Meeting the Vitamin A Requirement: The Efficacy and Importance of ß-Carotene in Animal Species.

Vitamin A is essential for life in all vertebrate animals. Vitamin A requirement can be met from dietary preformed vitamin A or provitamin A carotenoids, the most important of which is ß-carotene. The metabolism of ß-carotene, including its intestinal absorption, accumulation in tissues, and conversion to vitamin A, varies widely across animal species and determines the role that ß-carotene plays in meeting vitamin A requirement. This review begins with a brief discussion of vitamin A, with an emphasis on species differences in metabolism. A more detailed discussion of ß-carotene follows, with a focus on factors impacting bioavailability and its conversion to vitamin A. Finally, the literature on how animals utilize ß-carotene is reviewed individually for several species and classes of animals. We conclude that ß-carotene conversion to vitamin A is variable and dependent on a number of factors, which are important to consider in the formulation and assessment of diets. Omnivores and herbivores are more efficient at converting ß-carotene to vitamin A than carnivores. Absorption and accumulation of ß-carotene in tissues vary with species and are poorly understood. More comparative and mechanistic studies are required in this area to improve the understanding of ß-carotene metabolism.

Enhanced insulin secretion responsiveness and islet adrenergic desensitization after chronic norepinephrine suppression is discontinued in fetal sheep.

Intrauterine growth-restricted (IUGR) fetuses experience prolonged hypoxemia, hypoglycemia, and elevated norepinephrine (NE) concentrations, resulting in hypoinsulinemia and ß-cell dysfunction. Previously, we showed that acute adrenergic blockade revealed enhanced insulin secretion responsiveness in the IUGR fetus. To determine whether chronic exposure to NE alone enhances ß-cell responsiveness afterward, we continuously infused NE into fetal sheep for 7 days and, after terminating the infusion, evaluated glucose-stimulated insulin secretion (GSIS) and glucose-potentiated arginine-induced insulin secretion (GPAIS). During treatment, NE-infused fetuses had greater (P < 0.05) plasma NE concentrations and exhibited hyperglycemia (P < 0.01) and hypoinsulinemia (P < 0.01) compared with controls. GSIS during the NE infusion was also reduced (P < 0.05) compared with pretreatment values. GSIS and GPAIS were approximately fourfold greater (P < 0.01) in NE fetuses 3 h after the 7 days that NE infusion was discontinued compared with age-matched controls or pretreatment GSIS and GPAIS values of NE fetuses. In isolated pancreatic islets from NE fetuses, mRNA concentrations of adrenergic receptor isoforms (α1D, α2A, α2C, and ß1), G protein subunit-αi-2, and uncoupling protein 2 were lower (P < 0.05) compared with controls, but ß-cell regulatory genes were not different. Our findings indicate that chronic exposure to elevated NE persistently suppresses insulin secretion. After removal, NE fetuses demonstrated a compensatory enhancement in insulin secretion that was associated with adrenergic desensitization and greater stimulus-secretion coupling in pancreatic islets.

Hypoxaemia-induced catecholamine secretion from adrenal chromaffin cells inhibits glucose-stimulated hyperinsulinaemia in fetal sheep.

Abstract Hypoxaemia elicits adrenergic suppression of fetal glucose-stimulated hyperinsulinaemia. We postulate that this effect is mediated by catecholamines, exclusively, from fetal adrenal chromaffin cells. To investigate this hypothesis, square-wave hyperglycaemic clamp studies were performed under normoxaemic (26 ± 0.9 mmHg) and hypoxaemic (14 ± 0.3 mmHg) steady-state conditions in near-term fetal sheep that had undergone either surgical sham or bilateral adrenal demedullation (AD), values mentioned are ± SEM. Under normoxaemic conditions plasma noradrenaline concentrations were lower in AD fetuses than in sham-operated fetuses (457 ± 122 versus 1073 ± 103 pg ml(-1), P < 0.05). Plasma insulin concentrations were not different at euglycaemia between shams (0.46 ± 0.07 ng ml(-1)) and AD fetuses (0.44 ± 0.04 ng ml(-1)) and increased (P < 0.05) with hyperglycaemia in both groups although to a lesser extent in AD fetuses (0.94 ± 0.19 ng ml(-1)) compared to shams (1.31 ± 0.15 ng ml(-1); P < 0.05). Hypoxaemia increased plasma adrenaline (26-fold) and noradrenaline (5-fold) in shams but elicited no change in AD fetuses. Under hypoxaemic conditions, euglycaemic plasma insulin concentrations were reduced (P < 0.05) in both sham and AD fetuses to 0.30 ± 0.05 ng ml(-1) and 0.27 ± 0.01 ng ml(-1) respectively, and the insulin response to hyperglycaemia was abolished in shams but not affected in AD fetuses (0.33 ± 0.06 versus 0.73 ± 0.02 ng ml(-1), P < 0.05). Hypoxaemia also induced hyperlactacaemia and hypocarbia to a greater extent in shams than in AD fetuses, indicating that catecholamines potentiate reductions in oxidative metabolism independently of insulin. These findings demonstrate that the fetal adrenal chromaffin cells are the source for acute hypoxaemia-induced elevations in fetal plasma catecholamines and suppression of glucose-stimulated hyperinsulinaemia, but other factors reduce plasma insulin at euglycaemia.

Chronic pulsatile hyperglycemia reduces insulin secretion and increases accumulation of reactive oxygen species in fetal sheep islets.

Children from diabetic pregnancies have a greater incidence of type 2 diabetes. Our objective was to determine if exposure to mild-moderate hyperglycemia, by modeling managed diabetic pregnancies, affects fetal ß-cell function. In sheep fetuses, ß-cell responsiveness was examined after 2 weeks of sustained hyperglycemia with 3 pulses/day, mimicking postprandial excursions, and compared to saline-infused controls (n = 10). Two pulsatile hyperglycemia (PHG) treatments were studied: mild (mPHG, n = 5) with +15% sustained and +55% pulse; and moderate (PHG, n = 10) with +20% sustained and +100% pulse. Fetal glucose-stimulated insulin secretion and glucose-potentiated arginine insulin secretion were lower (P < 0.05) in PHG (0.86 ± 0.13 and 2.91 ± 0.39  ng/ml plasma insulin) but not in mPHG fetuses (1.21 ± 0.08 and 4.25 ± 0.56  ng/ml) compared to controls (1.58 ± 0.25 and 4.51 ± 0.56  ng/ml). Islet insulin content was 35% lower in PHG and 35% higher in mPHG vs controls (P < 0.01). Insulin secretion and maximally stimulated insulin release were also reduced (P < 0.05) in PHG islets due to lower islet insulin content. Isolated PHG islets also had 63% greater (P < 0.01) reactive oxygen species (ROS) accumulation at 11.1  mmol/l glucose than controls (P < 0.01), but oxidative damage was not detected in islet proteins. PHG fetuses showed evidence of oxidative damage to skeletal muscle proteins (P < 0.05) but not insulin resistance. Our findings show that PHG induced dysregulation of islet ROS handling and decreased islet insulin content, but these outcomes are independent. The ß-cell outcomes were dependent on the severity of hyperglycemia because mPHG fetuses had no distinguishable impairments in ROS handling or insulin secretion but greater insulin content.

Characterization of glucose-insulin responsiveness and impact of fetal number and sex difference on insulin response in the sheep fetus.

GSIS is often measured in the sheep fetus by a square-wave hyperglycemic clamp, but maximal ß-cell responsiveness and effects of fetal number and sex difference have not been fully evaluated. We determined the dose-response curve for GSIS in fetal sheep (0.9 of gestation) by increasing plasma glucose from euglycemia in a stepwise fashion. The glucose-insulin response was best fit by curvilinear third-order polynomial equations for singletons (y = 0.018x(3) - 0.26x(2) + 1.2x - 0.64) and twins (y = -0.012x(3) + 0.043x(2) + 0.40x - 0.16). In singles, maximal insulin secretion was achieved at 3.4 ± 0.2 mmol/l glucose but began to plateau after 2.4 ± 0.2 mmol/l glucose (90% of maximum), whereas the maximum for twins was reached at 4.8 ± 0.4 mmol/l glucose. In twin (n = 18) and singleton (n = 49) fetuses, GSIS was determined with a square-wave hyperglycemic clamp >2.4 mmol/l glucose. Twins had a lower basal glucose concentration, and plasma insulin concentrations were 59 (P < 0.01) and 43% (P < 0.05) lower in twins than singletons during the euglycemic and hyperglycemic periods, respectively. The basal glucose/insulin ratio was approximately doubled in twins vs. singles (P < 0.001), indicating greater insulin sensitivity. In a separate cohort of fetuses, twins (n = 8) had lower body weight (P < 0.05) and ß-cell mass (P < 0.01) than singleton fetuses (n = 7) as a result of smaller pancreata (P < 0.01) and a positive correlation (P < 0.05) between insulin immunopositive area and fetal weight (P < 0.05). No effects of sex difference on GSIS or ß-cell mass were observed. These findings indicate that insulin secretion is less responsive to physiological glucose concentrations in twins, due in part to less ß-cell mass.

Maternal amino acid supplementation for intrauterine growth restriction.

Maternal dietary protein supplementation to improve fetal growth has been considered as an option to prevent or treat intrauterine growth restriction. However, in contrast to balanced dietary supplementation, adverse perinatal outcomes in pregnant women who received high amounts of dietary protein supplementation have been observed. The responsible mechanisms for these adverse outcomes are unknown. This review will discuss relevant human and animal data to provide the background necessary for the development of explanatory hypotheses and ultimately for the development therapeutic interventions during pregnancy to improve fetal growth. Relevant aspects of fetal amino acid metabolism during normal pregnancy and those pregnancies affected by IUGR will be discussed. In addition, data from animal experiments which have attempted to determine mechanisms to explain the adverse responses identified in the human trials will be presented. Finally, we will suggest new avenues for investigation into how amino acid supplementation might be used safely to treat and/or prevent IUGR.

ß2-Adrenergic receptor desensitization in perirenal adipose tissue in fetuses and lambs with placental insufficiency-induced intrauterine growth restriction.

Placental insufficiency-induced intrauterine growth restriction (IUGR) fetuses have chronic hypoxaemia and elevated plasma catecholamine concentrations. In this study, we determined whether adrenergic responsiveness becomes desensitized in the perirenal adipose tissue of IUGR fetuses and lambs by measuring adrenergic receptor (AR) mRNA and protein levels. We also tested the ability of adrenaline to mobilize non-esterified fatty acids (NEFAs) in young lambs. Perirenal adipose tissue was collected from IUGR and control fetuses at 133 days of gestational age (dGA) and lambs at 18 days of age (dA). ß(2)-AR mRNA concentrations were 59% and 74% lower (P < 0.05) in IUGR fetuses and lambs compared to controls, respectively, which also resulted in lower protein levels (P < 0.05). No treatment differences were detected for α(1A)-, α(1B)-, α(1D)-, α(2A)-, α(2B)-, α(2C)-, ß(1)- and ß(3)-AR expression. mRNA concentrations were also determined for hormone sensitive lipase (HSL), perilipin (lipid droplet-associated protein), and two adipokines, leptin and adiponectin. Adiponectin and HSL were not different between treatments at either age. Compared to controls, perilipin and leptin mRNA concentrations were lower (P < 0.05) in IUGR fetuses but not in lambs. Because of the ß(2)-AR results, we challenged a second cohort of lambs with exogenous adrenaline at 21 dA. The ability of adrenaline to mobilize NEFA was 55 ± 15% lower (P < 0.05) in IUGRs than controls. Collectively, our findings indicate that elevated catecholamine exposure in utero causes desensitization of adipose tissue by down-regulation of ß(2)-AR, and this persists in lambs. This impairment in adrenergic stimulated lipolysis might partially explain early onset obesity in IUGR offspring.

Consequences of a compromised intrauterine environment on islet function.

Low birth weight is an important risk factor for impaired glucose tolerance and diabetes later in life. One hypothesis is that fetal beta-cells inherit a persistent defect as a developmental response to fetal malnutrition, a primary cause of intrauterine growth restriction (IUGR). Our understanding of fetal programing events in the human endocrine pancreas is limited, but several animal models of IUGR extend our knowledge of developmental programing in beta-cells. Pathological outcomes such as beta-cell dysfunction, impaired glucose tolerance, and diabetes are often observed in adult offspring from these animal models, similar to the associations of low birth weight and metabolic diseases in humans. However, the identified mechanisms underlying beta-cell dysfunction across models and species are varied, likely resulting from the different methodologies used to induce experimental IUGR, as well as from intraspecies differences in pancreas development. In this review, we first present the evidence for human beta-cell dysfunction being associated with low birth weight or IUGR. We then evaluate relevant animal models of IUGR, focusing on the strengths of each, in order to define critical periods and types of nutrient deficiencies that can lead to impaired beta-cell function. These findings frame our current knowledge of beta-cell developmental programing and highlight future research directions to clarify the mechanisms of beta-cell dysfunction for human IUGR.

Cats are able to adapt protein oxidation to protein intake provided their requirement for dietary protein is met.

Cats require more dietary protein than noncarnivorous species. Earlier work showed that cats lack the ability to regulate hepatic urea cycle enzymes in response to dietary protein concentration. We thus hypothesized that cats are unable to fully adapt protein oxidation to protein intake, particularly at low-protein concentrations. We used indirect respiration calorimetry to assess cats' ability to adapt substrate oxidation to diets containing different concentrations of protein, including 1 below their protein requirement. Nine cats (5 males and 4 females; 2.7 +/- 0.5 y; 4.49 +/- 0.19 kg) consumed each of 4 semipurified diets containing 7.5% [low protein (LP(3))], 14.2% [adequate protein (AP)], 27.1% [moderate protein (MP)], and 49.6% [high protein (HP)] of metabolizable energy from protein in a modified crossover design, beginning with the MP diet and then consuming the remaining diets in random order. After adaptation to each diet, cats completed a 5-d nitrogen balance trial and at least 2 12-h indirect calorimetry measurements. There was a significant effect of diet on protein oxidation (P < 0.0001), which measured 10.4 +/- 0.5, 14.1 +/- 1.0, 25.0 +/- 1.7, and 53.2 +/- 1.7% of total energy expenditure for the LP, AP, M,P and HP diets, respectively. The ratio of protein oxidation:protein intake was higher with the LP diet (1.39 +/- 0.07) than the other 3 diets (AP, 1.00 +/- 0.07; MP, 0.93 +/- 0.06; HP, 1.07 +/- 0.03; P < 0.0001), indicating a net loss of protein with the LP diet. Thus, cats are able to adapt protein oxidation to a wide range of dietary protein concentrations, provided their minimum protein requirement is met.

Energy restriction results in a mass-adjusted decrease in energy expenditure in cats that is maintained after weight regain.

Dietary energy restriction (ER) is used to treat obesity in cats but it is often unsuccessful. The purpose of this study was to determine whether ER results in a sustained decrease in mass-adjusted energy expenditure (EE) that may oppose weight loss and promote weight regain. EE and body composition were measured in 10 adult neutered cats at 3 time points: baseline (obese cats), during weight loss (40% ER), and following weight regain. The cats started with a body weight (BW) of 6.1 +/- 0.30 kg, body condition score (BCS) of 7.6 +/- 0.14 (on a 9-point scale), and fat body mass (FM) of 38 +/- 1.0% of BW. After weight loss, BW was 5.0 +/- 0.19 kg, BCS was 5.5 +/- 0.07 kg, and FM was 31 +/- 1.6% (P < 0.01). After weight regain, BW was 6.2 +/- 0.30 kg, BCS was 7.7 +/- 0.16, and FM was 42 +/- 1.8% (P < 0.01). Total EE decreased from 1258 +/- 33.7 kJ/d to 1025 +/- 39.6 kJ/d during weight loss (P < 0.001). After weight regain, EE was still lower than baseline (1103 +/- 41.5 kJ/d, P < 0.001). Energy intake (EI) at baseline (1337 +/- 50.6 kJ/d) was higher than EI after weight loss and regain (1217 +/- 61.2 kJ/d), resulting in no differences in energy balance (78 +/- 30.4 and 104 +/- 35.4 kJ/d, respectively, P = 0.581). These results support the hypothesis that ER results in a mass-adjusted decrease in EE in cats that is maintained after weight regain.