Contractile responses of isolated equine digital arteries under hypoxic or hyperoxic conditions in vitro: role of reactive oxygen species and Rho kinase.

The underlying pathophysiological triggers for equine acute laminitis are unknown, although digital vasoconstriction, ischaemia, hypoxia and reperfusion injury may be involved. The contractile responses of isolated equine digital arteries (EDAs), harvested from the hindlimbs of normal horses postmortem at an abattoir, were studied acutely (up to 3 h) under hyperoxic (95% oxygen, 5% CO2 ) and hypoxic (95% nitrogen, 5% CO2 ) conditions in organ baths. Phenylephrine (PHE; 10(-6) m), 5-hydroxytryptamine (5-HT; 10(-7) m) and high potassium (K(+) ; 118 mm) caused contraction in EDAs which was significantly (P<0.0001) enhanced under hypoxic conditions. In contrast, contraction stimulated by 9,11-dideoxy-9α,11α-epoxymethanoprostaglandin F2α (U44069; 3 × 10(-8) m) was not significantly enhanced by hypoxia (P=0.75). Hypoxia-enhanced contraction in response to K(+) was greater (P<0.03) in vessels with a functional endothelium than in vessels in which the endothelium was removed by rubbing. Fasudil (10(-6) to 10(-5) m), a Rho kinase inhibitor, and apocynin (10(-3) to 3 × 10(-3) m), an NADPH oxidase inhibitor, significantly (P ≤ 0.05) inhibited hypoxia-enhanced contraction in response to PHE and 5-HT. In conclusion, hypoxia-enhanced contraction occurred in EDAs. This appears to be partially mediated by reactive oxygen species produced by NAPDH oxidase, which activate Rho kinase to increase calcium sensitisation and enhance smooth muscle contraction.

Effect of feeding glucose, fructose, and inulin on blood glucose and insulin concentrations in normal ponies and those predisposed to laminitis.

Identification of ponies (Equus caballus) at increased risk of pasture-associated laminitis would aid in the prevention of the disease. Insulin resistance has been associated with laminitis and could be used to identify susceptible individuals. Insulin resistance may be diagnosed by feeding supplementary water-soluble carbohydrate (WSC) and measuring blood glucose and insulin concentrations. The aim of this study was to assess the glycemic and insulinemic responses of 7 normal (NP) and 5 previously laminitic (PLP), mixed breed, native UK ponies fed glucose, fructose, and inulin [1 g/(kg·d) for 3 d] or no supplementary WSC (control) in spring and fall after a 7-d adaptation to a pasture or hay diet. Blood samples were taken for 12 h after feeding on each day, and baseline and peak concentrations and area under the curve (AUC) for glucose and insulin were recorded. Linear mixed models were used for statistical analysis. Differences between PLP and NP groups were most marked after glucose feeding with differences in peak glucose (P = 0.02) and peak insulin (P = 0.016) concentrations. Season and diet adaptation also affected results. Peak concentrations of glucose and insulin occurred 2 to 4 h after WSC feeding. Peak insulin concentration was greater and more variable in fall, particularly in PLP adapted to fall pasture. Baseline glucose and insulin concentrations varied between individuals and with season and diet adaptation but were not greater in PLP than NP. Insulin AUC was greater in PLP than NP after feeding both glucose and fructose (P = 0.017), but there were no differences between PLP and NP in glucose AUC. Glycemic and insulinemic changes were less (P ≤ 0.05) after feeding fructose than glucose, although differences between PLP and NP were still evident. Minimal changes in glucose and insulin concentrations occurred after inulin feeding. Measurement of peak insulin 2 h after feeding of a single dose of glucose (1 g/kg) may be a simple and practical way to aid identification of laminitis-prone ponies before the onset of clinical disease, particularly when ponies are adapted to eating fall pasture.

Effect of induction dose, lactation stage and body condition on tiletamine-zolazepam anaesthesia in adult female grey seals (Halichoerus grypus) under field conditions.

Tiletamine-zolazepam (TZ) was used at a mean (sd) dose of 1.18 (0.15) mg/kg administered intramuscularly to anaesthetise adult female grey seals (Halichoerus grypus) under field conditions at three different stages during their lactation period. A significant correlation was observed between the induction dose and time to induction (r=-0.582, P=0.011). Stage of lactation had a significant effect on condition index (CI), calculated as axial girth divided by length (P<0.001), and time to induction (P=0.009). No effect of CI on induction or recovery time was demonstrated. Respiratory rate decreased during induction and increased significantly (P<0.001) during surgical biopsy of blubber. Recovery occurred after 32.5 (11.9) minutes. Minor complications (tremor, vocalisation and mild dyspnoea) were observed in a small number of cases, none of which required treatment.

Twenty-four hour growth hormone and leptin secretion in active postpubertal adolescent girls: impact of fitness, fatness, and age at menarche.

CONTEXT: GH is strongly related to body composition, physical activity, and pubertal progression. Adolescent girls decrease physical activity during puberty, whereas their weight increases. Because leptin is a good index of energy balance in active young women, we hypothesized that leptin is related to GH secretion in this population while taking into account fitness, fatness, and age at menarche. METHODS: We measured body composition and maximal oxygen consumption (VO(2)max) in 37 postpubertal adolescent girls aged 16-21 yr. GH was sampled every 10 min and leptin hourly for 24 h. We first analyzed 6-h time blocks by repeated measures for GH and leptin, with body mass index (BMI), percent body fat, and VO(2)max as covariates for the entire group and a lean subgroup. The deconvolution method was used to characterize GH pulsatility from individual time points. RESULTS: GH varied through the day (P < 0.0001), with the highest concentrations overnight. BMI, percent body fat, and VO(2)max were related to GH concentrations in the entire group, whereas leptin predicted GH in the entire group as well as the lean subgroup of girls. Higher leptin was related to lower GH concentrations (P = 0.011), regardless of time. A log leptin level increase by 1 unit decreased GH by 27%. Pulsatility characteristics showed a 1-yr increase of age at menarche increasing total GH input by 20% (P = 0.0035) independently from BMI. CONCLUSION: In postpubertal adolescent girls, leptin is related to GH concentration across the lean to overweight BMI spectrum. GH pulsatile secretion was greater in girls with later age at menarche.

Does exercise affect female hamster gonadotropins by reducing estradiol negative feedback?

Exercise stimulates reproductive function in hamsters exposed to short-day photoperiod (SDP) in contrast to its inhibitory effects in women and rats. SDP inhibits hamster reproduction in part by increasing the sensitivity of the hypothalamo-pituitary-gonadal axis (HPGA) to the negative feedback of gonadal steroids. To determine whether EX facilitates reproduction in female hamsters by affecting this mechanism, we examined the influence of estradiol (E2) on basal LH and FSH concentrations in exercising and sedentary hamsters maintained on long-day photoperiod (LD 14:10, LDP) or SDP (LD 8:16). In the LDP, serum LH and FSH were unaffected or reduced by exercise in ovariectomized (OVX) nonhormone-replaced hamsters, and LH was increased after tonic E2 replacement compared to sedentary controls. In the SDP, serum LH and FSH were significantly higher in OVX exercising than in sedentary hamsters, whether the exercisers were injected with a high dose of E2 or not. Thus, the effects of exercise on basal gonadotropin secretion in female hamsters appear to depend on the level of estradiol negative feedback (ENF). When this feedback is low (LDP OVX condition), exercise is either ineffective or inhibitory. When the ENF is increased by exposure to SDP and/or by treatment with E2, exercise has a stimulatory effect on basal gonadotropin secretion. Exercise may stimulate hamster gonadotropin secretion by reducing the ENF either by lowering the sensitivity of the HPGA to steroid negative feedback or by other means.

The effects of exercise on growth.

The way in which exercise influences statural, hypertrophic and reparative growth is examined from the perspective of the human lifespan. Statural growth depends on a neuroendocrine programme which channels nutrient energy towards increments in lean body mass. Exercise can facilitate statural growth and is a necessary stimulus for reparative growth through its stimulatory effects on secretion of growth hormone (GH) and other anabolic hormones. An exercise-associated increase in GH secretion is a response to acute or prolonged exercise-induced fuel shortage that directs metabolism towards utilisation of lipids and promotes growth. Exercise can transiently block the expression of statural growth by competitively removing the necessary nutritional support for growth. Statural growth retardation can be corrected by catch-up growth, but stunting may also be permanent (depending on the timing and magnitude of the energy drain). Hypertrophic growth is less dependent on hormonal and nutritional support than statural growth, and exercise provides the necessary mechanical stress for growth and remodelling of the musculoskeletal system. Excessive mechanical strain may suppress hypertrophic growth. The intermittent nature of exercise provides temporal organisation that is necessary for the normal operation of cellular growth process. Exercise by pregnant women does not appear to influence fetal growth. Evaluation of the effect of exercise on growth of children and adolescents is complicated by nonrandom selection of individuals for participation in organised sports, and by lack of information on the magnitude of exercise-induced energy drain. Exercise is essential for regulation of body composition in adulthood. It provides mechanical and metabolic stimuli that are necessary for hypertrophy of the musculoskeletal system and increased GH secretion for reparative growth.

Effect of exercise and food restriction on selected markers of the acute phase response in hamsters.

Acute aerobic exercise has been shown to elicit physiological changes characteristic of the acute phase response (APR), a nonspecific host defense response. Regular evocation of these changes may prime the immune system to improve resistance to disease. Because food deprivation is associated with an impaired APR, food restriction may prevent these beneficial changes. We tested the hypotheses that voluntary exercise elicits an APR and that food restriction modifies this response in four groups of hamsters: ad libitum-fed sedentary, ad libitum-fed exercised, food-restricted sedentary, and food-restricted exercised. Five variables altered during an APR were examined: core temperature, serum iron, serum interleukin-6, serum amyloid A, and serum glucocorticoids measured by biotelemetry, colorimetric analysis, B-9 cell growth assay, indirect enzyme-linked immunosorbent assay, and radioimmunoassay, respectively. Blood was drawn during the hamsters' inactive period after 19-20 days of access to running wheels. Resting core temperature was elevated by exercise and depressed by food restriction (P < 0.01). Iron was depressed by food restriction (P < 0.01). Cortisol, but not corticosterone, was elevated by food restriction (P < 0.001). There were no significant differences among groups in interleukin-6 (P > 0.49) or serum amyloid A (P > 0.29). We conclude that there is little evidence that voluntary exercise or exercise combined with food restriction causes an APR in hamsters.

Effect of voluntary exercise and food restriction in response to lipopolysaccharide in hamsters.

We tested the hypothesis that voluntary running and moderate food restriction alter the acute phase response (APR), one index of nonspecific immune function. Hamsters were kept sedentary or permitted to run and were fed ad libitum or had food restricted for 20 days and were then injected intraperitoneally with saline or lipopolysaccharide (LPS). Fever and circulating interleukin-6, serum amyloid A (SAA), serum iron, and cortisol were measured by biotelemetry, B-9 cell growth assay, indirect enzyme-linked immunosorbent assay, colorimetric analysis, and radioimmunoassay, respectively. The febrile temperature; hypoferremia; and elevation of circulating interleukin-6, SAA, and cortisol after LPS injection were not altered by exercise. Because baseline temperatures were elevated in the exercised hamsters, the change in temperature in response to LPS was less than it was in the sedentary hamsters. Food restriction significantly decreased SAA and elevated cortisol after LPS injection and depressed the absolute temperature to which the core temperature rose in response to LPS in one trial but not in another. Because food restriction depressed baseline temperatures, it also affected the change in temperature after LPS injection. The hypoferremic response to LPS was inhibited in hamsters that were both food restricted and permitted to run. We conclude that exercise does not enhance the APR to a low dose of LPS, whereas food restriction and the combination of exercise and food restriction depress some portions of the APR in hamsters.

Control by light of the temperature rhythm in food-restricted hamsters.

To establish the relative importance of light and food in the control of core temperature (Tc) rhythm in food-restricted hamsters, mature female hamsters maintained in 14L:10D lighting were fed restricted amounts of food at the onset of light (n = 6) or at the onset of dark (n = 6) and were compared to ad lib-fed animals. After 21-25 days of this entrainment, light stimulus was shifted by 12 h, and animals were kept in shifted lighting for another 13 days. Food restriction led to a 0.6 degree decrease in the mean Tc, which was expressed entirely during the day in night-fed hamsters and was evenly divided between day and night in day-fed animals. Thus, Tc and general activity rhythms maintained the entrainment to light under both dietary conditions, with peak values for all occurring during the early night. During 13 days following the 12-h shift in lighting, Tc and activity rhythms shifted in all animals, regardless of nutritional status, from entrainment to preceding lighting, through double rhythm frequency, indicating entrainment to preceding as well as current lighting, to entrainment to current lighting. Thus, in food-restricted hamsters, light stimulus rather than predictable timing of food prevails as the entrainer of Tc and activity rhythms.

Neurohumoral mediation of exercise-induced growth.

To assess exercise effects on growth, other variables modulating growth need to be taken into account. Endogenous control of growth proceeds from local actions of growth factors and dependence on nutrition abundance through guidance by growth hormone (GH) and other anabolic hormones to neuroendocrine suppression of growth. Nutrient abundance controls the reparative growth of lean body mass in adulthood by coupling it to anabolic endocrine reflexes. Growth is blocked when catabolic endocrine reflexes govern energy expenditure. The relationship between exercise intensity and growth is nonlinear. Growth is an intermittent process. Its expression and stimulation are dependent on ultradian and circadian rhythms of energy metabolism and neurohumoral release. High-resistance exercise selectively stimulates growth of the musculoskeletal system through expression of growth factor genes in the challenged tissues and without the GH guidance or abundant nutritional support. Habitual endurance exercise stimulates reparative growth of lean body mass through the neuroendocrine adaptations including increased pulsatile GH secretion. These also facilitate oxidative utilization of storage lipids thereby contributing to the regulation of body composition in adulthood. In the absence of sufficient high-resistance and endurance exercise regulation of adult body mass is impaired: excess LBM is lost during energy deficit, and excess fat accumulates during energy surplus.

Tumor necrosis factor is not involved in exercise-induced elevation in core temperature.

Female Sprague-Dawley rats (12:12-h light-dark photoperiod) with access to running wheels have an elevated body temperature (BT) both during exercise (nighttime) and nonexercise periods (daytime). We studied whether the exercise-induced increase in BT is modulated by the release of the cytokine, tumor necrosis factor-alpha (TNF). Two weeks after the onset of exercise, nighttime temperatures of exercising rats were elevated approximately 0.5 degree C compared with preexercise values (P = 0.006). By 3 wk after the onset of exercise, daytime temperatures had increased 0.3 degree C (P = 0.03) above control levels. To confirm that endogenously produced TNF can modulate fever in female rats, we injected six rats with antiserum to TNF (300 microliters/rat) and six rats with control serum 24 h before intraperitoneal injection of lipopolysaccharide (50 micrograms/kg). As occurred in earlier studies on male rats, antiserum-treated female rats had significantly enhanced fevers (P = 0.017). To determine whether endogenously produced TNF was involved in modulating the daytime and nighttime increases in BT, antiserum to TNF (300 microliters/rat, n = 7) or control serum (300 microliters/rat, n = 5) was injected intraperitoneally in exercising rats. Neither injection of antiserum nor control serum had any effect on daytime or nighttime BTs. Because BTs of exercising female rats injected with antiserum against TNF were not affected, we conclude that TNF is not responsible for modulating their exercise-induced rise in BT.

Role of prostaglandins in exercise-induced core temperature elevation in female Sprague-Dawley rats.

Female Sprague-Dawley rats (12:12-h photoperiod; body temperature, BT, measured with biotelemetry) with access to running wheels for 6 wk have an elevated BT (compared with rats with no access to exercise wheels, i.e, sedentary) both during the period of voluntary exercise (nighttime) (0.5 degree C, P = 0.0001) and the nonexercise period (daytime) (0.3 degree C, P = 0.002). To determine whether prostaglandin (PG) E was responsible for any portion of this daytime rise in BT, we injected a dose of sodium salicylate (300 mg/kg), which was shown to produce complete antipyresis in rats injected with lipopolysaccharide (LPS), into exercised and sedentary rats 4 h after the onset of the lights-on period. The injections of sodium salicylate led to a fall in body temperature in both the exercised and sedentary rats of similar amounts (-0.88 degree C vs. -0.61 degree C at 2 h postinjection, P = 0.59). We conclude that the increase in daytime BT of exercised female rats is not mediated by prostaglandins.

Food restriction postpones, not prevents, exercise-induced growth in hamsters.

Voluntary exercise during ad libitum feeding accelerates growth in mature female hamsters. If food is restricted during the exercise period, growth is suppressed, but ad libitum access to food at the cessation of exercise permits rapid catch-up growth in length and weight during retirement. To see whether the exercise-induced stimulus to grow persists when food-restriction is continued during retirement, female golden hamsters were matched by weight and assigned to exercise (EX) and sedentary (SED) groups fed ad libitum and to corresponding activity groups (REST-EX, REST-SED) food-restricted to 80% of starting weight. At the end of each week for 3 weeks following retirement, one REST-EX and one REST-SED group were fed ad libitum. At 21-35 days post-refeeding, mean gains in length and weight of each exercise group were greater (p < 0.05) than the gains of the corresponding sedentary group. Thus, exercise under food restriction remained an effective stimulus for acceleration of growth throughout three weeks of continued food restriction beyond termination of exercise, and ad libitum feeding permitted the delayed expression of exercise-induced catch-up growth in hamsters.

Stimulation by voluntary exercise of adrenal glucocorticoid secretion in mature female hamsters.

The possibility that habitual voluntary running induces a chronic change in adrenal glucocorticoid synthesis and secretion was examined in freely running mature female hamsters, in whom this behavior accelerates growth, reduces body fat levels, and elevates core temperature. Hamsters were free to run on horizontal discs or in vertical wheels between 32 and 80 days, in 14L:10D or in 10L:14D photoperiods, and at the end of this period, corticosterone and cortisol steroidogenesis and serial plasma corticosterone concentrations during day and night were used as measures of the chronic stimulation of adrenal cortical activity. Habitual voluntary running significantly increased steroidogenesis of both glucocorticoids and plasma corticosterone concentrations and alone accounted for all the variance in enhanced synthesis and secretion of corticosterone. Acute exercise and/or the nocturnal phase of circadian period enhanced the chronic stimulatory effects of exercise on cortisol. Despite its voluntary and apparently stress-free nature, running induces chronic increases in basal glucocorticoid secretion in mature female hamsters. Putative oversecretion of corticotropin releasing factor in freely running hamsters could account for increased steroidogenesis, acceleration of growth, reduced body fat levels, and core temperature elevation.

Activity disc and cage for continuous measurement of running activity and core temperature in hamsters.

We describe a design for the modular horizontal activity disc and tandem cages suitable for continuous monitoring of spontaneous running and of core temperatures in golden hamsters. An acrylic disc is equipped with a short brass axle. It is mounted inside a brass rotation sleeve at a 15 degrees angle off the horizontal plane. The disc module fits firmly inside either half of the tandem cage when activity measurements are needed. Easy removal allows for alternative use of cages. Minor modifications of disc dimensions and of disc base permit the use of activity modules with juvenile hamsters. The short distance between disc surface and cage floor permits continuous measurement of core temperature as well as running activity.

Body temperature rhythm and response to pyrogen in exercising and sedentary hamsters.

In this study, we tested the hypothesis that daily voluntary exercise results in a chronic elevation in core temperature in the female golden hamster. Temperature and activity were measured by biotelemetry. Hamsters ran 6-7 km per night (12:12 L:D) when permitted access to wheels. No running occurred during the light periods. During the 3rd wk of running, temperatures of exercising hamsters were significantly elevated by 0.5 degree C (P less than 0.001) during the dark period and by 0.3 degree C (P less than 0.003) during the light period compared with sedentary hamsters. Cessation of running removed the difference between groups, and resumption of running restored it. Both the injection of endotoxin and the psychological stress of cage switch resulted in similar peak temperatures in exercising and sedentary hamsters despite higher pre-treatment temperatures in the exercise group. We interpret these results to support the hypothesis that regular exercise may cause an upward resetting of the set-point for body temperature.

Melatonin increases serum growth hormone and insulin-like growth factor I (IGF-I) levels in male Syrian hamsters via hypothalamic neurotransmitters.

In male Syrian hamsters daily evening melatonin injections resulted in increased circulating levels of growth hormone (GH), as well as a modest increase in body weight. A substantial increase in serum levels of insulin-like growth factor I (IGF-I) was observed in all hamsters receiving evening injections of melatonin for 10 weeks. The melatonin-induced increase in serum IGF-I levels was interpreted as a result of increased release of GH during the 10 week period of melatonin administration. The increase in serum GH and IGF-I was associated with significantly decreased hypothalamic turnover of norepinephrine (NE). Since blocking NE synthesis with alpha methyl-p-tyrosine reduced serum GH, the melatonin-induced increase in GH could not readily be attributed to decreased NE turnover. Highly significant increases in 5-hydroxyindole acetic acid (5HIAA) concentrations and in ratios of 5HIAA to serotonin (5HT) were noted in extracts of hypothalamus and in extracts of brain stem, suggesting a serotonergic component to melatonin-induced increase in GH-induced IGF secretion and subsequent growth.