Effects of macronutrient intake on thigh muscle mass during home-based walking training in middle-aged and older women.

We examined whether post-exercise macronutrient supplementation during a 5-month home-based interval walking training (IWT) accelerated exercise-induced increases in skeletal muscle mass and strength in healthy middle-aged and older women. Thirty-five women (41-78 years) were randomly divided into two groups: IWT alone (CNT, n = 18) or IWT plus post-exercise macronutrient (7.6 g protein, 32.5 g carbohydrate, and 4.4 g fat) supplementation (NUT, n = 17). For IWT, all subjects were instructed to repeat five or more sets of 3-min low-intensity walking at 40% peak aerobic capacity (Vo2 peak ), followed by a 3-min high-intensity walking above 70% Vo2 peak per day for 4 or more days per week. We determined Vo2 peak , thigh muscle tissue area by computer tomography, and thigh muscle strength in all subjects before and after IWT. We found that an increase in hamstring muscle tissue area was 2.8 ± 1.2% in NUT vs -1.0 ± 0.7% in CNT and that in isometric knee flexion force was 16.3 ± 3.7% in NUT vs 6.5 ± 3.0% in CNT; both were significantly higher in NUT than in CNT (both, P < 0.001). Thus, post-exercise macronutrient supplementation enhanced the increases in thigh muscle mass and strength, although partially, in home-based IWT in middle-aged and older women.

Branched-chain amino acid supplementation attenuates muscle soreness, muscle damage and inflammation during an intensive training program.

AIM: The aim of this paper was to assess the effects of branched-chain amino acid (BCAA) supplementation on muscle soreness, muscle damage and inflammation during an intensive training program. METHODS: Twelve long-distance runners (20 + or - 1 year-old) participated in a double-blinded crossover designed study conducted during two intensive training periods (three-day). The subjects were provided either a drink containing BCAA (0.8% BCAA in a 3.5% carbohydrate solution; 2,500 mL/day) or an isocaloric placebo drink during each training period. All subjects completed the same training program (total running distance: males: 86 km, females: 64 km), and ate the same meals during the training period. Whole body muscle soreness and fatigue sensation were measured in the morning before and during the training period by Visual Analogue Scale method. Plasma creatine kinase (CK), lactate dehydrogenase (LDH), and granulocyte elastase (GEL) levels were measured as indicators of muscle damage and inflammation before and after the training period. RESULTS: Muscle soreness and fatigue sensation during the training period in the BCAA trial were lower than those in the placebo trial (-32% and -24%, respectively; P<0.05). The plasma CK, LDH, and GEL levels after the training program in the BCAA trial were lower than those in the placebo trial (-21%, -6%, and -15%, respectively; P<0.05). CONCLUSIONS: These results demonstrate that BCAA supplementation during an intensive training program effectively reduces the muscle soreness and fatigue sensation, and that the perceived changes could be attributed to the attenuation of muscle damage and inflammation.

Branched-chain amino acids supplementation attenuates the accumulation of blood lactate dehydrogenase during distance running.

AIM: We investigated the effect of branched-chain amino acids (BCAA) supplementation on tissue damage during distance running. METHODS: Eight male distance runners (mean +/- standard deviation; age: 20.4+/-1.2 years, body weight: 58.4+/-4.2 kg) participated in a double blinded cross over designed study conducted during training camp. During each intervention period, the subjects were asked to participate in a 25-km run, and the blood BCAA and lactate dehydrogenase (LDH) level, an index of tissue damage, were measured pre- and post-run. Either a drink containing BCAA (0.4% BCAA in a 4% carbohydrate solution) or an iso-calorie placebo drink was provided to the subjects 5 times during the run without any restriction in the volume. RESULTS: The total volume of the drink consumed by the subjects did not differ substantially between the trials: 591+/-188 (2.36 g BCAA) vs 516+/-169 mL in BCAA and placebo trial, respectively. During the run, the blood BCAA concentration was maintained in the BCAA trial. However, the blood BCAA concentration level tended to decrease in the placebo trial (P<0.1). The extent of the blood LDH increase in the BCAA trial was significantly less than that of the placebo trail (48% vs 58%, P<0.05). CONCLUSION: Maintaining the blood BCAA level throughout a long distance run contributes to a reduction in the LDH release and, therefore, the effect of BCAA supplementation is suggested to reduce the degree of muscle damage.

Branched-chain amino acids and arginine supplementation attenuates skeletal muscle proteolysis induced by moderate exercise in young individuals.

This study aimed at evaluating the effect of a single oral intake of branched-chain amino acids (BCAA) with Arg on skeletal muscle protein metabolism during moderate exercise in young individuals. Eight healthy volunteers (4 males and 4 females, means +/- SEM, 26 +/- 1 yrs, 177.8 +/- 3.7 cm, 72.6 +/- 3.9 kg) were studied in a randomized double-blind placebo-controlled cross-over trial. The subjects performed 3 bouts of 20-min cycling exercise (5-min break between each bout) at 126 +/- 13 W corresponding to 50 % of the maximal work intensity. A single oral supplement of either a BCAA drink containing 2 g of BCAA and 0.5 g of Arg or an isocaloric placebo drink was given at 10 min of the 1st exercise bout. Both arterial and venous blood samples were simultaneously taken from the radial artery and the femoral vein, respectively. Blood flow in the femoral artery was determined using the ultrasound Doppler technique. The blood sampling and blood flow measurements were performed at rest, every 10 min during each exercise bout. Net balance of BCAA and Phe across the leg muscles were measured by the arteriovenous difference method. The BCAA ingestion resulted in increases in both the plasma BCAA concentration and BCAA uptake into the working leg. The Phe release from the leg during exercise significantly increased as compared to the basal level in the placebo trial (0.97 +/- 0.28 vs. 0.23 +/- 0.22 micromol/min, p < 0.05). In the BCAA trial, the cumulative Phe release from the leg during the 3rd exercise bout was significantly lower than that in the placebo trial (5.0 +/- 7.4 vs. 35.9 +/- 13.2 micromol/25 min, p < 0.05). These results suggest that endurance exercise at moderate intensity enhances proteolysis in working muscles, and a single oral intake of 2 g of BCAA with Arg at onset of exercise effectively suppresses exercise-induced skeletal muscle proteolysis.

New approach for weight reduction by a combination of diet, light resistance exercise and the timing of ingesting a protein supplement.

We have reported that ingesting a meal immediately after exercise increased skeletal muscle accretion and less adipose tissue accumulation in rats employed in a 10 week resistance exercise program. We hypothesized that a possible increase in the resting metabolic rate (RMR) as a result of the larger skeletal muscle mass might be responsible for the less adipose deposition. Therefore, the effect of the timing of a protein supplement after resistance exercise on body composition and the RMR was investigated in 17 slightly overweight men. The subjects participated in a 12-week weight reduction program consisting of mild energy restriction (17% energy intake reduction) and a light resistance exercise using a pair of dumbbells (3-5 kg). The subjects were assigned to two groups. Group S ingested a protein supplement (10 g protein, 7 g carbohydrate, 3.3 g fat and one-third of recommended daily allowance (RDA) of vitamins and minerals) immediately after exercise. Group C did not ingest the supplement. Daily intake of both energy and protein was equal between the two groups and the protein intake met the RDA. After 12 weeks, the bodyweight, skinfold thickness, girth of waist and hip and percentage bodyfat significantly decreased in the both groups, however, no significant differences were observed between the groups. The fat-free mass significantly decreased in C, whereas its decrease in S was not significant. The RMR and post-meal total energy output significantly increased in S, while these variables did not change in C. In addition, the urinary nitrogen excretion tended to increase in C but not in S. These results suggest that the RMR increase observed in S might be associated with an increase in body protein synthesis.

Effect of amino acids and glucose on exercise-induced gut and skeletal muscle proteolysis in dogs.

The effect of amino acid and/or glucose administration before and during exercise on protein metabolism in visceral tissues and skeletal muscle was examined in mongrel dogs. The dogs were subjected to treadmill running (150 minutes at 10 km/h and 12% incline) and intravenously infused with a solution containing amino acids and glucose (AAG), amino acids (AA), glucose (G) or saline (S) in randomized order. The infusion was started 60 minutes before exercise and continued until the end of the exercise period. An arteriovenous-difference technique was used to estimate both tissue protein degradation and synthesis. When S was infused, the release of leucine (Leu) from the gut and phenylalanine (Phe) from the hindlimb significantly increased during exercise, thus indicating that exercise augmented proteolysis in these tissues. The balance of Leu across the gut during exercise demonstrated a net uptake with both AAG and AA, whereas a net release was observed for G and S. In addition, Leu uptake in the gut during the last 90 minutes of the exercise period tended to be greater with AAG versus AA (P = .06). Phe balance across the hindlimb during the late exercise period showed a significant release with S, AA, and G, whereas the balance with AAG did not show a significant release. These results suggest that exercise-induced proteolysis in the gut may be reduced by supplementation with AA, and this effect may be enhanced by concomitant G administration. However, in skeletal muscle, both AA and G may be required to prevent net protein degradation during exercise. G provided without AA did not achieve net protein synthesis in either tissue.

Crystal structure of Serratia protease, a zinc-dependent proteinase from Serratia sp. E-15, containing a beta-sheet coil motif at 2.0 A resolution.

The crystal structure of Serratia protease from Serratia sp. E-15 was solved by the single isomorphous replacement method supplemented with anomalous scattering effects from both the Zn atom in the native crystal and the Sm atom in the derivative crystal, and refined at 2.0 A resolution to a crystallographic R-factor of 0.194. The enzyme consists of N-terminal catalytic and C-terminal beta-sandwich domains, as observed in alkaline protease from Pseudomonas aeruginosa IFO3080. The catalytic domain with a five-stranded antiparallel beta-sheet and five alpha-helices shares a basically common folding topology with those of other zinc metalloendoproteases. The catalytic zinc ion at the bottom of the active site cleft is ligated by His176, His180, His186, Tyr216, and a water molecule in a distorted trigonalbipyramidal manner. The C-terminal domain is a beta-strand-rich domain containing eighteen beta-strands and a short alpha-helix, and has seven Ca2+ ions bound to calcium binding loops. An unusual beta-sheet coil motif is observed in this domain, where the beta-strands and calcium binding loops are alternately incorporated into an elliptical right-handed spiral so as to form a two-layer untwisted beta-sandwich structure. The Ca2+ ions in the C-terminal domain seem to be very important for the folding and stability of the beta-sheet coil structure.

Biphasic and differential expression of cytosolic glutamine synthetase genes of radish during seed germination and senescence of cotyledons.

Three structurally distinct cDNA clones for cytosolic glutamine synthetase (GS1) were isolated from libraries prepared from senescing radish cotyledons. Northern blot analysis showed that transcripts from two of the three genes encoding GS1, Gln1;1 and Gln1;3, accumulated in the cotyledons during both dark-induced and natural senescence. Transcripts from the last gene, Gln1;2, remained at a low level during both processes. Transcripts from all three Gln1 genes accumulated in cotyledons of germinating seeds. We infer from these findings that GS1 enzymes function in both germination and senescence to convert ammonium to glutamine to remobilize nitrogen from source to sink organs. We have also examined the pattern of expression of these genes in different tissues. All three genes are expressed in roots. A large amount of transcripts from Gln1;1 accumulated in hypocotyls. Whereas none were transcribed in flowers. During dark-induced senescence of cotyledons, application of inorganic nitrogen delayed chlorophyll degradation. Inorganic nitrogen enhanced the accumulation of Gln1;1 transcripts, but decreased those of Gln1;3. In contrast, application of glutamine promoted yellowing of cotyledons during the dark treatment, and slightly increased the amounts of transcripts from Gln1;3 but decreased those of Gln1;1. Transcription of the three Gln1 genes appears, therefore, to be differentially regulated in radish cotyledons during senescence and germination.

[Effects of sodium hyaluronate on the nociceptive response of rats with experimentally induced arthritis].

Antinociceptive effects of sodium hyaluronate (Na-HA) were studied on the basis of improvement in the graded abnormal gait elicited by arthritis induced by intra-articular administration of monosodium urate crystal (MSU) to rats. One hour before MSU injection, intra-articular administration of a 1.0% solution of Na-HA with different molecular weights, ranging from 4.70 x 10(5) to 2.02 x 10(6) (HA-200), improved the score of abnormal gait in a molecular weight-dependent manner in the experimental arthritis model. Similarly, administrations of HA-200 at concentrations ranging from 0.1 to 1.0% prior to MSU treatment resulted in improvement of the score in abnormal gait in a dose-dependent manner. To elucidate the antinociceptive mechanisms of Na-HA, effects of pretreatment with Na-HA (1.0%) of different molecular weights on prostaglandin E2 (PGE2) and bradykinin (BK) releases in synovial fluid 3 hr after MSU injection were studied. Increases in PGE2 and BK concentration in the synovial fluid were depressed in a molecular weight-dependent manner by Na-HA (1.0%) pretreatment. These results indicate that Na-HA attenuates the nociceptive responses inflicted by the MSU-induced arthritis. Such an antinociceptive effect may be due to the inhibition of PGE2 and BK synthesis in the synovial joint of rats.

Anticonvulsive effect of dapsone (4,4'-diaminodiphenyl sulfone) on amygdala-kindled seizures in rats and cats.

Dapsone (4,4'-diaminodiphenyl sulfone; DDS), an established anti-leprosy drug, showed anticonvulsive effects in the amygdaloid kindling model of epilepsy. Single doses of the drug in rats (6.25-12.5 mg/kg, i.p.) suppressed the kindled seizures in a dose-dependent manner without overt behavioral toxicity. With repeated oral administration in cats, relatively higher initial doses (13-23 mg/kg) were required to obtain seizure suppression, and neurotoxic signs occurred within a few days with serum drug levels of approximately 20 micrograms/ml. Although dapsone showed anticonvulsive effects in both animal species, the effective serum levels overlapped the toxic levels reported in the clinical treatment of leprosy. In the majority of the cats, however, seizure suppression was maintained even after the discontinuation of dapsone with lower serum levels than those observed at the beginning of the seizure suppression. Therefore, dapsone would be useful as an antiepileptic drug only when long-term anticonvulsive efficacy is demonstrated using smaller doses comparable to those used in the treatment of leprosy.

Distinct localization of prostaglandin D2, E2 and F2 alpha binding sites in monkey brain.

Autoradiographic localization of binding sites for [3H]prostaglandin D2, E2 and F2 alpha in the monkey brain was investigated by using in vitro labeling and an image processing system. Specific binding sites were distinctly localized in various nuclei in the hypothalamus, thalamus, and limbic system and their localization well correlated to the known functions of PGs in the brain by pharmacological and neurophysiological experiments. Furthermore, no direct relation was observed between the localization of PG binding sites and that of muscarinic cholinergic, alpha 1-adrenergic, and mu-opioid receptors in the preoptic area and hypothalamus.

Structure determination of polysaccharides in Aloe saponaria (Hill.) Haw. (Liliaceae).

Neutral polysaccharides that inhibit carrageenin-induced edema in rats were isolated from the nondialysate of the pulp of Aloe saponaria by gel filtration. These were shown to be a linear polymer of a 1,4-linked beta-D-mannopyranose (mol. wt. 15,000) containing 18% acetyl groups (As mannan 1), and a 1,4-linked alpha-D-mannopyranose polymer containing a single branch on the principal chain consisting of D-glucose residues linked at C-2 and C-4 (mol. wt. 66,000), with 10% acetyl groups (As mannan 2). As mannan 1 inhibited carrageenin-induced hind paw edema at 50 mg/kg ip in rats; As mannan 2 was not tested for pharmacological activity. A crude preparation of both As mannans was effective when given intraperitoneally, but was ineffective when given orally.

Hepatic tumors of rabbits induced by cycad extract.

Fifteen rabbits were administered cycad extract by gastric intubation, 1 ml/rabbit, once a week for 27 or 33 weeks. The extract contained 16.6 mg of cycasin/ml. Out of 9 rabbits surviving over 200 days, 7 animals had malignant neoplasms developing in the liver. Histological and electron microscopic examinations of the developed tumors revealed that they were malignant hemangioendotheliomas. The animals showed no proliferation of hepatic cells or any tumor development in other organs.

Induction of pleomorphy and calcium ion deficiency in Lactobacillus bifidus.

The induction of pleomorphism of Lactobacillus bifidus by NaCl was completely inhibited by CaCl(2). When the organism was cultivated in calcium-free medium, growth of the bifid form was exclusively observed. Supplementation of calcium ion in the medium caused bacilloid growth. Chemical analyses indicated that calcium content of the bifid form organisms was significantly less than that of the bacilloid form; i.e., in the former type, there was an approximately 30% decrease of calcium in the whole cells, and an 82% decrease in the cell wall, as compared with the respective content of the latter. These results indicate a suppressing role of calcium ion in the induction of pleomorphism of L. bifidus. Besides calcium content, sugar and amino acid compositions were shown to be different between the bifid and bacilloid forms. In the cell wall especially, the content of glucose in the bifid form was larger than that in the bacilloid form. Methionine and phenylalanine were present in the bifid form, but not in the bacilloid form. Cell walls of the bifid form organisms lacked a larger molecular weight peptidoglycan (7.5S) which was clearly detected in the bacilloid form. Evidence has been given for the relationships of calcium ion and cell wall components to the pleomorphism in L. bifidus.