Urea Infusion Research Articles

Noninvasively differentiating acute and chronic nephropathies via multiparametric urea-CEST, nuclear Overhauser enhancement-CEST, and quantitative magnetization transfer MRI.

Standardized blood tests often lack adequate sensitivity and specificity to capture the gradual progression of renal injuries. We suggest a multiparametric molecular MRI approach as a noninvasive tool for monitoring renal function loss and distinguishing different types of renal injuries. CEST and quantitative magnetization transfer (qMT) imaging were performed on cisplatin (n=16) and aristolochic acid (AA)-induced nephropathy (n=22) mouse models at 7T with an infusion of either saline or urea. Seven-pool Lorentzian fitting was applied for the analysis of CEST Z-spectra, and the T1 -corrected CEST contrast apparent exchange-dependent relaxation (AREX) from urea (+1ppm) and two nuclear Overhauser enhancement (NOE) pools (-1.6 and -3.5ppm) were measured. Similarly, qMT spectra were fitted into two-pool Ramani equation and the relative semi-solid macromolecular pool-size ratio was measured. Histology of mouse kidneys was performed to validate the MR findings. AA model showed disrupted spatial gradients of urea in the kidney and significantly decreased NOE CEST and qMT contrast. The cisplatin model showed slightly decreased qMT contrast only. The orrelation of MR parameters to histological features showed that NOE CEST and qMT imaging are sensitive to both acute and chronic injuries, whereas urea CEST shows a significant correlation only to acute injuries. These results indicate that our multiparametric approach allows comprehensive and totally noninvasive monitoring of renal function and histological changes for distinguishing different nephropathies.

Analysis of nano material based Zeolite catalytic converter and urea injection in single cylinder engine fuelled with diesel and plastic pyrolysis oil

The consistent state movement of Zeolite 4A and 5A with urea infusion framework has been researched tentatively for Nitrogen oxide, Hydrocarbon, Carbon monoxide & smoke with exceptionally oxidizing catalysts, for ordinary lean consumed motor exhaust. The ZSM powder is added to 8 % of bentonite earth, 5 % of carboxymethyl cellulose and 34 % of refined water & blended continually to shape like a trim sand. Carboxymethyl cellulose is used which has good binding property. The mold is placed in the oven at 150 °C with an increment of 75 °C each 45 mins until the temperature arrived at 450 °C then it has been saved for 60 mins for calcination. Most of the developing countries depend of crude oil because diesel is used as a main transportation fuel. Finding an alternative fuel to replace diesel is an important need, plastic pyrolysis oil is suitable for compression ignition engines. The property of plastic oil is similar to diesel fuel, Emission test has been performed with a combination of 50 % Diesel + 50 % Plastic Oil in direct injection engines and compared with Diesel. Our objective is to investigate the possibility of reducing NOx emissions in tail pipe. Reduction of oxides of nitrogen (NOx) was carried out by injecting urea in the tail pipe of a diesel engine. Urea has been infusion to accomplish NOx decrease with Zeolite impetus powered with Diesel fuel and Diesel + Plastic Pyrolysis Oil.

Urea supplementation in rumen and post-rumen for cattle fed a low-quality tropical forage.

We evaluated the differences between the supplementation of urea in rumen and/or abomasum on forage digestion, N metabolism and urea kinetics in cattle fed a low-quality tropical forage. Five Nellore heifers were fitted with rumen and abomasum fistulas and assigned to a Latin square design. The treatments were control, continuous infusion of urea in the abomasum (AC), continuous infusion of urea in the rumen, a pulse dose of urea in the rumen every 12 h (PR) and a combination of PR and AC. The control exhibited the lowest (P < 0·10) faecal and urinary N losses, which were, overall, increased by supplementation. The highest urinary N losses (P < 0·10) were observed when urea was either totally or partially supplied as a ruminal pulse dose. The rumen N balance was negative for the control and when urea was totally supplied in the abomasum. The greatest microbial N production (P < 0·10) was obtained when urea was partially or totally supplied in the abomasum. Urea supplementation increased (P < 0·10) the amount of urea recycled to the gastrointestinal tract and the amount of urea-N returned to the ornithine cycle. The greatest (P < 0·10) amounts of urea-N used for anabolism were observed when urea was totally and continuously infused in the abomasum. The continuous abomasal infusion also resulted in the highest (P < 0·10) assimilation of microbial N from recycling. The continuous releasing of urea throughout day either in the rumen or abomasum is able to improve N accretion in the animal body, despite mechanism responsible for that being different.

Influence of forage level and corn grain processing on whole-body urea kinetics, and serosal-to-mucosal urea flux and expression of urea transporters and aquaporins in the ovine ruminal, duodenal, and cecal epithelia.

The objectives were to determine the effects of forage level and grain processing on whole-body urea kinetics, N balance, serosal-to-mucosal urea flux (Jsm-urea), and messenger ribonucleic acid (mRNA) abundance of urea transporter-B (UT-B; SLC14a1) and aquaporins (AQP) in ovine ruminal, duodenal, and cecal epithelia. Thirty-two wether lambs were blocked by body weight into groups of four and assigned to one of four diets (n = 8) in a 2 × 2 factorial design. Dietary factors were forage level (30% [LF] vs. 70% [HF]) and corn grain processing (whole-shelled [WS] vs. steam-flaked [SF]). Four blocks of lambs (n = 4) were used to determine urea kinetics and N balance using 4-d [15N15N]-urea infusions with concurrent fecal and urine collections. Lambs were killed after 23 d of dietary adaptation. Ruminal, duodenal, and cecal epithelia were collected to determine Jsm-urea and mRNA abundance of UT-B and AQP. Lambs fed LF had greater intakes of dry matter (DMI; 1.20 vs. 0.86 kg/d) and N (NI; 20.1 vs. 15.0 g/d) than those fed HF (P < 0.01). Lambs fed SF had greater DMI (1.20 vs. 0.86 kg/d) and NI (20.6 vs. 14.5 g/d) than those fed WS (P < 0.01). As a percentage of NI, total N excretion was greater in lambs fed HF compared with those fed LF (103% vs. 63.0%; P < 0.01) and was also greater in lambs fed WS compared with those fed SF (93.6% vs. 72.1%; P = 0.02). Retained N (% of NI) was greater in lambs fed LF compared with those fed HF (37.0% vs. -2.55%; P < 0.01). Lambs fed SF had a greater (P = 0.02) retained N (% of NI; 28.0% vs. 6.50%) compared with those fed WS. Endogenous urea production (UER) tended (P = 0.09) to be greater in lambs fed HF compared with those fed LF. As a proportion of UER, lambs fed HF had a greater urinary urea-N loss (0.38 vs. 0.22) and lower urea-N transferred to the gastrointestinal tract (GIT; 0.62 vs. 0.78) or urea-N used for anabolism (as a proportion of urea-N transferred to the GIT; 0.12 vs. 0.26) compared with lambs fed LF (P < 0.01). Ruminal Jsm-urea was unaffected by diet. Duodenal Jsm-urea was greater (P < 0.01) in lambs fed HF compared with LF (77.5 vs. 57.2 nmol/[cm2 × h]). Lambs fed LF had greater (P = 0.03) mRNA expression of AQP3 in ruminal epithelia and tended (P = 0.06) to have greater mRNA expression of AQP3 in duodenal epithelia compared with lambs fed HF. Expression of UT-B mRNA was unaffected by diet. Our results showed that feeding more ruminally available energy improved N utilization, partly through a greater proportion of UER being transferred to the GIT and being used for anabolic purposes.

Noninvasive imaging of renal urea handling by CEST‐MRI

Renal function is characterized by concentration of urea for removal in urine. We tested urea as a CEST-MRI contrast agent for measurement of the concentrating capacity of distinct renal anatomical regions. The CEST contrast of urea was examined using phantoms with different concentrations and pH levels. Ten C57BL/6J mice were scanned twice at 7 T, once following intraperitoneal injection of 2M 150 µL urea and separately following an identical volume of saline. Kidneys were segmented into regions encompassing the cortex, outer medulla, and inner medulla and papilla to monitor spatially varying urea concentration. Z-spectra were acquired before and 20 minutes after injection, with dynamic scanning of urea handling performed in between via serial acquisition of CEST images acquired following saturation at +1 ppm. Phantom experiments revealed concentration and pH-dependent CEST contrast of urea that was both acid- and base-catalyzed. Z-spectra acquired before injection showed significantly higher CEST contrast in the inner medulla and papilla (2.3% ± 1.9%) compared with the cortex (0.15% ± 0.75%, P = .011) and outer medulla (0.12% ± 0.58%, P = .008). Urea infusion increased CEST contrast in the inner medulla and papilla by 2.1% ± 1.9% (absolute), whereas saline infusion decreased CEST contrast by -0.5% ± 2.0% (absolute, P = .028 versus urea). Dynamic scanning revealed that thermal drift and diuretic status are confounding factors. Urea CEST has a potential of monitoring renal function by capturing the spatially varying urea concentrating ability of the kidneys.

103 Post-ruminally delivered urea improves fiber utilization in ruminants.

Ruminants are able to transform non-protein nitrogen (NPN) into animal protein, but efficiency may depend on the site of delivery in the gastrointestinal tract. We hypothesized that post-ruminal NPN supply can be utilized in the rumen, via urea recycling. Four rumen fistulated Holstein heifers (561 ± 42Kg BW) were used in a Latin square design with ruminal and abomasal infusions. Treatments consisted of ammonium hydroxide solution constantly infused into the rumen (AR), or the abomasum (AA), urea solution constantly infused into the abomasum (UA) or urea supplied into the rumen as daily pulse dose (UR). Experimental periods consisted of 12 d for treatment administration and sampling followed by 2 d rest. Treatments were isonitrogenous (59 g N/d) and all heifers were fed hay (6% CP, 70% NDF) ad libitum with 1kg of compound feed (21.7% CP) supplied in the morning through rumen cannula. Indigestible NDF (iNDF) was used as faecal marker. Ruminal pH was analysed to fit a logistic model resulting in two parameters: slope corresponding to the range of cumulative pH and inflection point corresponding to the statistical median. Dry matter intake was highest (P=0.03) for heifers receiving UR or UA while AR led to lowest intake. Higher NDF digestibility was observed with UA compared to UR (P=0.05). The pH inflection point was higher (P=0.05) with AR compared to UR, AA, and UA. The pH range with UR was wider (lower slope; P=0.04), due to the rise in pH following urea infusion, in which ammonia concentration reached 63 mg/dL 3 hours after UR supply, while concentrations for AR, AA and UA were held constant (18.5 ± 3.1, 4.4 ± 0.8, 4.6 ± 1.0 mg/dL, respectively). Our study indicates that urea delivered post-ruminally improves fiber digestibility and rumen stability and offers a unique approach towards improving N utilization and efficiency in ruminant production systems.

Removal of urea by electro-oxidation in a miniature dialysis device: a study in awake goats.

The key to success in developing a wearable dialysis device is a technique to safely and efficiently regenerate and reuse a small volume of dialysate in a closed-loop system. In a hemodialysis model in goats, we explored whether urea removal by electro-oxidation (EO) could be effectively and safely applied in vivo. A miniature dialysis device was built, containing 1 or 2 "EO units," each with 10 graphite electrodes, with a cumulative electrode surface of 585 cm2 per unit. The units also contained poly(styrene-divinylbenzene) sulfonate beads, FeOOH beads, and activated carbon for respective potassium, phosphate, and chlorine removal. Urea, potassium, and phosphate were infused to create "uremic" conditions. Urea removal was dependent on total electrode surface area [removal of 8 mmol/h (SD 1) and 16 mmol/h (SD 2) and clearance of 12 ml/min (SD 1) and 20 ml/min (SD 3) with 1 and 2 EO units, respectively] and plasma urea concentration but not on flow rate. Extrapolating urea removal with 2 EO units to 24 h would suffice to remove daily urea production, but for intermittent dialysis, additional units would be required. EO had practically no effects on potassium and phosphate removal or electrolyte balance. However, slight ammonium releasewas observed, and some chlorine release at higher dialysate flow rates. Minor effects on acid-base balance were observed, possibly partly due to infusion of chloride. Mild hemolysis occurred, which seemed related to urea infusion. In conclusion, clinically relevant urea removal was achieved in vivo by electro-oxidation. Efficacy and safety testing in a large-animal model with uremia is now indicated.

Urea, a true uremic toxin: the empire strikes back.

Blood levels of urea rise with progressive decline in kidney function. Older studies examining acute urea infusion suggested that urea was well-tolerated at levels 8-10× above normal values. More recent invitro and invivo work argue the opposite and demonstrate both direct and indirect toxicities of urea, which probably promote the premature aging phenotype that is pervasive in chronic kidney disease (CKD). Elevated urea at concentrations typically encountered in uremic patients induces disintegration of the gut epithelial barrier, leading to translocation of bacterial toxins into the bloodstream and systemic inflammation. Urea induces apoptosis of vascular smooth muscle cells as well as endothelial dysfunction, thus directly promoting cardiovascular disease. Further, urea stimulates oxidative stress and dysfunction in adipocytes, leading to insulin resistance. Finally, there are widespread indirect effects of elevated urea as a result of the carbamylation reaction, where isocyanic acid (a product of urea catabolism) alters the structure and function of proteins in the body. Carbamylation has been linked with renal fibrosis, atherosclerosis and anaemia. In summary, urea is a re-emerging Dark Force in CKD pathophysiology. Trials examining low protein diet to minimize accumulation of urea and other toxins suggest a clinical benefit in terms of slowing progression of CKD.

Influence of milk urea concentration on fractional urea disappearance rate from milk to blood plasma in dairy cows

The relationship between milk urea nitrogen (MUN; mg of N/dL) and urinary N excretion is affected, among others, by diurnal dynamics in MUN, which in turn is largely influenced by feed intake pattern and characteristics of urea transfer from blood plasma to milk and vice versa. This study aimed to obtain insight in urea transfer characteristics within the mammary gland and from the mammary gland to blood plasma in dairy cows at various concentrations of plasma urea nitrogen (PUN; mg of N/dL) and MUN. Urea transfer from milk to blood plasma and urea transfer within the mammary gland itself was evaluated in a 4×4 Latin square design using 4 lactating multiparous Holstein-Friesian cows (milk production of 39.8±4.70kg/d and 90±3.9 d in milk). Treatments consisted of 4 primed continuous intravenous urea infusions of 0, 5, 10, and 15g of urea/h. Boluses of [15N15N]urea were injected in cistern milk at 20, 60, and 100 min before the 1700h milking. Milk was collected in portions of approximately 2 L at the 1700h milking. Milk samples were analyzed for urea and enrichment of 15N-urea. Results from one cow were discarded because of leakage of milk from the teats after injection of boluses of [15N15N]urea. Increasing urea infusion rate linearly increased PUN from 11.4 (0g of urea/h) to 25.9mg/dL (15g of urea/h) and MUN from 10.3 (0g of urea/h) to 23.5 (15g of urea/h) mg of N/dL. The percentage of injected [15N15N]urea recovered from milk at the time of injection was not affected by urea infusion rate and varied between 65.1 and 73.0%, indicating that a substantial portion of injected [15N15N]urea was not accounted for by collected milk. The estimated fractional disappearance rate of 15N-urea from milk to blood (Kurea; per hour) linearly increased from 0.429 (0g of urea/h) to 0.641 per hour (15g of urea/h). Cistern injected [15N15N]urea diffused within 20 min after injection toward alveoli milk. Calculations with the average Kurea estimated in this study show that 89% of an initial difference between PUN and MUN will have disappeared after 4 h. In conclusion, urea disappearance from milk in the mammary gland is substantial, as well as the intramammary urea exchange between cistern, duct, and alveoli milk. However, results have to be interpreted with caution given the lack of full recovery of dosed 15N urea at time of injection. Information on Kurea is useful to quantify the effects of diurnal variation in PUN on MUN, which enhances the utility of MUN as an indicator for N excretion in urine.

Effect of ruminal ammonia supply on lysine utilization by growing steers1

Six ruminally cannulated Holstein steers (202 ± 15 kg) were used to study the effects of ruminal ammonia loading on whole-body lysine (Lys) utilization. Steers were housed in metabolism crates and used in a 6 × 6 Latin square design. All steers received 2.52 kg DM/d of a diet (10.1% CP) containing 82% soybean hulls, 8% wheat straw, 5% cane molasses, and 5% vitamins and minerals, and 10 g/d of urea (considered to be part of the basal diet) was ruminally infused continuously to ensure adequate ruminal ammonia concentrations. All steers were ruminally infused continuously with 200 g/d of acetic acid, 200 g/d of propionic acid, and 50 g/d of butyric acid and abomasally infused with 300 g/d of glucose continuously to increase energy supply without increasing microbial protein supply. Steers were also abomasally infused continuously with an excess of all essential AA except Lys to ensure that Lys was the only limiting AA. Treatments were arranged as a 3 × 2 factorial with 3 levels of urea (0, 40, or 80 g/d) continuously infused ruminally to induce ammonia loading and 2 levels of Lys (0 or 6 g/d) continuously infused abomasally. Treatments did not affect fecal N output ( = 0.37). Lysine supplementation decreased ( < 0.01) urinary N excretion from 51.9 g/d to 44.3 g/d, increased ( < 0.01) retained N from 24.8 to 33.8 g/d, increased ( < 0.01) plasma Lys, and decreased ( ≤ 0.05) plasma serine, tyrosine, valine, leucine, and phenylalanine. Lysine supplementation also tended ( = 0.09) to reduce plasma urea-N. Urea infusions linearly increased ( = 0.05) retained N (27.1, 29.3, and 31.5 g/d) and also linearly increased ( < 0.01) urinary N excretion (31.8, 48.1, and 64.4 g/d), urinary urea (21.9, 37.7, and 54.3 g/d), urinary ammonia (1.1, 1.4, and 1.9 g/d), and plasma urea (2.7, 4.0, and 5.1 mM), and linearly decreased plasma alanine ( = 0.04) and plasma glycine ( < 0.01). Assuming that retained protein is 6.25 × retained N and contains 6.4% Lys, the incremental efficiencies of infused Lys utilization were 51%, 59%, and 69% for steers receiving 0, 40, and 80 g/d of urea, respectively, indicating that ruminal ammonia loads may improve the efficiency of Lys utilization. This is supported by observed increases in whole body-protein deposition in response to ammonia loading of our steers that were, by design, Lys deficient.

Nitrogen Absorbed from the Large Intestine Increases Whole-Body Nitrogen Retention in Pigs Fed a Diet Deficient in Dispensable Amino Acid Nitrogen1–3

Background: Nitrogen absorption from the large intestine is considered of limited value for supporting body protein synthesis in animals and humans, but it may be of benefit when the dietary supply of nitrogen for synthesis of dispensable amino acids (DAAs) is deficient.Objective: A whole-body nitrogen balance study was conducted to evaluate the impact of nitrogen absorption from the large intestine of pigs fed a diet deficient in DAA nitrogen.Methods: Nine cecally cannulated barrows were fed a cornstarch and casein-based diet with a high indispensable amino acid (IAA) nitrogen to total nitrogen ratio (IAA:TN; 0.75). Pigs were randomly assigned to saline or 1 of 2 urea nitrogen infusion rates into the cecum (low and high, 1.5 and 3.0 g/d, respectively) following a 3 × 3 Latin square design. At the high urea nitrogen infusion rate, IAA:TN was 0.55. At slaughter, liver samples were taken to measure activity of carbamoyl phosphate synthetase I (CPS-I), glutamate dehydrogenase (GDH), and Gln synthetase (Gln-S).Results: Whole-body nitrogen retention improved with urea infusion (4.86 ± 0.20 g/d, 6.40 ± 0.21 g/d, and 7.75 ± 0.19 g/d for saline and low and high infusion rates, respectively; P < 0.05), as well as body weight gain. The marginal efficiency of using nitrogen absorbed from the large intestine for improving nitrogen retention was not affected by urea nitrogen infusion rate (P > 0.10). Enzyme activity of CPS-I or Gln-S was not different between treatments (P > 0.10), but GDH showed a trend for a positive linear response with increasing urea nitrogen infusion rate (P = 0.06).Conclusion: These results indicate that urea nitrogen absorbed from the large intestine is efficiently used for increasing body protein deposition when feeding pigs a diet deficient in DAA nitrogen.

Nonprotein Nitrogen Is Absorbed from the Large Intestine and Increases Nitrogen Balance in Growing Pigs Fed a Valine-Limiting Diet1–3

Nitrogen absorption from the large intestine, largely as ammonia and possibly as amino acids (AAs), is generally thought to be of little nutritional value to nonruminant animals and humans. Ammonia-nitrogen absorbed from the large intestine, however, may be recycled into the small intestine as urea and incorporated into microbial AAs, which may then be used by the host. A cecal infusion study was performed to determine the form in which nitrogen is absorbed from the large intestine and the impact of large intestine nitrogen supply on nitrogen balance in growing pigs. Eighteen cecally cannulated barrows (initial body weight: 22.4 ± 1.2 kg) were used to determine the effect of supplying nitrogen into the large intestine from either casein or urea on whole-body nitrogen retention and urea kinetics. Treatments were cecal infusions of saline (control), casein, or urea with nitrogen infused at a rate of 40% of nitrogen intake. In a subsample of 9 pigs, 15N15N-urea was infused via i.v. during the nitrogen-balance period to determine urea kinetics. All pigs were fed a valine-limiting cornstarch-soybean meal–based diet. More than 80% of infused nitrogen was apparently absorbed. Urea flux and urinary nitrogen excretion increased (P ≤ 0.05) by the same amount for both nitrogen sources, but this increase did not fully account for the increase in nitrogen absorption from the large intestine. Whole-body nitrogen retention improved with nitrogen infusions (129 vs. 114 g/d; P < 0.01) and did not differ (P > 0.05) between nitrogen sources. Absorption of nitrogen from the large intestine appears to be in the form of nonprotein nitrogen, which appears to be returned to the small intestine via urea and used there for microbial AA production and should therefore be considered when determining nitrogen and AA supply and requirements.

The rate and pattern of urea infusion into the rumen of wethers alters nitrogen balance and plasma ammonia

Changes in N balance, urinary excretion of purine derivative (PD), urea, creatinine and ammonia and plasma ammonia, glucose, urea, insulin and IGF-1 were examined in four wethers (37 ± 2.6 kg BW). The animals were fitted with permanent ruminal catheters, fed lucerne hay (9.4 MJ/day; 23 g N/day; 7 g soluble N/day, 6 equal meals/day) and treated with contrasting rates of urea infusion into the rumen: first, a continuous infusion (CT), at 3.2 mg urea-N/min for 10 days and then a discontinuous infusion (DT) at 156 mg urea-N/min for 4 min; in 6 daily doses with the meals for 7 days. N balance was calculated from pooled samples of faeces and urine. Jugular blood samples were collected before and 1.5 h after the morning meal (M1) on days CT10, DT2, DT4 and DT6. N retention decreased during DT (p = 0.01) due to a significant increase of N excretion in urine (4 g/day; p = 0.009) and faeces (1 g/day; p = 0.02). Dry matter (p < 0.001) and N digestibility in vivo (p = 0.01) decreased significantly during DT. Urinary urea and PD excretion were not altered by treatment. Significant linear (p = 0.004) and quadratic (p = 0.001) effects were observed for plasma ammonia in M1 (from 170 CT10 to 235 μm DT2 and returned to 120 μm DT6). No changes were observed in plasma glucose, urea, insulin and IGF-1. Results indicate that changes from CT to DT reduced N retention in sheep due to enhanced urinary N excretion, but it was not associated with changes in urinary urea or PD excretion; or plasma concentrations of insulin and IGF-1. As the dry matter (DM) an N digestibility could account a 0.23 of the decrease in N retention; the largest fraction of the reduction in N retention remained unexplained by the results.

Intracarotid hypertonic sodium chloride differentially modulates sympathetic nerve activity to the heart and kidney

Hypertonic NaCl infused into the carotid arteries increases mean arterial pressure (MAP) and changes sympathetic nerve activity (SNA) via cerebral mechanisms. We hypothesized that elevated sodium levels in the blood supply to the brain would induce differential responses in renal and cardiac SNA via sensors located outside the blood-brain barrier. To investigate this hypothesis, we measured renal and cardiac SNA simultaneously in conscious sheep during intracarotid infusions of NaCl (1.2 M), sorbitol (2.4 M), or urea (2.4 M) at 1 ml/min for 4 min into each carotid. Intracarotid NaCl significantly increased MAP (91 ± 2 to 97 ± 3 mmHg, P < 0.05) without changing heart rate (HR). Intracarotid NaCl was associated with no change in cardiac SNA (11 ± 5.0%), but a significant inhibition of renal SNA (-32.5 ± 6.4%, P < 0.05). Neither intracarotid sorbitol nor urea changed MAP, HR, central venous pressure, cardiac SNA, and renal SNA. The changes in MAP and renal SNA were completely abolished by microinjection of the GABA agonist muscimol (5 mM, 500 nl each side) into the paraventricular nucleus of the hypothalamus (PVN). Infusion of intracarotid NaCl for 20 min stimulated a larger increase in water intake (1,100 ± 75 ml) than intracarotid sorbitol (683 ± 125 ml) or intracarotid urea (0 ml). These results demonstrate that acute increases in blood sodium levels cause a decrease in renal SNA, but no change in cardiac SNA in conscious sheep. These effects are mediated by cerebral sensors located outside the blood-brain barrier that are more responsive to changes in sodium concentration than osmolality. The renal sympathoinhibitory effects of sodium are mediated via a pathway that synapses in the PVN.

Urea-N recycling in lactating dairy cows fed diets with 2 different levels of dietary crude protein and starch with or without monensin

Rumensin (monensin; Elanco Animal Health, Greenfield, IN) has been shown to reduce ammonia production and microbial populations in vitro; thus, it would be assumed to reduce ruminal ammonia production and subsequent urea production and consequently affect urea recycling. The objective of this experiment was to determine the effects of 2 levels of dietary crude protein (CP) and 2 levels of starch, with and without Rumensin on urea-N recycling in lactating dairy cattle. Twelve lactating Holstein dairy cows (107±21d in milk, 647kg ± 37kg of body weight) were fed diets characterized as having high (16.7%) or low (15.3%) CP with or without Rumensin, while dietary starch levels (23 vs. 29%) were varied between 2 feeding periods with at least 7d of adaptation between measurements. Cows assigned to high or low protein and to Rumensin or no Rumensin remained on those treatments to avoid carryover effects. The diets consisted of approximately 40% corn silage, 20% alfalfa hay, and 40% concentrate mix specific to the treatment diets, with 0.5kg of wheat straw added to the high starch diets to enhance effective fiber intake. The diets were formulated using Cornell Net Carbohydrate and Protein System (version 6.1), and the low-protein diets were formulated to be deficient for rumen ammonia to create conditions that should enhance the demand for urea recycling. The high-protein diets were formulated to be positive for both rumen ammonia and metabolizable protein. Rumen fluid, urine, feces, and milk samples were collected before and after a 72-h continuous jugular infusion of 15N15N-urea. Total urine and feces were collected during the urea infusions for N balance measurements. Milk yield and dry matter intake were improved in cows fed the higher level of dietary CP and by Rumensin. Ruminal ammonia and milk and plasma urea nitrogen concentrations corresponded to dietary CP concentration. As has been shown in vitro, Rumensin reduced rumen ammonia concentration by approximately 23% but did not affect urea entry rate or gastrointestinal entry rate. Urea entry rate averaged approximately 57% of total N intake for cattle with and without Rumensin, and gastrointestinal rate was similar at 43 and 42% of N intake for cattle fed and not fed Rumensin, respectively. The cattle fed the high-protein diet had a 25% increase in urea entry rate and no effect of starch level was observed for any recycling parameters. Contrary to our hypothesis, Rumensin did not alter urea production and recycling.

Short communication: Assessing urea transport from milk to blood in dairy cows

The concentration of urea in milk (MUC) has emerged as a potentially useful tool to predict urinary N excretion. Various factors may affect the relationship between MUC and urinary N excretion, including transport characteristics of urea from blood to milk and vice versa. The main objective of this study was to test whether substantial transport of urea from milk to blood exists in lactating dairy cattle. The subobjectives were (1) to assess the effects of various urea gradient levels between blood and milk on urea transport from milk to blood and (2) to test the occurrence of urea transport between different compartments of the mammary gland such as the cistern and the alveoli. Urea transport was studied in 2 multiparous lactating Holstein-Friesian cows (36.0±6.18kg of milk/d; mean ± SD). In 3 separate trials, boluses of [15N15N]urea were injected in the cisterns via the teat canals at 20, 60, and 120min before the 1700-h milking at various levels of MUC and of blood plasma urea concentration (PUC). In trial 1, a primed continuous infusion of urea (105g at the start, continuing with 20g/h) into the jugular vein started at 0500 h and stopped at 0, 1, 2, and 3h before the 1700-h milking on d 1, 2, 3, and 4, respectively. In trial 2, 5.5g of urea was injected into the cisterns at 20, 60, and 120min before the 1700-h milking on d 5, 6, and 7, respectively. In trial 3, urea fluxes were measured without an experimentally induced gradient between MUC and PUC on d 8, 9, and 10, respectively. During milking, successive milk samples were taken from first to last milk. Blood and milk were analyzed for 15N-urea enrichment. Levels of 15N-urea in blood increased after injection of a [15N15N]urea bolus in milk, indicating urea transport from milk to blood. Between 21.0 and 35.3% of injected [15N15N]urea in milk was recovered after 2h. The fractional [15N15N]urea decline rate in milk varied between 0.0076 and 0.0096/min. The level of MUC, rather than the concentration gradient between MUC and PUC, appeared to affect this fractional rate of decline. Enrichment levels of 15N-urea in milk samples within a single milking showed that urea was transported from cistern milk to alveoli milk. In conclusion, the results indicate that transport of urea from milk to blood in lactating dairy cattle occurs and that urea is transported from cistern milk to alveoli milk.

Manucher Javid, Urea, and the Rise of Osmotic Therapy for Intracranial Pressure

Therapy with hypertonic solutions is one of the mainstays of neurosurgical treatment for all types of neurological injury. Although the initial research with hypertonic agents in the early decades of the 20th century showed great promise for these agents to lower intracranial pressure, this research also showed a considerable rate of adverse effects and complications. By the 1940s and 1950s, hypertonic therapy had been discounted as unsafe and was rarely used in neurosurgery. In the late 1950s, Manucher J. Javid and Paul Settlage at the University of Wisconsin began experimenting with infusions of urea as an agent to control intracranial pressure. Their experiments were wildly successful, and urea became a drug of major importance to neurosurgeons worldwide in only a few years. This article chronicles the work of Javid and Settlage, including a discussion of the early research on hypertonic agents, the initial difficulty the Wisconsin researchers had in disseminating their results, the widespread acceptance that followed, and the impact that these discoveries had on the neurosurgical community. The prominent place that hypertonic agents now hold in the armamentarium of neurosurgeons is owed to the work of Dr Javid, as illustrated in this historical analysis.

Effect of time duration of ruminal urea infusions on ruminal ammonia concentrations and portal-drained visceral extraction of arterial urea-N in lactating Holstein cows

The effects of a 6 versus 24h ruminal urea infusion in lactating dairy cows fed a basal diet deficient in N on ruminal ammonia concentration, arterial urea-N concentration, net portal-drained viscera (PDV) urea-N flux, arterial urea-N extraction across the PDV, and renal urea-N kinetics were investigated. Three Danish Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in major splanchnic blood vessels were randomly allocated to a 3 × 3 Latin square design with 21-d periods. Treatments were ventral ruminal infusion of water for 24h (water INF), 24-h infusion of 15g of urea/kg of dry matter intake (DMI; 24-h INF), and 6-h infusion of 15g of urea/kg of DMI (6-h INF). The 6-h INF was initiated 0.5h after the afternoon feeding, and ran until 2230h. Eight sample sets of arterial, portal, and hepatic blood, ruminal fluid, and urine were obtained at 0.5h before the morning feeding and 0.5, 1.5, 2.5, 3.5, 4.5, 5.5, and 6.5h after feeding (i.e., 9 to 15.5h after the 6h infusion was terminated). A substantial decrease in DMI for 6-h INF compared with 24-h INF and water INF was observed, and it has to be recognized that DMI may have confounding effects. However, the experimental setting plan was met (i.e., to cause changes in the daily pattern of ruminal ammonia and blood urea-N concentrations). The arterial urea-N concentration for 24-h INF and 6-h INF were greater than the arterial urea-N concentration with water INF throughout the sampling window. However, the arterial urea-N concentration for 6-h INF decreased steadily with sampling time reflecting a carryover effect from the ruminal urea infusion. The ruminal ammonia concentration and net portal flux of ammonia for 6-h INF were not different from water INF; hence, no carryover effect on ruminal ammonia concentration was observed. The portal flux of urea-N was not affected by treatment (i.e., even the combination of low ruminal ammonia and high arterial urea-N concentration with 6-h INF was not used by the cow to increase the uptake of urea-N across the PDV). Arterial urea-N extraction across the PDV was increased with water INF especially from 0.5 to 3.5h postprandial relative to the urea infusion treatments, reflecting increased epithelial permeability for urea-N. This indicates that daily ruminal peak of ammonia or blood urea-N concentrations overruled potential signals from low ruminal ammonia concentration observed during the sampling window. In conclusion, dairy cows appear unable to increase transport of urea-N from blood to gut in periods with low ruminal ammonia concentrations, even in a situation with infrequent N supply and apparent carryover effects on blood urea-N. It is speculated that mechanisms responsible for downregulation of epithelial urea-N transport based on daily maximum concentrations of ammonia in the rumen or urea-N in the blood suppresses any short-term signal from low ruminal ammonia during periods with low ruminal N supply.

Effects of nitrogen supply on inter-organ fluxes of urea-N and renal urea-N kinetics in lactating Holstein cows

The effects of decreasing ruminal urea infusion in lactating dairy cows fed a basal diet deficient in rumen degradable protein on inter-organ urea-N fluxes, epithelial urea-N extraction, and renal urea-N kinetics were investigated. Eight Danish Holstein cows fitted with a ruminal cannula and permanent indwelling catheters in the major splanchnic blood vessels and the gastrosplenic vein were used. The cows were randomly allocated to a triplicate incomplete 3×3 Latin square design with 14-d periods. Treatments were continuous ventral ruminal infusion of water, 4.1g of feed urea/kg of dry matter intake, and 8.5g of feed urea/kg of dry matter intake. Dry matter intake and milk yield decreased linearly with decreasing urea infusion. Arterial blood urea-N and ruminal ammonia concentrations decreased linearly with decreasing urea infusion. In absolute amounts, the urea-N recycling did not increase when urea infusion was decreased. Arterial urea-N extraction across the portal-drained viscera and rumen wall increased linearly with decreasing urea infusion (2.46, 3.65, and 4.32±0.31% and 7.5, 11.5, and 16.9±0.9%, respectively), indicating that cows responded to the changes in N supply. The relative urea-N extraction across the ruminal wall increased compared with the total portal-drained viscera extraction. We observed a postprandial decrease in ruminal extraction of arterial urea-N that might reflect that the activity of the protein, presumably facilitating urea-N transport, is regulated by ruminal ammonia. The urea-N clearance by the kidneys decreased (35, 30, and 25±2L/h) and the urea-N reabsorbed by the kidney increased (42, 51 and 56±3%) with decreasing urea infusion, indicating that the kidneys salvaged urea-N with low-N supply. The urea transporter B mRNA abundance in rumen papillae (papillae harvested at sampling days) was not affected by dietary N supply. The study showed, that rumen wall extraction of arterial urea-N is subjected to both long- and short-term regulation. Extraction increases with decreasing N supply long-term; however, a short-term postprandial decrease in extraction was observed. No association between long-term adaptation of urea-N extraction across the rumen wall and urea transporter B mRNA abundance could be demonstrated.

Effects of ruminal infusion of a slow-release polymer-coated urea or conventional urea on apparent nutrient digestibility, in situ degradability, and rumen parameters in cattle fed low-quality hay

Four ruminally cannulated beef steers were used in a 4 × 4 Latin square design experiment conducted to compare ruminal infusion effects of different sources of non-protein nitrogen (NPN) on intake; digestibility; ruminal dry matter (DM) and neutral detergent fiber (NDF) degradability; ruminal pH; and ammonia nitrogen (N-NH 3) of beef cattle fed Brachiaria humidicola cv. Llanero hay [CP = 24.9 g/kg DM; ash-free NDF corrected for contamination of CP (NDFom,cp) = 782.4 g/kg DM]. Treatments were: Control (hay only); U [hay + conventional urea (204.2 g/d)]; SRU [hay + slow-release polymer-coated urea (297.5 g/d)]; U + SRU [hay + a combination of 1:1 by weight of both urea types (130.4 g/d U + 130.4 g/d SRU)]. Supplements were infused into the rumen through the rumen cannula. Treatments SRU and U + SRU had an increasing of hay dry matter intake (DMI) (9.44 and 9.02 kg/d, respectively; P<0.01, compared with 7.62 kg/d for the control), as well as total NDF (P<0.01) and crude protein (CP, P<0.01). The CP digestibility was higher (P<0.01) for all treatments with NPN when compared to control. There were differences on DM and NDF degradability curves (P<0.05) between rumen exposure times, in which the disappearance was increased up to 120 h after incubation. On the first day of collections, the lowest pH values (6.89 and 6.91) were observed on SRU and U + SRU 6 h after the afternoon infusion, and the highest pH was observed immediately after the afternoon infusion of U (7.39). High levels of ammonia-N were kept for 8 h after the last infusion daily of SRU, which resulted in the most constant values of NH 3-N in the rumen. In general, our data suggested that SRU had a more adequate concentration of N during the fermentation of low-quality hay, and also had a proper pH for rumen activity.