Hypersensitivity to insulin during remissions in cyclosporin-treated IDDM patients.

OBJECTIVE: To test the sensitivity to insulin in recent-onset IDDM patients, its course according to treatment, and the advent of remissions. RESEARCH DESIGN AND METHODS: The euglycemic hyperinsulinemic clamp was used in 54 recent-onset IDDM patients and 14 healthy control subjects. Patients were tested after 1,2, and 4 wk of treatment with either insulin or insulin plus cyclosporin A, during cyclosporin A-associated long-lasting remissions, and during relapses. RESULTS: Insulin sensitivity was markedly decreased in all patients at onset. It was rapidly restored by insulin therapy, whether immunosuppression was associated with it or not. Insulin sensitivity was even higher than normal in the remission patients, who also were characterized by the reappearance of some endogenous insulin secretion and the sustained normalization of blood glucose profiles. During relapses, the deterioration of the blood glucose profiles was associated with some loss of insulin sensitivity. CONCLUSIONS: Cyclosporin A-associated remissions represent an original situation that associates euglycemia with the persistence of low endogenous insulin secretion. Cyclosporin A by itself had no influence on sensitivity to insulin, but allowed the reappearance of some insulin secretory capacity that contributed, with the improvement of insulin sensitivity, to the development of the diabetes honeymoon. The secretion of endogenous insulin, although lower than normal, was sufficient to secure a high sensitivity to insulin and the maintenance of normal blood glucose profiles, presumably because of the fact that insulin was released directly into the portal vein in these conditions. This metabolic state was precarious: the optimal sensitivity to insulin disappeared in patients who relapsed. These results have important clinical consequences: the preservation of islet residual secretory capacity by the use of newer nontoxic immunosuppressive protocols, combined with a minimal supportive insulin therapy in remission patients, may prolong remissions and maintain an optimal insulin sensitivity.

Contribution of hepatic fatty acid oxidation and exogenous galactose supply to the regulation of glucose homeostasis in the newborn rabbit.

The inhibition of hepatic gluconeogenesis by 3-mercaptopicolinic acid (3-MPA) leads to a profound hypoglycemia in both suckling and fasting 24-hour-old rabbits. This hypoglycemia is totally reversed 1 h after the intragastric injection of an amount of galactose corresponding to the one ingested daily by the suckling newborns. This results from an active gluconeogenesis from galactose, which bypasses the site of inhibition by 3-MPA. However, this amount of galactose is not sufficient to maintain a normal blood glucose concentration for a long time, since 3 h after galactose injection, the blood glucose concentrations of newborn rabbits return to hypoglycemic values. When hepatic fatty acid oxidation is inhibited by 2-[6-(4-chlorophenoxy)hexyl]oxirane-2-carboxylate (POCA), 24-hour-old fasting rabbits become rapidly hypoglycemic secondary to a decrease in liver gluconeogenesis. The rate of hepatic gluconeogenesis is totally restored by giving medium-chain triglycerides, and the 24-hour-old rabbits become normoglycemic.

Insulin action during fasting and refeeding in rat determined by euglycemic clamp.

To further characterize the role of insulin in glucose metabolism during fasting and refeeding, euglycemic-hyperinsulinemic clamps were performed in control, 3 day-fasted, and 3 day-fasted then 3 day-refed rats. Glucose production and utilization were measured by using [3-3H]glucose. In control and refed rats, hepatic glucose production was totally suppressed at insulin concentration higher than 500 microU/ml; by contrast, during fasting, hepatic glucose production was not suppressed even at insulin concentration tenfold higher. Maximal increment of glucose utilization was lower in fasted than in control rats. Three days of refeeding restored almost entirely normal responses to insulin for glucose utilization. Blood glucose concentration was clamped at a different level in fasted and in control and refed rats; however, increment in glucose clearance in response to insulin was lower in fasted rats than in the two other groups. Thus fasting produces a state of insulin unresponsiveness both at the hepatic and peripheral levels, normal responsiveness being restored after 3 days of refeeding.

Effects of gestational hyperglycemia on glucose metabolism and its hormonal control in the fasted, newborn rat during the early postnatal period.

To evaluate the effects of gestational hyperglycemia on glucose metabolism and its regulation in the fasted rat during the early postnatal period, unrestrained rats were continuously infused with glucose during the last week of pregnancy. Control rats were infused with distilled water. Newborns were studied during the first six postnatal hours. At birth, newborns from glucose-infused rats, compared with controls, showed higher plasma glucose levels, increased plasma insulin, and lower plasma glucagon and catecholamine concentrations. Between birth and 2 h postpartum, newborn rats from both groups exhibited a marked hypoglycemia, which was, however, more severe in newborns from glucose-infused rats (15 mg/dl) than in controls (26 mg/dl). During the first four postnatal hours, plasma insulin concentration remained higher, while plasma glucagon and catecholamine concentrations remained lower in newborns from hyperglycemic rats. At 6 h, the glycemia reached normal values and the concentrations of the different hormones were similar in controls and newborns from glucose-infused mothers. Concurrently, in the newborns from glucose-infused rats, hepatic glucose production was altered, as they were unable to mobilize liver glycogen stores during the six postnatal hours. Despite slightly delayed phosphoenolpyruvate carboxykinase induction, the rate of gluconeogenesis from 10 mmol/L lactate estimated on isolated hepatocytes was higher in newborns from hyperglycemic mothers than in controls. These results show that gestational hyperglycemia compromises the metabolic and hormonal adaptation of the newborn rat to early extrauterine life; the striking feature of these neonates is the absence of mobilization of liver glycogen stores, which can probably be explained by fetal and neonatal hyperinsulinism associated with the defect of counterregulatory hormones.

Growth of fetal guinea pig: physical and chemical characteristics.

The growth of the fetal guinea pig was studied in 32 fetuses in 12 litters, ranging from 39 days gestation to full term (67 days). The wet weight of the fetus was well approximated by an exponential function of gestational age (GA) in days [fetal weight (g) = 0.993 e(0.068 X GA), r = 0.94]. Dry weight increased more rapidly than wet weight [dry weight (g) = 0.039 e(0.102 X GA); r = 0.97], resulting in an increase in percent dry weight from approximately 10% at 40 days gestation to 30% at term. Fat content increased even more rapidly than dry weight [body fat (g) = 0.00123 e(0.136 X GA), r = 0.97], accounting for 33% of dry weight and 11.7% of wet weight at term. Using bomb calorimetric projections of caloric value of 9.3 kcal X g fat-1 X day-1 and 4.6 kcal X g nonfat dry wt-1 X day-1, we estimate that growth of the fetal guinea pig requires 220 kcal X kg fetal wt-1 X day-1 near term. Carbon and nitrogen contents of the fetus increased at different rates, reflecting the changes in fat and nonfat tissues. Amino acids contributed 80% of total body nitrogen and 41% of total body carbon near term. Cysteine concentrations increased and lysine concentrations decreased with gestational age; the concentrations of the other measured amino acids did not change with gestational age. These studies represent the first systematic study of the chemical growth of the fetus in a nonhuman species.

Evidence that hepatic mitochondrial mass decreases during the first sixteen hours following birth in starved newborn rats.

Fatty acid oxidation increases in newborn rats between 0 and 16 h after birth. We have tested the hypothesis that such a rise might be due to an increase in hepatic mitochondrial mass. The ratio total activity/specific activity for cytochrome c oxidase and citrate synthase was used as an index that reflected the changes in mitochondrial mass. This ratio was decreased by about 25% 16 h after delivery, indicating that hepatic mitochondrial mass was lower 16 h after birth than at birth in starved rats. We conclude that changes in the mitochondrial mass are not responsible for the increase in liver capacity to oxidize fatty acids.

Hyperglycaemia induced by glucose infusion in the unrestrained pregnant rat: effect on body weight and lipid synthesis in post-mature fetuses.

Mild hyperglycaemia was induced in unrestrained pregnant rats from day 20.5 to day 23.5 of pregnancy, using a continuous glucose infusion. Control rats were infused with distilled water. In post-mature fetuses from glucose-infused rats, raised plasma glucose and insulin concentrations were related to increased body weight (6.03 +/- 0.07 g) and total carcass fat (2.02 +/- 0.04% of fresh weight) compared with control fetuses of the same age (5.35 +/- 0.07 and 1.5 +/- 0.04 g, respectively). Concurrently, the rate of lipogenesis in the carcass, estimated from the incorporation of tritium from tritiated water into fatty acids, was significantly increased in fetuses from glucose infused rats compared with control rats (6.00 +/- 0.34 versus 2.62 +/- 0.27 and 3H2O X h-1 X g tissue-1, respectively.

[Maternal hyperglycemia and fetal development in the rat: effects of a continuous glucose perfusion in the rat at the end of gestation]. / Hyperglycémie maternelle et développement foetal chez le rat: conséquences d'une perfusion continue de glucose à la rate en fin de gestation.

Continuous glucose infusion was used to induce mild hyperglycemia in unrestrained pregnant rats during the last three days of pregnancy. Control rats were infused with distilled water. Compared with the controls, fetuses from glucose-infused rats showed higher plasma glucose levels, increased plasma insulin and lower plasma glucagon concentrations. Pregnancy prolonged until day 23.5 resulted in a sharp decrease in plasma insulin concentrations and a dramatic increase in plasma glucagon concentrations. In 23.5-day old fetuses from both groups, plasma insulin concentration rose when phentolamine was injected but not when propanolol was injected. Plasma glucagon concentration in 23.5-day old fetuses from glucose-infused rats dropped with propanolol injection. In fetuses from control rats, liver phosphoenolpyruvate activity increased markedly and liver glycogen stores decreased sharply. In fetuses from glucose-infused rats, liver phosphoenolpyruvate carboxykinase activity rose and glycogen content decreased, but to a lesser degree. Moreover, in postmature fetuses from glucose-infused rats, elevated plasma glucose and insulin concentrations were related to increased body weight and total carcass fat. Concurrently, the rate of lipogenesis in the carcass of these fetuses (estimated from the incorporation of 3H from 3H2O into fatty acids) was significantly increased.

Evidence that medium-chain fatty acid oxidation can support an active gluconeogenesis in the suckling newborn rat.

Long-chain fatty acid oxidation was inhibited in 1-day-old suckling rats by 2-tetradecylglycidic acid. Within 6 h it induced profound hypoketonemia and hypoglycemia together with a decreased rate of gluconeogenesis estimated from [U-14-C]-lactate incorporation into glucose. Medium-chain triglyceride feeding restored, within 3 h, normal blood concentrations of ketones and glucose as well as a high gluconeogenic rate from [U-14-C]-lactate. It is concluded that in the suckling newborn rat 2-tetradecylglycidic acid is effective in inhibiting selectively long-chain fatty acid oxidation and that medium-chain fatty acid oxidation can support an active gluconeogenesis.

Hyperglycaemia induced by glucose infusion in the unrestrained pregnant rat during the last three days of gestation: metabolic and hormonal changes in the mother and the fetuses.

Continuous glucose infusion was used to induced mild hyperglycaemia in unrestrained pregnant rats during the last three days of pregnancy. Control pregnant rats were infused with distilled water. Fetuses were studied after normal or prolonged pregnancy. Fetuses from glucose-infused rats, compared with controls, showed higher plasma glucose levels, increased plasma insulin and lower plasma glucagon concentrations. Pregnancy prolonged until day 23.5 resulted in a rise in the glucagon/insulin ratio from 6.5 to 67 in fetuses from control rats and from 1.3 to 13 in fetuses from glucose-infused rats. Concurrently in fetuses from control rats, liver phosphoenolpyruvate carboxykinase activity increased markedly and liver glycogen stores decreased sharply. In fetuses from glucose-infused rats, liver phosphoenolpyruvate carboxykinase activity rose and glycogen content decreased, but to a lesser extent. These results show that both the A and B cells of the rat fetal pancreas are sensitive to chronic glucose stimulation.

Evidence that stimulation of glucose metabolism by insulin is not altered in isolated soleus muscle of pregnant rats.

Various concentrations of insulin stimulated to the same extent glucose transport and metabolism in incubated soleus muscle of virgin and 19-day-pregnant rats. This suggests that the resistance to insulin that occurs during pregnancy in vivo does not result from an intrinsic defect in skeletal muscles.

An estimate of the caloric requirements of the human fetus.

The caloric requirements of the human fetus have been calculated from the reported values of the fetal oxygen consumption and the composition of the new tissue deposited, both expressed as their caloric equivalents. It is estimated that the deposition of fat represents over half the caloric accretion from the 27th week of gestation until term, and approximately 90% of the caloric accretion at term. The total caloric requirement of the human fetus at term is estimated at approximately 90-100 kcsl/kg/day.

Fuel metabolism in fasted newborn rabbits.

Newborn rabbits delivered by Caesarean section at term were fasted for 72 h at 36 degrees C. Despite the abrupt interruption of maternal supply of energy substrates, glycaemia remains stable for 4 h after birth. This can be related to glucose production via rapid liver glycogenolysis; however, indirect evidence suggests that gluconeogenesis could also contribute to glucose production during this period. There is a selective decrease in the concentrations of gluconeogenic substrates and a suitable hormonal environment for gluconeogenesis as decreased insulin and increased glucagon concentration just after birth. The relative hypoglycaemia which develops after 6 h of life (2.6 mM at 72 h), despite high blood concentrations of non-esterified fatty acids and ketone bodies is not due to a deficient gluconeogenesis per se, as injection of gluconeogenic substrates to 72 h fasted newborns produces a three-fold increase in plasma glucose concentration. It is suggested that this relative hypoglycaemia is secondary to limited gluconeogenic substrate availability in the form of low circulting concentrations of gluconeogenic amino acids.

Alterations in the rate of lipogenesis in vivo in maternal liver and adipose tissue on premature weaning of lactating rats: a possible regulatory role of prolactin.

Removal of pups for 24 h from rats at peak lactation decreased 3H2O incorporation into lipid in vivo in mammary gland by 95%, whereas it was increased in liver (77%) and adipose tissue (330%). These increases were prevented by administration of prolactin. Plasma insulin increased 3-fold on weaning and this was partially prevented by prolactin.