Splenic marginal zone macrophages expressing macrophage receptor with collagenous structure (MARCO) contribute to the clearance of blood-borne pathogens. We determined a splenic adherent cell fraction abundantly containing cells expressing a higher level of MARCO by flow cytometry, and examined the effects of daily administration of an anabolic dose of β2-agonist clenbuterol on the phagocytic capacity of the cells in mice. After 6 weeks of clenbuterol (1.0 mg/kg body weight/d) or vehicle administration to the mice, splenic adherent cells were isolated. These cells were separated into three cell-size subpopulations. Among them, the small-cell subpopulation contained abundantly the cells with markedly higher levels of MARCO and exhibited more intense phagocytic capacity against Escherichia coli, as compared with the other subpopulations. The phagocytic capacity of the small cells was significantly reduced after clenbuterol administration. These results suggest that the utilization of clenbuterol as doping drug impairs bacterial clearance in the spleen.

Objective Folate (vitamin B-9) plays key roles in cell growth and proliferation through regulating the synthesis and stabilization of DNA and RNA, and its deficiency leads to lymphocytopenia and granulocytopenia. However, precisely how folate deficiency affects the distribution of a variety of white blood cell subsets, including the minor population of basophils, and the cell specificity of the effects remain unclear. Therefore, we examined the effects of a folate-deficient diet on the circulating number of lymphocyte subsets [T-lymphocytes, B-lymphocytes, and natural killer (NK) cells] and granulocyte subsets (neutrophils, eosinophils, and basophils) in rats.
Methods Rats were divided into two groups, with one receiving the folate-deficient diet (FAD group) and the other a control diet (CON group). All rats were pair-fed for 8 weeks.
Results Plasma folate level was dramatically lower in the FAD group than in the CON group, and the level of homocysteine in the plasma, a predictor of folate deficiency was significantly higher in the FAD group than in the CON group. The number of T-lymphocytes, B-lymphocytes, and NK cells was significantly lower in the FAD group than in the CON group by 0.73-, 0.49-, and 0.70-fold, respectively, indicating that B-lymphocytes are more sensitive to folate deficiency than the other lymphocyte subsets. As expected, the number of neutrophils and eosinophils was significantly lower in the FAD group than in the CON group. However, the number of basophils, the least common type of granulocyte, showed transiently an increasing tendency in the FAD group as compared with the CON group.
Conclusion These results suggest that folate deficiency induces lymphocytopenia and granulocytopenia in a cell-specific manner.

Histone lysine methylation (HKM) is an epigenetic change that establishes cell-specific gene expression and determines cell fates. In this study, we investigated the expression patterns of histone H3 lysine 9 methyltransferases (H3K9MTases) G9a (euchromatic histone lysine N-methyltransferase 2, Ehmt2), GLP (euchromatic histone lysine N-methyltransferase 1, Ehmt1), SETDB1 (SET domain, bifurcated 1), PRDM2 (PR domain containing 2), SUV39H1 (suppressor of variegation 3-9 homolog 1), and SUV39H2, as well as the distribution of 3 types of HKM at histone H3 lysine 9: mono- (H3K9me1), di- (H3K9me2), or tri-methylation (H3K9me3), during mouse growth plate development. In the forelimb cartilage primordial at embryonic day 12.5 (E12.5), none of the H3K9MTases were detected and H3K9me1, H3K9me2, and H3K9me3 were scarcely detected. At E14.5, the H3K9MTases were expressed at low levels in proliferating chondrocytes and at high levels in prehypertrophic and hypertrophic chondrocytes. Among the H3K9 methylations, H3K9me1 and H3K9me3 were markedly noted in these chondrocytes. At E16.5, G9, GLP, SETDB1, PRDM2, SUV39H1, and SUV39H2, as well as H3K9me1, H3K9me2, and H3K9me3, were detected in prehypertrophic and hypertrophic chondrocytes in the growth plate. Western blotting and real-time quantitative polymerase chain reaction analysis revealed the distributions of G9 and GLP proteins and the expression of all the H3K9MTase mRNAs in prehypertrophic and hypertrophic chondrocytes. These data suggest that H3K9 methyltransferases are predominantly expressed in prehypertrophic and hypertrophic chondrocytes, and that they could be involved in the regulation of gene expression and progression of chondrocyte differentiation by affecting the methylation state of histone H3 lysine 9 in the mouse growth plate.© 2013 Elsevier B.V. All rights reserved.

The acute effects of oral administration of diallyl disulfide (DADS), the major organosulfur compound of garlic, on plasma glucose and free fatty acid (FFA) concentrations were examined in rats. Male, 10-week-old Sprague-Dawley rats were divided into DADS-free and DADS-administered (dose =10, 20, and 40 mg/kg body weight [BW]) groups. Plasma samples were prepared from whole blood drawn from the tail vein 0, 1,2, 4, and 6 hr after administration. The stomachs were isolated, and the contents were measured 8 hr after administration. In DADS-administered groups, plasma glucose concentrations were increased in a dose-dependent manner 1 hr after the administration. The increase was transient, except in groups administered 40 mg/kg BW of DADS, in which plasma glucose levels remained significantly higher than the DADS-free levels throughout the experimental period. Similar patterns were observed in the plasma FFA concentrations, although the significant differences were lower than those observed in the plasma glucose concentrations. The gastric contents were dose-dependently elevated after DADS administration. The increase was significant when 20 or 40 mg/kg BW of DADS was administered. These results suggest that oral administration of DADS can mobilize energy substrates into the blood, although a higher dose of DADS slows gastric emptying.

Objective The age-related effects of fasting on lipolysis, the production of ketone bodies, and plasma insulin levels were studied in male 3-, 8-, and 32-week-old Sprague- Dawley rats. Methods The rats were divided into fasting and control groups. The 3-, 8- and 32-week-old rats tolerated fasting for 2, 5, and 12 days, respectively. Results Fasting markedly reduced the weights of perirenal and periepididymal white adipose tissues in rats in the three age groups. The mean rates of reduction in both these adipose tissue weights during fasting periods were higher in the order of 3&gt
8&gt
32-week-old rats. Fasting transiently increased plasma free fatty acid (FFA), total ketone body, β-hydroxybutyrate, and acetoacetate concentrations in the rats in the three age groups. However, plasma FFA, total ketone body, β-hydroxybutyrate, and acetoacetate concentrations in the 3-week-old rats reached maximal peak within 2 days after the onset of fasting, although these concentrations in the 8- and 32-week-old rats took more than 2 days to reach the maximal peak. By contrast, the augmentation of plasma FFA, total ketone body, β-hydroxybutyrate, and acetoacetate concentrations in the rats in the three age groups had declined at the end of each experimental period. Thus, the capacity for fat mobilization was associated with tolerance to fasting. Plasma insulin concentrations in the rats in the three age groups were dramatically reduced during fasting periods, although basal levels of insulin were higher in the order of 32&gt
8&gt
3 week-old rats. Conclusion These results suggest that differences in fat metabolism patterns among rats in the three age groups during prolonged fasting were partly reflected the metabolic turnover rates, plasma insulin levels, and amounts of fat storage. © The Japanese Society for Hygiene 2011.

Effects of hindlimb suspension (HS) and ambulation recovery on hippocampal neurogenesis of newly weaned rats were studied by using immunohistochemical techniques. The number of proliferating cell nuclear antigen-positive (PCNA(+)) cells in the subgranular zone (SGZ) markedly decreased during normal growth. However, neither HS nor subsequent recovery caused additional changes in the number of PCNA. cells. The number of doublecortin-positive (DCX+) neurons decreased gradually during normal growth. HS resulted in a further decrease in these neurons. However, DCX+ cell numbers became identical to the levels in age-matched controls after 14 days of recovery. PCNA and DCX-double positive cells in the SGZ were also observed, and their cell numbers were not affected by HS and 14-day ambulation. Thus, HS suppressed the generation of DCX+ neurons without affecting PCNA(+) cells in the SGZ of weaned rats. Taken together, hippocampal neurogenesis in weaned rats was not severely affected by HS while it decreased significantly as they had grown. (C) 2011 Elsevier Ireland Ltd. All rights reserved.

Aims: Glucocorticoids bind to the glucocorticoid receptor (GR) and increase catabolism of muscle proteins via the ubiquitin-proteasome pathway. Activation of beta(2)-adrenergic receptor (beta(2)-AR) in skeletal muscle has been shown to induce muscle hypertrophy by promoting muscle protein synthesis and/or attenuating protein degradation. The aim of this study was to investigate the correlation between disuse-induced muscle atrophy, and expression of GR and beta(2)-AR.
Methods: Rats were subjected to casted-immobilization (knee and foot arthrodesis), a model for muscle disuse, for 10 days. Fast-twitch (extensor digitorum longus: EDL) and slow-twitch (soleus: SOL) muscles were isolated and subsequently used for analysis. The expression of GR and beta(2)-AR was analyzed by real-time RT-PCR and western blotting. In addition, we analyzed plasma catecholamine and corticosterone concentrations by ELISA.
Key findings: Casted-immobilization-induced muscle atrophy was much greater in the SOL muscle than in the EDL muscle. Casted-immobilization decreased the expression of GR mRNA and protein in the SOL muscle but not in the EDL muscle. Although the expression of beta(2)-AR protein in the cytosol and membrane-rich fractions was not changed by casted-immobilization in either muscle, casted-immobilization decreased the expression of beta(2)-AR mRNA in the SOL muscle. Plasma catecholamine and corticosterone concentrations, however, were largely unaffected by casted-immobilization during the experimental period.
Significance: This study provides evidence that casted-immobilization-induced muscle disuse downregulates GR expression in slow-twitch muscle. These results suggest that muscle disuse suppresses glucocorticoid signals, such as muscle protein breakdown and transcription of the beta(2)-AR gene, via downregulation of GR expression in slow-twitch muscle. (C) 2011 Elsevier Inc. All rights reserved.

The aim of this study was to elucidate whether dietary zinc-deficiency and its recovery play a role in controlling autonomic thermoregulation. We investigated the effects of dietary zinc-deficiency and its recovery on autonomic thermoregulation by measuring the rectal temperature, an index of deep body temperature. The weaned male Sprague Dawley rats were randomly divided into the dietary zinc-deficient diet (0.6 mg zinc/kg diet) group and the control diet (35.2 mg zinc/kg diet) group, and were fed for 4 weeks. In the recovery period, the rats of two groups were fed with the control diet for 3 weeks. The rectal temperature was significantly decreased throughout the period of zinc-deficiency and the hypothermic responses during the experimental period were recovered to the control group levels at least within 1 week in the recovery process from dietary zinc-deficiency. These results suggest that the deep temperature in rats was clearly associated with the dietary zinc intake levels.

Acute single administration effects of ethanol on the distribution of total white blood cells (WBCs), neutrophil, basophil, eosinophil, monocyte and lymphocytes were studied in rats. Acute single administration effects of ethanol on the number of red blood cells (RBCs), hemoglobin concentration and hematocrit were also examined. Male 8-week-old Sprague Dawley rats were randomly divided into the ethanol-administered (ETA) group and the control (CON) group. Two parts of an experiment, 1) 1st experiment : (ethanol dose : 1.0 g/kg body weight), and 2) 2nd experiment : (ethanol dose : 2.0 g/kg body weight) were carried out in rats. The rats were starved to 19:00, and deprived of diet for 12 hr and water for 1 hr before the single administration of ethanol. 1.0 or 2.0 g/kg body weight of ethanol (in 20% (w/w) ethanol) was orally administered to ETA group rats via a stainless stomach tube. In the CON group rats, 0.9% NaCl solution was orally given with the solution volume being equal, in the same manner. Single administration of 1.0 or 2.0 g/kg body weight of ethanol did not change the number of RBCs, hemoglobin concentration and hematocrit. Single administration of 1.0 g/kg body weight of ethanol did not also change the number of WBCs. However, administration of 2.0 g/kg body weight of ethanol increased significantly the number of neutrophil, basophil, monocyte and total WBCs without changing the number of eosinophil and lymphocytes. These results suggest that single administration of 2.0 g/kg body weight of ethanol to rats increased markedly the number of the natural immunity cells without changing the number of acquired cells.

The purpose of this study was to elucidate the effects of the recovery from dietary zinc-deficiency on the number of total white blood cells (WBCs), neutrophils, eosinophils and basophils, and plasma zinc and corticosterone concentrations in weanling male Sprague Dawley rats. Rats (n=34) of the zinc-deficient diet (0.6 mg zinc/kg diet) and control diet (35.2 mg zinc/kg diet) groups were fed for 4 wk, and then rats of both groups were fed with the control diet for 3 wk. Zinc-deficiency increased duration-dependently and clearly the number of total WBCs, neutrophils and eosinophils, and the increased numbers of these cells recovered to the control levels in week 2 of the recovery. On the other hand, the number of basophils increased by the zinc-deficiency recovered to the control levels in week 1 of the recovery. Zinc-deficiency significantly decreased plasma zinc concentrations by 85%, and markedly increased plasma corticosterone concentrations by 317%, as compared with the control group. In the recovery period, plasma zinc and corticosterone concentrations recovered to the control levels in week 2 of the recovery. These results suggest that zinc-deficiency and its recovery responses in the number of granulocytes and total WBCs are reversible, and their recovery rates depend on the subsets of granulocytes in rats.

Red peppers are used as a spice for enhancing the palatability of foods. Two major capsaicinoids, dihydrocapsaicin (DHC) and capsaicin (CAP) are responsible for up to 90% of the total pungency of pepper fruits. These capsaicinoids are known to enhance energy metabolism and thermogenesis. However, there is a little information on the effects of capsaicinoids on the lipolysis and carbohydrate metabolism. We studied the effects of DHC and CAP on plasma glucose, free fatty acid (FFA) and glycerol concentrations in rats. Male six-week-old Sprague Dawley rats were divided into the DHC, CAP and control groups. Each capsaicinoid (dose = 3 mg/kg BW/day) was subcutaneously administered to rats for 10 days. DHC increased markedly plasma glucose, FFA and glycerol concentrations on day 1-10 by 14-35%, 61-103% and 108-174%, respectively, as compared with those of the control group. CAP increased relatively plasma glucose concentrations on day 1-3 by 15-17%, as compared with the control group. However, there were no significant differences in plasma glucose concentrations on day 7-10 among three groups. On the contrary, CAP did not change plasma FFA and glycerol concentrations on day 1-3. However, CAP increased markedly plasma FFA and glycerol concentrations on day 7-10 by 54-89% and 92-98%, respectively, as compared with the control group. DHC and CAP did not change the weights of white (perirenal and periepididymal) and brown (interscapular) adipose tissues. In conclusion, the effects of capsaicinoids on plasma glucose, FFA and glycerol concentrations were relatively higher in the DHC than in the CAP, and capsaicinoids did not change the weight of white and brown adipose tissues.

The purpose of the present study was to elucidate the effects of dietary zinc-deficient feeding and its recovery on liver cytosolic alcohol dehydrogenase (ADH; alcohol: NAD+ oxidoreductase, EC1.1.1.1) activities and plasma zinc levels in rats. The weaned male Sprague Dawley rats were randomly divided into the zinc-deficient diet (ZDF: 1.9 mg zinc/kg diet) group and the control diet (53.5 mg zinc/kg diet) group, and were fed for 4 weeks. In the recovery periods, the rats of two groups were fed with the control diet for 3 weeks. Liver cytosolic protein content per body weight in the zinc-deficiency and its recovery period showed no significant changes between both groups. However, zinc-deficiency decreased significantly liver cytosolic ADH specific activity, total liver cytosolic ADH activity and total liver cytosolic ADH activity/body weight by 50%, 76% and 53%, respectively, as compared with the control diet group. Zinc-deficiency also decreased significantly plasma zinc concentration by 84%, as compared with the control diet group. On the contrary, no significant changes in liver cytosolic ADH specific activity, total liver cytosolic ADH activity and total liver cytosolic ADH activity/body weight in the recovery period were observed between both groups. Plasma zinc concentration in the recovery period was almost recovered to the control level. These results suggest that rat liver cytosolic ADH activity was clearly related to dietary zinc intake levels.

The main pungent component of wasabi (Eutrema japonica) is known to be isothiocyanate and its derivatives, volatile substances. Allyl isothiocyanate (AITC) accounts for more than half of isothiocyanate derivatives. However, there is a little information on the effects of AITC on the immune system by analyzing the number of white blood cells (WBCs) over the course of days of AITC administration. In the present study, we studied the effects of AITC (dose=20 mg/kg body weight/day for 10 days, subcutaneous: s.c.) on the number of WBCs (total WBCs, lymphocytes, monocyte, neutrophil, basophil and eosinophil) and plasma corticosterone concentrations in adult male rats. Administration of AITC decreased significantly the number of total WBCs on days 1-4 post s.c. injection by 25-27%. Administration of AITC also decreased the number of lymphocytes on days 1-10 by 21-36% and monocyte on days 1-8 by 28-78%. However, administration of AITC increased the number of neutrophil on days 8-10 by 61-112%. AITC did not change the number of eosinophil and basophil. Plasma corticosterone concentrations during the experimental period were 4.7-8.4 times significantly higher in the AITC group than in the control group, indicating that AITC induced stress-responses. The relative weights of thymus and adrenals per body weight were significantly lower and clearly higher in the AITC group than in the control group, respectively. These results suggest that AITC-mediated stress-responses are at least in part attributable to changes in the number of circulating WBCs.

The purpose of this study was to elucidate the effects of a main pungent component of wasabi, allyl isothiocyanate (AITC), on the number of lymphocytes, T-lymphocyte, B-lymphocyte and natural killer (NK) cells and plasma corticosterone concentrations in rats. AITC was given as either single dosage [20 mg/kg body weight per day; subcutaneous (s.c.) and oral] or as a daily dosage (10 mg and 20 mg/kg body weight per day; s.c.) for 4 days for the hourly and daily assessment of changes in the numbers of lymphocytes, respectively. A single s.c. injection or oral administration of AITC significantly reduced the number of lymphocytes at 4 hr to approximately 0.68 times, indicating that decreased effects of AITC on the number of lymphocytes are independent on the administration route. Administration of AITC for 4 days reduced dose-dependently the number of lymphocytes, and significantly reduced the number of T-lymphocyte and B-lymphocyte to 0.79 and 0.60 times, respectively. However, the number of NK cells did not change by AITC. Administration of AITC increased plasma corticosterone concentrations at 4 day of post s.c. injection to 5.4-6.0 times. These results suggest that AITC-mediated immunosuppresion is at least in part attributable to changes in the number and distribution of lymphocyte and its subsets.

The purpose of this study was to elucidate the effects of dexamethasone, a synthetic glucocorticoid, on the immune system by analyzing the number of white blood cells (WBCs) over the course of hours and days of dexamethasone administration. Dexamethasone was given as either a single dosage [1.0 mg/kg body weight (BW); subcutaneous injection (s.c.)] or as a daily dosage (1.0 mg/kg BW per day; s.c.) for 10 days for the hourly and daily assessment of changes in the number of white blood cells, respectively. A single administration of dexamethasone markedly decreased the number of total WBCs, as well as the number of lymphocyte, monocyte, neutrophil and eosinophil subsets with a nadir at 8 hr post-injection. The number of these cells recovered to the control levels at 24 hr. The numbers of total WBCs, lymphocytes, monocyte, eosinophil and basophil were reduced by the daily administration of dexamethasone. However, the number of neutrophil was significantly higher at days 2 and 8 after the injection. These results suggest that glucocorticoid-mediated immunosuppressions are at least partly attributable to quantitative changes in the number of circulating WBCs.

The subacute effects of dihydrocapsaincin (DHC) and capsaicin (CAP) (dose = 3 mg/kg body weight per day for 10 days, subcutaneous) on the number and distribution of white blood cells (WBCs) were studied in male adult rats. The administration of DHC and CAP for 10 days decreased significantly the number of total WBCs, lymphocytes and monocyte, and increased significantly the number of the neutrophil and eosinophil without changing the number of basophil. The administration of DHC significantly decreased thymus weight and increased adrenal weight, showing that DHC induced thymus atrophy and adrenal hypertrophy. These results suggest that capsaicinoids induced the decrease of acquired immunity responses and these phenomena may have in part participated in capsaicinoids-induced stress-responses in rats.

Distribution and total number of myonuclei in single soleus muscle fibers, sampled from tendon to tendon, were analyzed in mdx and wild-type (WT) mice. Apoptotic myonuclei and the microscopic structure around the myonuclei were also analyzed. Three types of muscle fibers of mdx mice with myonuclear distribution at either central, peripheral, or both central and peripheral regions were observed in the longitudinal analyses. All of the myonuclei were located at the peripheral region in WT mice. The total number of myonuclei counted in the whole length of fibers with peripheral myonuclei only was 17% less in mdx than in WT mice (p < 0.05). But the total myonuclear numbers in mdx mouse fibers with different distribution (peripheral vs. central) of myonuclei were identical, and the peripheral nucleus was noted where the central nucleus was missing. Myonuclei located between the center and peripheral regions were also seen in the cross-sectional analyses of muscle fibers. The cross-sectional area and length of fibers, sarcomere number, myonuclear size, myosin heavy chain expression, satellite cell number and neuromuscular junction were identical between each type of fiber. Apoptosis was not detected in any myonuclei located either in central or peripheral regions of muscle fibers. Thus, it was suggested that apoptosis-related loss of central myonuclei and regeneration-related new accretion at the peripheral region is not the cause of different distribution of myonuclei seen in muscle fibers in mdx mice. However, it was speculated that cross-sectional migration of myonuclei from central to peripheral regions may be induced in response to regeneration, because the total myonuclear numbers in fibers with different distribution of myonuclei were identical, and the peripheral nucleus was noted where the central nucleus was missing. Further, myonuclei located between the center and peripheral regions were also seen. However, the question remains as to how or why nuclei might migrate to the periphery in a regenerating muscle fiber, since there was no microscopic evidence of any structural changes around the myonuclei that may be responsible for the movement of the nucleus. Copyright (C) 2009 S. Karger AG, Basel

Increased oxidative stress in adipocytes causes dysregulated expression of inflammation-related adipokines. We have examined the effects of exercise training on oxidative stress in rat white adipose tissue (WAT), especially focusing on inflammation-related adipokines. The levels of lipid peroxidation in WAT of exercise-trained (TR) rats were lower than those ill Control (C) rats. The content of manganese-containing Superoxide dismutase in WAT of TR rats was increased as compared with those in C rats. In contrast, the expression of the NADPH oxidase NOX2 protein in WAT was downregulated by exercise training. Moreover, the levels of inflammation-related adipokines, Such as tumor necrosis factor-alpha and monocyte chemoattractant protein-1, in WAT of TR rats were lower than those in C rats. The effects of exercise training were more remarkable in visceral WAT than in subcutaneous. These results Suggest that exercise training decreases the expression of inflammation-related adipokines by reducing oxidative stress in WAT. (C) 2008 Elsevier Inc. All rights reserved.

We have previously reported the preventive effects of light resistance exercise (voluntary tower climbing exercise) after ingestion of a high-protein snack (HPS) on muscle loss in glucocorticoid-injected rats. However, such studies have not been performed in humans. In this cross-over study, we examined the effect of light resistance exercise after ingestion of HPS on plasma branched-chain amino acid (BCAA) concentrations in humans. Seven healthy young adult females (aged 22.1±1.2 y) participated in this study. They were assigned to either an exercise group or a control group. Seven days after the first experiment, they were crossed over to the opposite intervention. The subjects ingested HPS (15 g protein, 18 g sugar) 3 h after breakfast (basal meal). The plasma BCAA concentrations increased at 30 min after HPS ingestion. The subjects in the exercise group performed light resistance exercise (15 min dumbbell exercise using 300 g brown-rice-filled fabric dumbbells) when the plasma BCAA concentrations were increased (60 min after the snack ingestion). The control group maintained a resting position during the experiment. Changes in the plasma BCAA concentrations between 60 and 90 min after HPS ingestion increased continuously in the control group (+ 27 μmol/L) but decreased in the exercise group (- 37 μmol/L). Therefore, light resistance exercise after HPS ingestion may be effective for utilization of plasma BCAA in humans.

Zinc is known to play an important role for immune-functions. However, the effects of zinc-deficiency on the immune response system from the point of view of the distribution changes of the number of total white blood cells (WBCs) are still primarily unknown. Therefore, the effects of zinc-deficiency on the number of total WBCs, neutrophil, eosinophil, basophil, monocyte and lymphocytes (T lymphocyte, B lymphocyte and NK cell) were studied in rats. The weaned male rats were randomly divided into the zinc deficient diet (ZDD: 0.7 mg zinc/kg diet) group and the control diet (CON: 34.8 mg zinc/kg diet) group, and were pair-fed for 4 wk. The number of lymphocyte subsets, visceral organ weights, serum zinc, corticosterone and IL-6 concentrations were also determined. Zinc-deficiency increased duration-dependently the number of total white blood cells, granulocytes (neutrophil, eosinophil and basophil) and monocytes in 2-4 wk without changing the number of lymphocytes, T lymphocytes, B lymphocytes or NK cells. The relative weights of thymus and adrenals were 0.63 times (p&lt
0.01) lower and 1.60 times (p&lt
0.001) higher in ZDD group than in CON group, respectively. Zinc-deficiency increased serum corticosterone concentration to 1.48 times (p&lt
0.05) without changing serum IL-6 concentration, as compared with those of CON group. From these results, zinc-deficiency increases markedly the number of granulocytes and monocytes without changing the number of lymphocytes, T lymphocytes, B lymphocytes or NK cells. These results also suggest that zinc-deficiency induces stress responses and the responses may have in part participated in increased actions of the number of granulocytes and monocytes during zinc-deficiency, and induce thymus atrophy and adrenal hypertrophy.

The acute effects of dihydrocapsaicin (DHC) and capsaicin (CAP) on the number of white blood cells (WBCs), neutrophils, eosinophils, basophils, monocytes, lymphocytes, T lymphocytes, B lymphocytes and NK cells, and serum corticosterone levels were studied in rats. Male 7-wk-old SD rats were divided into DHC (3.0 mg/kg BW), CAP (3.0 mg/ kg BW) and control (CON) groups. The number of total WBCs was 1.30-1.42 times significantly higher in the DHC group than in the CON group at 6-12 h. The number of neutrophils was 1.62 times significantly higher in the DHC group than in the CON group at 12 h. The number of total WBCs and neutrophils, however, showed no significant changes between the CAP and CON groups. The number of lymphocytes was 0.61 and 0.70 times significantly lower in the DHC and CAP groups than in the CON group at 3 h. The number of T lymphocytes and B lymphocytes was 0.74 and 0.54 times lower in the DHC group than in the CON group, respectively. CAP, however, did not significantly change the number of T lymphocytes or B lymphocytes. No significant changes in the number of NK cells were observed among the three groups. CAP and DHC did not change the number of monocytes, eosinophils or basophils. No significant changes of the serum corticosterone levels were observed among the three groups. In conclusion, capsaicinoids decreased the number of acquired immunity cells, and increased the number of total WBCs and neutrophils without changing the number of monocytes, eosinophils or basophils. The magnitidue of these effects was relatively higher in DHC than in CAP.

Oligonol is a new lychee fruit-derived polyphenol converted into a low-molecular-form, which has improved absorption and strong antioxidative activity. The aim of the current study was to investigate the effects of Oligonol supplementation on perceived subjective mood states in addition to oxidative stress in 47 undergraduate athletes during 52 days of track-and-field training. This was a prospective single blind crossover study. The ratings of perceived exertion (RPE) responses were significantly (P<0.05) lower following Oligonol supplementation, suggesting that Oligonol intake caused the subjects to feel less fatigued during regular training. The results of our own questionnaire on fatigue and pain were as follows: Oligonol supplementation significantly (P<0.05) improved all the fatigue scores and tended to attenuate the feeling of three kinds of pains (muscular/anticular pain, lumbago, and menstrual pain), followed by a change fot the worse after the discontinuance of Oligonol intake. Unexpectedly, however, there were no definite changes in the Profile of Moods States (POMS) scores or oxidative stress markers (8-hydroxy-2'-deoxyguanosine and hexanoyl lysine) in urine after Oligonol supplementation. The results obtained suggest that Oligonol supplementation in young athletes shows significant subjective positive effects particularly on the feeling of fatigue during regular track-and-field training, possibly contributing to the maintenance of good conditioning.

The aim of the present study was to determine whether the antiobesity effects of tea catechins (TCs) are associated with the expression of uncoupling protein 1 (UCP1) in brown adipose tissue (BAT). Male Sprague-Dawley rats were fed a high-fat (HF; 35% fat) diet for 5 weeks, then divided into four groups and fed an HF, HF with 0.5% TC (HFTC), normal-fat (NF; 5% fat) or NF with 0.5% TC (NFTC) diet for 8 weeks. At the end of the experimental period, perirenal and epididymal white adipose tissues (WATs) and interscapular BAT were isolated. The NFTC group had significantly lower perirenal WAT weights than the NF group (NF: 12.7 +/- 0.53 g; NFTC: 10.2 +/- 0.43 g; P &lt;.01), but the HF and HFTC groups did not differ significantly. TC intake had no effects on epididymal WAT weights. The NFTC and HFTC groups had significantly lower BAT weights than the NF and HF groups, respectively. The NFTC group had significantly higher UCP1 mRNA levels in BAT than the NF group (NF: 0.35 +/- 0.02; NFTC: 0.60 +/- 0.11; P &lt;.05), but the HF and HFTC groups did not differ significantly. Thus, TC intake in the context of the NF diet reduced perirenal WAT weight and up-regulated UCP1 mRNA expression in BAT. These results suggest that the suppressive effect of TC on body fat accumulation is associated with UCP1 expression in BAT. (c) 2008 Elsevier Inc. All rights reserved.

CD31(-) CD45(-) side population (SP) cells are a minor SP subfraction that have mesenchymal stem cell-like properties in uninjured skeletal muscle but that can expand on muscle injury. To clarify the role of these SP cells in muscle regeneration, we injected green fluorescent protein (GFP)positive myoblasts with or without CD31(-) CD45(-) SP cells into die tibialis anterior muscles of immunodeficient NOD/scid mice or dystrophin-deficient mdx mice. More GFP positive fibers were formed after co-transplantation than after transplantation of GFP positive myoblasts atone in both mdx and NOD/scid muscles. Moreover, grafted myoblasts were more widely distributed after co-transplantation than after transplantation of myoblasts atone. Immunohistochemistry with anti-phosphorylated histone H3 antibody revealed that CD31(-) CD45(-) SP cells stimulated cell division of co-grafted myoblasts. Genome-wide gene expression analyses showed that these SP cells specifically express a variety of extracellular matrix proteins, membrane proteins, and cytokines. We also found that they express high levels of matrix metalloproteinase-2 mRNA and gelatinase activity Furthermore, matrix metal loproteinase-2 derived from CD31(-) CD45(-) SP cells promoted migration of myoblasts in vivo. Our results suggest that CD31(-) CD45(-) SP cells support muscle regeneration by promoting proliferation and migration of myoblasts. Future studies to further define the molecular and cellular mechanisms of muscle regeneration will aid in the development of cell therapies for muscular dystrophy.

The aim of the present study was to investigate the mechanism of apoptosis in human multiple myeloma cell line, U266, caused by 2-aminophenoxazine-3-one (Phx-3). Flow-cytometrical and morphological analyses showed that Phx-3 increased the population of annexin V-positive cells including early stage apoptotic cells and late stage apoptotic cells and induced DNA fragmentation or apoptotic body formation in U266 cells, indicating that Phx-3 induced the apoptosis of U266 cells. Activity of caspase-3 was extensively increased in U266 cells treated with Phx-3 time-dependently within 24 h, but this Phx-3-stimulated activity of the enzyme in the cells was completely cancelled by the addition of N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (z-VAD-fmk), a pan-caspase inhibitor. The addition of z-VAD-fmk almost blocked the apoptotic effect of Phx-3 against U266 cells, indicating that Phx-3-induced apoptosis of U266 cells was dependent on a caspase signaling pathway. Moreover, the apoptosis of U266 cells occurred after the induction of cell cycle arrest of the cells in the S and G2/M phase, the loss of mitochondrial membrane potential, and activation of caspase-3 reached maximum, which were caused by Phx-3 within 24 h. These results support the views that the apoptosis of U266 cells caused by Phx-3 may be preceded by the cell cycle arrest, depolarization of mitochondria and activation of caspase-3. These results support the view that Phx-3 may be utilized in future as chemotherapeutic agent against multiple myeloma which is extremely refractory to chemotherapy. © 2008 Pharmaceutical Society of Japan.

We examined whether phenoxazine derivatives such as 2-amino-4,4 alpha-dihydro-4 alpha,7-dimethyl-3H-phenoxazine3-one (Phx-1) and 2-aminophenoxazine-3-one (Phx-3) may have anticancer effects on the human gastric cancer cell lines, MKN45, MKN74, MKN7 and KATO III in vitro. Phx-1 inhibited the growth of these cancer cells in a dose- and time-dependent manner. The IC50 was approximately 65, 25, 100 and 70 mu M for MKN45, MKN74, MKN7 and KATO III respectively, after 72 h. Phx-3 exerted stronger antiproliferative effects against these cancer cells (lC(50): approximately 5, 1, 10 and 10 mu M for MKN45, MKN74, MKN7 and KATO 111, respectively, after 72 h) than Phx-1. Phx-1 and Phx-3 increased the population of TUNEL-positive cells in MKN45 and KATO III time-dependently from 24 to 72 h, suggesting that Phx-1 and Phx-3 have apoptotic activity against these gastric cancer cells. The activity of effector caspase-3 significantly increased in MKN45 treated with Phx-3 for 24 h, but did not altered in the cells treated with Phx-1 for 24 h. When z-VAD-ftnk, a pan-caspase inhibitor, was co-treated for 24 h, Phx-3-stimulated caspase-3 activity in MKN45 was reversed to the levels of normal activity, while the antiproliferative and apoptotic effects of Phx-3 against the cells were maintained. The activity of caspase-3 was not activated in KATO III by 24 h exposure for Phx-1 or Phx-3. In conclusion, both phenoxazines prevent the growth of the human gastric cancer cell lines, MKN45 and KATO III in vitro, and cause the apoptosis of these cell lines via a caspase-independent pathway. Although the intracellular action mechanisms of Phx-1 and Phx-3 are still unclear, these phenoxazines may be useful for the treatment of gastric cancer in the future.

Prolonged inactivity is known to induce changes in responses of many physiological defense systems such as the hypothalamo-hypophyseal-adrenocortical axis, the sympathetic nervous system, and immuno-responsive systems. However, effects of various types of inactivity on immuno-responsive systems are still unknown. Therefore, the effects of two types of inactivity (immobilization: IMM and whole body suspension: WBS) on the number of white blood cells were studied in rats. Rats were divided into the control group and each inactivity group to compare the number of total white blood cells, lymphocytes, monocyte, neutrophil, eosinophil, and basophil during the experimental periods. Both IMM and WBS were maintained for 11 days. IMM markedly increased the number of total white blood cells, monocyte, neutrophil, and eosinophil in the 1st to 10th day. However, the number of total white blood cells, monocyte, neutrophil, and eosinophil during the experiment of WBS were characterized by the presence of a lag phase followed by the significant increased actions. IMM did not change the number of basophil during the experimental period. However, WBS increased the number of basophil in the 1st to 8th day to 2.8-4.8 times, compared with the values of the control. Both IMM and WBS did not change the number of lymphocytes. From these results, WBS increases the number of natural immunity cells without changing acquired immunity cells, and there are different responses in the number of total white blood cells, monocyte, neutrophil, eosinophil, and basophil between IMM and WBS.

The present article briefly overviews the principal pathway of ethanol metabolism in the liver cell and hormonal regulations of ethanol metabolism, and a relationship between alcohol drinking volume (=alcohol consumption) and the degree of alcohol intoxication. This article also focuses on a possible mechanism of sex difference in rat liver cytosolic alcohol dehydrogenase (ADH) activity, and the effects of bile, bile acids, vegetable oils and unsaturated fatty acids on plasma ethanol levels and gastric emptying in ethanol-administered rats. Finally, the recent findings concernig a relationship between alcohol drinking and human health are described.

The difference was studied between use of the dominant hand and non-dominant hand in the cutting performance and control by women of a kitchen knife. Each subject was instructed to slice a radish to a thickness of 5 mm, the number of radish slices cut in 15 sec and the actual thickness of each being measured. The number of radish slices cut by the kitchen knife held in the dominant hand was significantly greater than when held in the non-dominant hand. The actual thickness of radish slices cut by the dominant hand was almost the same as the specified 5 mm, while those by the non-dominant hand tended to be thicker. The percentage of radish slices which were not fully cut was significantly higher by the non-dominant hand than by the dominant hand. These results indicate that the cutting performance and control of a kitchen knife held in the non-dominant hand were inferior to the levels achieved by the dominant hand.

Rat liver cytosolic alcohol dehydrogenase (ADH) is the principal enzyme which catalyzes the oxidation of ethanol. ADH activity is known to be significantly higher in females than in males. However, the precise mechanism of the sex-difference in ADH activity is uncertain. Recently we have shown that the inhibitory action of androgen and the slight facilitatory actions of progestin and estrogen are involved in the mechanism of sex-difference in adult rat liver ADH activity In the present study we studied the difference of the postnatal developmental changes of ADH activity between males and females. ADH activity increased rapidly after birth up to about 12 d of age, and was not different in the two sexes. In female rats, ADH activity peaked at about 30 d of age and then increased gradually to plateau levels. However, the ADH activity of male rats fell markedly between 30 and 45 d old, and the reduced enzyme activity was observed until 104 d old. In conclusion, the ADH activity of male rats fell between 30 and 45 d after birth. At the ages over the turning point, the ADH activity was significantly lower in male than in female rats.

Rat liver cytosolic alcohol dehydrogenase (ADH) activity is known to be significantly higher in females than in males. To elucidate a possible mechanism of sex difference in the ADH activities, we studied the in vivo effects of the administrations of beta-estradiol, progesterone or testosterone, and castrations (orchiectomy and ovariectomy) on the ADH activities in male and female Sprague-Dawley adult rats. Furthermore: we studied the ADH activities in six liver portions of sham-operated male, orchiectomized male, and female rats. The ADH activities were higher in orchiectomized male rats than in sham-operated male rats. These results were observed in all liver portions. The administration of testosterone (5 mg/kg BW, twice a day for 7 d) to orchiectomized male rats significantly decreased ADH activities. The ADH activities in ovariectomized female rats were comparable with those in sham-opepated female rats. The administration of beta-estradiol (50 mu g/kg BW, once every 2 d for 20 d) or progesterone (75 mu g/kg BW, once every 2 d for 20 d) to rats increased the ADH activities in males more than in females. Those results suggest that the inhibitory action of androgen and the slight facilitatory actions of progestin and estrogen are involved in the mechanism of sex difference in adult rat liver cytosolic ADH activity.

The difference between the dominant hand and the non-dominant hand in overarm throwing motions and ball-throwing distances were studied in student women. The subjects were divided into a trained group and an untrained group. A switch thrower, categorized in the training group, was also examined. The throwing time, moving distance and mean velocity of the ball in the overall throwing phase, back-swing phase and acceleration phase were determined by a high-speed video analysis system. Seven empirical parameters estimated from the overall throwing motion were also introduced. These physical quantities and parameters were compared between the dominant and non-dominant hands. The ball-throwing distances in the trained and untrained groups were 2.58 and 1.73 times higher for the dominant hand than for the non-dominant hand, respectively. The difference in these values for both the hands of the switch thrower, however, were very small. The throwing time, moving time, and mean ball velocity in the back-swing phase in the trained group were 1.16, 1.65 and 1.35 times higher for the dominant hand than for the non-dominant hand, respectively. These phenomena were not observed in the untrained group and the switch thrower. The mean ball velocities in the acceleration phase for the trained and untrained groups were 2.0 and 1.5 times higher for the dominant hand than for the non-dominant hand, respectively. These values were more highly correlated with the ball-throwing distance in the trained group than in the untrained group. Significant correlations between seven parameters and ball-throwing distance were all observed for the dominant hand. However, only three of these parameters showed significant correlations for the non-dominant hand. These results show that the relationship between ball-throwing distance and throwing motion is closer for the dominant hand than for the non-dominant hand. In the present paper, possible roles of the dominant and non-dominant hands in the relationship between throwing motion and ball-throwing distance are also discussed.

The effects of whole body suspension and recovery on the wet weights of visceral organs (thymus, heart, liver, spleen, adrenals, kidney, and testis) and skeletal muscles (soleus [SOL] and gastrocnemius [GAS]) were studied in adult male rats. Whole body suspension and recovery in pair-fed conditions were maintained for 12 days and 10 days, respectively. The rats were divided into three groups: cage control (CON), whole body suspended (WBS), and suspension harness-fitted/non-suspended constraint control (SHF). The relative wet weights (as a function of body weight) of the adrenals, kidney, heart, and testis showed a marked increase with the advance of suspension. In the recovery period, the relative wet weights of the adrenals and kidney gradually returned to control levels in 7-10 days. However, the relative wet weights of the testis and heart did not decrease to normal within 10 days. Conversely, the relative wet weights of the thymus, spleen, and liver were decreased significantly by suspension, and the weights of these three visceral organs returned to control levels completely in 2-7 days. Similar phenomena were observed in the SHF group. The order of the degree of suspension-induced size change of visceral organs was CON < WBS∼__=SHF group. Further, the recovery rate of the antigravity slow-twitch SOL muscle was markedly slower than that of the first-twitch GAS muscle. Muscle mass loss of 35% and 10% of body weight was observed in SOL and GAS muscles, respectively, of the WBS group at the 12th day of suspension. The relative wet weight of the GAS muscle decreased by suspension returned to control levels by the 4th day of recovery. That of the SOL muscle, however, did not, even after 10 days. The relative wet weights of the GAS and SOL muscles in the SHF group were similar to those of the CON group, suggesting that the unloading of body weight from hindlimbs induces muscle atrophy. We conclude that visceral organ size change may be induced by suspension and/or restraint stresses, and the dagree and rates of recovery differ from organ to organ

ラットの体重, 血清コレステロールレベルおよび腎臓と精巣周囲の脂肪細胞に及ぼす乾燥ビール酵母 (DY) の影響を明確にするため, 成長期と高齢期のラットを用いて検討した。その結果, 以下の知見を得た。<BR>1) 生後23～83日齢の期間自由摂餌下で飼育したラットの体重は, 生後24～100日齢の期間85%制限給餌下で飼育した場合と同様に, DY群が対照より低値を示した。このDYによる体重の変動作用は, 日齢で異なっていた。また, 性差がみられ, DYを多く含むほど大きかった。体重低下の程度と摂餌量または摂水量との間には, 相関がみられなかった。高齢期ラットの体重は, 40日間DY飼料で飼育しても25% DYで徐々に減少する傾向がみられたが, 有意な変化を示さなかった。したがって, DYによる体重の変動作用は成長期ラットで認められるが, 成長が停止した高齢期ラットでは大きな変化がみられない。<BR>2) 血清中の総コレステロールとHDL-コレステロールの各濃度は, 成長期ラットではDYによって大きな影響がみられなかったが, 高齢期ラットではDY量に比例して有意に低かった。<BR>3) 脂肪細胞の半径と体積は精巣周囲部でDYの含有量に比例して増大したが, 腎臓周囲部ではDYの影響がみられず, 部位差がみられた。25% DY群の場合, 精巣と腎臓周囲の脂肪細胞数は, 対照に比べて54～72%有意に少なかった。

To clarify the cellular changes occuring during muscle atrophy, biochemical and histological properties in the hindlimb muscle of 5- and 9-week-old male dystrophic (C57BL/6J-dy: dy/dy, homozygous) mice were compared with those of control (dy/?, heterozygous) mice. The muscle protein was fractionated into sarcoplasm, myofibril, and stroma. Muscle weight in dystrophic mice was 31-37% (p<0.001) of that in the controls, while DNA and RNA concentrations in dystrophic mice were 2.5-2.8 (p<0.001) and 1.5-1.8 times (p<0.001) higher, respectively, than the controls. However, total DNA and RNA contents in muscle were 71% (p<0.001) and 56% (p<0.001), respectively, of the control values in 9-week-old dystrophic mice, representing significant reductions. Protein concentration in dystrophic muscle was 81-83% of the control values. The order of decrease of each muscle protein fraction in dystrophic mice was myofibril > stroma > sarcoplasm, indicating that the degree of the reduction in protein level differs between fractions. The apparent cell volume (protein/DNA ratio) in dystophic mice was 30-33% (p<0.001) of the control animals. From these results, muscle atrophy in dystrophic mice may be induced partly by (a) decreased ribosomal capacity (decrease of RNA/protein ratio), and (b) decreases in DNA transcription rate and protein synthesis (decrease of RNA/DNA ratio). The number of nuclei/mm^2 in dystrophic mice was 1.6 times (p<0.001) higher than that in controls. However, the number of fibers/mm^2 in dystrophic mice was 53% (p<0.001) of the control animals. The ratio of number of centronuclei to number of nuclei in dystrophic mice was 9.8 times (p<0.001) higher than that in controls. The mean value of the frequency distribution of the cross-sectional area of fibers per unit area of muscle tissue in dystrophic mice was significantly lower than that of the controls. In conclusion, these biochemical and histological properties in hindlimb muscles of C57BL/6J-dy mice may provide a basis on which to pursue the study of disuse muscle atrophy.

The effects of vegetable oils (soybean oil and coconut oil) and C18-unsaturated fatty acids (oleic acid, C18:1; linoleic acid, C18:2; linolenic acid, C18:3) on plasma ethanol levels in male rats (6 weeks old) were investigated. Vegetable oils decreased and delayed the peak of plasma ethanol concentration: a dose-response to vegetable oils was observed in the concentration and time to maximum concentration of plasma ethanol but no change in disappearance time. These phenomena were observed in two conditions: 1) oral administration of vegetable oils before oral intubation of ethanol and 2) simultaneous oral administration of vegetable oils and ethanol. Similar responses were obtained in three C18-unsaturated fatty acids. No changes in hepatic alcohol and aldehyde dehydrogenase isozyme (high K(m) and low K(m)) activities were observed. The remaining rate of ethanol in stomach was significantly higher with administration of vegetable oils or linoleic acid. A high negative correlation between the maximum plasma ethanol concentration and the remaining rate of ethanol in stomach was found. These results suggest that the slowing of the gastric emptying is a major mechanism for the decreasing and delaying effects on plasma ethanol levels by vegetable oils. The present paper also suggests that fatty acids may participate in the decreasing and delaying actions on the peak of plasma ethanol concentration by vegetable oils.

唾液分泌現象に対する料理に添える吸い口やつま等の香りや盛り付けの影響を体系的に明らかにするため, ほぼ日常の生理レベルの状態にあるヒトの唾液量を脱脂綿を用いて簡便に測定する方法を検討した。そのために, 健康な女子の学生を被験者としてしらべた。その結果, 唾液量を脱脂綿を用いてほぼ生理レベルの状態で測定する至適条件は, 次のようであった: 脱脂綿のサイズ=3×3～4×4cmのサイズの正方形; 唾液採取時間=2分; 採取間隔=4分。本法は, 簡便で, しかも再現性があり, 同一個人の唾液量を1シリーズの実験で種々の条件において測定するのに適していることを示した。また, 本法を用いて, 唾液が『出る』または『出ない』と意識したときには, 唾液量が変動することを併せて報告した。

The effects of bovine bile (50-400 mg.kg-1 BW) on plasma ethanol levels in male rats (6-8 weeks old) were examined. Bovine bile decreased and delayed the peak of plasma ethanol concentration: a dose response to bovine bile was observed in the concentration and time to maximum concentration of ethanol but no change in disappearance rate. These phenomena were observed in two conditions: 1) oral administration of bovine bile before oral intubation of ethanol (1.0 g.kg-1 BW) and 2) simultaneous oral administration of bovine bile and ethanol. Similar responses were obtained in taurocholic acid. No changes in hepatic alcohol and aldehyde (low K(m) and high K(m)) dehydrogenase activities were observed. The remaining rate of ethanol in stomach was significantly higher with administration of bovine bile. A negative correlation between the maximum ethanol concentration and the remaining rate of ethanol in stomach was found. The intestinal absorption rate of ethanol decreased significantly in the presence of bovine bile. These results suggest that the delay of the gastric emptying and/or the decrease of the intestinal absorption rate of ethanol are major mechanisms for the decreasing and delaying effects on plasma ethanol by bovine bile. The present paper also suggests that bile acids such as taurocholic acid may participate in the lowering and delaying actions on the peak of plasma ethanol concentration by bovine bile.

S.D. 系ラットの肝臓におけるアルコール脱水素酵素 (ADH) の比活性値とタンパク質濃度に対する部位差, 性差および発育差を定量的に検討した。<BR>1) 肝ADHの比活性値とタンパク質濃度は, 左臓側葉等の容積の大きい部位で高値をとる。一方, 尾状突起や乳頭突起等の容積の小さい部位では, 左臓側葉における両値に比べて約30～40%低い。したがって, 肝におけるADHの比活性値は部位差がある。<BR>2) 同一週齢のラットにおけるADHの比活性値は, 雄に比べて雌が約2.0倍大きい。また, みかけの肝ADH総活性値も, 雌が雄に比べて約1.5倍大きい。したがって, 肝におけるADHの両活性は, 性差が著明に認められる。<BR>3) 肝ADHの比活性値は, 生後3～12日目まで直線的に増加し, その後はプラトーに達する。したがって, 比活性値は生後約2週間でほぼ成体レベルに達する。みかけの肝ADH総活性は, 生後日数に対してS字形に増加する。ラットの肝ADHのK_m値は, NADおよびエタノールに対して生後3～215日の範囲で変化が認められない。以上より, 発育に伴う肝ADHの活性の変化は, 両基質に対する親和性によらず, 酵素の比活性の変化によると考えられる。

NO₂-による食肉の詳細な発色機構を明確にするため, ホルスタイン種去勢牛の大腿筋の組織内ヘムタンパク質の酸化および還元の各過程を種々の条件で分光学的に測定し, 解析した。酸化反応過程については, NO₂-によるHbO₂の酸化反応と比較した。また, 筋組織内メト型ヘムの還元後の分光学的特性をしらべた。<BR>1) 大腿筋の組織内ヘムタンパク質は, ポリアクリルアミドゲル・ディスク電気泳動の分析から, Mbが約75%, Hbが約20%, 残りの約5%がその他のヘムタンパク質であった。<BR>2) NO₂-による筋組織内ヘムとHbO₂の酸化反応過程は, おのおのみかけの一次反応とS字形で進行した。したがって, 両反応系には著明な差が認められる。HbO₂の酸化反応は, H₂O₂添加で促進, カタラーゼ添加で遅延した。しかし, 筋組織内ヘムの酸化反応は, H₂O₂やカタラーゼの添加による影響が認められなかった。一方, 両反応系ともにSOD添加による影響がみられなかった。これらの反応性の差異に対する理由を考察した。<BR>3) 筋組織内メト型ヘムの還元速度は, pH 6.0とpH 7.4ともにNO₂-濃度の低いほうがはやかった。同一のNO₂-濃度では, その速度はpH 7.4のほうがpH 6.0に比べて約20%大きかった。[NO₂-] / [heme] =1～2では, pH 7.4のほうがpH 6.0より約10時間はやく還元された。[NO₂-] / [heme] =1 (pH 7.4, 5℃) のとき, 筋組織メト型ヘムがほぼ完全に還元されるまでに約16日を必要とした。[NO₂-] / [heme] =10では, まったく還元されなかった。還元後の筋組織内ヘムは, 544nmと577nmに吸収極大があった。これは, 2価鉄ヘム (Fe₂+) にNOが配位したNO型ヘムの誘導体と類似していた。

無制限給餌 (水) 下で飼育したラットにエタノール (EtOH) を経口投与し, そのEtOH代謝に対する日内変動の有無を定量的に明らかにする目的で研究を行なった。そのため, 8: 00と20: 00にEtOHを投与し, その血漿EtOH濃度 (p [EtOH]) の経時変化を測定し解析した。また, EtOH投与前12時間にラットが摂取した餌と水の重量を測定し, これらの各量とp [EtOH] の関係をしらべた。さらに, p [EtOH] が消失した直後に, 胃内 [EtOH] と胃内容物重量を測定した。SpragueDawley系雌ラット (184±10g) を温度25±2℃, 湿度50～60%の条件で飼育し, 20: 00に消灯し, 8: 00に点灯するようにセットした。ラットにはEtOH投与直前まで任意に餌と水を与えた。投与EtOH量は100mg/100g体重 (20%EtOH溶液) とした。その結果, 以下の結論を得た。<BR>1) 20: 00から8: 00まで (暗期) と8: 00から20: 00まで (明期) の摂餌量は, おのおの5.7±1.6, 1.6±0.7mg・g^-1B.W. ・hr^-1である。前者は後者の約3.6倍である。摂水量は暗期が明期に比べて約2.2倍大きい。したがって, ラットの摂餌量と摂水量はともに暗期が明期に比べて有意に大きい。<BR>2) 20: 00にEtOHを投与して得られたp [EtOH] の最大値 (A), Aに達するまでの時間 (B), 血漿EtOHの消失時間 (C) および血漿EtOHの積分値 (D) は, 8: 00のEtOH投与で得られたA, B, C, Dの値に比べおのおの2.0, 1.5, 1.6, 3.2倍となり, 有意に高い。したがって, 無制限給餌下で飼育したラットのp [EtOH] の経時過程に, みかけ上の日内変動が明らかに認められる。しかし, EtOHの消失速度定数 (k) 値には暗期と明期で差がみられない。この現象が, 代謝レベルによるのか, または他の生理的特性と関係があるのかを明らかにするため, さらに解析した。<BR>3) 肝細胞質内の肝ADH比活性値は, 8: 00と20: 00にEtOHを投与したグループとの間に有意な差が認められず, 日内変動はみられない。したがって, EtOH分解能は両群の間に差がない。<BR>4) 胃内容物重量と胃内 [EtOH] から求めた胃内Et-OH残存率は, 8: 00投与の実験で25.8±10.7%, 20: 00投与の場合で0.3±0.4%である。前者が後者に比べて約86倍大きく, 日内変動が明らかにみられる。<BR>5) p [EtOH] の経時過程から得られた各パラメータ (A～D) 値と胃内容物重量または胃内EtOH量との間には, 高い負の相関が認められ, ほぼ逆比例の関係が成立する。<BR>以上の結果より, ラットのp [EtOH] の経時過程に著しい日内変動が認められるのは, 胃内容物量の多少により, 投与されたEtOHの胃内残存率が大きく変化し, 血漿へのEtOHの移行も大きく変動するためと推定できる。

個体曝露チェンバーを用いて100PPmおよび200PPm一酸化窒素 (NO) に対するラット (Sprague-DawleySPF, 雄;体重320±10g) の急性曝露実験を行った。血中のNO化ヘモグロビン (且b-NO) およびメトヘモグロビン (MetHb) の時間推移を調べ, 簡略化した反応モデルにより計算機シミュレーションを行った。<BR>(1) 個体曝露チェンバー (内容積, 約1400ml) を作製し, NOと02との気相反応を最小限にとどめ, チェンバー内でエーテル麻酔ができるよう工夫して実験した。(2) 血中Hb-NOおよびMetHb濃度は30～60分後に定常状態となり, MetHb量はHb-NO量の7～15倍であった。引き続き, 新鮮空気に曝露すると, 且b-NO, MetHbともに約20分の半減期で消失した。(3) Hb-NOおよびMetHbの時間的推移を計算機シミュレーションし, 各傷害段階の反応速度定数を半定量的 (多数の不明な過程があるため) に求め, NOのヘモグロビン傷害に及ぼす影響を議論した。(4) 本実験より次の結論を導いた。(i) NOの定常的吸入状態 (100PPm) で形成されるHb-NO量はかなり低値 (～0.4%) を維持する。(ii) MetHb量もMetHb還元酵素系が活性を維持する限りさほど上昇しない (<10%)。(iii) MctHb還元酵素活性は吸入気中のNO濃度に依存し, 高濃度のNO吸入は当酵素活性を抑制すると考えざるを得ない。