Postabsorptive Phase Research Articles

Physiologically Based Biopharmaceutics Model of Vildagliptin Modified Release (MR) Tablets to Predict In Vivo Performance and Establish Clinically Relevant Dissolution Specifications.

The objective of the study was to predict pharmacokinetic (PK) and pharmacodynamic (PD) parameters of matrix-based modified release (MR) drug product of vildagliptin. Physiologically based biopharmaceutics modeling (PBBM) was developed using GastroPlus™ based on the available data including immediate-release (IR) drug product of vildagliptin. In vitro-in vivo correlation (IVIVC) was developed using mechanistic deconvolution to predict plasma concentration-time profile and PK parameters for a MR drug product planned for clinical use. Both methods, i.e., PBBM and IVIVC, were compared for the predicted PK parameters. Integration of DDDPlus™ and GastroPlus™ modeling was performed to explore clinically relevant dissolution specifications for vildagliptin MR tablets. The bioequivalence (BE) between batches with different dissolution specifications was evaluated using virtual clinical trials. The PD effect of dipeptidyl peptidase-IV (DPP-IV) inhibition was simulated utilizing PDPlus™ model in GastroPlus™. The results indicated that IVIVC best correlated the simulated PK parameters with those observed with the clinical study. The outcomes highlight the importance of integration of in vitro and in silico tools towards predictability of PK and PD parameters for a MR drug product. However, the post absorptive phase was found to be more dependent on the demographics of the healthy subjects.

Renal Tubular Handling of Glucose and Fructose in Health and Disease.

The proximal tubule of the kidney is programmed to reabsorb all filtered glucose and fructose. Glucose is taken up by apical sodium-glucose cotransporters SGLT2 and SGLT1 whereas SGLT5 and potentially SGLT4 and GLUT5 have been implicated in apical fructose uptake. The glucose taken up by the proximal tubule is typically not metabolized but leaves via the basolateral facilitative glucose transporter GLUT2 and is returned to the systemic circulation or used as an energy source by distal tubular segments after basolateral uptake via GLUT1. The proximal tubule generates new glucose in metabolic acidosis and the postabsorptive phase, and fructose serves as an important substrate. In fact, under physiological conditions and intake, fructose taken up by proximal tubules is primarily utilized for gluconeogenesis. In the diabetic kidney, glucose is retained and gluconeogenesis enhanced, the latter in part driven by fructose. This is maladaptive as it sustains hyperglycemia. Moreover, renal glucose retention is coupled to sodium retention through SGLT2 and SGLT1, which induces secondary deleterious effects. SGLT2 inhibitors are new anti-hyperglycemic drugs that can protect the kidneys and heart from failing independent of kidney function and diabetes. Dietary excess of fructose also induces tubular injury. This can be magnified by kidney formation of fructose under pathological conditions. Fructose metabolism is linked to urate formation, which partially accounts for fructose-induced tubular injury, inflammation, and hemodynamic alterations. Fructose metabolism favors glycolysis over mitochondrial respiration as urate suppresses aconitase in the tricarboxylic acid cycle, and has been linked to potentially detrimental aerobic glycolysis (Warburg effect). © 2022 American Physiological Society. Compr Physiol 12:2995-3044, 2022.

Renal gluconeogenesis in insulin resistance: A culprit for hyperglycemia in diabetes.

Renal gluconeogenesis is one of the major pathways for endogenous glucose production. Impairment in this process may contribute to hyperglycemia in cases with insulin resistance and diabetes. We reviewed pertinent studies to elucidate the role of renal gluconeogenesis regulation in insulin resistance and diabetes. A consensus on the suppressive effect of insulin on kidney gluconeogenesis has started to build up. Insulin-resistant models exhibit reduced insulin receptor (IR) expression and/or post-receptor signaling in their kidney tissue. Reduced IR expression or post-receptor signaling can cause impairment in insulin’s action on kidneys, which may increase renal gluconeogenesis in the state of insulin resistance. It is now established that the kidney contributes up to 20% of all glucose production via gluconeogenesis in the post-absorptive phase. However, the rate of renal glucose release excessively increases in diabetes. The rise in renal glucose release in diabetes may contribute to fasting hyperglycemia and increased postprandial glucose levels. Enhanced glucose release by the kidneys and renal expression of the gluconeogenic-enzyme in diabetic rodents and humans further point towards the significance of renal gluconeogenesis. Overall, the available literature suggests that impairment in renal gluconeogenesis in an insulin-resistant state may contribute to hyperglycemia in type 2 diabetes.

Coordinated regulation of miR-27 by insulin/CREB/Hippo contributes to insulin resistance

MicroRNA-27 is a critical non-coding metabolic gene that is often aberrantly overexpressed in non-alcoholic fatty livers (NAFLD). However, the pathogenic role of miR-27 in NAFLD remains unknown. In this study, we attempted to identify the mechanism by which miR-27 was regulated in the context of insulin resistance, a predisposed metabolic disorder in NAFLD. Our data from cell culture and animal studies showed that insulin, CREB, and Hippo signalings coordinately regulated miR-27. First, miR-27 was upregulated in palmitate-treated cells and high fat diet-fed mouse livers, which exhibited insulin resistance and CREB overexpression. Second, miR-27 peaked in the mouse liver at the post-absorptive phase when CREB activity was increased. Also, miR-27 was increased rapidly in cell lines when CREB was deactivated by insulin treatment. Third, miR-27 was decreased in cultured cells when CREB was downregulated by siRNA or metformin treatment. In contrast, Forskolin-mediated activation of CREB promoted miR-27 expression. Fourth, Hippo signaling repressed miR-27 in a CREB-independent manner: miR-27 was reduced in cells at full confluence but was inhibited in cells transfected with siRNA against Lats2 and Nf2, which were two positive regulators of Hippo signaling. Lastly, bioinformatics and luciferase assay showed that miR-27 inhibited Akt phosphorylation by targeting Pdpk1 and Pik3r1. Overexpression of miR-27 impaired Akt phosphorylation in cell lines and primary mouse hepatocytes upon insulin stimulation. In conclusion, our data suggest that insulin, CREB, and Hippo signalings contribute to aberrant miR-27 overexpression and eventually lead to insulin resistance in NAFLD.

Renal gluconeogenesis: an underestimated role of the kidney in systemic glucose metabolism.

Glucose levels are tightly regulated at all times. Gluconeogenesis is the metabolic pathway dedicated to glucose synthesis from non-hexose precursors. Gluconeogenesis is critical for glucose homoeostasis, particularly during fasting or stress conditions. The renal contribution to systemic gluconeogenesis is increasingly recognized. During the post-absorptive phase, the kidney accounts for ∼40% of endogenous gluconeogenesis, occurring mainly in the kidney proximal tubule. The main substrate for renal gluconeogenesis is lactate and the process is regulated by insulin and cellular glucose levels, but also by acidosis and stress hormones. The kidney thus plays an important role in the maintenance of glucose and lactate homoeostasis during stress conditions. The impact of acute and chronic kidney disease and proximal tubular injury on gluconeogenesis is not well studied. Recent evidence shows that in both experimental and clinical acute kidney injury, impaired renal gluconeogenesis could significantly participate in systemic metabolic disturbance and thus alter the prognosis. This review summarizes the biochemistry of gluconeogenesis, the current knowledge of kidney gluconeogenesis, its modifications in kidney disease and the clinical relevance of this fundamental biological process in human biology.

Glucose transporters in the kidney in health and disease.

The kidneys filter large amounts of glucose. To prevent the loss of this valuable fuel, the tubular system of the kidney, particularly the proximal tubule, has been programmed to reabsorb all filtered glucose. The machinery involves the sodium-glucose cotransporters SGLT2 and SGLT1 on the apical membrane and the facilitative glucose transporter GLUT2 on the basolateral membrane. The proximal tubule also generates new glucose, particularly in the post-absorptive phase but also to enhance bicarbonate formation and maintain acid-base balance. The glucose reabsorbed or formed by the proximal tubule is primarily taken up into peritubular capillaries and returned to the systemic circulation or provided as an energy source to further distal tubular segments that take up glucose by basolateral GLUT1. Recent studies provided insights on the coordination of renal glucose reabsorption, formation, and usage. Moreover, a better understanding of renal glucose transport in disease states is emerging. This includes the kidney in diabetes mellitus, when renal glucose retention becomes maladaptive and contributes to hyperglycemia. Furthermore, enhanced glucose reabsorption is coupled to sodium retention through the sodium-glucose cotransporter SGLT2, which induces secondary deleterious effects. As a consequence, SGLT2 inhibitors are new anti-hyperglycemic drugs that can protect the kidneys and heart from failing. Recent studies discovered unique roles for SGLT1 with implications in acute kidney injury and glucose sensing at the macula densa. This review discusses established and emerging concepts of renal glucose transport, and outlines the need for a better understanding of renal glucose handling in health and disease.

Emerging Role of Lymphatics in the Regulation of Intestinal Lipid Mobilization.

Intestinal handling of dietary triglycerides has important implications for health and disease. Following digestion in the intestinal lumen, absorption, and re-esterification of fatty acids and monoacylglycerols in intestinal enterocytes, triglycerides are packaged into lipoprotein particles (chylomicrons) for secretion or into cytoplasmic lipid droplets for transient or more prolonged storage. Despite the recognition of prolonged retention of triglycerides in the post-absorptive phase and subsequent release from the intestine in chylomicron particles, the underlying regulatory mechanisms remain poorly understood. Chylomicron secretion involves multiple steps, including intracellular assembly and post-assembly transport through cellular organelles, the lamina propria, and the mesenteric lymphatics before being released into the circulation. Contrary to the long-held view that the intestinal lymphatic vasculature acts mainly as a passive conduit, it is increasingly recognized to play an active and regulatory role in the rate of chylomicron release into the circulation. Here, we review the latest advances in understanding the role of lymphatics in intestinal lipid handling and chylomicron secretion. We highlight emerging evidence that oral glucose and the gut hormone glucagon-like peptide-2 mobilize retained enteral lipid by differing mechanisms to promote the secretion of chylomicrons via glucose possibly by mobilizing cytoplasmic lipid droplets and via glucagon-like peptide-2 possibly by targeting post-enterocyte secretory mechanisms. We discuss other potential regulatory factors that are the focus of ongoing and future research. Regulation of lymphatic pumping and function is emerging as an area of great interest in our understanding of the integrated absorption of dietary fat and chylomicron secretion and potential implications for whole-body metabolic health.

5334000-53340000Optimal Salinity for Head-Starting Northern River Terrapins (Batagur baska Gray, 1831).

Simple SummaryNorthern river terrapins can adapt physiologically or behaviorally across a salinity gradient, so that they can move between freshwater habitats and brackish water or estuaries. However, there is no available information on the optimal salinity of this species for supporting captive husbandry programs before release to natural habitat. In the current study, the optimal salinity for the terrapins was investigated to serve the well-being of turtles. At the end of the rearing period, various observed parameters related to growth, feed utilization, and health indicated that optimal salinity for rearing the terrapins is about 4 ppt, while rearing in freshwater (0 ppt) or other brackish (8 and 12 ppt) water conditions had negative effects on some key parameters that reflect well-being. A polynomial regression fit to terrapin weight gain as function of salinity predicts the optimal salinity as 4.35 ppt. Findings from the current study could be directly used in ex situ conservation programs of northern river terrapins before release to natural habitat. Additionally, concurrent conservation and restoration of the wild habitats with preferred salinity is essential for flourishing terrapin population.Northern river terrapins (Batagur baska Gray, 1831) are Asia’s largest turtles living in both freshwater and brackish water. In the current study, the optimal salinity for head-starting programs of this critically endangered species was investigated in order to serve the well-being of turtles before release to natural habitat. Forty-eight terrapins (54.64 ± 0.18 g initial body weight) were randomly distributed to four salinity levels (0, 4, 8, and 12 ppt) and reared for eight weeks, using three replicates with four terrapins each. At the end of rearing trial, growth performance and feed utilization parameters were superior in terrapins reared at 4 ppt, followed by 8 ppt in the rank order of treatments. Negative stress responses were observed in terrapins reared at 12 ppt, as the fecal activity of amylase-to-trypsin ratio was changed significantly, but not that of proteolytic enzymes. The fecal thermal transition properties indicated an abundance of nutrients in the post-absorptive phase for terrapins reared at 4 ppt, followed by the 8 ppt treatment group. The preferred 4 ppt salinity had no negative effects on the health status of the terrapins in terms of carapace elemental composition or hematological parameters. Second-order polynomial regression suggests 4.35 ppt as the optimal salinity for maximal weight gain. Findings from the current study could be directly used in ex situ conservation programs of northern river terrapins before release to natural habitat.

Significant effects of late evening snack on liver functions in patients with liver cirrhosis: A meta-analysis of randomized controlled trials.

Reducing post-absorptive (fasting) phase by eating late evening snacks (LESs) is a potential intervention to improve substrate utilization and reverse sarcopenia. This study analyzed the results of published randomized controlled trials and controlled clinical trials to evaluate the effects of LES on liver function of patients with cirrhosis. A meta-analysis was conducted. The search strategy included electronic database searches, and 300 articles were searched. Eight of these articles provided qualified data for pooling and analysis. Outcomes assessments included serum albumin, total bilirubin, alanine aminotransferase, prothrombin time, and aspartate aminotransferase, complications of cirrhosis, severity of liver disease, and blood glucose levels. Our analysis included eight studies comprising 341 patients (167 in LES groups and 174 in control groups). The results showed that LES intervention helped to maintain liver reserves. These eight studies demonstrated that LES intervention had significant effects for liver biochemical parameters on albumin, ammonia, and prothrombin time, with respective effect sizes of 0.233, -0.425, and -0.589; liver enzymes include aspartate aminotransferase and alanine aminotransferase, with respective effect sizes of -0.320 and -0.284. Studies on clinical signs of liver dysfunction showed lower occurrence rates of ascites and hepatic encephalopathy than in the control group. LES had no significant effect on Child-Pugh score. The overall results of the meta-analysis indicated that having LES can improve liver function reserve for patients with liver cirrhosis, with or without hepatocellular carcinoma. LES is a promising intervention for reversing anabolic resistance and the sarcopenia of cirrhosis, resulting in an improved quality of life for patients with cirrhosis.

Modeling the effect of the cephalic phase of insulin secretion on glucose metabolism

The nervous system has a significant impact in glucose homeostasis and endocrine pancreatic secretion in humans, especially during the cephalic phase of insulin release (CPIR); that is, before a meal is absorbed. However, the underlying mechanisms of this neural-pancreatic interaction are not well understood and therefore often neglected, despite their significance to achieving an optimal glucose control. As a result, the dynamics of insulin release from the pancreas are currently described by mathematical models that reproduce the behavior of the β cells using exclusively glucose levels and other hormones as inputs. To bridge this gap, we have combined, for the first time, metabolic and neural mathematical models in a unified system to reproduce to a great extent the ideal glucoregulation observed in healthy subjects. Our results satisfactorily replicate the CPIR and its impact during the post-absorptive phase. Furthermore, the proposed model gives insight into the physiological interaction between the brain and the pancreas in healthy people and suggests the potential of considering the neural information for restoring glucose control in people with diabetes.Graphical (a) Physiological scenario. Diagram of the biological interaction among the most important organs involved in glucose control during meal intake. (b) Scheme of the unified bio-inspired neural-metabolic model. Each of the boxes represents one subsystem of the model. The pink shades boxes depicts the novel subsystems introduced to the current metabolic models (grey shaded boxes). Insulin-related action and mass fluxes (solid black lines) and glucose-related action and mass flux (dotted black lines) are depicted to show the relationship among the blocks. I(t), Ic(t), G(t) and SI related to plasma insulin, plasma cephalic insulin, plasma glucose and insulin sensitivity, respectively.

A central role for hypoxia-inducible factor (HIF)-2α in hepatic glucose homeostasis.

Hepatic glucose production is regulated by hormonal and dietary factors. At fasting, 80% of glucose released into the circulation is derived from the liver, among which gluconeogenesis accounts for 55% and the rest by glycogenolysis. Studies suggest a complex mechanism involved in the regulation of hepatic glucose metabolism during fasting and post-absorptive phase. Oxygen plays a key role in numerous metabolic pathways such as TCA cycle, gluconeogenesis, glycolysis and fatty acid oxidation. Oxygenation of the gastrointestinal tract including liver and intestine is dynamically regulated by changes in the blood flow and metabolic activity. Cellular adaptation to low oxygen is mediated by the transcription factors HIF-1α and HIF-2α. HIF-1α regulates glycolytic genes whereas HIF-2α is known to primarily regulate genes involved in cell proliferation and iron metabolism. This review focuses on the role of the oxygen sensing signaling in the regulation of hepatic glucose output with an emphasis on hypoxia inducible factor (HIF)-2α. Recent studies have established a metabolic role of HIF-2α in systemic glucose homeostasis. Understanding the HIF-2α dependent mechanism in hepatic metabolism will greatly enhance our potential to utilize the oxygen sensing mechanisms to treat metabolic diseases.

Cephalic phase of insulin secretion in response to a meal is unrelated to family history of type 2 diabetes.

The pre-absorptive cephalic phase of insulin secretion is elicited during the first ten min of a meal and before glucose levels rise. Its importance for insulin release during the post-absorptive phase has been well documented in animals but its presence or importance in man has become increasingly controversial. We here examined the presence of an early cephalic phase of insulin release in 31 well matched individuals without (n = 15) or with (n = 16) a known family history of type 2 diabetes (first-degree relatives; FDR). We also examined the potential differences in individuals with or without impaired fasting (IFG) and impaired glucose tolerance (IGT). We here demonstrate that a cephalic phase of insulin secretion was present in all individuals examined and without any differences between control persons and FDR or IFG/IGT. However, the overall importance of the cephalic phase is conjectural since it was unrelated to the subsequent post-absorptive insulin release or glucose tolerance. One of the best predictors of the incremental cephalic phase of insulin release was fasting insulin level and, thus, a relation to degree of insulin sensitivity is likely. In conclusion, an early pre-absorptive and cephalic phase of insulin release is robustly present in man. However, we could not document any relation to family history of Type 2 diabetes nor to the post-absorptive phase and, thus, confirm its importance for subsequent degree of insulin release or glucose tolerance.

Evaluation of Acute Alcohol Intoxication as the Primary Cause of Death: A Diagnostic Challenge for Forensic Pathologists.

Deaths caused by acute alcohol intoxication (AAI) remain a major public health issue. This study is retrospective and descriptive: an 8-year case analysis of deaths due to AAI in Maryland. Study showed that of 150 AAI deaths, the death rate among Hispanics (10.41/100,000 population) was significantly higher than all the non-Hispanics combined (1.88/100,000 population). The majority of individuals were young adults, overweight, and binge drinkers. The obese group showed significantly lower mean heart and peripheral blood alcohol concentration (BAC) (0.36%, 0.37%) than the normal weight group (0.45%, 0.42%). Based on the PBAC and urine AC ratio, 49.6% deaths likely occurred close to peak phase, followed by postabsorptive phase (31.6%) and absorptive phase (18.8%). Our results indicate that forensic pathologists should evaluate postmortem BAC in the light of individual's age, drinking history, body weight, possible phase of alcohol intoxication, and other autopsy findings when certifying AAI as primary cause of death.

Effects of polydextrose with breakfast or with a midmorning preload on food intake and other appetite-related parameters in healthy normal-weight and overweight females: An acute, randomized, double-blind, placebo-controlled, and crossover study

Polydextrose (PDX) reduces subsequent energy intake (EI) when administered at midmorning in single-blind trials of primarily normal-weight men. However, it is unclear if this effect also occurs when PDX is given at breakfast time. Furthermore, for ecological validity, it is desirable to study a female population, including those at risk for obesity. We studied the effects of PDX, served as part of a breakfast or midmorning preload, on subsequent EI and other appetite-related parameters in healthy normal-weight and overweight females. Per earlier studies, the primary outcome was defined as the difference in subsequent EI when PDX was consumed at midmorning versus placebo. Thirty-two volunteers were enrolled in this acute, double-blind, placebo-controlled, randomized, and crossover trial to examine the effects of 12.5 g of PDX, administered as part of a breakfast or midmorning preload, on subsequent EI, subjective feelings of appetite, well-being, and mood. Gastric emptying rates and the blood concentrations of glucose, insulin, cholecystokinin, ghrelin, glucagon-like peptide 1 (GLP-1), and peptide tyrosine-tyrosine were measured in the group that received PDX as part of their breakfast. There were no differences in EI between volunteers who were fed PDX and placebo. PDX intake with breakfast tended to elevate blood glucose (P = 0.06) during the postabsorptive phase, significantly lowered insulin by 15.7% (P = 0.04), and increased GLP-1 by 39.9% (P = 0.02); no other effects on blood parameters or gastric emptying rates were observed. PDX intake at midmorning reduced hunger by 31.4% during the satiation period (P = 0.02); all other subjective feelings of appetite were unaffected. Volunteers had a uniform mood profile during the study. PDX was well tolerated, causing one mild adverse event throughout the trial.

Effect of heat stress on the serum concentrations of free amino acids and some of their metabolites in growing pigs.

Exposure to heat stress (HS) may affect the intestinal epithelia of pigs, resulting in impaired digestive and absorptive capacity. The serum concentration (SC) of free AA in pigs can be used as indicators of their availability. This study was conducted with 12 crossbred (Landrace × Hampshire × Duroc) pigs (29.0 ± 2.8 kg initial BW) distributed into 2 groups to analyze the SC of free AA and some AA metabolites in pigs exposed to HS conditions. The treatments were pigs housed under natural HS conditions in a room with no ambient temperature control (23.6 to 37.6°C, HS) and pigs housed at thermoneutral conditions (24 ± 2°C), feed restricted to a level similar to that of their HS counterparts. All pigs received a wheat-soybean meal diet. Blood samples were collected at both the absorptive (2.5 h after a meal) and postabsorptive (10.0 h after a meal) phase. At the absorptive phase, the SC of free Arg, Leu, Lys, Phe, Thr, and Trp were lower ( < 0.05) and the SC of His, Val, Ala, Pro, Ser, and Tyr tended to be lower ( < 0.10) in HS pigs. At the postabsorptive phase, the SC of free Arg, His, Met, Asn, Gln, and Tyr were higher ( < 0.05) but the SC of Ala was lower ( < 0.01) and the SC of Pro tended to be lower ( < 0.10) in HS pigs. The absorptive SC of carnosine, ornithine (Orn), and Tau were lower ( < 0.05) and of citrulline (Cit), cystathionine, and urea tended to be lower ( < 0.10) in HS pigs. The postabsorptive SC of 3-methyl-His, homo-Cys, OH-Lys, and OH-Pro increased ( = 0.05) and of Cit tended to increase ( = 0.10) but that of carnosine and sarcosine ( < 0.05) decreased in HS pigs. The results of this study show a marked and differential effect of HS on the SC of AA. These data indicate that HS negatively affects the digestive and absorptive capacity of pigs and that the metabolism of some AA is modified in pigs to counteract the negative effects of the HS.

Sustained effects of a protein and lipid preload on glucose tolerance in type 2 diabetes patients

BackgroundSmall amounts of nutrients given as a ‘preload’ can reduce post-meal hyperglycaemic peaks in type 2 diabetes (T2D) patients by activating a number of mechanisms involved in glucose homoeostasis. This study was undertaken to ascertain whether this positive effect extends to the late absorptive phase and to identify the main mechanisms involved. Material and methodsEight well-controlled T2D patients, aged 40–70 years, were randomized to consume a ‘preload’ of either water or non-glucidic nutrients (50g of Parmesan cheese, one boiled egg) 30min before a 300-min oral glucose tolerance test. ResultsAfter the nutrient preload, significant reductions were observed in peak glucose (−49%; P<0.02), total plasma glucose (iAUC: −28%; P<0.03), exogenous glucose (iAUC: −30%; P<0.03) and insulin clearance (−28%; P<0.04), with enhancement of insulin secretion (iAUC: +22%; P<0.003). These effects were associated with higher plasma levels of GLP-1 (iAUC: +463%; P<0.002), GIP (iAUC: +152%; P<0.0003) and glucagon (iAUC: +144%; P<0.0002). ConclusionIn T2D patients, a protein and lipid preload improves glucose tolerance throughout the whole post-absorptive phase mainly by reducing the appearance of oral glucose, and improving both beta-cell function and insulin bioavailability.

Effect of dietary excess of branched-chain amino acids on performance and serum concentrations of amino acids in growing pigs.

Depressed performance and availability of some amino acids (AA) in pigs fed excess Leu diets appear to be related to lower feed intake. Surplus Ile and Val may help to overcome this effect. An experiment was conducted with 24 pigs (31.8 ± 1.2 kg initial BW) to evaluate the effect of dietary excess of either Leu alone or with surplus Ile and Val on performance and serum concentration (SC) of essential AA. Treatments were as follows: T1, basal diet; T2, basal plus 0.43% L-Leu (excess Leu); T3, basal added with 0.43% L-Leu, plus 0.20% L-Ile and 0.25% L-Val (excess LIV). The basal diet was formulated to contain 0.90% standardized ileal digestible Lys and added with crystalline L-Lys, L-Thr, DL-Met, L-Trp, L-Leu, L-Ile, L-His and L-Val to create essential AA:Lys ratios close to an ideal protein for growing pigs. All pigs were fed the same amount of feed twice a day (average, 3.42× the requirement of NEm). Blood samples were collected at 2.5 (absorptive) and 11.0 h (post-absorptive) post-prandial to analyse SC of AA. Excess of either Leu or LIV did not affect growth rate nor feed conversion. Excess Leu increased Leu SC and decreased Ile and Val SC (p < 0.05) at both absorptive and post-absorptive phases, but excess LIV restored the SC of Ile and Val. The SC of other essential AA was not affected by excess of either Leu or LIV. The SC of all AA during absorptive, on average, was about two times higher than that of post-absorptive phase. These results suggest that the reduced availability (SC) of Ile and Val in pigs consuming excess Leu diets is attributed to a reduced absorption and increased cellular degradation rates of them.

Cause and manner of death and phase of the blood alcohol curve

In a large number of forensic autopsies (N=28,184) the concentrations of ethanol in femoral blood and bladder urine were determined and the urine-to-blood concentration ratios of ethanol were calculated. Based on the differences in ethanol concentration between urine and blood, the deaths were classified as having occurred during the absorptive, the peak or the post-absorptive phase of the blood–alcohol curve. Most people died in the post-absorptive phase, N=24,223 (86%), whereas 1538 individuals (5.5%) were still absorbing alcohol and 2423 (8.6%) were at or close to the peak BAC at time of death. Both blood–alcohol concentration (BAC) and urine–alcohol concentration (UAC) were significantly higher in the post-absorptive phase (p<0.001). The proportions of people dying in the absorptive and peak phases increased with advancing age. The cause of death (CoD) and manner of death (MoD) according to death certificates were compared with phase of the blood–alcohol curve using a multinomial regression model with and without making adjustment for possible effects of age, gender and BAC. The relative risk (RR) and relative risk ratios (RRR) showed some associations between CoD and phase of the blood–alcohol curve. Undetermined MoD was significantly higher in the absorptive phase compared with the post-absorptive phase (RRR=2.12). Deaths related to esophagus, stomach and duodenum (RRR=2.04) and alcoholic liver diseases (RRR=1.85) were significantly higher at or close to peak phase compared to the post-absorptive phase. Road-traffic fatalities were more prevalent in the peak BAC phase (RRR=1.33) and deaths by accidental falls were less in the absorptive phase (RRR=0.58) compared with the post-absorptive phase. The phase of alcohol intoxication seems relevant to consider by forensic experts when alcohol-related deaths are investigated.

Neural regulation of pancreatic islet cell mass and function.

Intracellular glucose signalling pathways control the secretion of glucagon and insulin by pancreatic islet α- and β-cells, respectively. However, glucose also indirectly controls the secretion of these hormones through regulation of the autonomic nervous system that richly innervates this endocrine organ. Both parasympathetic and sympathetic nervous systems also impact endocrine pancreas postnatal development and plasticity in adult animals. Defects in these autonomic regulations impair β-cell mass expansion during the weaning period and β-cell mass adaptation in adult life. Both branches of the autonomic nervous system also regulate glucagon secretion. In type 2 diabetes, impaired glucose-dependent autonomic activity causes the loss of cephalic and first phases of insulin secretion, and impaired suppression of glucagon secretion in the postabsorptive phase; in diabetic patients treated with insulin, it causes a progressive failure of hypoglycaemia to trigger the secretion of glucagon and other counterregulatory hormones. Therefore, identification of the glucose-sensing cells that control the autonomic innervation of the endocrine pancreatic and insulin and glucagon secretion is an important goal of research. This is required for a better understanding of the physiological control of glucose homeostasis and its deregulation in diabetes. This review will discuss recent advances in this field of investigation.

Ethanol elimination kinetics following massive ingestion in an ethanol naive child

At low-to-moderate concentrations, ethanol elimination follows zero-order kinetics. It is unknown whether renal, pulmonary or other first-order processes become significant in patients with very high serum ethanol concentrations. Additionally, it is unclear whether ethanol naive subjects induce their metabolism during acute intoxication. We present the toxicokinetic analysis in a child with a massive ingestion of ethanol. A 15-year-old girl without significant medical history presented to the Emergency Department after drinking 24 ounces of tequila. She was found unresponsive at home with a Glasgow Coma Score of 3. Her presenting vitals were as follows: 118/69 mmHg blood pressure; pulse rate was 88 beats per minute; respiratory rate of 20 breaths per minute; pulse-oximetry is 96% on room air. Other than obtundation, her physical examination was normal. She was intubated for airway protection and admitted to the ICU. Her initial serum ethanol concentration was 543 mg/dL. A repeat level 3 h later was 722 mg/dL. Post-absorptive ethanol concentrations decreased from 693 mg/dL to 291 mg/dL over the following 15.5 h. The patient had spontaneous eye opening 24 h after presentation. Her projected serum ethanol concentration at that time was 215 mg/dL. She was extubated 2 h later and had an uneventful recovery. The elimination of ethanol in the post-absorptive phase remained zero-order at a rate of 26.3 mg/dL/h (5.7 mmol/L/h) with a Pearson's correlation coefficient (R (2)) of 0.9968 (p < 0.01). There was no evidence of acute induction in metabolism although pharmacodynamic tolerance likely occurred. Even at very high ethanol concentrations in ethanol naive subjects, elimination of ethanol follows a zero-order toxicokinetic model.