Prolonged administration of secretin to normal rats increases biliary proliferation and secretin-induced ductal secretory activity

Background and aim

Cholangiocyte proliferation is coordinately regulated by a number of gastrointestinal hormones/peptides, some of which display stimulatory effects and some have inhibitory actions on cholangiocyte proliferation. Enhanced biliary proliferation [for example after bile duct ligation (BDL) and partial hepatectomy] is associated with increased expression of secretin receptor (SR), cystic fibrosis transmembrane conductance regulator (CFTR) and Cl^–/HCO₃^– anion exchanger 2 and secretin-stimulated ductal secretion, whereas loss/damage of bile ducts [for example after acute carbon tetrachloride (CCl₄) administration] is associated with reduced secretin-stimulated ductal secretory activity. There is growing information regarding the role of gastrointestinal hormones the regulation of biliary growth. For example, while gastrin, somatostatin and serotonin inhibit bile duct hyperplasia of cholestatic rats by downregulation of cAMP signaling, secretin has been shown to stimulate the proliferation of normal mice by activation of cyclic adenosine 3'',5''-monophosphate (cAMP)-dependent signaling. However, no information exists regarding the stimulatory effects of secretin on biliary proliferation of normal rats. Thus, we evaluated the in vivo and in vitro effect of secretin on biliary proliferation, the expression of markers key of ductal secretion and secretin-stimulated ductal secretion.

Methods

Normal male rats were treated with saline or secretin (2.5 nmoles/kg BW/day by osmotic minipumps for one week). We evaluated: (I) intrahepatic bile duct mass (IBDM) in liver sections and PCNA expression in purified cholangiocytes; (II) SR and CFTR mRNA expression and secretin-stimulated cAMP levels in purified cholangiocytes; and (III) secretin-stimulated bile and bicarbonate secretion in bile fistula rats. In vitro, normal rat intrahepatic cholangiocyte lines (NRIC) were treated with BSA (basal) or secretin (100 nM) for 24 to 72 hours in the absence/presence of a PKA or a MEK inhibitor before evaluating proliferation by MTS assays.

Results

Prolonged administration of secretin to normal rats increased IBDM and PCNA expression in purified cholangiocytes compared to saline-treated normal rats. Also, secretin increased the expression of proteins (SR and CFTR) that are key in the regulating ductal secretion and enhanced secretin-stimulated cAMP levels and bile and bicarbonate secretion. In vitro, secretin increased the proliferation of NRIC, increase that was prevented by PKA and MAPK inhibitors.

Conclusions

We have demonstrated that secretin stimulates both in vivo and in vitro biliary proliferation and secretin-stimulated ductal secretory activity in normal rats. We suggest that the stimulatory effect of secretin on biliary proliferation and secretion may be important for preventing biliary dysfunction during ductopenic disorders.     

Beta thalassemia minor is a beneficial determinant of red blood cell storage lesion

Blood donor genetics and lifestyle affect the quality of red blood cell (RBC) storage. Heterozygotes for beta thalassemia (bThal⁺) constitute a non-negligible proportion of blood donors in the Mediterranean and other geographical areas. The unique hematological profile of bThal⁺ could affect the capacity of enduring storage stress, however, the storability of bThal⁺ RBC is largely unknown. In this study, RBC from 18 bThal⁺ donors were stored in the cold and profiled for primary (hemolysis) and secondary (phosphatidylserine exposure, potassium leakage, oxidative stress) quality measures, and metabolomics, versus sex- and age-matched controls. The bThal⁺ units exhibited better levels of storage hemolysis and susceptibility to lysis following osmotic, oxidative and mechanical insults. Moreover, bThal⁺ RBC had a lower percentage of surface removal signaling, reactive oxygen species and oxidative defects to membrane components at late stages of storage. Lower potassium accumulation and higher uratedependent antioxidant capacity were noted in the bThal⁺ supernatant. Full metabolomics analyses revealed alterations in purine and arginine pathways at baseline, along with activation of the pentose phosphate pathway and glycolysis upstream to pyruvate kinase in bThal⁺ RBC. Upon storage, substantial changes were observed in arginine, purine and vitamin B6 metabolism, as well as in the hexosamine pathway. A high degree of glutamate generation in bThal⁺ RBC was accompanied by low levels of purine oxidation products (IMP, hypoxanthine, allantoin). The bThal mutations impact the metabolism and the susceptibility to hemolysis of stored RBC, suggesting good post-transfusion recovery. However, hemoglobin increment and other clinical outcomes of bThal⁺ RBC transfusion deserve elucidation by future studies.     

Glutamine attenuates endotoxin-induced lung metabolic dysfunction: potential role of enhanced heat shock protein 70

OBJECTIVE: Septic shock leads to derangement of cellular metabolism. Enhanced heat shock protein 70 (HSP-70) can preserve cellular metabolism after other forms of cellular stress. Glutamine (GLN) can enhance lung HSP-70 expression after lethal endotoxemia. However, it is unknown whether GLN can enhance HSP-70 expression and attenuate lung metabolic dysfunction after sublethal endotoxemia. Our aim was to determine whether GLN could upregulate HSP-70 and attenuate metabolic dysfunction in lung tissue after sublethal endotoxemia. METHODS: Sprague-Dawley rats were assigned to one of five groups. The first two groups were treated with Escherichia coli lipopolysaccharide (LPS; 1 mg/kg intravenously). GLN (0.75 g/kg intravenously) or balanced salt solution as a control was administered 5 min after LPS administration. The next two groups of rats were treated with quercetin (HSP-70 inhibitor; 400 mg/kg intraperitoneally) 6 h before LPS administration. The final group received no treatment. Lung tissue was harvested 24-h after LPS and analyzed with immunofluorescence and western blot for HSP-70. Tissue metabolites were quantified by 1H and 31P nuclear magnetic resonance spectroscopy. RESULTS: GLN compared with balanced salt solution (BSS) administration in LPS-treated animals led to significant increases in lung HSP-70. Increased HSP-70 expression was observed in lung epithelial cells and macrophages. GLN significantly improved the ratio of adenosine triphosphate to adenosine diphosphate in the lung after LPS. Quercetin inhibited a GLN-mediated increase in lung HSP-70 and blocked a beneficial effect of GLN on the ratio of adenosine triphosphate to adenosine diphosphate after LPS. CONCLUSIONS: A single dose of GLN can enhance HSP-70 in pulmonary epithelial cells and macrophages after sublethal endotoxemia. Further, GLN can attenuate endotoxin-induced lung metabolic dysfunction. GLN's beneficial effect on lung tissue after metabolic dysfunction caused by sublethal endotoxemia may be mediated in part by enhanced HSP-70.     

Dietary conjugated linoleic acid positively affects immunologic variables in lactating sows and piglets

We studied the effects of conjugated linoleic acid (CLA) on metabolic and immunologic variables in lactating sows and piglets. Gestating sows (n = 16) were assigned to 1 of 2 weight- and parity-matched groups supplemented with 0% (C) or 0.5% (T) of a CLA preparation containing 50% CLA isomers. Supplementation started in late pregnancy and continued throughout lactation. At weaning, 80 piglets, half from each group of sows, were assigned to 0% CLA (C) or 0.5% CLA (T). Thus, there were four groups of 20 piglets: C-C, C-T, T-T, and T-C. Body weight and the number of piglets per litter at birth and weaning, and the chemical composition of colostrum did not differ among the groups. CLA affected the fatty acid composition of colostrum fat; palmitoleic and gamma-linolenic acid were significantly lower compared with controls, whereas eicosenoic and eicosatrienoic acids were significantly higher. Feeding CLA increased (P < 0.05) colostrum IgG in sows. Sows fed CLA had higher (P < 0.05) serum leptin, IgG, and lysozyme. Nursing piglets from CLA-fed sows had significantly higher (P < 0.01) serum lysozyme and IgG. Consumption of CLA did not affect postweaning growth. Postweaning piglets fed CLA (T-T, C-T) had a higher IgG titer at 25 d (P < 0.05) and 35 d (P < 0.01) after weaning. Serum lysozyme was also higher at 25 d (P < 0.05) in CLA-fed piglets (T-T, C-T). At 35 d, serum alpha-1 acylglicoprotein was lower (P < 0.05) in piglets fed CLA. Dietary CLA had a positive effect on immunologic variables in lactating sows and piglets.     

Feeding and urine cotinine values in babies whose mothers smoke

In Canada, 8% to 20% of infants are breast-fed by mothers who smoke. To determine whether breast-feeding increases infants' exposure to tobacco smoke products, urinary cotinine excretion was measured in 172 babies, 33 of whom were breast-fed. A milk sample was taken from the mothers who were breast-feeding, and cotinine was measured with gas chromatography. The breast-fed babies had a median cotinine to creatinine ratio of 433 ng/mg, whereas the bottle-fed babies' median was 200 ng/mg (P less than 10(-4)). Similar differences were observed after adjustment for the number of cigarettes smoked by the mother and by other smokers in the home in the previous 24 hours. The correlation coefficient between the number of cigarettes smoked by the mother and the breast milk cotinine concentration was significant (r = .69, P = 2 X 10(-4)). Moreover, urine cotinine values from the breast-fed babies increased with higher concentrations of cotinine in the mother's milk (r = .56, P = .02). These results provide evidence that breast-feeding increases exposure to tobacco smoke components in infants whose mothers smoke. This is yet another argument for strongly encouraging women who smoke to stop smoking during pregnancy and lactation.     

Assessing opioid administration in pediatric burn patients with nonsurgical management

BackgroundDespite the vast literature studying the opioid crisis, sparse data describe this in the pediatric burn population. This study sought to assess patient-level characteristics and their potential effects on opioid administration in nonsurgical pediatric burn inpatients.MethodsAdmitted burn patients from 2013 to 2018 with nonsurgical management at an American Burn Association (ABA) verified pediatric burn center were retrospectively identified. Morphine milligram equivalents by weight (MME/kg) per admission were evaluated through a multiple loglinear regression with race, sex, age, total body surface area burned (TBSA), and burn depth as predictors. Simple linear regression was used to evaluate the temporal trend of median opioid utilization.ResultsA total of 806 patients (55% White, 35% Black, 5% Hispanic, 5% Other) were included. In an adjusted analysis, no differences in opioid administration were seen by sex, burn degree, or for Blacks and Hispanics when compared with Whites. Increased MME/kg was associated with older age (10–18 years; p < 0.0001) and larger burns (>5% TBSA burned; p < 0.0001). From 2013 to 2018, median MME/kg per admission declined significantly (2013:0.21, 2018:0.09; p = 0.0103).ConclusionsNonsurgical burn patients who were older and presented with larger TBSA experienced marked increases in opioid utilization. Overall, opioid administration decreased over time.     

Positive cumulative fluid balance at 72 h is associated with adverse outcomes following acute pediatric thermal injury

Objective

To determine the association between fluid resuscitation volume following pediatric burn injury and impact on outcomes.