Standing balance of vehicle passengers: The effect of vehicle motion, task performance on post-drive balance.

BACKGROUND: Motion platforms and driving simulators have been shown to contribute to motion sickness and a short-term increase in standing postural sway. However, no studies to date have investigated how the motion of a passenger vehicle and the performance of a task during a drive on a closed test track affects post-drive standing balance. RESEARCH QUESTIONS: What are the effects of (1) a continuous, scripted drive on a closed test track, and (2) the performance of a handheld tablet-based task during the scripted drive, on post-drive standing balance? METHODS: Fifty adults (23 males, 27 females; 40.0 ± 20.6 yr) rode in the front passenger seat of a midsized sedan on a scripted drive. Participants were assigned to one of the acceleration levels (Low, Moderate) and completed both Task and No-Task test conditions, involving a visual-based task on a handheld tablet device. Before and after each scripted drive, participants completed two standing balance exercises: 1) feet tandem, eyes open, on firm support, and 2) feet together, eyes closed, on foam support. An inertial measurement unit (IMU) captured estimates of postural trunk sway. Root-mean-square (RMS) of angular position and velocity in the anteroposterior (A/P) and mediolateral (M/L) directions, and elliptical fit and path length of sway trajectory were computed. A nonparametric analysis was performed on the balance metrics. RESULTS: Exposure to a scripted drive in a vehicle affected participants' postural sway, especially after using a handheld device during the drive. M/L RMS sway velocity and path length increased for both exercises following the scripted drive with task. Additionally, M/L RMS sway increased for the more challenging balance exercise, during which participants stood with feet together on foam support with eyes closed. SIGNIFICANCE: This study is the first to explore balance following a scripted drive on a closed test track. Changes in post-drive balance introduces potential risks to vehicle passengers; concurrent performance of a task on a handheld device further increases the likelihood that post-drive balance will be negatively affected.

Automated detection of fluorescent cells in in-resin fluorescence sections for integrated light and electron microscopy.

Integrated array tomography combines fluorescence and electron imaging of ultrathin sections in one microscope, and enables accurate high-resolution correlation of fluorescent proteins to cell organelles and membranes. Large numbers of serial sections can be imaged sequentially to produce aligned volumes from both imaging modalities, thus producing enormous amounts of data that must be handled and processed using novel techniques. Here, we present a scheme for automated detection of fluorescent cells within thin resin sections, which could then be used to drive automated electron image acquisition from target regions via 'smart tracking'. The aim of this work is to aid in optimization of the data acquisition process through automation, freeing the operator to work on other tasks and speeding up the process, while reducing data rates by only acquiring images from regions of interest. This new method is shown to be robust against noise and able to deal with regions of low fluorescence.

Mineral Composition of Uroliths Obtained from Sheep and Goats with Obstructive Urolithiasis.

BACKGROUND: Knowledge of the mineral composition of the causative urolith is important to develop preventative strategies. Advances in analytic techniques have led to detection of urolith components not previously recognized. HYPOTHESIS/OBJECTIVES: The objectives of this study were to characterize uroliths in sheep and goats structurally and clinically. We hypothesized that amorphous magnesium calcium phosphate (AMCP) would be a naturally occurring urolith type in sheep and goats. ANIMALS: Forty-nine sheep and goats presenting for obstructive urolithiasis from June 15, 2014 through June 14, 2016 were reviewed along with the demographic data of all small ruminants admitted during that same period. METHODS: Medical records were reviewed for demographic and historical data, and 36 uroliths obtained from these cases during diagnostic or therapeutic procedures were analyzed by infrared spectroscopy to determine chemical composition. RESULTS: AMCP is a naturally occurring urolith type in obstructed male sheep and goats and was the most common urolith type in this study, where it occurred as a majority component with struvite (39% of uroliths) or as a pure component (11%). Pure struvite was found in 1 urolith (2%). Calcium carbonate was the second most frequent urolith with 31% of uroliths being pure calcium carbonate. CONCLUSIONS AND CLINICAL IMPORTANCE: This study demonstrates that uroliths, which appear consistent with struvite, could actually be calcium-containing AMCP. Urolith analysis is critical in determining mineral content of uroliths to guide dietary recommendations for prevention.

Immunosuppressive human anti-CD83 monoclonal antibody depletion of activated dendritic cells in transplantation.

Current immunosuppressive/anti-inflammatory agents target the responding effector arm of the immune response and their nonspecific action increases the risk of infection and malignancy. These effects impact on their use in allogeneic haematopoietic cell transplantation and other forms of transplantation. Interventions that target activated dendritic cells (DCs) have the potential to suppress the induction of undesired immune responses (for example, graft versus host disease (GVHD) or transplant rejection) and to leave protective T-cell immune responses intact (for example, cytomegalovirus (CMV) immunity). We developed a human IgG1 monoclonal antibody (mAb), 3C12, specific for CD83, which is expressed on activated but not resting DC. The 3C12 mAb and an affinity improved version, 3C12C, depleted CD83(+) cells by CD16(+) NK cell-mediated antibody-dependent cellular cytotoxicity, and inhibited allogeneic T-cell proliferation in vitro. A single dose of 3C12C prevented human peripheral blood mononuclear cell-induced acute GVHD in SCID mouse recipients. The mAb 3C12C depleted CMRF-44(+)CD83(bright) activated DC but spared CD83(dim/-) DC in vivo. It reduced human T-cell activation in vivo and maintained the proportion of CD4(+) FoxP3(+) CD25(+) Treg cells and also viral-specific CD8(+) T cells. The anti-CD83 mAb, 3C12C, merits further evaluation as a new immunosuppressive agent in transplantation.

Lactobacillus fermentum NCIMB 5221 and NCIMB 2797 as cholesterol-lowering probiotic biotherapeutics: in vitro analysis.

Cardiovascular and coronary artery disease risk are correlated with cholesterol levels and are significant health concerns. Current cholesterol-lowering approaches includes lifestyle and diet modifications, as well as statins which presents numerous shortcomings. The probiotic bacteria, Lactobacillus fermentum NCIMB 5221 and NCIMB 2797, have demonstrated cholesterol-lowering potential in animal studies. However, there is a lack in understanding the mechanism(s) behind these observed effects. The goal of this work is to investigate, in vitro, the cholesterol-lowering mechanisms of these two strains. To determine the cholesterol-lowering mechanisms, probiotic cholesterol assimilation, colon epithelial adhesion and inhibition of cholesterol uptake by colon epithelial (Caco-2) cells were investigated. L. fermentum NCIMB 2797 (P=0.012) and NCIMB 5221 (P=0.003) assimilated cholesterol and their cell surface hydrophobicity was 70.30±8.85% and 55.60±2.59%, respectively. Both L. fermentum strains showed no significant impact (P>0.05) on Caco-2 cell viability. Of most interest, Caco-2 pre-exposure to L. fermentum NCIMB 5221 significantly decreased (P=0.015) cholesterol uptake, with 85.98±2.07% uptake compared to the untreated cells. Similarly, L. fermentum NCIMB 2797 probiotic cells significantly decreased (P=0.019) cholesterol uptake by Caco-2 cells, with 86.45±1.71% uptake observed compared to the control cells. The results demonstrate that L. fermentum NCIMB 5221 and L. fermentum NCIMB 2797 have the potential via various modes of action to lower cholesterol. Additional studies are required to understand the mechanism(s) of action behind probiotic cholesterol assimilation and behind the cholesterol uptake inhibition by colon epithelial cells.

Impact of a yogurt matrix and cell microencapsulation on the survival of Lactobacillus reuteri in three in vitro gastric digestion procedures.

The goal of this study was to assess the interaction between microencapsulation and a yogurt food matrix on the survival of Lactobacillus reuteri NCIMB 30242 in four different in vitro systems that simulate a gastric environment. The four systems were: United States Pharmacopeia (USP) solutions, a static two-step (STS) procedure which included simulated food ingredients, a constantly dynamic digestion procedure (IViDiS), as well a multi-step dynamic digestion scheme (S'IViDiS). The pH profiles of the various procedures varied between systems with acidity levels being: USP > STS > IViDiS = S'IVIDiS. Addition of a food matrix increased the pH in all systems except for the USP methodology. Microencapsulation in alginate-based gels was effective in protecting the cells in model solutions when no food ingredients were present. The stability of the probiotic culture in the in vitro gastric environments was enhanced when (1) yoghurt or simulated food ingredient were present in the medium in sufficient quantity, (2) pH was higher. The procedure-comparison data of this study will be helpful in interpreting the literature with respect to viable counts of probiotics obtained from different static or dynamic in vitro gastric systems.

Use of next-generation sequencing and candidate gene analysis to identify underlying defects in patients with inherited platelet function disorders.

BACKGROUND: Inherited platelet function disorders (PFDs) are heterogeneous, and identification of the underlying genetic defects is difficult when based solely on phenotypic and clinical features of the patient. OBJECTIVE: To analyze 329 genes regulating platelet function, number, and size in order to identify candidate gene defects in patients with PFDs. PATIENTS/METHODS: Targeted analysis of candidate PFD genes was undertaken after next-generation sequencing of exomic DNA from 18 unrelated index cases with PFDs who were recruited into the UK Genotyping and Phenotyping of Platelets (GAPP) study and diagnosed with platelet abnormalities affecting either Gi signaling (n = 12) or secretion (n = 6). The potential pathogenicity of candidate gene defects was assessed using computational predictive algorithms. RESULTS: Analysis of the 329 candidate PFD genes identified 63 candidate defects, affecting 40 genes, among index cases with Gi signaling abnormalities, while 53 defects, within 49 genes, were identified among patients with secretion abnormalities. Homozygous gene defects were more commonly associated with secretion abnormalities. Functional annotation analysis identified distinct gene clusters in the two patient subgroups. Thirteen genes with significant annotation enrichment for 'intracellular signaling' harbored 16 of the candidate gene defects identified in nine index cases with Gi signaling abnormalities. Four gene clusters, representing 14 genes, with significantly associated gene ontology annotations were identified among the cases with secretion abnormalities, the most significant association being with 'establishment of protein localization.' CONCLUSION: Our findings demonstrate the genetic complexity of PFDs and highlight plausible candidate genes for targeted analysis in patients with platelet secretion and Gi signaling abnormalities.

Rare platelet GPCR variants: what can we learn?

Platelet-expressed GPCRs are critical regulators of platelet function. Pharmacological blockade of these receptors forms a powerful therapeutic tool in the treatment and prevention of arterial thrombosis associated with coronary atherosclerosis and ischaemic stroke. However, anti-thrombotic drug therapy is associated with high inter-patient variability in therapeutic response and adverse bleeding side effects. In order to optimize the use of existing anti-platelet drugs and to develop new therapies, more detailed knowledge is required relating to the molecular mechanisms that regulate GPCR and therefore platelet function. One approach has been to identify rare, function-disrupting mutations within key platelet proteins in patients with bleeding disorders. In this review, we describe how an integrated functional genomics strategy has contributed important structure-function information about platelet GPCRs with specific emphasis upon purinergic and thromboxane A2 receptors. We also discuss the potential implications these findings have for pharmacotherapy and for understanding the molecular basis of mild bleeding disorders.

How should we test for nonsevere heritable platelet function disorders?

Heritable platelet function disorders (HPFD) are a heterogeneous group of bleeding disorders with diverse clinical and laboratory characteristics. In contrast to the severe phenotype disorders, Glanzmann thrombasthenia and Bernard-Soulier syndrome, most nonsevere HPFD are incompletely characterized. This is a consequence of the poor standardization of diagnostic tests and of the lack of consensus about diagnostic criteria for the different subgroups of nonsevere HPFD. Distinguishing patients who have a nonsevere HPFD from those who do not is an essential first step in diagnosis which may be aided by bleeding assessment tools and screening tests such as the Platelet Function Analyser-100. However, high diagnostic accuracy can only be achieved with both light transmission aggregation (LTA) and secretion tests, for which streamlined agonist panels may be of similar utility to extended panels. Standardization of the methodology of these tests and quality assurance are essential for robust diagnosis. Identification of which platelet pathway is defective in patients with nonsevere HPFD is also usually possible with LTA and secretion tests. This strategy also sometimes enables exact diagnosis by implicating a single candidate protein and gene. Next-generation sequencing may offer a rapid approach to diagnosis of nonsevere HPFD, although rigorous strategies must be adopted to distinguish causative gene defects from bystander variations.

Effect of castration method on neurohormonal and electroencephalographic stress indicators in Holstein calves of different ages.

As public concern for food animal welfare increases, a need to validate objective pain assessment tools exists in order to formulate animal welfare policies and facilitate regulatory approval of compounds to alleviate pain in livestock in the United States. The aims of this study were (1) to compare the physiological response to pain induced by surgical and nonsurgical (band) castration in calves and (2) to elucidate age-related differences in pain response of calves subjected to different castration methods. Seventy-six Holstein bull calves were blocked by age (≤8-wk and ≥6-mo-old) and randomly assigned to 1 of 4 treatment groups: control (n=20), castration by banding (n=18), cut-and-clamp surgical castration (n=20), and cut-and-pull surgical castration (n=18). Measurements included electroencephalogram, heart rate variability, infrared thermography, electrodermal activity, and concentrations of serum cortisol and plasma substance P before, during, and within 20min following castration. Electroencephalogram recordings showed desynchronization for all treatments, consistent with increased arousal; yet the magnitude of desynchronization was greatest for 6-mo-old calves castrated by cut-and-clamp. Additionally, older calves in the cut-and-pull group showed greater desynchronization than younger calves in the same group. Based on the heart rate variability analysis, 6-mo-old calves in the control or cut-and-pull castration groups showed greater sympathetic tone than younger calves in the same treatment groups. Overall, younger calves showed lower electrodermal activity than older calves. Regardless of treatment, concentrations of cortisol and plasma substance P were greater in 6-mo-old calves relative to their younger counterparts, indicating a more robust response to all treatments in older calves. In summary, neurohormonal and electroencephalographic stress responses of calves to castration were age-specific. Castration by cut-and-clamp showed the most pronounced stress response in 6-mo-old calves. These findings provide evidence that support welfare policies recommending castration at an early age and the use of analgesic compounds at the time of surgical castration especially in older calves. However, the potential long-term negative consequences of early untreated pain must be considered and warrant further investigation.

Kiss1 and Kiss1r mRNA expression in the rat placenta: changes with gestational age and regulation by glucocorticoids.

INTRODUCTION: Kisspeptin, the neuropeptide product of the KISS1 gene, is synthesized by neurons within the hypothalamus and is critical for fertility. Human placenta also expresses KISS1 and kisspeptin receptor (KISS1R) mRNA within the trophoblast compartment, where it is thought to act as a physiological invasion inhibitor. METHODS: We determined the expression of Kiss1 mRNA in rat placenta and examined the effect of gestational age and feto-placental growth restriction, achieved through excess maternal glucocorticoid exposure. RESULTS: Dexamethasone induced fetal growth restriction at both day 16 and day 22 of gestation, but placental growth restriction only at day 22. Real-time quantitative RT-PCR revealed an increase in Kiss1 and Kiss1r mRNA from day 16-22 in the labyrinth and junctional zones of the rat placenta. Immunolocalization confirmed kisspeptin expression in the placenta and was prominent in trophoblast tissue. Dexamethasone exposure elevated the expression of Kiss1 mRNA in the labyrinth and junctional zones of day 16 placentas. In contrast, Kiss1 mRNA in the labyrinth zone was reduced following dexamethasone-treatment at day 22. Kiss1r expression was increased in both placental zones at day 16 and 22 in response to dexamethasone-treatment. CONCLUSIONS: We confirm the presence of Kiss1 and Kiss1r mRNA in the rat placenta with expression increasing over the final third of pregnancy, suggestive of a role in restricting placental growth. Furthermore, the effects of dexamethasone on placental Kiss1/Kiss1r suggest glucocorticoid-induced placental growth retardation could be mediated, in part, via early stimulation of Kiss1 and the subsequent inhibition of trophoblast proliferation and invasion.

The inflammatory state of the rat placenta increases in late gestation and is further enhanced by glucocorticoids in the labyrinth zone.

INTRODUCTION: Inflammation plays central roles in key aspects of successful reproduction: ovulation, implantation and parturition. In this study we characterised the inflammatory profile of the rat placenta in late gestation with and without maternal glucocorticoid (dexamethasone) treatment. METHODS: Placentas (n = 6/group) were collected from untreated (Con) rats at days 16 and 22 (term = day 23) and from dexamethasone-treated (Dex) rats at day 22. mRNA and protein expression was determined for enzymes of prostaglandin synthesis and metabolism (Ptgs-1, Ptgs-2, 15-Pgdh), pro-inflammatory cytokines (Tnf-α, Il-1ß, Il-6), and the macrophage marker Emr-1 in the junctional (JZ) and labyrinth (LZ) zones of the placenta. RESULTS: Tnf-α, Il-1ß and Il-6 mRNAs all increased (2- to 4-fold) in both placental zones between days 16 and 22 (P < 0.01). Ptgs-2 mRNA (30-fold; P < 0.01) and PTGS-2 protein (2.4-fold; P < 0.05) similarly increased in LZ. In contrast, 15-Pdgh expression increased in JZ but decreased in LZ; these changes were accompanied by decreased levels of PGE2 in the JZ and a trend towards increased LZ levels. Dex treatment inhibited fetal and placental growth, but had minimal effects on expression of Ptgs-1, Ptgs-2 or 15-Pdgh. Nevertheless, Dex treatment increased LZ PGE2 levels (5-fold, P < 0.01) at the end of gestation. Dex treatment increased Tnf-α mRNA expression in LZ (40%; P < 0.05), but modestly suppressed cytokine protein expression in JZ. CONCLUSIONS: These data demonstrate that the inflammatory state of the LZ increases near term coincident with the known increase in local glucocorticoid levels. This suggests the classic anti-inflammatory actions of glucocorticoids do not occur in the placental LZ.

Platelet dysfunction associated with the novel Trp29Cys thromboxane A2 receptor variant.

BACKGROUND: Genetic variations that affect the structure of the thromboxane A2 receptor (TP receptor) provide insights into the function of this key platelet and vascular receptor, but are very rare in unselected populations. OBJECTIVES: To determine the functional consequences of the TP receptor Trp29Cys (W29C) substitution. PATIENTS/METHODS: We performed a detailed phenotypic analysis of an index case (P1) with reduced platelet aggregation and secretion responses to TP receptor pathway activators, and a heterozygous TP receptor W29C substitution. An analysis of the variant W29C TP receptor expressed in heterologous cells was performed. RESULTS: Total TP receptor expression in platelets from P1 was similar to that of controls, but there was reduced maximum binding and reduced affinity of binding to the TP receptor antagonist [(3) H]SQ29548. HEK293 cells transfected with W29C TP receptor cDNA showed similar total TP receptor expression to wild-type (WT) controls. However, the TP receptor agonist U46619 was less potent at inducing rises in cytosolic free Ca(2+) in HEK293 cells expressing the W29C TP receptor than in WT controls, indicating reduced receptor function. Immunofluorescence microscopy and cell surface ELISA showed intracellular retention and reduced cell surface expression of the W29C TP receptor in HEK293 cells. Consistent with the platelet phenotype, both maximum binding and the affinity of binding of [(3) H]SQ29548 to the W29C TP receptor were reduced compared to WT controls. CONCLUSION: These findings extend the phenotypic description of the very rare disorder TP receptor deficiency, and show that the W29C substitution reduces TP receptor function by reducing surface receptor expression and by disrupting ligand binding.

Bilateral buphthalmia in a 4-month-old Texas longhorn steer.

Congenital ocular disease occurs uncommonly in cattle, with multiple abnormalities reported only sporadically in the literature. This report describes a case of anterior segment dysgenesis resulting in glaucoma in a 4-month-old Texas Longhorn steer. On clinical exam, bilateral buphthalmia was present and intraocular pressures exceeded 47 mm Hg in both eyes. On histopathologic examination, the iridocorneal angle and filtration apparatus were distorted due to collapse of the ciliary cleft and anterior displacement of the anterior portion of the ciliary body. No evidence of inflammation or other causes of glaucoma were recognized.

Cholesterol lowering and inhibition of sterol absorption by Lactobacillus reuteri NCIMB 30242: a randomized controlled trial.

BACKGROUND/OBJECTIVES: The percentage of hypercholesterolemic individuals not reaching their LDL-cholesterol (LDL-C) goal remains high and additional therapeutic strategies should be evaluated. The objective of this study was to evaluate the cholesterol-lowering efficacy and mechanism of action of bile salt hydrolase-active Lactobacillus reuteri NCIMB 30242 capsules in hypercholesterolemic adults. SUBJECTS/METHODS: A total of 127 subjects completed a randomized, double-blind, placebo-controlled, parallel-arm, multicenter study. Subjects were randomized to consume L. reuteri NCIMB 30242 capsules or placebo capsules over a 9-week intervention period. The primary outcome was LDL-C relative to placebo at the study end point. RESULTS: L. reuteri NCIMB 30242 capsules reduced LDL-C by 11.64% (P<0.001), total cholesterol by 9.14%, (P<0.001), non-HDL-cholesterol (non-HDL-C) by 11.30% (P < 0.001) and apoB-100 by 8.41% (P = 0.002) relative to placebo. The ratios of LDL-C/HDL-cholesterol (HDL-C) and apoB-100/apoA-1 were reduced by 13.39% (P = 0.006) and 9.00% (P = 0.026), respectively, relative to placebo. Triglycerides and HDL-C were unchanged. High-sensitivity C-reactive protein and fibrinogen were reduced by 1.05 mg/l (P = 0.005) and 14.25% (P = 0.004) relative to placebo, respectively. Mean plasma deconjugated bile acids were increased by 1.00 nmol/l (P=0.025) relative to placebo, whereas plasma campesterol, sitosterol and stigmasterol were decreased by 41.5%, 34.2% and 40.7%, respectively. CONCLUSIONS: The present results suggest that the deconjugation of intraluminal bile acids results in reduced absorption of non-cholesterol sterols and indicate that L. reuteri NCIMB 30242 capsules may be useful as an adjunctive therapy for treating hypercholesterolemia.

Relationship of whole body nitrogen utilization to urea kinetics in growing steers.

Urea kinetics were measured in 2 experiments, with treatments designed to change protein deposition by the animal. Our hypothesis was that increased protein deposition by cattle (Bos taurus) would reduce urea production and recycling to the gastrointestinal tract. Urea kinetics were measured by continuous intravenous infusion of (15)N(15)N-urea followed by measurement of enrichment in urinary urea at plateau. In Exp. 1, 6 steers (139 kg) were maintained in a model in which leucine was the most limiting AA. Treatments were arranged as a 2 × 3 factorial and were provided to steers in a 6 × 6 Latin square design. Leucine treatments included 0 or 4 g/d of abomasally supplemented L-leucine, and energy treatments included control, abomasal glucose infusion (382 g DM/d), or ruminal VFA infusion (150 g/d of acetic acid, 150 g/d of propionic acid, and 50 g/d of butyric acid). Leucine supplementation increased (P < 0.01) N retention, and energy supplementation tended to increase (P = 0.09) N retention without differences between glucose and VFA supplements (P = 0.86). Energy supplementation did not strikingly improve the efficiency of leucine utilization. Although both leucine and energy supplementation reduced urinary urea excretion (P ≤ 0.02), treatments did not affect urea production (P ≥ 0.34) or urea recycling to the gut (P ≥ 0.30). The magnitude of change in protein deposition may have been too small to significantly affect urea kinetics. In Exp. 2, 6 steers (168 kg) were maintained in a model wherein methionine was the most limiting AA. Steers were placed in 2 concurrent 3 × 3 Latin squares. Steers in one square were implanted with 24 mg of estradiol and 120 mg trenbolone acetate, and steers in the other square were not implanted. Treatments in each square were 0, 3, or 10 g/d of L-methionine. Implantation numerically improved N retention (P = 0.13) and reduced urea production rate (P = 0.03), urinary urea excretion (P < 0.01), and urea recycling to the gastrointestinal tract (P = 0.14). Effects of methionine were similar to implantation, but smaller in magnitude. When protein deposition by the body is increased markedly, ruminally available N in the diet may need to be increased to offset reductions in urea recycling.

Effects of ruminal casein and glucose on forage digestion and urea kinetics in beef cattle.

Effects of supplemental glucose and degradable intake protein on nutrient digestion and urea kinetics in steers (Bos taurus) given ad libitum access to prairie hay (4.7% CP) were quantified. Six ruminally and duodenally cannulated steers (initial BW 391 kg) were used in a 4 × 4 Latin square with 2 extra steers. Treatments were arranged as a 2 × 2 factorial and included 0 or 1.2 kg of glucose and 240 or 480 g of casein dosed ruminally once daily. Each period included 9 d for adaptation, 4 d for total fecal and urine collections, and 1 d for ruminal and duodenal sampling. Jugular infusion of (15)N(15)N-urea with measurement of enrichment in urine was used to measure urea kinetics. Glucose reduced forage intake by 18% (P < 0.01), but casein did not affect forage intake (P = 0.69). Glucose depressed (P < 0.01) total tract NDF digestion. Glucose supplementation decreased ruminal pH 2 h after dosing, but the effect was negligible by 6 h (treatment × time; P = 0.01). Providing additional casein increased the ruminal concentration of NH(3), but the increase was less when glucose was supplemented (casein × glucose; P < 0.01). Plasma urea-N was increased (P < 0.01) by additional casein but was reduced (P < 0.01) by glucose. Microbial N flow to the duodenum and retained N increased (P ≤ 0.01) as casein increased, but neither was affected by glucose supplementation. Urea-N entry rate increased (P = 0.03) 50% with increasing casein. Urinary urea-N excretion increased (P < 0.01) as casein increased. The proportion of urea production that was recycled to the gut decreased (P < 0.01) as casein increased. Glucose supplementation decreased (P < 0.01) urinary urea excretion but did not change (P ≥ 0.70) urea production or recycling. The amount of urea-N transferred to the gut and captured by ruminal microbes was less for steers receiving 480 g/d casein with no glucose than for the other 3 treatments (casein × glucose interaction, P = 0.05), which can be attributed to an excess of ruminally available N provided directly to the microbes from the supplement. Overall, the provision of supplemental glucose decreased forage intake and digestibility. Increasing supplemental casein from 240 to 480 g/d increased urea production but decreased the proportion of urea-N recycled to the gut.

Effects of supplemental energy and protein on forage digestion and urea kinetics in growing beef cattle.

Effects of supplemental energy sources on nutrient digestion and urea kinetics at 2 levels of degradable intake protein were evaluated in cattle (Bos taurus). Six ruminally and duodenally cannulated steers (208 ± 17 kg) were used in a 6 × 6 Latin square with treatments arranged as a 3 × 2 factorial. Energy treatments included a control, 600 g glucose dosed ruminally once daily, and 480 g VFA infused ruminally over 8 h daily. Casein (120 or 240 g) was dosed ruminally once daily. Steers had ad libitum access to prairie hay (5.8% CP). Jugular infusion of (15)N(15)N-urea with measurement of enrichment in urine was used to measure urea kinetics. Infusing VFA decreased (P < 0.01) forage intake by 27%. Supplementing glucose decreased (P < 0.01) total tract NDF digestibility and tended to decrease ruminal NDF digestibility; depressions in response to glucose tended to be greater at the lower level of casein. Increasing casein decreased (P < 0.02) ruminal pH. Infusing VFA decreased pH only during infusions, whereas glucose decreased pH 2 h after dosing. Ruminal concentrations of NH(3), acetate, and propionate decreased and butyrate concentration increased when glucose was supplemented. Increasing casein supplementation increased (P < 0.01) ruminal concentrations of NH(3), acetate, and propionate. Supplemental energy decreased (P = 0.03) plasma urea-N concentration, but casein level did not affect it (P = 0.16). Microbial N flow was greater (P < 0.04) for 240 than for 120 g/d casein but was not affected by supplemental energy (P = 0.23). Urea-N entry rate and gut entry of urea-N were not affected (P ≥ 0.12) by supplemental energy or casein, but the proportion of urea production that was recycled to the gut was less (P = 0.01) when 240 g/d rather than 120 g/d casein was provided. Compared with VFA, glucose tended (P = 0.07) to increase the proportion of urea-N entry rate that was recycled to the gut. Supplementation with glucose led to more (P = 0.01) microbial uptake of recycled urea than did supplementation with VFA. Urea recycling did not differ greatly among treatments despite impacts on ruminal pH and NH(3) and on plasma urea-N that were expected to alter urea transport across ruminal epithelium. Lack of treatment effects on urea production indicate that the complete diets did not provide excessive amounts of N and that increases of intestinally available AA were used efficiently by cattle for protein deposition.

Effect of nitrogen supplementation and zilpaterol-HCl on urea kinetics in steers consuming corn-based diets.

We studied effects of zilpaterol-HCl on steers consuming corn-based diets with nitrogen (N) supplementation provided by dried distillers grains with solubles (DDGS) or urea. Two sets of six steers (approximately 350 kg) were used in two replicates of similarly designed trials. Within each replicate, three steers were fed 60 mg/day of zilpaterol-HCl throughout the trial and three steers received no zilpaterol-HCl. Within zilpaterol treatment, three corn-based dietary N treatments were offered in Latin square designs: control (9.6% crude protein), urea (UREA; 12.4% crude protein) or DDGS (13.7% crude protein). Total feed intake was unexpectedly greater (p < 0.01) with zilpaterol feeding but was not affected by dietary N (p = 0.76). Nitrogen intake was greater (p < 0.01) when zilpaterol was fed and was greater (p < 0.05) for DDGS and UREA than for control. Despite greater N intake, zilpaterol did not affect urea entry rate (p = 0.80) or urea-N recycled to the gastrointestinal tract (GER; p = 0.94). As a percentage of N intake, urea entry rate (p = 0.19) tended to be less when zilpaterol was fed (91 vs. 123% of N intake), and GER was numerically (p = 0.34) less (72 vs. 92% of N intake). Microbial N flow was greater (p = 0.02) for zilpaterol than for control but did not differ (p = 0.78) among dietary N treatments. As a percentage of N intake, microbial N flow was unaffected by zilpaterol (p = 0.97), but was greater (p < 0.05) for control than DDGS or UREA. The lack of change in urea entry and GER in response to zilpaterol, despite greater N intake, as well as lower urea entry and GER when expressed as proportions of N intake provide some evidence that the amount of N available for urea production and recycling was reduced by zilpaterol.