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Disorders of glucose homeostasis are common in chronic kidney disease (CKD) and are associated with increased mortality, but the mechanisms of impaired insulin secretion in this disease remain unclear. Here, we tested the hypothesis that defective insulin secretion in CKD is caused by a direct effect of urea on pancreatic β cells. In a murine model in which CKD is induced by 5/6 nephrectomy (CKD mice), we observed defects in glucose-stimulated insulin secretion in vivo and in isolated islets. Similarly, insulin secretion was impaired in normal mouse and human islets that were cultured with disease-relevant concentrations of urea and in islets from normal mice treated orally with urea for 3 weeks. In CKD mouse islets as well as ureaexposed normal islets, we observed an increase in oxidative stress and protein O-GlcNAcylation. Protein O-GlcNAcylation was also observed in pancreatic sections from CKD patients. Impairment of insulin secretion in both CKD mouse and urea-exposed islets was associated with reduced glucose utilization and activity of phosphofructokinase 1 (PFK-1), which could be reversed by inhibiting O-GlcNAcylation. Inhibition of O- GlcNAcylation also restored insulin secretion in both mouse models. These results suggest that insulin secretory defects associated with CKD arise from elevated circulating levels of urea that increase islet protein O-GlcNAcylation and impair glycolysis., Introduction Disorders of glucose homeostasis affect approximately 50% of patients suffering from chronic kidney disease (CKD) (1-4) and play a major role in their mortality (5, 6). Glucose homeostasis relies [...]

Islet amyloid polypeptide (IAPP) has been shown to form amyloid deposits in pancreatic islets, thereby furthering type 2 diabetes disease progression. Further discovery of new molecules is needed to create a diverse set of molecules that impede pancreatic amyloidosis. We have recently designed and synthesized N-phenyl-N'- (2-ethyl)ureas (EU) that are non-cytotoxic small molecules, to evaluate the role of the aryl-substituted moiety on the inhibition of hIAPP fibrillization. Several EUs were tested in vitro for their anti-amyloidogenic activity using the fluorometric ThT assay, the photo-induced cross-linking (PIUCP) assay, and cell survival assay in pancreatic MIN-6 cells. EU-362 and EU-418 were able to significantly inhibit the formation of hIAPP fibrils and protected cells from amyloid cytotoxic effects. Our results suggest that increasing the nucleophilic potency of the aryl moiety significantly enhances the anti-amyloidogenic activity of the molecules. Key words: N-phenyl-N'-(2-ethyl)urea (EU), amyloid, IAPP, diabetes, inhibitors of fibrillization. Le polypeptide amyloide des ilots pancreatiques humains (hIAPP) peut changer de conformation et former des depots dans les ilots pancreatiques, contribuant ainsi a la progression du diabete de type 2. De nouvelles molecules antiamyloidogeniques plus puissantes, presentant a la fois des proprietes adequates de biodisponibilite et une efficacite in vivo, sont a developper. Les N-phenyle-N'-(2-ethyle) urees (EU) sont de petites molecules non cytotoxiques recemment developpees dans notre laboratoire. Notre programme de decouverte en chimie medicinale implique revaluation du role des groupements aryles substitues sur l'inhibition de la formation de fibrilles provenant de l'hIAPP. Quelques EUs ont ete testees in vitro pour leur activite antiamyloidogene en utilisant la fluorescence de la thioflavine T, la liaison croisee photo-induite et la viabilite cellulaire sur les cellules pancreatiques MIN-6. EU-362 et EU-418 etaient capables d'inhiber significativement la formation de fibrilles et de proteger les cellules contre les effets toxiques de l'amyloide. De plus, EU-362 a abroge l'oligomerisation de l'hIAPP. Nos resultats suggerent qu'une augmentation du potentiel nucleophile sur le groupement aryle augmente significativement l'activite antiamyloidogenes de ces molecules. Mots-cles: N-phenyl-N'-(2-ethyl)urea (EU), amyloide, IAPP, diabete, inhibiteurs de la fibrillisation., Introduction Amyloidosis is a progressive and still incurable disease caused by various misfolded proteins that aggregate into toxic oligomers and fibrils (Sipe et al. 2014). Amyloid research over the past [...]

Byline: Sonali. Vadi, Kenneth. Yim Hypernatremia secondary to urea-induced solute diuresis is due to the renal excretion of electrolyte-free water. This concept is explained here step-wise physiologically with the help [...]

Hypertonicity increases urea transport independently of, as well as synergistically with, vasopressin in the inner medullary collect duct (IMCD). We previously showed that hypertonicity does not increase the level of cAMP in the IMCD, but it does increase the level of intracellular calcium. Since we also showed that hypertonicity increases both the phosphorylation and biotinylation of the urea transporters UT-A1 and UT-A3, this would suggest involvement of a calcium-dependent protein kinase in the regulation of urea transport in the inner medulla. In this study, we investigated whether protein kinase C (PKC), which is present in the IMCD, is a regulator of urea permeability. We tested the effect of PKC inhibitors and activators on urea permeability in the isolated, perfused rat terminal IMCD. Increasing osmolality from 290 to 690 mosmol/kg[H.sub.2]O significantly stimulated (doubled) urea permeability; it returned to control levels on inhibition of PKC with either 10 [micro]M chelerythrine or 50 [micro]M rottlerin. To determine the potential synergy between vasopressin and PKC, phorbol dibutyrate (PDBu) was used to stimulate PKC. Vasopressin stimulated urea permeability 247%. Although PDBu alone did not change basal urea permeability, in the presence of vasopressin, it significantly increased urea permeability an additional 92%. The vasopressin and PDBu-stimulated urea permeability was reduced to AVP alone levels by inhibition of PKC. We conclude that hypertonicity stimulates urea transport through a PKC-mediated phosphorylation. Whether PKC directly phosphorylates UT-A1 and/or UT-A3 or phosphorylates it as a consequence of a cascade of activations remains to be determined. tubule perfusion; calcium signaling; urine concentration; vasopressin; hypertonicity doi: 10.1152/ajprenal.00322.2010

Although supraphysiological concentrations of urea are known to increase oxidative stress in cultured cells, it is generally thought that the elevated levels of urea in chronic renal failure patients have negligible toxicity. We previously demonstrated that ROS increase intracellular protein modification by O-linked β-N-acetyl-glucosamine (O-GlcNAc), and others showed that increased modification of insulin signaling molecules by O-GlcNAc reduces insulin signal transduction. Because both oxidative stress and insulin resistance have been observed in patients with end-stage renal disease, we sought to determine the role of urea in these phenotypes. Treatment of 3T3-L1 adipocytes with urea at disease-relevant concentrations induced ROS production, caused insulin resistance, increased expression of adipokines retinol binding protein 4 (RBP4) and resistin, and increased O-GlcNAc-modified insulin signaling molecules. Investigation of a mouse model of surgically induced renal failure (uremic mice) revealed increased ROS production, modification of insulin signaling molecules by O-GlcNAc, and increased expression of RBP4 and resistin in visceral adipose tissue. Uremic mice also displayed insulin resistance and glucose intolerance, and treatment with an antioxidant SOD/catalase mimetic normalized these defects. The SOD/catalase mimetic treatment also prevented the development of insulin resistance in normal mice after urea infusion. These data suggest that therapeutic targeting of urea-induced ROS may help reduce the high morbidity and mortality caused by end-stage renal disease., Introduction Elevated urea in chronic renal failure (CRF) is considered to have negligible toxicity because Johnson et al. found in 1972 that in patients with far-advanced renal failure, blood urea [...]

Urea kinetics were measured in normal women after 5 d consuming a low protein diet [LP, 67 mg N/(kg[multiplied by]d), 0.42 9 protein/(kg [multiplied by] d)]. To determine whether the availability of methionine limits the utilization of nonessential nitrogen from low protein diets, the study was repeated on four further occasions with the addition of dietary supplements of L-methionine, 9 mg N/(kg [multiplied by] d) (LP-M); urea, 52 mg N/(kg [multiplied by] d) (LP-U); urea and methionine (LP-UM); or urea, 26 mg N/(kg [multiplied by] d), and glycine, 26 mg N/(kg [multiplied by] d), (LP-UG). Urea kinetics were derived after prime and intermittent oral doses of [15N15N]urea from the measurements of enrichment by isotope ratio mass spectrometry in urea isolated from urine. Nitrogen balance was significantly improved when the women consumed LP-U and LPUG, but not LP-M or LP-UM. The urinary excretion of 5-L-oxoproline was measured as a marker of glycine availability and was significantly lower when women consumed LP-U and LP-UG compared with either LP or LPM and LP-UM. There was a significant correlation between urinary 5-L-oxoproline and urinary sulfate excretion (r = 0.68, P = 0.00003). The availability of methionine was not limiting for nitrogen metabolism when women consumed these diets, whereas the response to supplementation with urea alone or urea with glycine showed that the availability of nonessential nitrogen was limiting. Glycine is consumed in the detoxification of excess methionine, and supplementation with methionine appeared to place a competitive demand on the availability of glycine for other metabolic processes. KEY WORDS: glycine; serine; cysteine; sulfur; women

We conducted two growth trials to evaluate the effects of monensin on amino acid sparing. When Holstein steers were fed a 90% concentrate diet supplemented with soybean meal (13.5% CP), the DMI, ADG, and efficiencies of feed and nitrogen utilization were greater than with urea (P < .10). Monensin improved ADG with both nitrogen supplements (P < .01), but the positive effects of monensin on efficiencies of feed (P = .12) and nitrogen (P = .26) utilization were greater for soybean meal than for urea. Increasing amounts of monensin (0, 11, or 22 mg/kg of DM) caused a linear increase in DMI with urea. Diets with soybean had greater intakes than diets with urea (P < .01); the greatest intake was of a soybean diet with monensin at 11 mg/kg of DM. Holstein steers fed soybean meal at 13.5% CP had lower DMI and greater efficiencies of feed and nitrogen utilization than steers fed 16.7% CP (P < .10). Crude protein level had no effect on ADG (P > .10). Monensin always increased the efficiencies of feed and nitrogen utilization (P < .05), but these trends were greater for diets with 16.7 than for those with 13.5% CP. Overall, monensin decreased DMI (P < .01), but this effect was greater for 16.7% than for 13.5% CP. Because the positive effects of monensin on diet N[E.sub.g] (P = .16) and efficiency of nitrogen utilization (P = .26) were greater for soybean meal than for urea, it seemed that monensin was sparing amino acids. Key Words: Monensin, Protein, Feed Intake, Feed Conversion, Steers

In Exp. 1, 88 yearling steers (332 kg) were fed dry-rolled corn finishing diets to evaluate effects of dietary urea level on performance and carcass characteristics. Diets contained 0, .5, 1.0, or 1.5% urea (DM basis), which supplied all supplemental N, and 10% chopped prairie hay. Gains (P = .10) and gain efficiency (G/F; P < .05) were increased by .5% urea, with little improvement by additional urea. Regression analysis estimated optimal dietary urea at .9% of DM for ADG and G/F. Fat thickness (P < .05) and yield grade (P < .10) increased linearly with dietary urea level. In Exp. 2, four ruminally and duodenally cannulated steers (557 kg) were fed the diets used in Exp. 1 to evaluate effects of dietary urea on site and extent of digestion. True ruminal OM and starch digestion were increased 25 and 37%, respectively, by .5% urea, but higher urea levels did not differ from .5%. Flows of total N and microbial N to the duodenum were not affected by urea level. In Exp. 3, 100 yearling steers (347 kg) were fed dry-rolled corn finishing diets that contained 10% alfalfa hay as the dietary roughage to evaluate effects of dietary urea level on performance and carcass characteristics. Urea levels were 0, .35, .70, 1.05, or 1.40% urea (DM basis), with no other supplemental N provided. Dry matter intake (P = .10), ADG (P < .05), and G/F (P < .05) increased with intermediate concentrations of urea but decreased with the highest concentration. Regression analysis indicated that the optimal dietary urea level was .5% of DM for ADG and G/F. Urea increased dietary energy utilization but not metabolizable protein supply. Key Words: Digestion, Animal Feeding, Growth, Urea

Effects of level and method of urea supplementation on N utilization and characteristics of digestion of a N-deficient (.85% N), citrus pulp-based diet were evaluated in a 4 x 4 Latin square design experiment, using four ruminally and intestinally cannulated crossbred (D'man x Sardi) rams. Two levels of urea (15.1 and 8.8 g/d, providing total N intakes of 12.0 and 9.1 g/d, respectively) and two methods of supplementation (urea mixed with the diet or continuously infused in the rumen) were compared in a 2 x 2 factorial arrangement of treatments. Ruminal pH varied between 6.2 and 7.0 across treatments and tended (P > .10) to be lower in animals receiving urea infusion. Ruminal ammonia concentrations were lower (P < .01) and more stable with urea infusion than with urea feeding. Ruminal VFA concentrations and profiles were unaffected (P > .10) by treatments. Site and extent of OM and fiber digestion were not influenced (P > .10) by either level of urea in the diet or method of supplementation. Average ruminal OM and ADF digestion were 53.1 and 64.5%, respectively. Total tract digestion values averaged 75.9 and 73.5%, respectively. With the exception of N efficiency (duodenal nonammonia N/ total N intake) that was 31% lower (P < .10) for the high- than for the low-urea treatment, there were no treatment responses or interactions (P > .10) on characteristics of N utilization. Microbial N flow to the small intestine averaged (across treatments) 9.0 g/d and constituted 87% of the nonammonia N leaving the abomasum. It was concluded that neither OM digestion nor microbial net synthesis or efficiency is enhanced by high levels of ruminal ammonia or by continuous N release in the rumen. Key Words: Nitrogen, Supplementation, Digestion, Sheep