IL4-10 Fusion Protein Shows DMOAD Activity in a Rat Osteoarthritis Model.

OBJECTIVE: Ideally, disease-modifying osteoarthritis (OA) drugs (DMOAD) should combine chondroprotective, anti-inflammatory, and analgesic effects in a single molecule. A fusion protein of interleukin-4 (IL-4) and IL-10 (IL4-10 FP) possesses these combined effects. In this study, the DMOAD activity of rat IL4-10 FP (rIL4-10 FP) was tested in a rat model of surgically induced OA under metabolic dysregulation. DESIGN: rIL4-10 FP was produced with HEK293F cells. Bioactivity of purified rIL4-10 FP was determined in a whole blood assay. Male Wistar rats (n = 20) were fed a high-fat diet (HFD) to induce metabolic dysregulation. After 12 weeks, OA was induced according to the Groove model. Two weeks after OA induction, rats were randomly divided into 2 groups and treated with 10 weekly, intra-articular injections of either rIL4-10 FP (n = 10) or phosphate buffered saline (PBS; n = 10). Possible antibody formation was evaluated using ELISA, cartilage degeneration and synovial inflammation were evaluated by histology and mechanical allodynia was evaluated using the von Frey test. RESULTS: Intra-articular injections with rIL4-10 FP significantly reduced cartilage degeneration (P = 0.042) and decreased mechanical allodynia (P < 0.001) compared with PBS. Only mild synovial inflammation was found (nonsignificant), limiting detection of putative anti-inflammatory effects. Multiple injections of rIL4-10 FP did not induce antibodies against rIL4-10 FP. CONCLUSION: rIL4-10 FP showed chondroprotective and analgesic activity in a rat OA model with moderate cartilage damage, mild synovial inflammation, and pain. Future studies will need to address whether less frequent intra-articular injections, for example, with formulations with increased residence time, would also lead to DMOAD activity.

Diet as a risk factor for antimicrobial resistance in community-acquired urinary tract infections in a middle-aged and elderly population: a case-control study.

OBJECTIVES: There is an ongoing debate as to what extent antimicrobial resistance (AMR) can be transmitted from animals to humans via the consumption of animal products. Because epidemiological data on the role of diet in AMR in humans are lacking, we investigated this association between diet and AMR for different antimicrobial drugs in Escherichia coli (E. coli) in urinary tract infections (UTIs). METHODS: Susceptibility of E. coli in urinary cultures and information on diet (with food frequency questionnaires) were obtained from participants of the Rotterdam study, a population-based prospective cohort study. The association between intake of several food groups (meat, seafood, eggs, dairy products, crops) and resistance of E. coli to several antimicrobial drugs (amoxicillin, amoxicillin-clavulanic acid, trimethoprim, sulfamethoxazole-trimethoprim, first-generation cephalosporins, cefotaxime, nitrofurantoin, norfloxacin) was studied. RESULTS: Urinary cultures with E. coli were obtained from 612 individuals, of whom 481 (78.6%) were women. Resistance rates varied from 246/611 (40.3%) for amoxicillin and 167/612 (27.3%) for trimethoprim to only 29/612 (4.7%) for nitrofurantoin and 16/462 (3.5%) for cefotaxime. A higher intake of chicken was associated with cefotaxime resistance (OR 2.18; 95% CI 1.05-4.51 per tertile increase); a higher intake of pork was associated with norfloxacin resistance (OR 1.42; 95% CI 1.04-1.95 per quartile increase). In contrast, a higher intake of cheese was associated with lower AMR to amoxicillin (OR 0.84; 95% CI 0.72-0.99 per quartile increase) and amoxicillin-clavulanic acid (OR 0.67; 95% CI 0.53-0.86 per quartile increase). CONCLUSIONS: These findings support the hypothesis that diet may play a role in the AMR of E. coli in UTIs.

Data on a rat infection model to assess porous titanium implant coatings.

A model is needed to study the effectiveness of different anti-bacterial coatings on complex metal implants in a bone environment. This article shares data on the design of porous titanium implants for intramedullary implantation in the proximal rat tibia. The implant length, diameter and porosity were optimized after testing on cadaveric specimens. This article shares data on which parameters are critical to establish a chronic implant infection in Sprague Dawley rats when using the new implant design. To this end, different strains of Staphylococcus aureus and inoculation doses were investigated.

Controlled release of celecoxib inhibits inflammation, bone cysts and osteophyte formation in a preclinical model of osteoarthritis.

Major hallmarks of osteoarthritis (OA) are cartilage degeneration, inflammation and osteophyte formation. COX-2 inhibitors counteract inflammation-related pain, but their prolonged oral use entails the risk for side effects. Local and prolonged administration in biocompatible and degradable drug delivery biomaterials could offer an efficient and safe treatment for the long-term management of OA symptoms. Therefore, we evaluated the disease-modifying effects and the optimal dose of polyesteramide microspheres delivering the COX-2 inhibitor celecoxib in a rat OA model. Four weeks after OA induction by anterior cruciate ligament transection and partial medial meniscectomy, 8-week-old female rats (n = 6/group) were injected intra-articular with celecoxib-loaded microspheres at three dosages (0.03, 0.23 or 0.39 mg). Unloaded microspheres served as control. During the 16-week follow-up, static weight bearing and plasma celecoxib concentrations were monitored. Post-mortem, micro-computed tomography and knee joint histology determined progression of synovitis, osteophyte formation, subchondral bone changes, and cartilage integrity. Systemic celecoxib levels were below the detection limit 6 days upon delivery. Systemic and local adverse effects were absent. Local delivery of celecoxib reduced the formation of osteophytes, subchondral sclerosis, bone cysts and calcified loose bodies, and reduced synovial inflammation, while cartilage histology was unaffected. Even though the effects on pain could not be evualated directly in the current model, our results suggest the application of celecoxib-loaded microspheres holds promise as novel, safe and effective treatment for inflammation and pain in OA.

Local and systemic inflammatory lipid profiling in a rat model of osteoarthritis with metabolic dysregulation.

OBJECTIVE: Bioactive oxidised lipids (oxylipins) are important signalling mediators, capable of modulating the inflammatory state of the joint and anticipated to be of importance in joint homeostasis and status of osteoarthritis. The aim of this study was to quantify oxylipin levels in plasma and synovial fluid from rats with experimentally induced osteoarthritis to investigate the potential role of oxylipins as a marker in the disease process of early osteoarthritis. DESIGN: Forty rats were randomly allocated to a standard or high-fat diet group. After 12 weeks, local cartilage damage was induced in one knee joint in 14 rats of each diet group. The remaining 6 rats per group served as controls. At week 24, samples were collected. Oxylipin levels were quantified by liquid chromatography-mass spectrometry. RESULTS: Overall, 31 lipid-derived inflammatory mediators were detected in fasted plasma and synovial fluid. Principal component analysis identified four distinct clusters associated with histopathological changes. Diet induced differences were evident for 13 individual plasma oxylipins, as well as 5,6-EET in synovial fluid. Surgical-model induced differences were evident for three oxylipins in synovial fluid (15-HETE, 8,9-DHET and 17R-ResolvinD1) with a different response in lipid concentrations for synovial fluid and plasma. CONCLUSIONS: We demonstrate the quantification of oxidised lipids in rat plasma and synovial fluid in a model of early experimental osteoarthritis. Oxylipins in the synovial fluid that were altered as consequence of the surgically induced osteoarthritis were not represented in the plasma. Our findings suggest differential roles of the oxylipins in the local versus peripheral compartment.

Risk factors of recurrent hamstring injuries: a systematic review.

BACKGROUND: Although recurrent hamstring injury is a frequent problem with a significant impact on athletes, data on factors determining the risk for a recurrent hamstring injury are scarce. OBJECTIVE: To systematically review the literature and provide an overview of risk factors for re-injury of acute hamstring muscle injuries. STUDY DESIGN: Prospective studies on risk factors for re-injury following acute hamstring injuries were systematically reviewed. Medical databases and reference lists of the included articles were searched. Two reviewers independently selected potential studies and assessed methodological quality; one reviewer extracted the data. A best-evidence synthesis of all studied risk factors was performed. RESULTS: Of the 131 articles identified, five prospective follow-up studies fulfilled our inclusion criteria. These studies reported a recurrence incidence of 13.9-63.3% in the same playing season up to 2 years after initial injury. Limited evidence for three risk factors and one protective factor for recurrent hamstring injury was found; patients with a recurrent hamstring injury had an initial injury with a larger volume size as measured on MRI (47.03 vs 12.42 cm(3)), more often had a Grade 1 initial trauma (Grade 0: 0-30.4%; Grade 1: 60.9-100%; Grade 2: 8.7%) and more often had a previous ipsilateral anterior cruciate ligament (ACL) reconstruction (66.6% vs 17.1%) independent of graft selection. Athletes in a rehabilitation programme with agility/stabilisation exercises rather than strength/stretching exercises had a lower risk for re-injury (7.7% vs 70%). No significant relationship with re-injury was found for 11 related determinants. There was conflicting evidence that a larger cross-sectional area is a risk factor for recurrent hamstring injury. CONCLUSIONS: There is limited evidence that athletes with a larger volume size of initial trauma, a Grade 1 hamstring injury and a previous ipsilateral ACL reconstruction are at increased risk for recurrent hamstring injury. Athletes seem to be at lower risk for re-injury when following agility/stabilisation exercises.

Abdominal Palpation Haptic Device for Colonoscopy Simulation Using Pneumatic Control.

In this paper, we describe the development of a haptic device to be used in a simulator aiming to train the skills of gastroenterology assistants in abdominal palpation during colonoscopy, as well as to train team interaction skills for the colonoscopy team. To understand the haptic feedback forces to be simulated by the haptic device, we conducted an experiment with five participants of varying BMI. The applied forces and displacements were measured and hysteresis modeling was used to characterize the experimental data. These models were used to determine the haptic feedback forces required to simulate a BMI case in response to the real-time user interactions. The pneumatic haptic device consisted of a sphygmomanometer bladder as the haptic interface and a fuzzy controller to regulate the bladder pressure. The haptic device showed good steady state and dynamic response was adequate for simulating haptic interactions. Tracking accuracy averaged 94.2 percent within 300 ms of the reference input while the user was actively applying abdominal palpation and minor repositioning.

Are coupled rotations in the lumbar spine largely due to the osseo-ligamentous anatomy?--a modeling study.

Prior studies have found that primary rotations in the lumbar spine are accompanied by coupled out-of-plane rotations. However, it is not clear whether these accompanying rotations are primarily due to passive (discs, ligaments and facet joints) or active (muscles) spinal anatomy. The aim of this study was to use a finite element (FE) model of the lumbar spine to predict three-dimensional coupled rotations between the lumbar vertebrae, due to passive spinal structures alone. The FE model was subjected to physiologically observed whole lumbar spine rotations about in vivo centres of rotation. Model predictions were validated by comparison of intra-discal pressures and primary rotations with in vivo measurements and these showed close agreement. Predicted coupled rotations matched in vivo measurements for all primary motions except lateral bending. We suggest that coupled rotations accompanying primary motions in the sagittal (flexion/extension) and transverse (axial rotation) planes are primarily due to passive spinal structures. For lateral bending the muscles most likely play a key role in the coupled rotation of the spine.

A robotic testing facility for the measurement of the mechanics of spinal joints.

A robotic testing facility for the measurement of joint mechanics was used to determine the significance of tears in the intervertebral disc on the mechanics of the spinal joint. Ten lumbar joints of sheep were dynamically loaded and manipulated. Comparisons were made between the behaviour of the intervertebral disc in flexion and extension at two test speeds. The influence of the posterior elements and of rim lesions was assessed by testing the joint immediately before and after removal of the posterior elements and after the creation of a 4 mm x 10 mm rim lesion in the disc. Stiffness of the spinal joint dropped significantly upon removal of the posterior elements, from 0.81 to 0.23 N m/deg for flexion and from 0.65 to 0.40 N m/deg for extension. Maximum moments dropped 37 per cent for flexion and 63 per cent for extension. The rim lesion caused a further significant stiffness reduction to 0.21 and 0.31 N m/deg respectively. Maximum moments reduced a further 12 per cent and 16 per cent respectively. A higher test speed (2 deg/s instead of 0.5 deg/s) usually did not change these results significantly. The novel six-degrees-of-freedom robotic testing facility used in this study was demonstrated to be an effective system for studying the mechanics of complex biological joints.

Slip and damage properties of jaws of laparoscopic graspers.

BACKGROUND: The optimal jaws of laparoscopic graspers can be used to manipulate the tissue properly with minimal damage. The criteria jaws should satisfy are investigated. METHODS: The cecum of pigs was clamped between 13 pairs of jaws differing in size and profile. Various pinch and pull forces were applied. At 5 N pull force, the minimally required pinch force to prevent slip and the pinch force that is maximally allowable without causing damage were obtained. RESULTS: With increasing size of the jaws, the contact area with the tissue increases. With increasing contact area, the pinch force leading to tissue damage was increased from 15 to 37 N. A profile of the jaws decreased the pinch force needed to prevent slip of tissue from 22 to 3 N. CONCLUSION: An optimal jaw possesses a large contact area to prevent tissue damage and a slight profile to prevent tissue slip.

Inter- and intraindividual variabilities of perforation forces of human and pig bowel tissue.

BACKGROUND: A laparoscopic bowel grasper should be suitable for safely grasping the bowel in a wide variety of patients. Therefore, the inter- and intraindividual variabilities in the strength of bowel tissue to resist perforation force should be analyzed. METHODS: The large and small bowels of pigs ( n = 14) and the human small bowel ( n = 7) were clamped between two hemispheres 1.5 mm in diameter. The pinch force was increased until the tissue was perforated. RESULTS: The perforation force for the pig large bowel was higher than for the small bowel (13.5 +/- 3.7 vs 11.0 +/- 2.5 N; p = 0.014). No difference was found between the human and pig small bowel (10.3 +/- 2.9 vs 11.0 +/- 2.5 N). The intercoefficient of variation varied between 22% and 28%, and the intracoefficient of variation varied between 14% and 18%. CONCLUSIONS: The strength of the pig bowel is approximately comparable to the strength of the human bowel, and, therefore, testing of graspers on pig bowel is justified. However, due to the large interindividual variation, large safety margins should be taken into account.

Forces and displacements in colon surgery.

BACKGROUND: This study aims to find criteria for designing safer laparoscopic graspers for manipulation of delicate tissues. Since the experiment was performed in an open-surgery setup without any movement restrictions, the results are relevant to general surgery as well as laparoscopic. METHOD: Four surgeons were asked to apply via two clamps enough force to a pig's colon to stretch the mesocolon for dissection. Directions, magnitudes, and application points of the forces were recorded. RESULTS: The surgeons applied per clamp on average 2.5 N and maximally 5 N to the colon. These forces were at an angle of about 40 degrees to 70 degrees relative to the horizontal plane and approximately 60 degrees relative to each other. CONCLUSIONS: Safe laparoscopic graspers should be able to transmit at least a 5 N pull force to the tissue without damaging it. To minimize the forces used, the angles mentioned should be taken into account when placing the trocars.

Force-directed design of a voluntary closing hand prosthesis.

This paper presents the design of a body-powered voluntary closing prosthetic hand. It is argued that the movement of the fingers before establishing a grip is much less relevant for good control of the object held than the distribution of forces once the object has been contacted. Based on this notion, the configurations of forces on the fingers and the force transmission through the whole mechanism were taken as a point of departure for the design, rather than movement characteristics. For a good distribution of pinching forces on the object and a natural behavior, the prosthesis is made adaptive and flexible. To achieve good force feedback, the disturbing influences of the cosmetic glove are strongly reduced by a compensation mechanism. To further improve the transmission of forces, friction is reduced by furnishing the whole mechanism with rolling links. This force-directed design approach has led to a simple mechanism with low operating force and good feedback of the pinching force.

Different techniques to study rumen fermentation characteristics of maturing grass and grass silage.

Grass samples were harvested during the 1993 growing season after a precut on April 27, 1993 and were stored frozen or left to ensile in 30-L buckets. Effects on chemical composition and fermentation kinetics of the maturation of the grass and of ensiling were investigated. Chemical composition and fermentation kinetics were determined using the gas production technique, in vitro techniques, and the nylon bag technique. Two silage samples were also investigated in vivo. Maturation caused a decrease in crude protein content and organic matter degradability and an increase in neutral detergent fiber, acid detergent fiber, and lignin contents. Degradation rates were highest for the youngest samples and decreased as grass and silage matured. This result could be observed from the first derivative of the cumulative gas production curves (i.e., the rate of gas production) and from the gas production parameters. The rate of degradation of the nonsoluble fraction of the young samples, determined using the gas production technique, was relatively higher in rumen fluid from a cow fed silage from grass cut at a young stage. More mature samples were degraded relatively faster in rumen fluid from a cow fed mature grass silage, suggesting a specific adaptation of the rumen microorganisms to the grass properties. There was a good relationship among the second phase of gas production (i.e., fermentation of the nonsoluble fraction), maturity of the grass and grass silage samples, degradability determined with the Tilley and Terry technique, and degradability determined after 46 h of incubation in rumen fluid. Results obtained with both of the different in vitro techniques and the nylon bag technique were confirmed by the in vivo experiments involving the two silage samples.

Starch and stage of maturity of grass silage: site of digestion and intestinal nutrient supply in dairy cows.

The interaction between the quality of grass silage and starch supplementation on ruminal digestion was studied in an experiment with a 2 x 2 factorial design using four dairy cows. Treatment factors were grass silage harvested after either 21 or 37 d of regrowth and two concentrations of steam-flaked corn starch (0 or 4 kg/d). Ruminal volume and flow of duodenal digesta were estimated. When forage was harvested at a more mature stage, only minor effects were noted for silage composition and, consequently, ruminal and intestinal digestion. The addition of starch to the diet tended to reduce ruminal digestion of neutral detergent fiber. The reduction in ruminal digestion was not compensated by increased digestion in the large intestine. Starch increased duodenal nonammonia N flow because of an increase in bacterial N flow. The increase in bacterial N was accompanied by a reduction in the escape of feed N from the rumen. Results from this study indicate that the addition of ruminally available starch to diets based on grass silage reduced ruminally degradable neutral detergent fiber and increased the duodenal supply of protein. These effects have to be taken into account to predict production responses to extra starch.

Starch supplementation of grass harvested at two stages of maturity prior to ensiling: intake, digestion, and degradability by dairy cows.

The effects of the maturity of grass prior to ensiling and the supplementation of starch to grass silage on apparent digestibility, degradability, rumen content, and feed intake by dairy cows were investigated using a Latin square design. Treatments were silages from early or late cut grass with or without 4 kg of supplemental flaked corn starch. The silage from early cut grass contained more N and sugars than did the silage from late cut grass but was lower in neutral detergent fiber (NDF). Degradation characteristics were not different between the two silages. Apparent digestibilities of dry matter (DM), organic matter (OM), and N in the silage from late cut grass were lower than those in the silage from early cut grass, but NDF digestibility was not affected. Starch supplementation increased the lag phase for DM and OM in both silages, but the rate of degradation was decreased for NDF. Starch supplementation did not influence digestibilities of DM and OM in silage from early cut grass but decreased the digestibilities of DM and OM in silage from late cut grass. Crude protein and NDF digestibilities were decreased for silages from early and late cut grass. Starch supplementation increased NDF in the rumen of cows fed the silage from early cut grass, but NDF was not affected by starch supplementation for cows fed the silage from late cut grass. Rumen-degradable starch negatively influences degradability and apparent OM digestibility; the extent of the decrease is related to the maturity of the NDF.

Influence of maturity of grass silage and flaked corn starch on the production and metabolism of volatile fatty acids in dairy cows.

An experiment employing a Latin square design was used to quantify the effects of two stages of maturity of grass silage (early cut and late cut) and three concentrations of flaked corn starch (0, 2, and 4 kg) on the molar proportion of rumen volatile fatty acids (VFA), the production of rumen VFA, and the net fluxes of VFA in the splanchnic tissue of cows. The molar proportions of VFA in rumen fluid were similar for cows fed both silages. When the silage diets were supplemented with starch, the proportion of propionic acid increased for cows fed diets containing early cut grass silage, but no effects were found for cows fed diets containing late cut grass silage. Estimated gastrointestinal production of acetate, propionate, butyrate, and branched-chain fatty acids plus valerate was related to intake of metabolizable energy and organic matter fermented into VFA. The portal release of acetate was approximately 14% lower than the estimated production of acetate by cows fed diets containing early cut grass silage, but cows fed diets containing late cut grass silage showed a variable difference between estimated production and portal release (31, 24, and 15%, respectively) as starch supplementation increased. The portal release of butyrate plus beta-hydroxybutyrate and the release of branched-chain fatty acids plus valerate were approximately 70 and 25%, respectively, of the estimated gastrointestinal production. Propionate production was similar to the portal release of propionate. Net flux measurements in splanchnic tissue in combination with gastrointestinal digestion and kinetics provide information that increases the knowledge of pathways and metabolism and quantifies the availability of individual nutrients for milk production in dairy cows.

Effects of source of starch on net portal flux of glucose, lactate, volatile fatty acids and amino acids in the pig.

The ileal digestibilities of maize starch and native pea starch do not differ. However maize starch is digested faster than pea starch and the ileal amino acid digestibility of a diet containing pea starch is lower. In the present study, the net portal fluxes of glucose, lactate, volatile fatty acids (VFA) and amino acids were measured for diets including 650 g maize starch or pea starch/kg. The diets were fed at a level 870 kJ digestible energy/kg0.75 twice daily (06.00 and 18.00 hours) to four female pigs in a crossover design. Portal vein blood flow did not differ between maize and pea starches (1620 and 1484 ml/min respectively; SED 100; P = 0.23). For maize starch portal glucose flux was significantly higher during the first 6 h after feeding, was not different 8 h after feeding and was significantly lower thereafter. Net portal glucose flux was higher for maize starch than for pea starch (1759 and 1265 mmol/12 h respectively; SED 182; P = 0.054). Net portal lactate flux was not significantly different between maize and pea starches (36.5 and 67.2 mmol/12 h respectively; SED 24.1; P = 0.27) and net portal VFA flux was lower for maize starch than for pea starch (169 and 218 mmol/12 h respectively; SED 18; P = 0.054). Net portal fluxes of valine, isoleucine, phenylalanine, tryptophan, arginine, serine, cystine, tyrosine, lysine, histidine and the sum of essential amino acids tended to be or were higher (P < 0.1 or P < 0.05) and net portal flux of aspartic acid tended to be lower for pea starch (P < 0.1). It can be concluded that, although ileal digestibility of both starches is equal, the rate of appearance of glucose in the portal vein was higher for maize starch, influencing the net portal flux of amino acids.

Comparison and evaluation of mechanistic rumen models.

Mechanistic rumen models of Baldwin (1995), Danfaer (1990) and Dijkstra et al. (1992) were compared on identical inputs that were derived from trials with lactating dairy cows fed on grass herbage. Consistent differences were detected between models and between predicted and observed outputs. None of the models seemed to predict all nutrient flows best. The models particularly differed in the representation of microbial metabolism: degradation of insoluble substrate, fermentation of substrate into volatile fatty acids, and incorporation of substrate into microbial matter. Differences amongst models in the prediction of these processes compensated for each other and consequently all models predicted the duodenal flow of non-NH3 N, microbial N and organic matter reasonably well. Large differences remained in the prediction of individual nutrient flows, however, and it was stressed that in order to enhance prediction of the profile of nutrient flows, the mechanisms of microbial metabolism need to be tested on their ability to describe the intraruminal transactions. However, this requires more-detailed information on individual nutrient flows and on the microbial or non-microbial origin of duodenal contents. Parameter inputs for physical and chemical feed properties were identified that are improperly defined in extant models or susceptible to error. The description of these feed characteristics needs to be developed further and become identifiable for a wide range of dietary conditions.

Effect of resistant starch on net portal-drained viscera flux of glucose, volatile fatty acids, urea, and ammonia in growing pigs.

Net portal-drained viscera (PDV) flux of glucose, VFA, ammonia, and urea was determined in pigs fed diets with or without resistant starch. Diets consisted of 65% cornstarch (diet CS), 32.5% cornstarch and 32.5% raw potato starch (diet CPS), or 65% raw potato starch (diet PS); the remaining 35% supplied all amino acids, fat, fiber, minerals, and vitamins. The diets contained twice the maintenance requirement for energy and were fed twice daily to four barrows (initial BW 56 kg) in three periods in a crossover design. The pigs were fitted with catheters in a mesenteric vein, a mesenteric-artery, and the portal vein, and net PDV flux was calculated by multiplying portal-arterial concentration differences and corresponding portal vein flow. Net PDV flux of glucose was significantly less after feeding diets CPS and PS, and portal absorption of ileally digested glucose was 89, 66, and 41% for diets CS, CPS, and PS, respectively. Net PDV flux of VFA was lowest after feeding diet CS and three to four times higher after feeding diets CPS and PS. Net PDV flux of ammonia was highest for diet CS and almost halved after feeding diets CPS and PS. There was a small negative net PDV flux of urea for diets CS and CPS, which significantly increased after feeding diet PS. These results suggest that excretion of nitrogen is shifted from urine to feces primarily by reduction of the net PDV flux of ammonia when resistant starch is fed.