Browsing by Author "Denbow, D. Michael"
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- Aging, Physical Activity, and Energy Intake RegulationVan Walleghen, Emily Lynn (Virginia Tech, 2006-07-31)More than seventy percent of Americans over the age of sixty are classified as overweight or obese, and the future incidence of these conditions is expected to rise. Although it is unclear why older adults are predisposed to weight gain, decreased total energy expenditure may contribute to positive energy balance. It is also possible that age-related impairments in energy intake regulation result in the inability to appropriately adjust food intake to meet energy requirements with advancing age. The purpose of these investigations was to determine the influence of age and habitual physical activity on acute regulation of energy intake. Secondary objectives were to determine if there are sex differences in energy intake regulation, and to determine if pre-meal water consumption decreases meal energy intake in young and older adults. To achieve these objectives, the ability to spontaneously adjust energy intake at a meal under "preloading" conditions in which a yogurt shake or water was consumed prior to the meal was determined. We hypothesized that older adults would demonstrate less accurate energy intake regulation than younger adults, but that energy intake dysregulation would be attenuated in physically active older adults. We also expected that young men would have higher accuracy of energy intake regulation compared to young women matched for dietary cognitive restraint and cardiorespiratory fitness, and that pre-meal water consumption would decrease meal energy intake in young and older adults. Our main finding was that energy intake regulation is significantly impaired in older compared to younger adults, and that habitual physical activity improves short-term, but not acute, energy intake regulation. We also found that young men demonstrate significantly higher accuracy of energy intake regulation compared to young women. Lastly, we determined that pre-meal water consumption significantly decreases meal energy intake in older, but not young, adults. Overall, these results indicate that acute energy intake regulation is less accurate with advancing age, but that regular physical activity improves short-term energy intake regulation. Additionally, sex appears to influence energy intake regulation, and water consumption is a potential strategy to reduce energy intake in older adults.
- Alterations in Lipid Metabolism in Mouse Tissues and Hepatic Cell Lines in Response to the Trans10,Cis12-18:2 Isomer of Conjugated Linoleic AcidViswanadha, Srikant (Virginia Tech, 2003-07-15)Conjugated linoleic acid (CLA) reduces adipose mass in several species. Studies were conducted to determine: 1) the effect of dietary trans10,cis12-CLA on growth, tissue fatty acid profile, mRNA expression for stearoyl-CoA desaturase (SCD) in adipose and liver, and mRNA expression for fatty acid synthase (FAS) in adipose of mice, 2) the effect of a dietary combination of trans-vaccenic acid (TVA) and trans10,cis12-CLA on delta9- desaturation, and 3) the effect of cis9,trans11-CLA, trans10,cis12-CLA, and carnitine palmitoyltransferase-1 (CPT-1) inhibitors on expression of mRNA for CPT-1 and fatty acid profile in mouse hepatocytes (AML-12) and human hepatoma cells (HepG2). In the first study, male or female mice were fed diets containing 0, 0.15%, or 0.30% trans10,cis12-CLA for 6 wk. Epididymal adipose weights (males) and inguinal adipose weights (females) decreased by 81% and 52%, respectively, in response to 0.30% trans10,cis12-CLA. Dry carcass weights decreased from 4.75 g for the control to 3.62 g for mice fed 0.30% trans10,cis12-CLA and the decrease was due to a reduction in ether extract. Liver weights increased linearly from 0.55 g (control) to 0.65 g (0.30% trans10,cis12-CLA). Dietary trans10,cis12-CLA (0.30%) reduced FAS and SCD mRNA in adipose by 60 and 30 % respectively, compared with the control, suggesting reduced lipogenesis and desaturation might be primary factors responsible for reducing body fat. In the second study, adult male or female mice were fed diets containing 0.40% TVA in combination with 0, 0.15, or 0.30% trans10,cis12-CLA for 10 d. Both TVA and trans10,cis12-CLA were incorporated into plasma, liver, adipose, muscle, and bone lipids proportional to their concentrations in the diets. Desaturation ratios were not affected in adipose, liver, and bone. However, ratios of 16:0 to 16:1 and 18:0 to 18:1 increased from 0.81 to 0.86 and 0.15 to 0.19 respectively, in response to dietary trans10,cis12-CLA (0.30%), suggesting inhibition of delta9 desaturation in muscle. In the third study, AML-12 or HepG2 cells were incubated with control media or media containing 15 uM etomoxir (ETM), 30 uM ETM, 15 uM hemipalmitoylcarnitinium (HPC), 30 uM HPC, 100 uM cis9,trans11-CLA, or 100 uM trans10,cis12-CLA for 24 h. Half the cells were harvested for analysis of fatty acids, mRNA for CPT-1, and cholesterol after 24 h. The remaining cells were incubated for an additional 24 h in control medium. Incorporation (% of total fatty acids) of trans10,cis12-CLA was greater than cis9,trans11-CLA in AML-12 (34 vs 23.6) and HepG2 (28 vs 18) cells. Cells incubated with trans10,cis12-CLA had higher ratios of 16:0 to 16:1, 18:0 to 18:1, and 18:2n6 to 20:4n-6, suggesting inhibition of delta9, delta5 , and delta6 desaturation. Cis9,trans11-CLA also reduced ratio of 18:2n-6 to 20:4n-6 in both cell lines. Trans10,cis12-CLA increased mRNA for CPT-1 in both cell lines compared with the control, suggesting enhanced oxidation of fatty acids. In addition, trans10,cis12-CLA caused a 4-fold and 5-fold increase in free cholesterol content of AML-12 and HepG2 cells, respectively. Overall, results demonstrated that trans10,cis12-CLA modulated lipid metabolism in tissues in vivo and altered fatty acid metabolism, cholesterol synthesis, and CPT-1 mRNA in hepatic cell lines in vitro.
- Altered Autonomic Nervous System Function in Chickens Divergently Selected for Body WeightKuo, Alice Yi-Wen (Virginia Tech, 2000-08-04)Autonomic nervous system activity is related to body weight regulation. Based on the MONA LISA hypothesis it has been suggested that most obese subjects and animals have low sympathetic nervous system activity. The aim of this study was to investigate whether there are differences in autonomic nervous system activity between lines of chickens selected for either high (HWS) or low body weight (LWS). In Exp. 1, various pharmacological agents were injected intravenously, and the changes in blood pressure (BP) and heart rate (HR) of both HWS and LWS chickens were compared. The results showed that the HWS birds had a greater increase in BP and HR than the LWS following injection of atropine, a muscarinic receptor blocker, and LWS birds had a greater decrease in BP and HR to propranolol, a beta- adrenergic receptor blocker than the HWS birds. These results suggested that HWS chickens have higher parasympathetic tone, whereas LWS chickens have a higher sympathetic nervous system tone regulating the cardiovascular system. HWS and LWS chickens displayed a similar response in BP and HR following injection of the ganglion blocker tetraethylammonium chloride. These results suggest that there is no significant difference in the central autonomic nervous system in the cardiovascular regulation between HWS and LWS together. Since there does not appear to be any differences in the activity of the autonomic nervous system activity at the level of the central nervous system, these findings imply that the difference in response to atropine and propranolol could be caused by differences in adrenal activity. The ratio of heart rate and blood pressure after the injection of phenylephrine showed significant difference between these two lines of birds, but not when phenylephrine was injected following atropine. This result indicated that HWS are more dependent on the parasympathetic nervous system to regulate the baroreceptor reflex. The percentage of adrenal and sympathetic impact on the regulation of heart rate showed that LWS females required greater adrenal activity than the other birds. In Exp. 2, the body weight and food intake responses of HWS and LWS chickens to ip injections of reserpine were compared. Reserpine caused a transitory decrease in food intake and body weight in both lines of birds. However HWS chickens recovered more slowly from the depression caused by reserpine than the LWS chickens. This could be due to lower sympathetic nervous system activity. In conclusion, it appears that HWS may have lower sympathetic activity than LWS. Combining the results of both experiments, it appears that the HWS birds have lower sympathetic and higher parasympathetic activity. Furthermore central nervous system autonomic activity in BP and HR regulation is not different between HWS and LWS, but the activity of the adrenal gland may be different between these two lines of birds.
- Androgen aromatization and cytosol estradiol-receptors in the mediation of masculine sexual behavior in Japanese quailParsons, Martha Anne Cohen (Virginia Polytechnic Institute and State University, 1982)The neuroendocrinology of masculine sexual behavior was examined in Japanese quail selected bi-directionally for adult mating frequency. Experiments were conducted with brain tissues from functionally castrated, High (HM) and Low Mating (LM) line quail to compare (1) the in vivo aromatization of ³H-testosterone and (2) cytosolic estradiol-binding by in vitro saturation analyses. After in vivo incubation with ³H-testosterone, all radioactivity recovered in brain tissues was in the form of testosterone, dihydrotestosterone, or estradiol. Neither the total ³H nor ³H-testosterone metabolite radioactivity differed upon comparison of the two genetic lines. Of all ³H-testosterone metabolite radioactivity, ³H-estradiol represented 45 ± 6% in the HM line and 46 ± 6% in the LM line, indicating that the line difference in mating frequency was not due to a corresponding difference in aromatase activity. Inasmuch as both the HM and LM line birds actively converted testosterone to estradiol, these results implicated a neural mechanism involving estradiol-receptor interactions. Estradiol-receptor binding parameters were estimated in subsequent experiments by fitting a hyperbolic saturation curve to point measurements of total binding. Using this single-class binding site model, the apparent dissociation constant (Kd) for the estradiol-receptor interaction in the HM line was 0.40 ± .06 nM and the number of specific estradiol binding sites (Bmax) was 10.4±.4 fmoles/mg cytosol protein. For the LM line, an apparent Kd of 0.41 ± .20 nM and a B of 9.50 ± 1.0 fmoles/mg cytosol protein were obtained. The similarity between corresponding binding parameters for the divergent mating lines indicated that the number and/or estradiol-binding affinity of a single class of cytosol receptors was also not responsible for the line difference in mating frequency. Though these results confirm that interaction of estradiol with cytosol receptors was not the limiting neuroendocrinological mechanism in the differential expression of masculine sexual behavior in quail, the possible involvement of dynamic interrelationships between cytoplasmic and nuclear estrogen binding was discussed.
- Application of microbial phytase and its influencing factors in vivo and in vitroQian, Hao (Virginia Tech, 1995)Five experiments were conducted in vivo to investigate the efficacy of phytase in improving the availability of phytate P as influenced by dietary Ca:total P (tP) ratios for pigs, broilers, and turkey poults. In pigs and poultry, microbial phytase was effective in improving performance, P and Ca digestibility, bone mineralization, and in decreasing fecal P excretion by enhancing hydrolysis of phytate P for young pigs, broilers and turkey poults fed a corn-soybean meal diet. Maximum responses were achieved at supplemental phytase levels of 750 to 1,050 units (U)/kg diet for young pigs and 600 to 900 U/kg diet for poultry. Based on nonlinear and linear response equations generated from the phytase and available P (aP) data in young pigs and non phytate P (nP) in broilers and turkeys, P-equivalency functions for phytase were developed. For pigs, the P-equivalency equation was Y = .2622(1 - .9706e⁻⋅⁰⁰¹⁸⁵ˣ); for broilers, the equation was Y = .2330(1 - .9818e⁻⋅⁰⁰⁰⁷⁴ˣ); and for turkey poults, the equation was Y = .1220(1 - 1.7721e⁻⋅⁰⁰⁵³ˣ). For these three equations, X = added phytase (U/kg diet) and Y = P-equivalency values (%). Based on these equations, 1 g of P as inorganic defluorinated phosphate could be replaced by 300 and 208 U of phytase/kg of diet for pigs fed diets containing .07 and .16% aP, by 937 U of phytase for broilers fed with .27% nP diet, or by 340 and 511 U of phytase/kg diet for turkey poults fed diets containing .27 and .36% nP, respectively. Phosphorus-equivalency values of phytase were also obtained by generating P-equivalency functions at each P level and each Ca:tP ratio. The phytase efficacy was influenced by dietary Ca:tP ratios, P, and vitamin D₃ levels. In pigs and poultry, a wide Ca:tP ratio decreased phytase efficacy because all measurements were decreased as the dietary Ca:tP ratio became wider. In young pigs, widening the ratios from 1.2 to 2.0:1 resulted in a decrease in phytase efficacy of 21.1 and 12.1% for .07 and .16% aP diets, respectively. In poultry, widening the ratio from 1.4 to 2.0 led to a decrease in phytase efficacy by 7.3% for broilers fed diets containing .27% nP, and by 6.3 and 4.2% for turkey poults fed diets containing .27 and .36% nP, respectively. A synergistic effect of vitamin D₃ addition and phytase supplementation was observed for broilers. Addition of vitamin D₃ indicated a potential for improving utilization of phytate P and Ca in the presence and absence of microbial phytase. Average daily gain, apparent P digestibility and bone ash content were the most sensitive measurements to assess microbial phytase efficacy for the replacement of inorganic P for pigs and poultry. These measurements were also sensitive for assessing the effects of varying Ca:tP ratios and levels of P. In summary, 1 g of P from defluorinated phosphate could be replaced by 250 to 400, 600 to 950 and 340 to 550 U of phytase/kg diet, respectively for young pigs, broilers and turkey poults when they were fed a com-soybean meal diet. Dietary Ca:tP ratio of 1.2:1 for young pigs and 1.1 to 1.4:1 for poultry resulted in maximum phytase efficacy. An in vitro study was performed for the evaluation of effects of cations on the characteristics of microbial phytase from A. niger. A discontinuous assay was applied to assay A. niger phytase. The enzyme was observed to have a high affinity for sodium phytate with a Km of 62 µM and a Vmax of 139 U of specific activity per mg of phytase protein. Malachite green was used as the color reagent in the discontinuous assay, which increased the sensitivity 50 fold over molybdovanadate as the color reagent. All cations tested in vitro (Mg²⁺, Mn²⁺, Ca²⁺, Cr³⁺, Fe³⁺, Cu²⁺ and Zn²⁺) inhibited phytase activity, and imposed a competitive or mixed inhibition; a binding of cations with phytate also was involved in the inhibition by decreasing the effective substrate concentration. The inhibition by Ca²⁺ and Mg²⁺ caused only a partial inhibition because the enzymatic reaction rate was never reduced to zero and replots of slopes for Ca²⁺ and Mg²⁺ were hyperbolic. Cations of Zn²⁺, Cu²⁺, Fe³⁺, Cr³⁺ and Mn²⁺ gave a pure inhibition. A decreasing order of the inhibitory effect from cations was observed on the phytase activity: Zn²⁺ > Cu²⁺ > Fe³⁺ > Cr³⁺ > Ca²⁺ > Mn²⁺ > Mg²⁺ based on the Ki value that increased from a low value for Zn²⁺ to a high value for Mg²⁺. In summary, cations possess a potential for decreasing A. niger phytase activity by a competitive or mixed-type inhibition system; binding of cations with the phytate substrate also inhibited the activity of A. niger phytase.
- Aspects of lactation endocrinology: I. lactogenic receptors in bovine mammary tissue at different stages of lactation: II. growth hormone concentrations in Holstein cattle of differing genetic meritKazmer, Gary Wayne (Virginia Polytechnic Institute and State University, 1985)Mammary tissue from nine Holstein cows was collected within one week of parturition, at 60 and 180 days postpartum. Blood samples were collected at 6-hr intervals from two days prior to . until two days after surgery. A membrane-enriched fraction of tissue homogenates was prepared by differential centrifugation. Displacement curve data was analyzed by a microcomputer program. Mean prolactin (Prl) during the periparturient period was greater than either postpartum period, but not prior to biopsy. Dissociation constants (Kd) estimated with NIH-bPRL-6 as competitor were not different among stages of lactation, and averaged 8.97 x 10⁻⁸M. Receptor concentrations were less during _the periparturient period than later lactation. The Kd was 100-fold greater when estimated with human growth hormone as competitor. It is concluded that lactogenic hormone receptor concentrations in bovine mammary tissue increase with the onset of lactation, following a pattern similar to that observed in non-ruminants. Three experiments were conducted to investigate endocrine metabolic hormone profiles in Holstein cattle of differing genetic merit at several ages. Control animals were randomly bred to non-AI sires originating in the Virginia Tech Dairy herd. Selected animals were offspring of commercially available AI sires. In one experiment, mean plasma Prl was greater in control animals after feeding and insulin injection, while growth hormone (GH) was greater in selected animals at all ages. Free fatty acids were greater in selected animals at 6 and 24 months of age, while glucose (Glc) and urea were unaffected by genetic merit. In a second experiment, Holstein bull calves were administered Glc and thyrotropin releasing hormone (TRH) on different days. Plasma GH was greater in selected animals. Plasma Prl was greater in control animals after TRH. In the third experiment, Holstein cows received TRH at 30, 90 and 200 days postpartum (DPP). Net energy balance was negative at 30, while positive at 90 and 200 DPP. Plasma GH before and after TRH was greater in selected animals, and greater during early than later lactation. Thus, the results of the three experiments indicate that increased plasma GH may be associated with selection for increased milk yield.
- Availability of zinc from an amino acid chelate in Zn depleted pigsSwinkels, Johannes W. G. M. (Virginia Tech, 1992)This study was conducted to compare the availability of Zn from two Zn sources, an amino acid chelate and ZnSO₄. In three experiments, 78 Zn depleted and 24 Zn adequate pigs were used. Pigs were depleted of Zn by feeding an isolated soy protein, semi purified diet containing 17 ppm Zn. Of the 78 depleted pigs, 60 pigs were Zn repleted. During Zn repletion in Exp. 1, depleted pigs were fed the low Zn diet supplemented with 5, 15, or 45 ppm Zn either as ZnSO₄ or as Zn amino acid chelate (ZnAAC). In Exp. 2 and 3, low Zn diets were only supplemented with 45 ppm Zn. Zinc adequate pigs, used in Exp. 1 and 2, were fed the 45 ppm supplemental Zn diets. To evaluate differences in site and rate of apparent Zn absorption, chromic oxide was added to the diets of depleted pigs in Exp. 1 and 3. In all experiments, a 24-d period was sufficient to severely deplete the porcine body Zn stores, and to cause parakeratosis and growth retardation. Serum Zn concentrations and serum ALP-activities of depleted pigs dramatically decreased (P < .01) during the first 14 d of Zn depletion. At the end of Zn depletion, Zn contents in liver, kidney, pancreas, brain, and small intestine tissues of pigs fed the low Zn diet were reduced (P < .01) by 10 to 40 % compared with the adequate pigs fed the ZnSO₄ and ZnAAC diets. In Exp. 2, the growth retardation was associated with a low (P < .05) serum mitogenic activity and pituitary RNA content of depleted pigs compared with pair-fed adequate pigs. Moreover, the growth hormone mRNA fraction tended to be reduced (P < .10) for the Zn depleted pigs. In Exp. 1, the apparent absorption of Zn was higher (P < .01) for pigs fed ZnAAC compared with the ZnSO₄ group; however, this was not confirmed in Exp. 3 unless coefficients were corrected for Cr recovery. Furthermore, absorption of Zn occurred primarily within jejunal and distal segments of the small intestine. In the balance of Exp. 3, disappearance rates of Zn, Cu, Fe and DM were higher (P < .01) in depleted pigs fed ZnAAC compared with ZnSO₄. The recovery of Cr also was different (P < .01) between pigs fed the ZnSO₄ (87 %) and ZnAAC (70 %) diets. Moreover, the moisture content of the fecal matter was 11 % higher (P < .01) for the ZnAAC group compared with pigs fed ZnSO₄. In Exp. 1, depleted pigs fed the 15 ppm ZnSO₄ and ZnAAC diets regained their ability to grow, however, replenishment of body fluid and tissue Zn pools did not occur within the 24-d Zn repletion period. Both the 5 ppm ZnSO₄ and ZnAAC groups did not respond to Zn repletion within a 12-d period. In all experiments, the rate and degree of repletion of body fluid and tissue Zn stores was not different between pigs fed the 45 ppm ZnSOq and ZnAAC diets, although a higher (P < .05) serum mitogenic activity was observed for the adequate pigs fed ZnAAC compared with ZnSO4. In conclusion, an amino acid chelate did not improve growth, or rate and degree of replenishment of body fluid and tissue levels of Zn compared with pigs fed ZnSO₄. However, ZnAAC may have influenced intestinal luminal conditions since a higher rate of disappearance of Zn, Cu, Fe, Cr, and DM was measured.
- Biological, Nutritional, and Processing Factors Affecting Breast Meat Quality of BroilersSantiago, Hector Luis (Virginia Tech, 2002-02-07)A series of experiments were conducted to investigate the effects of certain biological, nutritional, and processing factors on breast meat quality of broilers. In the first experiment, the influence of genetic strain, plane of nutrition, and age at slaughter on breast meat quality was evaluated. Breast meat from a high yield line of broilers had lower pH at 24 h postmortem (PM), greater L*, a*, and b* values, lower water holding capacity (WHC), and higher expressible moisture (EM) than those of a line selected for rapid growth. Breast meat from birds processed at 42 d had lower WHC than those processed at 53 d, even though no differences in pH and L* values at 24 h were observed. No significant effects due to plane of nutrition on meat quality traits were observed. In Experiment two, the influence of strain and chilling methods (ice or air chilled) on breast meat quality was studied in broilers. Breast meat quality significantly differed among strains, with one of the strains evaluated having higher muscle pH, lower L* values, and higher WHC than the other strains. Ice-water chilling significantly reduced the rate and extent of PM pH decline, but had significantly lower WHC and higher EM than those from carcasses chilled by air. However, chilling conditions did not influence breast meat color. In Experiment three, the effects of strain and gender on breast muscle quality of broilers was studied. Meat quality traits were evaluated on both sexes of six genetic crosses of commercial strains. No significant differences in breast meat quality traits among strains were observed. However, differences between sexes were highly significant. The P. major muscles of females had lower pH values at all PM times, higher L*, a*, and b* values, and lower WHC than males. The P. minor of females had significantly lower pH, lower WHC, higher EM, but similar color L*, a*, and b* values than males. In Experiment four, the effects of strain, gender, and age at slaughter on breast meat quality were studied. Strain differences were observed in both sexes, but these differences did not show any specific relationship with the strain genotype. Breast muscles from a male pure line had superior meat quality, with higher muscle pH, and WHC, but higher L* values than the other strains. Significant differences in breast meat quality traits due to age at slaughter were also observed. Regardless of gender, breast muscle pH at 24 h PM and WHC decreased linearly with age, while breast muscle temperature and L* values increased in a linear fashion with advancing age at slaughter. No significant strain by age interactions were observed for any of the meat quality traits evaluated. The results of these studies indicate that commercial genotypes differ significantly in PM muscle metabolism and subsequent meat quality. The results also indicate that female broilers and older birds might be more susceptible to meat quality problems.
- Central administration of dipeptides, beta-alanyl-BCAAs, induces hyperactivity in chicksTsuneyoshi, Yousuke; Tomonaga, Shozo; Asechi, Mari; Morishita, Koji; Denbow, D. Michael; Furuse, Mitsuhiro (2007-05-31)Background Carnosine (β-alanyl-L-histidine) is a putative neurotransmitter and has a possible role in neuron-glia cell interactions. Previously, we reported that carnosine induced hyperactivity in chicks when intracerebroventricularly (i.c.v.) administered. In the present study, we focused on other β-alanyl dipeptides to determine if they have novel functions. Results In Experiment 1, i.c.v. injection of β-alanyl-L-leucine, but not β-alanyl-glycine, induced hyperactivity behavior as observed with carnosine. Both carnosine and β-alanyl-L-leucine stimulated corticosterone release. Thus, dipeptides of β-alanyl-branched chain amino acids were compared in Experiment 2. The i.c.v. injection of β-alanyl-L-isoleucine caused a similar response as β-alanyl-L-leucine, but β-alanyl-L-valine was somewhat less effective than the other two dipeptides. β-Alanyl-L-leucine strongly stimulated, and the other two dipeptides tended to stimulate, corticosterone release. Conclusion These results suggest that central β-alanyl-branched chain amino acid stimulates activity in chicks through the hypothalamus-pituitary-adrenal axis. We named β-alanyl-L-leucine, β-alanyl-L-isoleucine and β-alanyl-L-valine as Excitin-1, Excitin-2 and Excitin-3, respectively.
- Central cannabinoid regulation of food intake in chickensZhang, Jin (Virginia Tech, 2005-06-22)Marijuana has been used for medicinal and recreational purposes for thousands of years. Many people think of marijuana in the context of an illegal drug. Because of the antimarijuana attitude, research with cannabinoids was neglected for a long time. Although this substance is related to social problems, scientists are interested in its action and possible medicinal properties. Since the identification of the structure of Î 9--tetrahydrocannabinol, the main psychoactive ingredient of marijuana, there has been increased interest in this compound. Following the discovery of two cannabinoid receptors, CB1 and CB2 receptors, it was determined that CB1 receptors are in high density in the central nervous system while CB2 receptors are found primarily in the immune system. The endogenous cannabinoid ligands, anandamide and 2-arachidonoylglycerol, were observed in the central nervous system and peripheral tissues. Endocannabinoids differ from other "classical" neurotransmitters because they do not appear to be stored in synaptic vesicles, and they act as retrograde messengers within the brain. The endogenous cannabinoid signaling system includes cannabinoid receptors, their endogenous ligands called endocannabinoids, and the proteins for their synthesis and inactivation. The cannabinoid system appears to act as a neuromodulatory system. During the past ten years, the endogenous cannabinoid system has been implicated in a variety of physiological functions including pain reduction, motor regulation, learning, memory, and reward. Because obesity and eating disorders are prevalent, scientists are working at the molecular level to study the mechanisms controlling body weight and regulation of food intake. Several of the neuropeptides present in hypothalamic nuclei contribute to energy balance and food intake regulation. Endogenous cannabinoid and cannobinoid receptors are found in the hypothalamus and are associated with the regulation of food intake. Although the mechanisms whereby cannabinoids influence food intake remain unclear, results suggest that the cannabinoid system will be an important target in future studies in obesity. Most research on cannabinoids has focused on their role in food intake regulation in mammalian species. It is important to determine the role of endocannabinoids in other species. The effect of intracerebroventricular injection of agonists and antagonists of both CB1 and CB2 receptors in 8 to 11 week-old male Single Comb White Leghorn and 3 to 6 weeks old male broilers was investigated. It was found that agonists of both the CB1 and CB2 receptor increased food intake significantly; however, the CB2 receptor agonist had a stronger and longer lasting effect. Antagonists of both receptors decreased food intake significantly. The CB1 receptor antagonist appeared to block both cannabinoid receptors in birds, whereas the CB2 receptor antagonist did not block both receptors. Previous studies have indicated that the CB2 receptor is found only outside the brain and spinal cord, and is involved with the immune system. From the present results, it appears that both cannabinoid receptors are present in the chicken brain. Furthermore, the CB2 receptor may also be localize in the chicken brain. There are also differences in cannabinoid system between Leghorn and broilers.
- Central control of food intake in the domestic fowlSkewes, Peter Alan (Virginia Polytechnic Institute and State University, 1985)This study was initiated to determine if factors exist in the blood or cerebrospinal fluid (CSF) of the domestic fowl that act upon the central nervous system to control food intake. Plasma collected from free-feeding and 24-hour fasted leghorn cockerels was lyophilized, reconstituted to 2, 4, or 5 times the original concentration, and injected, via a stereotaxically implanted 23 gauge stainless steel guide cannula, into the lateral ventricle of free-feeding leghorn cockerels. Food intake was significantly reduced following injection of 2, 4, and 5 times normal concentration of plasma from free-feeding birds. Plasma from fasted birds did not alter food intake regardless of concentration, but did significantly reduce water intake when concentrated to five times normal. A similar study was conducted with fractions of plasma of different molecular weight ranges. Plasma collected from free-feeding cockerels was partitioned by gel filtration into the following molecular weight fractions: >5000 molecular weight, <5000 molecular weight, 1500-5000 molecular weight, and <1500 molecular weight. The fractions were lyophilized and reconstituted to four times the original concentration and injected into the lateral ventricle of free-feeding leghorn cockerels. Food intake was significantly decreased by the <5000 and <1500 molecular weight fractions, whereas water intake was not affected. The 1500-5000 molecular weight fraction and the fraction above 5000 did not affect food or water intake. To determine if this food intake inhibiting factor existed in the cerebrospinal fluid (CSF) of the domestic fowl, CSF was collected from free-feeding and 24-hour fasted broilers and injected into the lateral ventricle of leghorn and broiler cockerels. Food intake was not affected by either the normal or four-times normal concentration of CSF collected from free-feeding or 24-hour fasted broilers. Water intake was significantly increased in the leghorn and broiler birds receiving the four times normal concentration of CSF collected from 24-hour fasted birds, but was not affected in the birds receiving CSF collected from the free-feeding donors. It appears, therefore, that a food intake inhibiting factor exists in the plasma of the free-feeding domestic fowl that does not exist in the CSF.
- Central mechanisms of prolactin-releasing peptides orexigenic effect in chickensWang, Guoqing (Virginia Tech, 2015-06-29)Prolactin-releasing peptide (PrRP) is an endogenous hypothalamic neuropeptide that when exogenously injected increases food intake in chickens, but decreases it in rodents and goldfish. We designed three sets of experiments to elucidate the mechanisms of PrRP's orexigenic effect in chicks. In experiment one, food and water intake were evaluated in chicks after receiving intracerebroventricular (ICV) injection of the vehicle, 0.75, 3, 12, 47 or 188 pmol PrRP. The administration of 12 and 47 pmol PrRP increased food intake for up to 120 min after injection, and 188 pmol increased it for up to 180 min. The lowest effective dose was 3 pmol, which increased food intake for up to 60 min after injection. Water intake was not affected. To investigate the molecular mechanisms, c-Fos immunohistochemistry was performed and mRNA expression of some appetite-associated neurotransmitters was measured in chicks that received either vehicle or 188 pmol of PrRP. The rostral paraventricular nucleus (PVN) was activated which coincided with increased neuropeptide Y (NPY) mRNA expression in the whole hypothalamus. In experiment two, food and water intake were evaluated in chicks fed a high carbohydrate (HC), high fat (HF) or high protein (HP) diet after ICV injection of vehicle, 3 or 188 pmol PrRP. Chicks fed the HP diet increased food intake at a lower dose than chicks fed HF and HP diets after ICV PrRP injection. In addition, ICV injection of vehicle, 3 and 188 pmol PrRP were performed in chicks fed all three diets, and ICV PrRP injection induced preferential intake of the HP diet over HC and HF diets. The expression of some appetite-associated neuropeptides in the hypothalamus was also measured in chicks fed the HC, HF or HP diet after ICV injection of vehicle or 188 pmol PrRP. There was a diet effect on mRNA abundance of all appetite-associated genes measured (P < 0.05), with greater expression in chicks fed the HF or HP than HC diet. While neuropeptide Y (NPY) mRNA abundance was similar between vehicle and PrRP-injected chicks that consumed HP or HF diets, expression was greater (P < 0.05) in PrRP- than vehicle-injected chicks that consumed the HC. In experiment three, the orexigenic effect of PrRP was tested in chicks selected for low (LWS) and high (HWS) body weight after central administration of vehicle, 24, 94 and 375 pmol PrRP. The LWS chicks had a lower threshold and higher magnitude of food intake increase in response to PrRP injection. Results demonstrate that PrRP is a potent orexigenic factor in chickens and that effects are likely mediated through the hypothalamus. The orexigenic effect of PrRP was influenced by dietary macronutrient composition, and diet in turn influenced the food intake response to PrRP. These results may contribute to a novel understanding of appetite regulation.
- Cerebral Laterality, Emotion, and Cardiopulmonary Functions: An Investigation of Left and Right CVA PatientsComer, Clinton S. (Virginia Tech, 2014-05-05)Stroke, or cerebrovascular accident (CVA), is a prominent cause of long term disability in the United States. It has been evidenced that the outcome of a CVA patient differs as a function of the cerebral hemisphere that is damaged by the stroke, especially in terms of emotional changes. The Right Hemisphere Model of Emotion posits that the right hemisphere is specialized for processing emotional content, regardless of valence. In contrast, the Bi-Hemispheric Model of Emotion posits that each hemisphere has its own emotional specialization. The current experiment tested the competing predictions of the two theoretical perspectives in a mixed sample of left cerebrovascular accident (LCVA) patients and right cerebrovascular accident (RCVA) patients using a Dichotic Listening task and the Affective Auditory Verbal Learning Test (AAVLT). Heart Rate (HR) and Pulse Oxygen Saturation (SpO2) were also recorded as sympathetic measures. It was expected that the predictions of the Bi-Hemispheric Model would be supported. A series of mixed design ANOVAs were used to analyze the data. Results revealed that both groups may have exhibited decreased auditory detection abilities in the ear contralateral to CVA location. Additionally, CVA patients recalled significantly more positive words, than negative or neutral words, and exhibited a significant learning curve. LCVA patients exhibited a recency effect, while RCVA patients exhibited a heightened primacy effect. Findings from HR and SPO2 measures suggested a parasympathetic response to neutral information as well as an impaired sympathetic response to negative information in RCVA patients. Taken together these results lend partial support to the hypotheses drawn from the Bi-Hemispheric Model of Emotion, as evidenced by the diametrically opposite effects in these groups, which appears to reflect opposing cerebral processes.
- Characterization of Pituitary Protein Expression Patterns During Stages in the Reproductive Cycle of Turkey HensSpellerberg, Amy Marie (Virginia Tech, 2003-04-11)Improvements in turkey reproductive efficiency is a very desirable goal for the turkey industry. The ability to maintain turkey hens in the egg-laying (LAY) stage and produce one additional egg per hen a year is estimated to save the turkey industry approximately $1.5 million dollars per year. Overall protein expression generated by tissues of the hypothalamic-hypophyseal complex, namely the anterior pituitary, of the mature turkey hen have a profound impact on reproductive cycling (Scanes, 2000). One of the key physiological factors produced by the anterior pituitary and shown to play a significant role in the regulation of egg laying is the protein prolactin (Prl). The objectives for this study are to examine the overall protein expression patterns from turkey hen pituitary tissue during the nonphotostimulated (NPS), photostimulated (PS), and egg laying (LAY) stages. Attempts to isolate transcription factors that regulate the expression of Prl using an affinity chromatography technique or southwestern screening of a bacteriophage expression library were not successful. A global analysis of protein expression, using two-dimensional polyacrylamide gels (2D gels), was conducted using whole cell, cytoplasmic and nuclear protein extracts from pituitary tissue collected during the NPS, PS and LAY reproductive stages. Approximately 1,046 proteins ranging in pI from 4.6-8.2 and molecular weights between 100 kDa-6kDa were resolved. Protein expression patterns were replicated and verified using pituitaries harvested from NPS, PS and LAY stage turkey hens from another laboratory. Proteins showing considerable changes (563 proteins increased in expression and 98 proteins decreased in expression from the NPS to the LAY stage) in their expression between the reproductive stages were grouped in analysis sets for future identification. These proteins may prove to be important to the reproductive cycling of the turkey hen and warrant future investigation. The results of this study contribute to the overall understanding of the role that the pituitary, as a critical part of the hypothalamic-hypophyseal complex, plays in turkey hen reproductive cycling.
- Cloning, Expression, and Developmental and Dietary Regulations of a Chicken Intestinal Peptide Transporter and Characterization and Regulation of an Ovine Gastrointestinal Peptide Transporter Expressed in a Mammalian Cell LineChen, Hong (Virginia Tech, 2001-09-28)To study peptide absorption in chickens, an intestinal peptide transporter cDNA (cPepT1) was isolated from a chicken cDNA library. The cDNA was 2,914-bp and encoded a protein of 714 amino acid residues. Twenty-three di-, tri-, and tetra-peptides were used for functional analysis of cPepT1 in Xenopus oocytes and Chinese hamster ovary (CHO) cells. For most di- and tripeptides tested, the Kt was in the micromolar range, except Lys-Lys and Lys-Trp-Lys. Northern analysis demonstrated that cPepT1 is expressed strongly in the small intestine, and at lower levels in kidney and cecum. These results demonstrated the presence and functions of a peptide transporter in chickens. cPepT1 mRNA abundance was evaluated in response to developmental and dietary regulations. In Experiment 1, eggs at incubation day 18 (E18) and Cobb chicks after hatch (d 0) were sampled before treatments. Three groups of chicks were fed diets containing 12, 18, or 24% crude protein (CP). Feed intake of chicks fed the 18 or 24% CP diets was restricted to that of chicks fed the 12% CP diet. In Experiment 2, a fourth group with free access to the 24% CP diet was added. cPepT1 mRNA abundance was quantified from northern blots. By d 0, there was a 50-fold increase in cPepT1 mRNA abundance compared with E 18. In chicks fed the 12% CP diet, cPepT1 mRNA abundance decreased throughout the 35 d. Chicks fed 18 or 24% CP diets showed an increase in cPepT1 mRNA abundance with time. In chicks with free access to the 24% CP diet, cPepT1 mRNA decreased until d 14 but returned to an intermediate level at d 35. Our results indicate that cPepT1 mRNA is regulated by both dietary protein and developmental stage. To investigate the kinetics of an ovine peptide transporter (oPepT1), CHO cells were transfected with oPepT1 cDNA. Uptake of Gly-Sar by transfected cells was pH-dependent, concentration-dependent, and saturable. Competition studies showed that all di-, tri-, and tetra-peptides inhibited uptake of Gly-Sar. Pretreatment of the cells with staurosporine resulted in an increase in peptide transport. This increase was blocked by pretreatment with PMA. The results indicate that protein kinase plays a role in oPepT1 function.
- Constraints on the Evolution of Viviparity in the Lizard Genus SceloporusMathies, Tom (Virginia Tech, 1998-09-18)I evaluated possible constraints on the evolution of viviparity in the lizard genus Sceloporus by experimentally extending the length of egg retention past the normal time of oviposition for a number of oviparous species. Observations also included a representative of the genus Urosaurus, the sister genus to Sceloporus. I determined the effects of retention on embryonic development, hatchlings, and gravid females. Results indicated that the proximate constraints on longer retention times and viviparity are 1) embryonic development becomes arrested or severely retarded in utero, and 2) the ability to maintain gravidity past the normal time of oviposition is limited in some species. Observations on Urosaurus further showed that extended egg retention results in hatchlings with traits that are associated with lower fitness. I also tested the hypothesis that reproductive Sceloporus lower their body temperatures during activity because their normal body temperatures are detrimental to embryos. Observations on a viviparous species of Sceloporus indicated that the normal body temperature of the female was detrimental to embryonic development. This result is indicates that viviparity would be constrained in some squamate lineages if maternal body temperatures are too high for successful embryonic development. I also evaluated the hypothesis that the evolutionary transition from oviparity to viviparity involves a "reduction" of the eggshell concurrent with longer durations of egg retention. If this hypothesis is correct, then attributes of eggshells that should enhance exchange of respiratory gases (i.e., thickness, density, permeability to water vapor) would be correlated with the maximal developmental stage that embryos are able to attain in the oviducts (i.e., stage of developmental arrest). The results of this study indicated that these features of shells do not determine the stage at which development becomes arrested. Thus, the results do not support the hypothesis that shell reduction occurs concurrently with longer periods of egg retention. The results are consistent with the alternative hypothesis that reduction of the eggshell occurs after viviparity has evolved.
- Corticotrophin Releasing Hormone Modulation of Feed Intake, Gastric Motility, and Behavior in Low and High Body Weight Selected Lines of ChickensCline, Mark A. (Virginia Tech, 2005-05-04)The effect of intracerebroventricular (ICV) injection of corticotrophin releasing hormone (CRH) and related compounds on appetite, behavior, and gastric motility in lines of chickens, one selected for low body weight (LWS) and the other high body weight (HWS), was determined. Nucleotide sequence and expression patterns of the CRHr2 receptor, involved in appetite regulation, were also determined. Some individuals of the LWS line are anorexics and many die simply from not eating while some individuals in the HWS line are compulsive eaters and exhibit obesity. CRH is a 41 residue peptide that initiates an organism's stress response and is a potent inhibitor of appetite. An ICV injection of CRH dose-dependently decreased feed intake in both lines but did not effect water intake. When CRH receptor antagonists were ICV injected an increase in feed intake in the LWS line but not in the HWS line was observed, however the appetite reducing effect of CRH was attenuated in the HWS line but not in the LWS line. The LWS line has higher concentration of corticosterone than does the HWS line. In both lines at all times treatment with CRH caused an increase in locomotion and no CRH-treated chicks from either line slept post injection. Chicks from the LWS line that were treated with CRH exhibited other anxiety related behaviors sooner than the HWS line. The LWS line showed a liner increase in crop emptying time as the dose of ICV CRH increased. The HWS line responded with a quadratic dose response to CRH treatment. Polymorphisms in the CRHr2 receptor were found in both lines in the same positions, thus we concluded these differences do not significantly contribute to body weight differences. However, differences detected in expression patterns between lines for the CRHr2 receptor may contribute to their different body weights. We conclude that differences in the CRH system, its concentrations and differential receptor action, of these two lines may be partly responsible for their altered body weight phenotype.
- Developmental and Growth Hormone Regulation of the Expression of Liver-Enriched Transcription Factors in Bovine LiverEleswarapu, Satyanarayana Venkata (Virginia Tech, 2004-05-11)Liver gene expression changes during development and is affected by growth hormone (GH). These changes in gene expression may be due to the differential expression of the liver-enriched transcription factors (LETFs). To study the potential involvement of LETFs in the regulation of gene expression in the bovine liver, we cloned the cDNA fragments of nine bovine LETFs, including hepatocyte nuclear factor (HNF)-1Æ Ã , 1Æ Ã , 3Æ Ã , 3Æ Ã , 3Æ Ã , 6, albumin D-element binding protein (DBP), and CCAAT/enhancer-binding proteins (C/EBP) -Æ Ã and Æ Ã , and compared the expression levels of them between adult and fetal bovine liver and between GH-treated and untreated adult bovine liver. The mRNA abundance of the LETFs was determined by ribonuclease protection assay (RPA). The cloned bovine LETF cDNA sequences showed high degrees of similarity (79 % to 99 %) to the LETF sequences of other species. The mRNA levels of HNF-1Æ Ã , HNF-3Æ Ã , and HNF-6 were significantly higher (P < 0.05) in the fetal liver (n=3) than in the adult liver (n=7). There were significant increases (P < 0.05) in the mRNA expression of HNF-3Æ Ã and HNF-6 in the liver of cows 24 h (n=6) and 1w (n=6) after GH administration. The results of this study suggest that HNF-1Æ Ã , HNF-3Æ Ã , and HNF-6 may play a role in differential regulation of gene expression between the fetal and adult bovine liver and that HNF-3Æ Ã and HNF-6 may be also involved in GH regulation of gene expression in the bovine liver.
- Developmental Gene Expression in the Small Intestine of Chickens from Lines Divergently Selected for High or Low Juvenile Body WeightMiller, Carin R. (Virginia Tech, 2007-09-10)Nutrient transporters in the small intestine are responsible for dietary nutrient assimilation and therefore the expression of these transporters can influence the overall nutrient status as well as the growth and development of the animal. This thesis examined correlated responses to selection in the developmental gene expression of the peptide transporter PepT1, the glutamate/aspartate transporter EAAT3, the sodium-dependent glucose transporter SGLT1, and the fructose transporter GLUT5 in the small intestine of chickens from lines divergently selected for high (HH) or low (LL) eight-week body weight and their reciprocal crosses, (HL and LH). Chicks were weighed and killed on embryonic day 20 (E20), day of hatch (DOH with no access to feed), and days 3 (D3), 7(D7), and 14 (D14) post hatch. Duodenum, jejunum, ileum and liver were collected. DNA extracted from liver was used to sex birds by PCR. RNA was extracted from the intestinal segments of four males and four females from each mating combination (MC) and time point except E20 HL males (n = 3) and D7 LL females (n = 2). Expression of nutrient transporters was assayed by real-time PCR using the relative quantification method. In comparing HH and LL males and females there was a line by segment interaction in PepT1 gene expression, with no segment difference in HH and greatest expression in the ileum of the LL (P < 0.05). There was also a MC by age by sex interaction for PepT1 gene expression (P < 0.0001) with peak gene expression occurring on DOH for LL females, on D7 for HH females, on D7 for LL males and D14 for HH males. Overall, females had greater EAAT3 expression (P < 0.03). Gene expression of EAAT3 was greatest in the ileum, intermediate in the jejunum, and least in the duodenum (P < 0.0007). There was an age by segment interaction for EAAT3 expression (P = 0.0002) and a MC by segment interaction (P < 0.02), with LL having greater expression than HH in the ileum. Females had greater SGLT1 expression than males (P < 0.0001). There was a sex by age interaction for the expression of SGLT1 (P < 0.0001). Females induced SGLT1 expression on DOH and maintained this level through D14, while males gradually increased expression through D7 and decreased expression by D14. These results indicate that expression of PepT1, EAAT3, SGLT1 are differentially expressed in male and female chickens regardless of selection for high or low juvenile body weight. These results also show a sexual dimorphism in the capacity to absorb peptides, anionic amino acids, and glucose from the intestine, which has implications for the poultry industry with regard to diet formulations for straight-run and sex-separate grow-out operations. In comparing male HH, HL, LH, and LL chicks, overall LL had the greatest level of expression (P <0.06), HH had the least level of expression (P < 0.006) and HL and LH had intermediate levels of expression (P < 0.06). Greatest PepT1 gene was expression in the ileum (P < 0.0003) and there was a MC by segment interaction with expression increasing from duodenum to ileum in LL, but there was no segment difference in any other MC (P < 0.08). Within each intestinal segment there was a MC difference (P < 0.02). There was an effect of sire for PepT1 expression, with progeny from low weight selected sires (LWS) having greater expression than progeny from high weight selected (HWS) sires (P = 0.0008). There was no difference between intestinal segments in progeny from HWS sires, however, greatest PepT1 gene expression was seen in the ileum of progeny from LWS sires (P < 0.0001). Overall, expression of EAAT3 was greatest in the ileum, intermediate in the jejunum and least in the ileum (P < 0.0001) and there was a segment by age interaction for EAAT3 expression (P < 0.0001). In all MCs except HH, EAAT3 gene expression increased from duodenum to ileum (P < 0.08). Within the ileum, the LL had greatest EAAT3 gene expression, LH and HL had intermediate gene expression, and HH had least expression (P < 0.08). Expression of SGLT1 gradually increased through D7 and decreased by D14 (P < 0.0001) and overall, was greatest in the distal small intestine (P < 0.0001). There was a MC by segment interaction, with SGLT1 gene expression being greatest in the distal small intestine in LL, LH, and HL, but greatest in the jejunum of HH (P < 0.04). Within the ileum, LL had greater SGLT1 gene expression than HH (P < 0.06). Overall, greatest GLUT5 expression was in the distal small intestine (P < 0.0001) and there was a MC by segment interaction, with expression being greatest in the distal small intestine in LL and HL (P < 0.02), greatest in the ileum of LH (P < 0.08), and greatest in the jejunum of HH (P < 0.09). Within the ileum there was a MC difference (P < 0.07). These results indicate that selection for high or low juvenile body weight may have influenced the gene expression pattern of these nutrient transporters in the small intestine, which may contribute to the overall differences in the growth and development of these lines of chickens.
- Diet-induced thermogenesis in the domestic chickenBlackman, James R. (Virginia Polytechnic Institute and State University, 1985)Four experiments were conducted to examine the energetic responses of chicks from lines divergently selected for 56-day body weight to caloric intake. Caloric overconsumption or calorie-protein imbalances were induced by providing glucose solutions (16% w/v) in lieu of water in the first experiment, and by force-feeding the diet to crop capacity in the subsequent trials. Feed restriction was accomplished by providing 75% of ad libitum intake each day. Both low-weight (LN) and high-weight (HN) chicks reduced their voluntary feed intake when glucose solutions were provided; caloric intake was reduced by 25% and 10% in LN and HN chicks, respectively. Weight gain and the deposition of protein and ash in the carcass were reduced by the glucose treatment, but carcass lipid deposition was increased. The efficiency of energy deposition was improved in HN but unchanged in LN chicks. Feed restriction reduced feed intake, energetic efficiency, and gain of body weight, carcass dry matter, protein, lipid and ash. The suppression of lipid deposition was more severe in the LN chicks than in the HNs. Overfeeding of a complete diet increased body weight, the deposition of all carcass constituents, and energetic efficiency. Feed restriction exerted an opposite effect, with the LN chicks exhibiting more pronounced responses than the HNs. Determined metabolizable energy values of the diet differed among feeding treatments and between lines, and line by feeding treatment interactions were observed. Body core and surface temperatures demonstrated that the LN chicks were less able than the HNs to maintain homeothermy under treatments imposed. Heat production tended to be increased by overfeeding and reduced by feed restriction. Metabolic rates were higher in LN than in HN chicks when the chicks were fed ad libitum or subjected to restricted feeding, but not when force-fed. The latter observations were attributed to differences in physical activity. Plasma triiodothyronine (T₃) of LN chicks was increased by force-feeding and reduced by restricted feeding, but no significant differences in plasma T₃ occurred in HNs. Thyroxine (T₄) and the T₃:T₄ ratios were unchanged by feed intake, although LNs exhibited higher plasma T₃ and T₃:T₄ ratios than HNs. Although endocrinological changes to caloric consumption analogous to those observed in mammals occurred, energy balance did not confirm the existence of diet-induced thermogenesis in the fowl.