1. Unsupplemented control animals (UCA);
2. Supplemented control animals (SCA);
3. Unsupplemented hypokinetic animals (UHA); and
4. Supplemented hypokinetic animals (SHA).

For the simulation of the hypokinetic effect, the hypokinetic animals were kept in small individual cages made of wood, which restricted their movements in all directions without hindering food and water intake. The control and hypokinetic supplemental animals receive 0.9 mg/mL Mg sulfate daily with their drinking water. Prior to and during the experimental period, urinary excretions of Mg, calcium, and phosphate along with their concentrations in serum, water intake, and urine excretion, and body weight were determined in the control and hypokinetic animals. In the supplemental and unsupplemental hypokinetic rats, urinary excretions and serum concentrations of electrolytes increased significantly, whereas serum concentration and urinary excretion thereof remained unchanged in the supplemented and unsupplemented control animals. It was concluded that a daily intake of large amounts of Mg supplementation cannot be used to prevent or normalize Mg deficiency in rats during prolonged exposure to HK.     

Daily copper supplement effects on copper balance in trained subjects during prolonged restriction of muscular activity

The aim of this study was to assess the effect of a daily intake of copper supplements on negative copper balance during prolonged exposure to hypokinesia (decreased number of kilometers per day). During hypokinesia (HK), negative copper balance is shown by increased, not by decreased, serum copper concentration, as it happens in other situations. Studies were done during a 30-d prehypokinetic period and a 364-d hypokinetic period. Forty male trained volunteers aged 22–26 yr with a peak oxygen uptake of 66.4 mL/min/kg and with an average of 13.7 km/d running distance were chosen as subjects. They were equally divided into four groups: unsupplemented ambulatory control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented hypokinetic subjects (SHKS), and supplemented ambulatory control subjects (SACS). The SACS and SHKS groups took 0.09 mg copper carbonate/kg body weight daily. The SHKS and UHKS groups were maintained under an average running distance of 1.7 km/d, whereas the SACS and UACS groups did not experience any modifications in their normal training routines. During the 30-d prehypokinetic period and the 346-d hypokinetic period, urinary excretion of copper, calcium, and magnesium and serum concentrations of copper, calcium, and magnesium were measured. Copper loss in feces and copper balance was also determined. In both UHKS and SHKS groups, urinary excretion of copper, calcium, and magnesium and concentrations of copper, magnesium, and calcium in serum increased significantly when compared with the SACS and UACS groups. Loss of copper in feces was also increased significantly in the SHKS and UHKS groups when compared with the UACS and SACS groups. Throughout the study, the copper balance was negative in the SHKS and UHKS groups, whereas in the SACS and UACS groups, the copper balance was positive. It was concluded that a daily intake of copper supplements cannot be used to prevent copper deficiency shown by increased copper concentration. Copper supplements also failed to prevent negative copper balance and copper losses in feces and urine in endurancetrained subjects during prolonged exposure to HK.     

Nuclear factor I (NF I) binds to an NF I-type site but not to the CCAAT site in the human alpha-globin gene promoter.

Methylation interference and missing contact analyses demonstrate that nuclear factor I (NF I) recognizes an NF I-like site (5'-GGG(N)6GCCAG-3') within the alpha-globin promoter rather than the adjacent CCAAT box. Consistent with this, mutations within the CCAAT box do not alter significantly the affinity and specificity of the interaction whereas elimination of the 5'-GGG-3' half-site of the recognition sequence reduces the DNA binding strength of NF I by 2 orders of magnitude down to the range of unspecific interaction. On the other hand, the mutated alpha-globin promoter sequence that is no longer bound by NF I, although it retains an intact CCAAT box, interacts specifically with a protein component from nuclear extracts of HeLa cells. From these results we conclude that NF I is not the factor that interacts with the CCAAT box and that the second half of the canonical 5'-TGG(N)6GCCAA-3' NF I binding site cannot be regarded as identical with the CCAAT promoter element, as suggested previously.