Blood amino acids concentration during insulin induced hypoglycemia in rats: the role of alanine and glutamine in glucose recovery

Summary. Our purpose was to determine the blood amino acid concentration during insulin induced hypoglycemia (IIH) and examine if the administration of alanine or glutamine could help glycemia recovery in fasted rats. IIH was obtained by an intraperitoneal injection of regular insulin (1.0 U/kg). The blood levels of the majority of amino acids, including alanine and glutamine were decreased (P < 0.05) during IIH and this change correlates well with the duration than the intensity of hypoglycemia. On the other hand, the oral and intraperitoneal administration of alanine (100 mg/kg) or glutamine (100 mg/kg) accelerates glucose recovery. This effect was partly at least consequence of the increased capacity of the livers from IIH group to produce glucose from alanine and glutamine. It was concluded that the blood amino acids availability during IIH, particularly alanine and glutamine, play a pivotal role in recovery from hypoglycemia.     

Impaired glucose tolerance (IGT) is not associated with disturbed homocysteine metabolism

Summary. Elevated plasma total homocysteine (tHcy) has been suggested to be an additional risk factor for cardiovascular disease in subjects with impaired glucose tolerance (IGT) and Type 2 diabetes (T2D). In order to investigate whether an insulin resistant/chronic hyperinsulinemic situation in male diabetic and prediabetic subjects directly influences the tHcy metabolism, fasting tHcy and post-methionine load tHcy plasma levels (PML-tHcy) were determined in 15 men with IGT, 13 men with newly dia-gnosed T2D, and 16 normoglycemic controls (NGT). Fasting tHcy (IGT, 13.1 ± 4.6; T2D, 12.8 ± 4.0; NGT, 10.7 ± 4.4 μmol/L) and PML-tHcy (IGT, 46.5 ± 17.39; T2D, 41.1 ± 6.8; NGT, 38.0 ± 9.7 μmol/L) showed no differences between the groups. Fasting tHcy and PML-tHcy correlated with fasting proinsulin (r = 0.395, p < 0.05; r = 0.386, p< 0.05) and creatinine (r = 0.489, p < 0.01; r = 0.339, p < 0.05), resp. Multiple regression analysis showed only a relationship between fasting tHcy and creatinine. No relationships have been found between fasting tHcy and PML-tHcy, resp., and indicators of an insulin resistant state, e.g., insulin and proinsulin, as well as serum cobalamin and folate concentrations. In conclusion, our data suggest that the degree of glucose intolerance has no direct impact on the metabolism of homocysteine. However, tHcy levels tend to be elevated with the development of nephropathy, indicating an association between tHcy and renal function in these subjects. Received May 11, 1999     

Effects of creatine supplementation on glucose tolerance and insulin sensitivity in sedentary healthy males undergoing aerobic training

Recent findings have indicated that creatine supplementation may affect glucose metabolism. This study aimed to examine the effects of creatine supplementation, combined with aerobic training, on glucose tolerance in sedentary healthy male. Subjects (n = 22) were randomly divided in two groups and were allocated to receive treatment with either creatine (CT) ( approximately 10 g . day over three months) or placebo (PT) (dextrose). Administration of treatments was double blind. Both groups underwent moderate aerobic training. An oral glucose tolerance test (OGTT) was performed and both fasting plasma insulin and the homeostasis model assessment (HOMA) index were assessed at the start, and after four, eight and twelve weeks. CT demonstrated significant decrease in OGTT area under the curve compared to PT (P = 0.034). There were no differences between groups or over time in fasting insulin or HOMA. The results suggest that creatine supplementation, combined with aerobic training, can improve glucose tolerance but does not affect insulin sensitivity, and may warrant further investigation with diabetic subjects.     

The application of a model of glucose and insulin dynamics to explain an observed effect of leptin administration in reversal of developmental programming

A dynamical model describing the glucose-insulin physiological system was applied to an experiment on the administration of the adipokine leptin between neonatal days 3 and 13 to rats whose dams were subject to different levels of nutrition during gestation. The effect of leptin treatment on the glucose-insulin equilibrium point and on the levels of other associated metabolites showed a significant change in direction that depended on the level of prenatal nutrition. Leptin has been shown to affect two factors that affect the equilibrium levels of glucose and insulin, gluconeogenesis and insulin sensitivity. Each effect is described by a parameter in the dynamical model. Mathematical analysis shows that changes in these parameters in the manner promoted by leptin would indeed increase or decrease the glucose-insulin equilibria depending on their initial equilibrium levels which might be induced by the level of prenatal nutrition. This analysis explains the results of the leptin experiment in the context of the dynamics of the glucocorticoid system. It also proposes a physiological mechanism for the expression of plasticity in the organism based on the status of the glucose and insulin equilibria.     

Glucose, insulin and glucagon in the diabetic goose.

The relationships between plasma glucose, insulin and glucagon were studied in geese made diabetic by subtotal pancreatectomy. As early as the first hours after the operation, the plasma glucose increases and a permanent diabetes develops. This diabetic state is characterized by two features: a very low plasma insulin level, which does not vary during the survival of the diabetic animals and a concentration of plasma glucagon (of pancreatic origin) which transiently diminishes then rises far above the normal level, and is correlated with the basal concentration of plasma glucose. The impaired glucose tolerance observed in diabetic animals is related to the suppression of the glucose-insulin and glucose-glucagon feedback mechanisms.     

Modeling the physiological glucose-insulin dynamic system on diabetics

A novel mathematical model is presented to describe the dynamic behavior of plasma glucose and insulin on diabetic subjects. Though various models have been proposed to simulate the short-term (a variety of intravenous glucose or insulin injection) glucose-insulin dynamics, it is intended to construct a modified delay differential equations (DDEs) model based on the human glucose-insulin metabolic system. Five specific adjustable parameters inside the model are defined as the factors of the major physiological functions. Then several clinical data sets (56 subjects) which includes the information of food ingestion and exogenous insulin injection are verified and the model could practically reflect the dynamics and oscillation behavior on diabetic subjects by varying the adjustable parameters. Moreover, the corresponding parameters are fairly helpful to identify the patient's conditions of major physiological functions. This generic glucose-insulin dynamic model can be expected to develop such advanced therapy strategies for diabetics in the future.     

Numerical bifurcation analysis of delay differential equations arising from physiological modeling

This paper has a dual purpose. First, we describe numerical methods for continuation and bifurcation analysis of steady state solutions and periodic solutions of systems of delay differential equations with an arbitrary number of fixed, discrete delays. Second, we demonstrate how these methods can be used to obtain insight into complex biological regulatory systems in which interactions occur with time delays: for this, we consider a system of two equations for the plasma glucose and insulin concentrations in a diabetic patient subject to a system of external assistance. The model has two delays: the technological delay of the external system, and the physiological delay of the patient's liver. We compute stability of the steady state solution as a function of two parameters, compare with analytical results and compute several branches of periodic solutions and their stability. These numerical results allow to infer two categories of diabetic patients for which the external system has different efficiency.     

Modeling the glucose-insulin regulatory system and ultradian insulin secretory oscillations with two explicit time delays

In the glucose-insulin regulatory system, ultradian insulin secretory oscillations are observed to have a period of 50-150 min. After pioneering work traced back to the 1960s, several mathematical models have been proposed during the last decade to model these ultradian oscillations as well as the metabolic system producing them. These currently existing models still lack some of the key physiological aspects of the glucose-insulin system. Applying the mass conservation law, we introduce two explicit time delays and propose a more robust alternative model for better understanding the glucose-insulin endocrine metabolic regulatory system and the ultradian insulin secretory oscillations for the cases of continuous enteral nutrition and constant glucose infusion. We compare the simulation profiles obtained from this two time delay model with those from the other existing models. As a result, we notice many unique features of this two delay model. Based on our intensive simulations, we suspect that one of the possibly many causes of ultradian insulin secretion oscillations is the time delay of the insulin secretion stimulated by the elevated glucose concentration.     

Elimination kinetics of L-alanyl-L-glutamine in ICU patients

Summary. A randomised, double blind, placebo-controlled study was performed giving 0.5 g · kg⁻¹ · day⁻¹ of undiluted alanyl-glutamine (20%) or saline in a peripheral vein during 4 hours in ICU patients (n = 20). During the infusion period a steady state in plasma concentration was reached for alanyl-glutamine, but not for alanine, glutamine or glutamate. On the other hand there was no accumulation of any of the amino acids, as the pre-infusion concentrations were reached within 8 hours after the end of infusion. The half-life of the dipeptide was 0.26 hours (range, 0.15–0.63 h). The distribution volume of alanyl-glutamine was larger than the extracellular water volume, indicating a rapid hydrolysis of the dipeptide. There was no detectable alanyl-glutamine in the urine of any of the patients. All patients had excretion of small amounts of amino acids in urine, but the renal clearance of alanine, glutamine and glutamate were not different between the two groups.     

The role of growth hormone and amino acids on brain protein synthesis in aged rats given proteins of different quantity and quality

Summary. The purpose of the present study was to determine whether the regulation of brain protein synthesis was mediated through changes in the plasma concentrations of insulin and growth hormone (GH), and whether the concentrations of amino acids in the brain and plasma regulate the brain protein synthesis when the quantity and quality of dietary protein is manipulated. Two experiments were done on three groups of aged rats given diets containing 20% casein, 5% casein or 0% casein (Experiment 1), and 20% casein, 20% gluten, or 20% gelatin (Experiment 2) for 1 d (only one 5-h period) after all rats were fed the 20% casein diet for 10 d (only 5-h feeding per day). The aggregation of brain ribosomes, the concentration in plasma GH, and the branched chain amino acids in the plasma and cerebral cortex declined with a decrease of quantity and quality of dietary protein. The concentration of plasma insulin did not differ among groups. The results suggest that the ingestion of a higher quantity and quality of dietary protein increases the concentrations of GH and several amino acids in aged rats, and that the concentrations of GH and amino acids are at least partly related to the mechanism by which the dietary protein affects brain protein synthesis in aged rats.     

Dietary γ-aminobutyric acid affects the brain protein synthesis rate in young rats

Summary. The purpose of this study was to determine whether the γ-aminobutyric acid (GABA) affects the rate of brain protein synthesis in male rats. Two experiments were done on five or three groups of young rats (5 wk) given the diets containing 20% casein administrated 0 mg, 25 mg, 50 mg, 100 mg or 200 mg/100 g body weight GABA dissolved in saline by oral gavage for 1 day (d) (Experiment 1), and given the diets contained 0%, 0.25% or 0.5% GABA added to the 20% casein diet (Experiment 2) for 10 d. The plasma concentration of growth hormone (GH) was the highest in rats administrated 50 mg and 100 mg/100 g body weight GABA. The concentration of serum GABA increased significantly with the supplementation groups. The fractional (Ks) rates of protein synthesis in brain regions, liver and gastrocnemius muscle increased significantly with the 20% casein + 0.25% GABA diet and still more 20% casein + 0.5% GABA compared with the 20% casein diet. In brain regions, liver and gastrocnemius muscle, the RNA activity [g protein synthesized/(g RNA·d)] significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg RNA/g protein) was not related to the fractional rate of protein synthesis in any organ. Our results suggest that the treatment of GABA to young male rats are likely to increase the concentrations of plasma GH and the rate of protein synthesis in the brain, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis.     

A deterministic size-structured population model for the worm Naidis elinguis

In this paper we model the population dynamics of the worm Nais elinguis, which reproduces by division into two unequal parts. By using renewal theory we derive the asymptotic behaviour of a Naidis elinguis population. In particular we prove a certain relation between the fraction of the population that was born small (respectively the fraction that was born large) and the inter-division times. Received 20 January 1999 / Revised version: 1 August 1999?Published online: 10 April 2001     

Asymptotics of the total net direct pharmacological effect for large drug doses

The direct pharmacological effect E is described by the E _max model relating E to the drug plasma concentration C _p . The area under the effect vs. time curve (AUC _E ) is used as the measurement of the total net pharmacological effect. The drug plasma concentrations are solutions of compartmental systems of ordinary differential equations with the input terminated after a finite time and controlled in a proportional manner by a single dose-like parameter. The asymptotics of the time derivative of C _p for large doses are derived and used as conditions which have to be satisfied by functions for which the asymptotics of the integral defining AUC _E are derived. The AUC _E is proportional to the time T _C>EC50 for which the drug concentration stays above the threshold level EC ₅₀. The threshold EC ₅₀ denotes the drug plasma concentration which elicits 50% of the maximum effect. The parameter T _C>EC50 is proportional to the logarithm of drug dose for large doses and its asymptotics is calculated up to the order o(1) as dose increases to infinity. The results are applied to basic pharmacokinetic systems. Received: 7 December 1999 / Revised version: 23 May 2000 / Published online: 23 October 2000     

Effects of Glucose, Insulin, and Supernatant from Pancreatic β-cells on Brain–Pancreas Relative Protein in Rat Hippocampus

Brain–pancreas relative protein (BPRP) is a novel protein that mainly expresses in brain and pancreas. In our previous study, we found that various stressors significantly decreased the expression of BPRP in pancreas in vivo, accompanied by changes in insulin and glucose levels, and that expression of BPRP in pancreas also decreased significantly in diabetic rats induced by Streptozocin (STZ). All these findings suggest that BPRP may be a glucose or insulin-sensitive protein. However, how the changes in insulin or glucose levels influence the expression of BPRP in hippocampus requires further study. Here, we investigated the effects of insulin or glucose on the expression of BPRP in primary cultured hippocampal neurons. We supplied hippocampal neurons with glucose, insulin, or supernatant from pancreatic β-cells, which secrete insulin into the supernatant. Our data showed that insulin had beneficial effect on the viability while no significant effect on the expression of BPRP in hippocampal neurons. On the contrary, 40 mM glucose or free glucose culture significantly decreased the expression of BPRP, while had no significant effect on the viability and apoptosis of hippocampal neurons. Further study showed that levels of insulin in the supernatant collected from pancreatic β-cells medium changed over days, and that supernatant increased the viability of hippocampal neurons, while it had no obvious effect on the expression of BPRP in hippocampal neurons. These results suggest that BPRP may be a glucose-sensitive protein.     

An alternative stochastic model of generation of oligodendrocytes in cell culture

According to our previous model, oligodendrocyte – type 2 (O-2A) astrocyte progenitor cells become competent for differentiation in vitro after they complete a certain number of critical mitotic cycles. After attaining the competency to differentiate, progenitor cells divide with fixed probability p in subsequent cycles. The number of critical cycles is random; analysis of data suggests that it varies from zero to two. The present paper presents an alternative model in which there are no critical cycles, and the probability that a progenitor cell will divide again decreases gradually to a plateau value as the number of completed mitotic cycles increases. In particular all progenitor cells have the ability to differentiate from the time of plating. The Kiefer-Wolfowitz procedure is used to fit the new model to experimental data on the clonal growth of purified O-2A progenitor cells obtained from the optic nerves of 7 day old rats. The new model is shown to fit the experimental data well, indicating that it is not possible to determine whether critical cycles exist on the basis of these experimental data. In contrast to the fit of the previous model, which suggested that the addition of thyroid hormone increased the limiting probability of differentiation as the number of mitotic cycles increases, the fit of the new model suggests that the addition of thyroid hormone has almost no effect on the limiting probability of differentiation. Received: 6 March 2000 / Revised version: 18 September 2000 / Published online: 30 April 2001     

Suppression of myofibrillar proteolysis in chick skeletal muscles by α-ketoisocaproate

Summary. We previously reported that L-leucine suppresses myofibrillar proteolysis in chick skeletal muscles. In the current study, we compared the effects of L- and D-enantiomers of leucine on myofibrillar proteolysis in skeletal muscle of chicks. We also assessed whether leucine itself or its metabolite, α-ketoisocaproate (α-KIC), mediates the effects of leucine. Food-deprived (24 h) chicks were orally administered 225 mg/100 g body weight L-leucine, D-leucine or α-KIC and were sacrificed after 2 h. L-Leucine administration had an obvious inhibitory effect on myofibrillar proteolysis (plasma N^τ-methylhistidine concentration) in chicks while D-leucine and α-KIC were much more effective. We also examined the expression of the proteolytic-related genes (ubiquitin, proteasome, m-calpain and cathepsin B) by real-time PCR of cDNA in chick skeletal muscles. Ubiquitin mRNA expression was decreased by D-leucine and α-KIC but not L-leucine. Proteasome and m-calpain mRNA expressions as well as cathepsin B mRNA expression were likewise decreased by L-leucine, D-leucine and α-KIC. These results indicate that D-leucine and α-KIC suppress proteolytic-related genes, resulting in an decrease in myofibrillar proteolysis while L-leucine is much less effective in skeletal muscle of chicks, may be explain by conversion of D-leucine to α-KIC.     

A dynamical model for plant cell wall architecture formation

We discuss a dynamical mathematical model to explain cell wall architecture in plant cells. The highly regular textures observed in cell walls reflect the spatial organisation of the cellulose microfibrils (CMFs), the most important structural component of cell walls. Based on a geometrical theory proposed earlier [A. M. C. Emons, Plant, Cell and Environment 17, 3–14 (1994)], the present model describes the space-time evolution of the density of the so-called rosettes, the CMF synthesizing complexes. The motion of these rosettes in the plasma membrane is assumed to be governed by an optimal packing constraint on the CMFs plus adherent matrix material, that couples the direction of motion, and hence the orientation of the CMF being deposited, to the local density of rosettes. The rosettes are created inside the cell in the endoplasmatic reticulum and reach the cell-membrane via vesicles derived from Golgi-bodies. After being inserted into the plasma membrane they are assumed to be operative for a fixed, finite lifetime. The plasma membrane domains within which rosettes are activated are themselves also supposed to be mobile. We propose a feedback mechanism that precludes the density of rosettes to rise beyond a maximum dictated by the geometry of the cell. The above ingredients lead to a quasi-linear first order PDE for the rosette-density. Using the method of characteristics this equation can be cast into a set of first order ODEs, one of which is retarded. We discuss the analytic solutions of the model that give rise to helicoidal, crossed polylamellate, helical, axial and random textures, since all cell walls are composed of (or combinations of) these textures. Received: 10 July 1999 / Revised version: 7 June 2000 / Published online: 16 February 2001     

The effect of thyroid hormone status on plasma glucose-insulin interrelationship in broiler chickens

The aim of the present experiments was to study the plasma glucose-insulin relationship in eu-, hypo- or hyperthyroid broiler chickens. None of the thyroid states modified the fasting plasma glucose and insulin levels. Hypothyroid chickens exhibited a normal glucose tolerance and a normal glucose-induced insulin release after oral glucose (2 g/kg body weight) administration compared to euthyroid chickens. In contrast, hyperthyroid chickens exhibited an improved glucose tolerance accompanied by a lower insulin release. Insulin injections at a concentration of 0.1 U/kg body weight was only hypoglycemic in hyperthyroid chickens, which confirms the observation that these chickens are more sensitive to insulin. From this study it can be suggested that alterations in body composition according to thyroid status are at least partly mediated by changes in the control of carbohydrate metabolism by pancreatic hormones.     

Decrease of plasma taurine in Gaucher disease and its sustained correction during enzyme replacement therapy

Summary. Gaucher disease is caused by an autosomal-recessive deficiency of glucocerebrosidase. Cells of monocytic/macrophagic origin accumulate glucosylceramide. This leads to hepatosplenomegaly, bone destruction, thrombocytopenia and anemia. Enzyme replacement therapy (ERT) with macrophage-targeted glucocerebrosidase leads to normalization of these parameters. The way of macrophage activation in Gaucher disease is not known. Recently, the osmolytes taurine, betaine and inositol were identified as important regulators of macrophage function in liver. Therefore, the role of plasma taurine in Gaucher disease as a primarily macrophage-derived disease was studied. Fasting plasma levels were measured from blood samples of healthy control subjects (n = 29, m : f = 11 : 18, mean age 37 ± 3 years), from un-treated Gaucher patients (n = 16, m : f = 7 : 9, mean age 44 ± 4 years) and those treated for 37 ± 2 months (n = 54, m : f = 19 : 35, mean age 47 ± 2 years). Amino acid analysis was carried out in a BioChrom amino acid analyzer. In the untreated patients, plasma taurine was 45 ± 3 μM, as compared to the controls with a plasma taurine of 63 ± 4 μM (p < 0.01). The aver-age increase of plasma taurine during the first year of ERT was 18 ± 8 μM (n = 10). Patients treated for an average of 37 months (range 1–9 years of ERT) had a plasma taurine of 65 ± 4 μM (n = 54), which was not different from the controls. It is concluded that Gaucher patients show decreased plasma taurine levels and that therapy of Gaucher disease might correct this. It has to be established, whether decreased taurine availability is a cofactor of the permanent activation of glucosylceramide-storing monocytes/macrophages in this disease. Received January 25, 2000/Accepted January 31, 2000     

Reactivity and oxidative potential of fructose and glucose in enkephalin-sugar model systems

Summary. The reactions of Leu- and Met-enkephalin (Tyr-Gly-Gly-Phe-Leu/Met) with fructose resulted in the parallel formation of Heyns compounds (N-peptidyl-d-mannosamine and -d-glucosamine) and sugar-peptide generated imidazolidinone diastereomers. Glucose showed higher level of reactivity than fructose with respect to the extent of glycated product formation. The presence of fructose in the incubation mixtures makes Met residue more susceptible to oxidation than glucose. Authors’ address: Dr. Štefica Horvat, Division of Organic Chemistry and Biochemistry, Ruđer Bošković Institute, POB 180, 10002 Zagreb, Croatia