GHRT-89HK Sigma-AldrichHuman Ghrelin (TOTAL) RIA

Background/Objectives:Postprandial lipaemia is an established risk factor for atherosclerosis. To investigate the acute effect of four milk-derived dietary proteins (alpha-lactalbumin, whey isolate, caseinoglycomacropeptide and whey hydrolysate) on postprandial lipaemia, we have conducted a randomized, acute, single-blinded clinical intervention study with crossover design.Subjects/Methods:A total of 11 obese non-diabetic subjects (age: 44-74, BMI: 30-41.4 kg m(-2)) were included. On 4 different days the subjects ingested a high-fat meal with the following energy distribution: 66% energy from fat (100 g of butter), 15% of energy from carbohydrate (90 g of white wheat bread) and 19% of energy from protein (45 g of pure protein). Our primary variable was plasma triglyceride measured in the 8-h postprandial period. Secondarily, retinyl palmitate, non-esterified free fatty acids, glucose, insulin, glucagon, GLP-1 and GIP, active and total grehlin and cholecystokinin were measured.Results:We observed no statistically significant (P=0.8) differences between meals on our primary variable that is, triglycerides. Whey hydrolysate was associated with a significantly (P=0.02) smaller postprandial suppression of non-esterified free fatty acids compared with the other dietary proteins.Conclusion:We did not observe significant differences in postprandial lipaemia to the four milk-derived dietary proteins. Whey hydrolysate caused less postprandial suppression of free fatty acids.European Journal of Clinical Nutrition advance online publication, 27 July 2011; doi:10.1038/ejcn.2011.142.

Reversed feeding (RF) is known to disrupt hormone rhythmicity and metabolism. Although these effects may be mediated in part by phase inversion of glucocorticoid secretion, the precise mechanism is incompletely characterized. In this study, we demonstrate that acute nocturnal food deprivation in male rats suppressed the amplitude of spontaneous GH secretion during the dark phase by 62% (P < 0.001), without affecting baseline secretion. Prolonged RF, which reduced pituitary weight (by 22%; P < 0.05), also suppressed GH pulse height sufficiently to reduce skeletal growth (by 4-5%; P < 0.01) and terminal liver weight (by 11%; P < 0.001). Despite this suppression of the GH axis, proportionate adiposity was not elevated, probably due to the accompanying 16% reduction in cumulative food intake (P < 0.01). We demonstrate that RF also resulted in phase inversion of core clock gene expression in liver, abdominal white adipose tissue (WAT) and skeletal muscle, without affecting their expression patterns in the suprachiasmatic nucleus. In addition, RF resulted in phase inversion of hepatic peroxisome proliferator-activated receptor γ2 mRNA expression, a 3- to 5-fold elevation in fatty acid synthase mRNA in WAT in both light- and dark-phase samples (P < 0.01) and an elevation in muscle uncoupling protein 3 mRNA expression at the beginning of the light phase (P < 0.01). Consumption of a high-fat diet increased inguinal (by 36%; P < 0.05) and retroperitoneal WAT weight (by 72%; P < 0.01) only in RF-maintained rats, doubling the efficiency of lipid accumulation (P < 0.05). Thus, RF not only desynchronizes central and peripheral circadian clocks, and suppresses nocturnal GH secretion, but induces a preobesogenic state.

The effects of feeding corn or rice, either raw or heat processed (HP), on apparent total tract digestibility (ATTD) and apparent ileal digestibility (AID) of nutrients and insulin and ghrelin concentrations in the serum were studied in young pigs. Pigs were weaned at approximately 23 ± 3 d of age and weighed 7.4 ± 1.2 kg. Each of the 4 treatments was replicated 9 times and the experimental unit was a pig individually housed. Pigs (5 males and 4 females/treatment) were fed their respective diets ad libitum from 23 to 47 d of age. At 37 d of age, the effects of dietary treatments on the fasting and postprandial concentrations of insulin and total and acylated ghrelin were studied. The ATTD of OM, GE, and ether extract (EE) was, respectively, 4.3, 5.4, and 3.6% greater (P < 0.05) for the rice than for the corn diets but CP digestibility was not affected. Similar results were observed for AID. Heat processing of the cereal increased (P < 0.05) the ATTD by 2.1% for OM, 3.2% for GE, 7.1% for EE, and 2.2% for CP and tended to increase the AID of CP (P = 0.06) and starch (P = 0.09). The postprandial serum insulin response was greater and prolonged in pigs fed raw rice than pigs fed raw corn (P < 0.05). Also, the effects of HP on serum insulin response were more pronounced with corn than with rice (cereal x HP, P < 0.05). Total ghrelin concentration was not affected by treatment but acylated ghrelin was greater (P < 0.05) at 6 h postprandial in pigs fed rice than pigs fed raw corn. Feeding rice and HP corn increased nutrient digestibility and insulin response in the early postprandial period and acylated ghrelin response in the late postprandial period compared with feeding raw corn.

This study examined how adiposity signals are related to adiposity during recovery from forced overweight (OW). Rats were rendered OW by chronic intragastric overfeeding (OW). Overfeeding was stopped when OW rats reached 126-129% of saline-infused normal-weight (NW) rats. Adipose tissue (AT) mass was estimated by computed tomography, and blood was drawn from chronic atrial cannulas throughout. Basal levels (i.e. after 2-3 h fasts late in the diurnal phase) of the hypothesized adiposity signals insulin, leptin, ghrelin, and amylin were assayed. OW rats gained approximately 130 g more body weight (BW) and approximately 100 g more AT mass during overfeeding. Plasma levels of insulin and leptin increased, whereas those of ghrelin decreased, linearly with AT mass; amylin did not change reliably. During recovery, OW rats' BW and AT mass decreased but were still elevated vs. NW rats after 39 d. OW rats' insulin returned to NW levels on d 1 of recovery and decreased below NW levels thereafter. Leptin was no longer elevated after d 8 of recovery. Ghrelin and amylin did not change reliably during recovery. Although AT mass decreased in OW rats during each intermeasurement interval between d 0 and d 23 of recovery, insulin and leptin did so during only the first interval (d 0-5). Insulin and leptin levels were exponentially related to AT mass during recovery. These data indicate that basal insulin, leptin, ghrelin, and amylin do not encode AT mass in rats dynamically regulating BW and adiposity during recovery from OW.

Aims: To compare the responses of GIP, GLP-1, ghrelin and IGFBP-1 between meals with different fat and energy content in adolescents with type 1 diabetes (T1DM) and to relate them to gastric emptying and glycaemia. Methods: On different days and in a random order, 7 adolescents with T1DM ingested a high- and low-fat meal (fat content: 38 and 2 g, energy content: 640 and 320 kcal, respectively). At normoglycaemia, the same prandial insulin dose was given at both meals and to all subjects. Postprandial blood samples were taken repeatedly over 4 hours. Gastric emptying was estimated by the paracetamol absorption method. Results: The area under the curve (AUC) for GIP(0-240 min) and for GLP-1(0-120 min) was larger, but smaller for relative ghrelin(0-240 min), after the high-fat meal (p = 0.002, 0.030 and 0.043, respectively). IGFBP-1 decreased significantly, but not differently, after the meals. Larger GLP-1 secretion correlated with slower gastric emptying (p = 0.029) and higher fasting ghrelin levels correlated with lower postprandial glycaemia (p = 0.007). Conclusion: In adolescents with T1DM, the postprandial responses of GIP, GLP-1 and ghrelin, but not that of IGFBP-1, depend more on meal size than on insulin.

This paper reports an investigation into metabolic and endocrine maturity in the neonate lamb and the relationships between litter size, birth weight, and maternal metabolic and endocrine variables on behavior at birth and survival over the first 72 h of life. Data were from multiparous, fine-wool Merino ewes (n = 150; equal numbers of single-lamb and twin-lamb bearing status) lambed on pasture after late gestational glucocorticoid treatments. Stepwise multiple regression analysis was used to investigate relationships between lamb survival, behavior, endocrinology, and physiology. Improved lamb viability at 72 h after birth was related to decreased chill indices at birth, singleton litter status, greater presuckling rectal temperature, increased ewe prelambing plasma ghrelin concentration, female sex, heavier birth weight, and decreased lamb presuckling plasma glucose concentration. Greater rectal temperatures were associated with heavier birth weight and gestation lengths shorter than 146 d, but no relationship with neonatal behavioral progression was evident. Presuckling glucose concentrations were greater in singletons and lambs born to ewes of greater BCS at d 95 of gestation, and lambs of heavier birth weight, but were also associated with decreased rectal temperatures. This might reflect a delay in glucose utilization during the adjustment from a fetal metabolic rate to a rate appropriate for cold external environments. Singleton lambs exhibited decreased presuckling plasma NEFA concentrations and were almost 8 times more likely to survive to 72 h than a twin-born lamb. Birth weight was lesser in lambs born to ewes with elevated plasma glucose and leptin concentrations before lambing and was positively related to ewe BW at d 95 of gestation and to length of gestation. Greater presuckling plasma ghrelin and leptin concentrations were measured for shorter gestation lengths. Neonate presuckling ghrelin concentrations above 650 pg/mL tended (P = 0.077) to be associated with improved lamb survival to 72 h. This was consistent with a curvilinear decline in neonate survival rates to 72 h after birth as time of latency to suckle increased. No relationship was observed between lamb plasma glucose concentrations and behavioral expression after lambing. Lambs exhibiting greater metabolic and endocrine maturity at birth had improved survival in a cold environment to 72 h after birth. The role of ghrelin in ovine fetal development warrants further investigation.

Context: Ghrelin, an endogenous ligand for the GH secretagogue receptor, is an orexigenic peptide hormone produced primarily by the stomach. Recent studies suggest significant differences in the specificity of currently available ghrelin assays. Objective: The aim of the study was to compare four ghrelin assays (two commercially available and two developed by our group) of differing specificity, each used on the same set of more than 800 plasma samples from a human study. Design: Thirteen volunteers were sampled every 20 min for 6 h after consumption of one of three isocaloric drinks consisting of either 80% fat, 80% carbohydrate, or 80% protein. The samples were assayed by RIA for total and active ghrelin, as well as by sandwich assays for acyl and des-acyl ghrelin. The ghrelin profiles for each individual were smoothed using a statistical algorithm to lessen the effects of pulsatility and noise. Results: The sandwich assays for acyl and des-acyl ghrelin yielded ghrelin values that were lower than those from the corresponding RIAs. The ghrelin profiles after nutrient ingestion were similar, yet key differences among the four assays were apparent; in particular, percentage changes were significantly greater in the sandwich assays. Conclusions: The lower levels and greater relative changes in ghrelin values reported by the sandwich assays are consistent with greater assay specificity. When applied to the nutrient study, the sandwich assays were better able to distinguish the different responses to different nutrients than were the RIAs.

CONTEXT: It is uncertain how between-meal variations in energy availability and physiological changes in ghrelin, leptin, and insulin affect appetite. Objective: The aim of the study was to examine the influence on human appetite of the meal size and its nutrient content or changes in energy availability and concentrations of ghrelin, leptin, and insulin. DESIGN: We conducted a crossover study manipulating meal size and energy availability through exercise energy expenditure and iv nutrient replacement (TPN). Setting: The study was performed at a Clinical Research Center. PARTICIPANTS: Ten healthy postmenopausal women (age, 59.7 +/- 1.5 yr; mean body mass index, 26 kg/m(2)) were studied. Interventions: We conducted trials based on different morning meal size (418 vs. 2090 KJ), presence or absence of exercise energy expenditure (2273 to 2361 KJ), energy replacement by TPN (1521 to 1538 KJ), and a midday ad libitum meal. Main Outcome Measures: Changes in hunger, fullness, midday ad libitum food consumption, and concentrations of ghrelin, leptin, insulin, and metabolic fuels were measured. We also performed midday meal tests for the presence of caloric compensation. RESULTS: Appetite was influenced by the size and energy content of the meals, but not by variation in energy availability which also did not trigger consummatory compensation. Exercise reduced hunger and increased fullness. Ghrelin, leptin, and insulin responded to changes in energy availability but not to meal size. Appetite was unaffected by physiological changes in ghrelin, leptin, or insulin. CONCLUSIONS: During rest, appetite is influenced by the size and energy content of meals, but it bears no homeostatic relationship to between-meal changes in energy availability due to small meals, exercise, or TPN, or concentrations of ghrelin, leptin, and insulin.

OBJECTIVES: The purpose of this study was to examine alterations in lipid profiles and in the serum concentrations of acylated and desacylated ghrelin, paraoxonase and arylesterase in psychiatric patients before and after treatment with 40 mg citalopram daily for 3 months. DESIGN AND METHODS: Samples were collected from 22 healthy controls and 24 psychiatric patients before and after citalopram treatment. Blood levels of acylated and desacylated ghrelin were measured by radioimmunoassay. Paraoxonase and arylesterase activities were determined spectrophotometrically. Lipid parameters were measured on the OLYMPUS-AU400. RESULTS: It was found that the levels of acylated, desacylated ghrelin, paraoxonase arylesterase, total cholesterol and triglyceride were lower in depressive patients before citalopram treatment than in the control group. Those parameters were not restored after citalopram treatment except for the arylesterase level. CONCLUSION: Decreased PON1 and ghrelin levels as well as fluctuations in lipid profiles may be involved in the etiology of depressive disorders.

Perinatal mortality is a major contributing factor to reproductive wastage in grazing sheep industries. Enhanced metabolic and endocrine maturity at birth may improve the behavioral competency and thermoregulatory ability of neonates, potentially improving lamb survival over the first 72 h of life. Maternal glucocorticoid treatment in late gestation was investigated as a mechanism for manipulating metabolic and endocrine maturity in the ovine neonate. Multiparous, fine-wool Merino ewes (n = 150) were divided into 3 groups to lamb on pasture. Within each group, 5 single-lamb and 5 twin-lamb bearing ewes were randomly allocated to 1 of 5 treatments. Treatments included a saline control (1 mL), or dexamethasone (2 mg/mL as the sodium phosphate) injected intramuscularly at 1 of 2 dose rates (1.5 or 3.0 mg) at d 130 or 141 of gestation. One-half of the control ewes were injected at d 130 and the remainder at d 141. Dexamethasone treatment had no effect on lamb survival to 72 h after birth, although there tended (P = 0.09) to be a smaller proportion of lambs dying due to dystocia than for control lambs. Heart girth at birth in singleton and twin lambs was reduced (P 0.01) at the greater dose rate. Further, treatment also reduced birth weight (by about 5%) and presuckling rectal temperatures in twin lambs, but not in singleton lambs. These reductions were also dependent on the sex of the lamb. Dexamethasone treatment did not alter gestation length or lamb presuckling plasma glucose, NEFA, urea, or leptin concentrations, but treatment at d 141 increased (P 0.05) ghrelin concentrations in singleton and male lambs. Behavioral interactions between ewes and neonatal lambs were generally unaffected, although treatment at d 130 produced lambs that took longer to bleat than lambs of untreated ewes (P 0.05). Treatment did not affect the concentration of measured blood metabolites or hormones at weaning. Although there were interactions between litter size, lamb sex, and the dose rate and time of treatment on weaning weight, BW recorded 73 d after weaning was unaffected by treatment. Despite changes in birth weight, rectal temperature, lamb behavior, and presuckling plasma ghrelin concentrations, survival in the first 72 h of life, and lamb growth performance were unaffected by periparturient maternal glucocorticoid treatment.