Resistance Training Threshold for Elevating Bone Mineral Density in Growing Female Rats.

The purpose of this study was to determine the minimum amount of resistance exercise that would stimulate bone formation yielding an elevation in bone mineral density (BMD) during the growth period in female rats. Female rats were randomly divided into: Control (Con, n=8), 3 ladder climb resistance-trained group (3LC, n=8), 4 ladder climb resistance-trained group (4LC, n=8), 5 ladder climb resistance-trained group (5LC, n=8), and 6 ladder climb resistance-trained group (6LC, n=8). All exercised groups were conditioned to climb a vertical ladder with weights appended to their tail 3 days/wk for a total of 6 wks. After 6 wks, left tibia BMD (g/cm2) was significantly greater for 4LC (0.197±0.003), 5LC (0.200±0.004) and 6LC (0.202±0.003) when compared to Con (0.185±0.006). Left femur BMD (g/cm2) was significantly greater for 4LC (0.260±0.005), 5LC (0.269±0.004) and 6LC (0.272±0.006) when compared to Con (0.244±0.006). There were no significant differences in tibia and femur BMD between 4LC, 5LC, and 6LC groups. The results suggest that during growth, a high volume of resistance exercise was required to elicit an elevation in BMD in females.

Better cognitive control of emotional information is associated with reduced pro-inflammatory cytokine reactivity to emotional stress.

Stress is strongly associated with several mental and physical health problems that involve inflammation, including asthma, cardiovascular disease, certain types of cancer, and depression. It has been hypothesized that better cognitive control of emotional information may lead to reduced inflammatory reactivity to stress and thus better health, but to date no studies have examined whether differences in cognitive control predict pro-inflammatory cytokine responses to stress. To address this issue, we conducted a laboratory-based experimental study in which we randomly assigned healthy young-adult females to either an acute emotional stress (emotionally evocative video) or no-stress (control video) condition. Salivary levels of the key pro-inflammatory cytokines IL-1ß, IL-6, and IL-8 were measured before and after the experimental manipulation, and following the last cytokine sample, we assessed participants' cognitive control of emotional information using an emotional Stroop task. We also assessed participants' cortisol levels before and after the manipulation to verify that documented effects were specific to cytokines and not simply due to increased nonwater salivary output. As hypothesized, the emotional stressor triggered significant increases in IL-1ß, IL-6, and IL-8. Moreover, even in fully adjusted models, better cognitive control following the emotional (but not control) video predicted less pronounced cytokine responses to that stressor. In contrast, no effects were observed for cortisol. These data thus indicate that better cognitive control specifically following an emotional stressor is uniquely associated with less pronounced pro-inflammatory cytokine reactivity to such stress. These findings may therefore help explain why superior cognitive control portends better health over the lifespan.

High volumes of resistance exercise are not required for greater bone mineral density during growth.

PURPOSE: The purpose of this study was to determine the minimum amount of resistance exercise that would stimulate bone formation, via an elevation in bone mineral density (BMD), during the growth period in male rats. METHODS: Forty male rats were randomly divided into control group (Con, n = 8), one ladder climb resistance-trained group (1LC, n = 8), two ladder climb resistance-trained group (2LC, n = 8), three ladder climb resistance-trained group (3LC, n = 8), and four ladder climb resistance-trained group (4LC, n = 8). All exercised groups were conditioned to climb a vertical ladder with weights appended to their tail 3 d·wk(-1) for a total of 6 wk. RESULTS: After 6 wk, left tibia BMD (mean ± SE) was significantly greater for 2LC, 3LC, and 4LC (0.233 ± 0.003 g·cm(-2)) when compared with Con (0.218 ± 0.003 g·cm). Left femur BMD was significantly greater for 2LC, 3LC, and 4LC (0.318 ± 0.003 g·cm(-2)) when compared with 1LC (0.299 ± 0.008 g·cm(-2)) and Con (0.289 ± 0.010 g·cm(-2)).There were no significant differences in BMD between 2LC, 3LC, and 4LC groups. CONCLUSION: The results suggest that during growth, a low amount of resistance exercise was just as effective as high volumes of strength training for stimulating bone modeling.

Prevalence of vitamin D3 deficiency in orange county residents.

With the prevalence of sunlight exposure in Orange County, California, one would expect it to be rare and unusual to find high incidence of Vitamin D deficiencies among its residents. This study evaluated the concentration of Vitamin D3 as part of a larger study to evaluate bone health in Orange County residents. Our preliminary data shows that 19.2% of the 151 subjects evaluated had low Vitamin D3 (<30 pg/ml) and illuminates a growing problem in the United States. We speculate that the widespread sensitivity to skin cancer and sun exposure, the increased use of sun-screens and the filtering of UV waves in automobile glass has put the public at risk for low Vitamin D.

Compromised bone health in non-obese, older women with low caloric intake.

Low calorie diets are often employed to reverse obesity and lower the risk of other diseases. Further, to maintain a certain physique and/or minimize the incidence of obesity, normal weight, older women may consume a diet low in calories. However, a low caloric intake can potentially compromise bone formation elevating the risk of osteoporosis. Thus, the purpose of this study was to compare the bone mineral density (BMD) from non-obese, peri- and postmenopausal women who were placed into one of three groups based upon their daily caloric intake to determine if bone health was compromised when caloric intake was low. Women (age range 44-80 years old) with a body mass index <25 were separated into one of three groups based upon a dietary analysis to assess nutritional intake: those who consumed 100% of the recommended minimum daily requirement (RMDR, n = 21), 80% RMDR (n = 16), or 55% RMDR (n = 15). BMD was determined via dual energy X-ray absorptiometer from the lumbar spine and femur (left & right). The 55% RMDR group demonstrated significantly lower femoral BMD (~10% lower, P < 0.05) and a trend toward lower BMD of the spine compared to both the 80% RMDR (P = 0.13) and 100% RMDR groups (P = 0.15). There were no significant differences in BMD between the 80% RMDR and 100% RMDR groups. The results suggest that bone health was maintained with moderately low caloric intake (i.e. 80%RMDR) whereas extremely low caloric intake (i.e. 55% RMDR) can compromise bone health in non-obese, older women.

Sex differences in hepatic gluconeogenic capacity after chronic alcohol consumption.

Alcohol-induced hypoglycemia has traditionally been attributed to the amount of ethanol consumed rather than any inherent decline in glucose output capacity by the gluconeogenic organs and/or an increase in skeletal muscle glucose uptake. Further, while the potential for sex differences that might impact glucose homeostasis following chronic alcohol consumption has been recognized, direct evidence has been noticeably absent. This paper will provide a brief review of past and present reports of the potential for sex differences in glucose homeostasis following chronic ethanol consumption. This paper will also provide direct evidence from our laboratory demonstrating sex differences from chronic alcohol consumption resulting in a decrement in glucose appearance and more importantly, a specific decline in hepatic gluconeogenic (HGN) capacity in the absence and presence of ethanol. All our studies involved 8 weeks of chronic alcohol consumption in male and female Wistar rats, as well as a 24 to 48 hour fast to deplete hepatic glycogen stores. Under the conditions of chronic alcohol consumption and an acute dose of ethanol, we provide in vivo evidence of an early decline in whole body glucose appearance in females fed an ethanol diet compared to controls. While the decline was also observed in males fed the alcohol diet, it occurred much later compared to ethanol fed females. The site for the decline in whole body glucose production (i.e., either the kidneys or the liver) was beyond the scope of our prior in vivo study. In a follow-up study using the in situ perfused liver preparation, we provide additional evidence for a specific reduction in HGN capacity from lactate in ethanol fed females compared to ethanol fed males in the absence of alcohol in the perfusion medium. Finally, employing the isolated hepatocyte technique, we report decrements in HGN from lactate in ethanol fed females compared to ethanol fed males in the presence of ethanol in the incubation medium. The mechanism for the specific decline in HGN within the liver of ethanol fed females remains to be determined. To the extent that our observations in animals can be extrapolated to humans, we conclude that alcoholic women are more susceptible to ethanol-induced hypoglycemia compared to alcoholic men.

Alcohol-induced suppression of gluconeogenesis is greater in ethanol fed female rat hepatocytes than males.

The impact of alcohol-induced suppression on hepatic gluconeogenesis (HGN) after chronic ethanol consumption between males and females is unknown. To determine the effects of chronic alcohol consumption (8 weeks) on HGN, the isolated hepatocyte technique was used on 24 h fasted male and female Wistar rats. Livers were initially perfused with collagenase and the hepatocytes were isolated. Aliquots of the cell suspension were placed in Krebs-Henseleit buffer and incubated for 30 min with lactate, [U -14C]lactate, and nine different concentrations of ethanol (EtOH). Dose-effect curves were generated for the determination of maximal and half-maximal alcohol-induced inhibition on HGN. There was no significant difference in HGN (lactate only and no EtOH) between males and females fed the control diet (88.5 +/- 5.1 nmol/mg protein/30 min). Similarly, the HGN (lactate only and no EtOH) in males fed the ethanol diet (ME) were not significantly different (82.8 +/- 3.5 nmol/mg protein/30 min) compared to controls. In contrast, the females chronically fed the ethanol diet (FE) had significantly (P < .05) lower HGN (67.8 +/- 4.6 nmol/mg protein/30 min) compared to both ME and controls. With alcohol in the incubation medium, the HGN significantly (P<.05) declined in all groups. While alcohol suppressed HGN to a larger (P < .05) extent in ME (45.8 +/- 3.7 nmol/mg protein/30 min) compared to controls (64.0 +/- 3.8 nmol/mg protein/30 min), the inhibition was even greater (P < .05) in FE (32.7 +/- 3.2 nmol/mg protein/30 min). The more pronounced effect of chronic alcohol consumption on HGN in the presence of ethanol in female hepatocytes was supported by the concomitant decreases (P < .05) in 14C-lactate incorporation into 14C-glucose, lactate uptake, and 14C-lactate uptake. The results suggest that chronic alcohol consumption elicits a greater reduction on HGN in the presence of ethanol in the hepatocytes of females compared to males.

Impact of flow rate on lactate uptake and gluconeogenesis in glucagon-stimulated perfused livers.

The impact of reduced hepatic flow on lactate uptake and gluconeogenesis was examined in isolated glucagon-stimulated perfused livers from 24-h-fasted rats. After surgical isolation, livers were perfused (single pass) for 30 min with Krebs-Henseleit (KH) bicarbonate buffer, fresh bovine erythrocytes (hematocrit approximately 20%), and no added substrate. After this "washout" period, steady-state perfusions were initiated with a second reservoir containing the KH buffer, bovine erythrocytes, [U-(14)C]lactate (10,000 dpm/ml), lactate (2.5 mM), and glucagon (250 microg/ml). Perfusion flow rate was adjusted to one of five rates (i.e., 1.8, 2.7, 3.9, 7.4, and 11.0 ml.min(-1).100 g body wt(-1)). After the perfusion, the liver was dissected out and weighed so as to establish the actual flow rate per gram of liver. The resulting flow rates ranged from 0.52 to 4.03 ml.min(-1).g liver(-1). As a function of flow rate, lactate uptake rose in a hyperbolic fashion to an apparent plateau of 2.34 micromol.min(-1).g liver(-1). Fractional extraction (FX) of lactate from the perfusate demonstrated an exponential decline with increased flow rates (r=0.97). At flow rates above 1.0 ml.min(-1).g liver(-1), adjustments in FX compensated for changes in lactate delivery, resulting in steady rates of lactate uptake and gluconeogenesis. Below 1.0.min(-1).g liver(-1) the increased FX was unable to compensate for the decline in lactate delivery and lactate uptake declined rapidly. Gluconeogenesis demonstrated similar kinetics to lactate uptake, reflecting its dominant role among pathways for lactate removal under the current conditions.

Lactate delivery (not oxygen) limits hepatic gluconeogenesis when blood flow is reduced.

The purpose of this study was to determine, using the isolated liver perfusion technique, whether the limiting factor for hepatic gluconeogenesis (GNG) from lactate was precursor delivery or oxygen availability during reduced flow rates of 0.85 or 0.60 ml.min(-1).g liver(-1). After a 24-h fast, three different experimental protocols were employed. Protocol 1 examined the impact on GNG when reservoir lactate concentration was maintained but oxygen delivery was elevated via increases in hematocrit (Hct). Elevating the Hct from 22.5+/- 0.8% to 30.9+/- 0.4% at a blood flow of 0.89+/- 0.01 ml.min(-1).g liver(-1) increased the oxygen consumption (Vo(2)) but did not augment GNG. Similarly, when the Hct was elevated from 22.5+/- 0.8% to 41.5+/- 0.7% at 0.59+/- 0.04 ml.min(-1).g liver(-1), Vo(2) was increased, but GNG was unaffected. Protocol 2 examined the impact on GNG when Hct was maintained but precursor delivery was elevated via increases in reservoir lactate concentration ([LA]). Specifically, elevating the [LA] from 2.31+/- 0.07 to 3.61+/- 0.33 mM at a flow rate of 0.82+/- 0.04 ml.min(-1).g liver(-1) significantly increased GNG. Similarly, elevating the [LA] from 2.31+/- 0.07 to 4.24+/- 0.37 mM at a flow rate of 0.58+/- 0.02 ml.min(-1).g liver(-1) increased GNG. Finally, we examined the impact of increasing both the oxygen and lactate delivery (Protocol 3). Again, Vo(2) was elevated with increased oxygen delivery, but GNG was not augmented beyond that observed with elevations in lactate delivery alone, i.e., Protocol 2. The results indicate that, during decrements in blood flow, GNG is limited primarily by precursor delivery, not oxygen availability.

Opposing effects of chronic alcohol consumption on hepatic gluconeogenesis for female versus male rats.

BACKGROUND: The impact of chronic alcohol consumption on hepatic gluconeogenesis (HGN) between males and females is unknown. To determine the effects of chronic alcohol consumption (8 weeks) on HGN, the isolated liver perfusion technique was used on 24-hr-fasted male and female Wistar rats. METHODS: After surgical isolation, livers were perfused (single pass) for 30 min with Krebs-Henseleit bicarbonate buffer and fresh bovine erythrocytes with no added substrate (washout period). After the washout period, livers were perfused with lactate (10 mM) and [U-14C]lactate (15,000 dpm/ml) using the recirculation method. RESULTS: There was no significant difference in HGN between males and females fed the control diet. In contrast, the females chronically fed the ethanol diet (FE) had significantly lower HGN rates (2.73 +/- 0.37 micromol/min x g liver protein(-1)), whereas males fed the ethanol diet (ME) had significantly higher HGN rates (4.99 +/- 0.45 micromol/min x g liver protein(-1)) than controls (3.83 +/- 0.34 micromol/min x g liver protein(-1)). Concomitant decreases were also observed for both 14C-lactate incorporation into 14C-glucose and rates of lactate uptake for FE, while corresponding increases were observed for 14C-lactate incorporation into 14C-glucose for ME. The livers from ME were able to convert a greater percentage of the lactate into glucose, resulting in the elevation in gluconeogenic capacity. CONCLUSION: Chronic alcohol consumption lowers the hepatic gluconeogenic capacity from lactate in females and elevates HGN in males.

Differential effects of alcohol upon gluconeogenesis from lactate in young and old hepatocytes.

The purpose of this study was to determine the effect of age upon hepatic gluconeogenesis (HGN) from lactate in the presence of various concentrations of alcohol from young (3 months) and old (24 months) male rats. After a 24-hour fast, livers were perfused with collagenase and the hepatocytes were isolated. Aliquots of the cell suspension were placed in Krebs-Henseleit buffer and incubated with lactate, [U-(14)C]lactate, and nine different concentrations of ethanol (EtOH) for 30 min. Dose-effect curves were generated for the determination of maximal and half-maximal alcohol-induced inhibition on gluconeogenesis. There were no significant differences in basal HGN (lactate only and no EtOH) between young and old hepatocytes, 86.9+/-6.3 nmol/mg protein/30 min. The addition of ethanol significantly reduced HGN from lactate in both groups. At the highest ethanol concentration (15 mM), the glucose production was inhibited more from old, 46.1+/-1.2 nmol/mg protein/30 min, compared to young hepatocytes, 56.0+/-1.6 nmol/mg protein/30 min. The greater age-related reduction in HGN was confirmed by the minimal glycogenolysis, and the concomitant decline in [U-(14)C]glucose production, lactate uptake, and [U-(14)C]lactate uptake. The results suggest that alcohol elicits a greater inhibition upon HGN from lactate in old compared to young liver cells.

Chronic alcohol consumption yields sex differences in whole-body glucose production in rats.

AIMS: The effects of chronic alcohol consumption (8 weeks) on glucose kinetics, in the absence (water, 4 g/kg) and presence of an acute ethanol dose (4 g/kg), were examined in 48 h fasted male and female Wistar rats. METHODS: Primed continuous infusions of [6-3H]- and [U-14C]glucose were employed to assess rates of glucose appearance (Ra), glucose disappearance (Rd), and apparent glucose carbon recycling. RESULTS: After injecting the male and female controls with water, there were no significant alterations in glucose kinetics. Compared to controls, chronic alcohol-fed female animals (injected with water) demonstrated significantly lower: glucose Ra, blood glucose concentration, and apparent glucose carbon recycling for a majority of the experimental period. In separate groups injected with ethanol, the glucose Ra fell by 31% for male rats fed the control diet (MC), 43% for male rats fed the ethanol diet (ME), 29% for female rats fed the control diet (FC), and 42% for female rats fed the ethanol diet (FE). Further, compared to controls (MC and FC), the blood glucose concentration was significantly lower prior to and following the ethanol injection for FE. In addition, FE animals had significantly lower rates of glucose Ra and glucose carbon recycling compared to controls prior to and after the ethanol injection. ME animals demonstrated similar declines in glucose Ra (compared to FE), but only after the ethanol injection. Conversely, ME were able to match the decrease in glucose Ra with comparable declines in glucose Rd resulting in blood glucose concentrations that did not differ from controls. CONCLUSIONS: Chronic alcohol consumption results in sex differences in whole-body glucose production and glucose regulation.

Effect of endurance training and fasting on renal gluconeogenic enzymes in the rat.

The purpose of this study was to examine the effects of chronic exercise training (running 30 m/min, 10% grade, 90 min/d for 8-10 weeks) on specific renal enzyme activities involved with the gluconeogenic pathway in the fed and 24-hr fasted state in rats. A portion of the kidney (containing the cortex and medulla) was homogenized from which cytosolic (c) and mitochondrial (m) fractions were separated. Maximal gluconeogenic enzyme activities were assessed for: phosphoenolpyruvate carboxykinase (cPEPCK), fructose 1,6-bisphosphatase (cFBP), pyruvate carboxylase (mPC), aspartate aminotransferase (cAspAT), alanine aminotransferase (cAlaAT), and lactate dehydrogenase (cLDH). In the fed state, there was no significant difference between groups in any of the enzymes examined (nmoles/min x mg protein-1): cPEPCK (25.8+/- 1.7), cFBP (106.8+/- 7.1), mPC (20.7+/- 1.8), cAspAT (1047.1 +/- 38.6), cAlaAT (52.3 +/- 4.3), and cLDH (1728.6+/- 163.2). After the 24-hr fast, there was a significant increase in cPEPCK (52.4+/- 2.9 and 52.0 +/- 2.1) and mPC (44.6 +/- 4.3 and 47.6 +/- 4.9), control and trained, respectively. These results suggest that the maximal enzyme activities for cPEPCK and mPC can be augmented as a result of fasting that was independent of the training status.

Effect of age and endurance training on the capacity for epinephrine-stimulated gluconeogenesis in rat hepatocytes.

The effects of endurance training on hepatic glucose production (HGP) from lactate were examined in 24-h-fasted young (4 mo) and old (24 mo) male Fischer 344 rats by using the isolated-hepatocyte technique. The liver cells were incubated for 30 min with 5 mM lactate ([U-14C]lactate; 25000 dpm/ml) and nine different concentrations of epinephrine (Epi). Basal HGP (with lactate only and no Epi) was significantly greater for young trained (T) (99.6 +/- 6.2 nmol/mg protein) compared with young controls (C) (78.2 +/- 6.0 nmol/mg protein). The basal HGP was also significantly greater for old T (97.3 +/- 5.9 nmol/mg protein) compared with old C (72.2 +/- 3.9 nmol/mg protein). After the incubation with the various concentrations of Epi, Hanes-Woolf plots were generated to determine kinetic constants (Vmax and EC50). Maximal Epi-stimulated hepatic glucose production (Vmax) was significantly greater for young T (142.5 +/- 6.5 nmol/mg protein) compared with young C (110.9 +/- 4.8 nmol/mg protein). Similarly, the Vmax was significantly greater for old T (138.2 +/- 5.0 nmol/mg protein) compared with old C (103.9 +/- 2.5 nmol/mg protein). Finally, there was an increase in the EC50 from the hepatocytes of old T (56.2 +/- 6.2 nM) compared with young T (32.6 +/- 4.9 nM). In like manner, there was an increase in the EC50 from the hepatocytes of old C (59.7 +/- 5.8 nM) compared with young C (33.1 +/- 2.7 nM). The results suggest that training elevates HGP in the basal and maximally Epi-stimulated condition, but with age there is a decline in EC50 that is independent of training status.

Hepatic gluconeogenic capacity from various precursors in young versus old rats.

Hepatic gluconeogenic capacity was studied in young (4 months of age) and old (24 months of age) male Fischer 344 rats fasted for 24 hours using the isolated hepatocyte technique. Following the isolation of liver cells, the following precursors were added to the cell suspensions and incubated for 30 minutes: lactate (5 mmol/L), pyruvate (5 mmol/L), alanine (5 mmol/L), glutamine (5 mmol/L), oxaloacetate (5 mmol/L), glycerol (5 mmol/L), dihydroxyacetone (10 mmol/L), fructose (10 mmol/L), or saline (no precursor addition). To confirm that glucose production reflects gluconeogenic capacity, there was significant depletion of hepatic glycogen after the 24-hour fast and minimal alterations in glycogen content once substrates were added. Adjusting the gluconeogenic rates to reflect 100% cell viability resulted in no difference between young and old animals for any substrate used with the sole exception of fructose. The hepatic glucose production from fructose was 34% greater for young versus old animals. The results suggest that following a period of starvation the basal glucose production rates from hepatocytes, incubated with precursors entering the gluconeogenic pathway prior to fructose-6-phosphate, are equivalent in young and old rats.