Effects of dietary lysine levels on the concentrations of selected nutrient metabolites in blood plasma of late-stage finishing pigs.

Lysine is the first-limiting amino acid (AA) in typical swine diets and plays very important roles in promoting growth performance of pigs. This research was conducted to study the effects of dietary lysine on blood plasma concentrations of protein, carbohydrate, and lipid metabolites of pigs. Eighteen crossbred finishing pigs (nine barrows and nine gilts; initial BW 92.3 ± 6.9 kg) were individually penned in an environment controlled barn. Pigs were assigned to three dietary treatments according to a randomized complete block design with gender as block and pig as experimental unit (6 pigs/treatment). Three corn and soybean meal-based diets were formulated to contain total lysine at 0.43%, 0.71%, and 0.98% (as-fed basis) for Diets I (lysine deficient), II (lysine adequate), and III (lysine excess) respectively. After 4 weeks on trial, jugular vein blood was collected and plasma was separated. The plasma concentrations of total protein, albumin, urea nitrogen (UN), triglyceride, total cholesterol, and glucose were determined using an ACE Clinical Chemistry System (Alfa Wassermann, Inc., West Caldwell, NJ, USA). Data were analysed using the GLM Procedure with PDIFF (adjust = T) option of SAS. No differences (p > 0.10) were found between barrows and gilts for any of the metabolites measured. While there were no differences (p > 0.10) between pigs fed Diets II and III in plasma concentrations of UN, albumin, and total cholesterol, the concentration of albumin in these pigs was higher (p < .05) than that of pigs fed Diet I, and the concentrations of UN and total cholesterol in these pigs were lower (p < .05) than that of pigs fed Diet I. There were no differences (p > 0.10) among the three dietary treatments in plasma concentrations of total protein, triglycerides, and glucose. These findings indicated that the plasma metabolite profile can be affected by changing dietary lysine content only. Thorough understanding how the plasma metabolite profile is alternated by dietary lysine will facilitate nutrient management for more sustainable swine production.

Feeding glycerol-enriched yeast culture improves lactation performance, energy status, and hepatic gluconeogenic enzyme expression of dairy cows during the transition period.

This study aimed to evaluate the effects of feeding glycerol-enriched yeast culture (GY) on feed intake, lactation performance, blood metabolites, and expression of some key hepatic gluconeogenic enzymes in dairy cows during the transition period. Forty-four multiparous transition Holstein cows were blocked by parity, previous 305-d mature equivalent milk yield, and expected calving date and randomly allocated to 4 dietary treatments: Control (no additive), 2 L/d of GY (75.8 g/L glycerol and 15.3 g/L yeast), 150 g/d of glycerol (G; 0.998 g/g glycerol), and 1 L/d of yeast culture (Y; 31.1 g/L yeast). All additives were top-dressed and hand mixed into the upper one-third of the total mixed ration in the morning from -14 to +28 d relative to calving. Results indicated that the DMI, NE intake, change of BCS, and milk yields were not affected by the treatments ( > 0.05). Supplementation of GY or Y increased milk fat percentages, milk protein percentages, and milk protein yields relative to the Control or G group ( < 0.05). Cows fed GY or G had higher glucose levels and lower ß-hydroxybutyric acid (BHBA) and NEFA levels in plasma than cows fed the Control ( < 0.05) and had lower NEFA levels than cows fed Y ( < 0.05). On 14 d postpartum, cows fed GY or G had higher enzyme activities, mRNA, and protein expression of cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C; < 0.05); higher enzyme activities ( < 0.05) and a tendency toward higher mRNA expression ( < 0.10) of glycerol kinase (GK); and a tendency toward higher enzyme activities of pyruvate carboxylase (PC) in the liver ( < 0.10) when compared with cows fed Control or Y. The enzyme activities, mRNA, and protein expression of PEPCK-C, PC, and GK did not differ between cows fed GY and G ( > 0.10). In conclusion, dietary GY or Y supplementation increased the milk fat and protein content of the cows in early lactation and GY or G supplementation improved the energy status as indicated by greater plasma glucose and lower plasma BHBA and NEFA concentrations and upregulated the hepatic gluconeogenic enzymes of dairy cows during the transition period. Feeding cows with a GY mixture in the peripartum period combined the effects of yeast on lactation performance and the effects of glycerol on energy status in dairy cows.

Gene expression profiling indicates an increased capacity for proline, serine, and ATP synthesis and mitochondrial mass by the liver of steers grazing high vs. low endophyte-infected tall fescue.

Grazing -infected forages results in a variety of reduced animal performance parameters, collectively known as "fescue toxicosis." The initial, limited evaluations of hepatic mechanisms affected by fescue toxicosis have used transcriptomic expression profiling of experimental phenotypes developed by short-term feeding of concentrated ergot alkaloids or fescue seeds to rodents and steers. To assess the effects of fescue toxicosis in growing cattle using a commercially relevant phenotype, we induced fescue toxicosis in beef steers by summer-long grazing (89 to 105 d) of a single high toxic endophyte-infected tall fescue pasture (HE; 0.746 µg/g ergot alkaloids; 5.7 ha; = 10; BW = 267 ± 14.5 kg) vs. a low toxic endophyte tall fescue-mixed pasture (LE; 0.023 µg/g ergot alkaloids; 5.7 ha; = 9; BW = 266 ± 10.9 kg). High toxic endophyte tall fescue-mixed pasture steers had decreased BW (313 vs. 338 kg) and an increased potential for hepatic gluconeogenesis from AA-derived carbons. To gain a greater perspective into fescue toxicosis-induced hepatic metabolism and identify candidate regulatory mechanisms, the goal of the current research was to examine liver samples for changes in gene (mRNA) expression profiles using a Bovine Affymetrix microarray and selected reverse-transcription PCR and immunoblot analyses. The expression (false discovery rate < 10%; < 0.01) of 147 genes was increased (7 to 268%) and that of 227 was decreased (4 to 87%) in livers of HE vs. LE steers. The top (1) functional gene category was cell-mediated immune response (33 genes; ≤ 0.012), (2) canonical cell signaling pathway was primary immunodeficiency signaling (8 genes; ≤ 0.0003), and (3) canonical metabolic pathways were oxidative phosphorylation (5 genes; ≤ 0.016) and purine metabolism (8 genes; ≤ 0.029). High toxic endophyte tall fescue-mixed pasture steers had increased ( ≤ 0.022) expression of genes critical for increased (1) Pro () and Ser () synthesis, (2) shunting of AA carbons into pyruvate () and ATP synthesis (, , , COX4, , and ), and (3) mitochondrial mass (COX4). Targeted reverse-transcribed PCR or immunoblot assays corroborated ( ≤ 0.035) these latter microarray findings for , , , , and COX4. Moreover, network analysis identified glucocorticoid receptor-mediated signaling as the most probable mechanism to coordinate the above findings. These results greatly extend our knowledge of the consequences of summer-long grazing of endophyte-infected tall fescue to the hepatic metabolism of growing steers.

Glutamine synthetase and alanine transaminase expression are decreased in livers of aged vs. young beef cows and GS can be upregulated by 17ß-estradiol implants.

Aged beef cows (≥ 8 yr of age) produce calves with lower birth and weaning weights. In mammals, aging is associated with reduced hepatic expression of glutamine synthetase (GS) and alanine transaminase (ALT), thus impaired hepatic Gln-Glu cycle function. To determine if the relative protein content of GS, ALT, aspartate transaminase (AST), glutamate transporters (EAAC1, GLT-1), and their regulating protein (GTRAP3-18) differed in biopsied liver tissue of (a) aged vs. young (3 to 4 yr old) nonlactating, nongestating Angus cows (Exp. 1 and 2) and (b) aged mixed-breed cows with and without COMPUDOSE (17ß-estradiol) ear implants (Exp. 3), Western blot analyses were performed. In Exp. 1, 12 young (3.62 ± 0.01 yr) and 13 aged (10.08 ± 0.42 yr) cows grazed the same mixed forage for 42 d (August-October). In Exp. 2, 12 young (3.36 ± 0.01 yr) and 12 aged (10.38 ± 0.47 yr) cows were individually fed (1.03% of BW) a corn-silage-based diet to maintain BW for 20 d. For both Exp. 1 and 2, the effect of cow age was assessed by ANOVA using the MIXED procedure of SAS. Cow BW did not change ( ≥ 0.17). Hepatic ALT (78% and 61%) and GS (52% and 71%) protein content (Exp. 1 and 2, respectively) was decreased ( ≤ 0.01), whereas GTRAP3-18 (an inhibitor of EAAC1 activity) increased ( ≤ 0.01; 170% and 136%) and AST, GLT-1, and EAAC1 contents did not differ ( ≥ 0.17) in aged vs. young cows. In Exp. 2, free concentrations (nmol/g) of Glu, Ala, Gln, Arg, and Orn in liver homogenates were determined. Aged cows tended to have less ( = 0.10) free Gln (15.0%) than young cows, whereas other AA concentrations did not differ ( 0.26). In Exp. 3, 14 aged (> 10 yr) cows were randomly allotted ( = 7) to sham or COMPUDOSE (25.7 mg of 17ß-estradiol) implant treatment (TRT), and had ad libitum access to alfalfa hay for 28 d. Blood and liver biopsies were collected 14 and 28 d after implant treatment. Treatment, time after implant (DAY), and TRT × DAY effects were assessed by ANOVA using the MIXED procedure of SAS. Cow BW was not affected ( ≥ 0.96). Implant increased ( ≤ 0.02) total plasma estradiol by 220% (5.07 vs. 1.58 pg/mL) and GS protein by 300%, whereas the relative content of other proteins was not altered ( ≥ 0.16). We conclude that hepatic expression of ALT and GS are reduced in aged vs. young cows, and administration of 17ß-estradiol to aged cows increases plasma estradiol and hepatic GS, but not that of other proteins that support hepatic Glu metabolism.

Effects of dietary supplementation of selenium-enriched probiotics on production performance and intestinal microbiota of weanling piglets raised under high ambient temperature.

This study was designed to evaluate the efficacy of selenium-enriched probiotics (SeP) on production performance and intestinal microbiota of piglets raised under high ambient temperature. Forty-eight cross-bred weanling piglets (28 days old), randomly allotted into 12 pens (four piglets/pen) and four dietary treatments (three pens/treatment group), were fed ad libitum for 42 days a basal diet (Con) or the basal diet supplemented with probiotics (Pro), sodium selenite (ISe) or a SeP preparation. Blood and faecal samples were collected on days 0, 14, 28 and 42 post-treatment. The SeP group had higher final BW (p < 0.05), greater ADG (p < 0.05) and lower FCR (p < 0.01) than the Pro, ISe or Con group. The diarrhoea incidence rate of either SeP or Pro group was lower (p < 0.01) than the ISe or Con group. Blood Se concentration and GSH-Px activity were both higher (p < 0.01) in the SeP than in the Pro, ISe or Con group. On days 28 and 42, the serum concentrations of T3 were higher (p < 0.01) and T4 lower (p < 0.01) in the SeP than in the ISe group, and the T3 and T4 concentrations in the ISe group, in turn, were higher (p < 0.05) and lower (p < 0.01), respectively, than in the Pro or Con group. Also on days 28 and 42, the faecal counts of lactobacillus bacteria were higher (p < 0.01) while Escherichia coli lower (p < 0.01) in the SeP or Pro group as compared to the ISe or Con group. The results of RFLP showed that the faecal microbial flora in the SeP group changed the most (numerically) as compared to the Pro or ISe group. These results suggest that the SeP product may serve as a better alternative to antibiotics than the solo probiotics for using as a growth promoter for weanling piglets.

Feeding glycerol-enriched yeast culture improves performance, energy status, and heat shock protein gene expression of lactating Holstein cows under heat stress.

This study was conducted to evaluate the effects of supplemental common yeast culture (CY) and glycerol-enriched yeast culture (GY) on performance, plasma metabolites, antioxidant status, and heat shock protein 70 (HSP70) mRNA expression in lactating Holstein cows under heat stress. During summer months, 30 healthy multiparous lactating cows (parity 3.25 ± 0.48; 60 ± 13 d in milk [DIM]; 648 ± 57 kg BW; an average milk yield of 33.8 ± 1.6 kg/d) were blocked by parity, previous milk yield, and DIM and randomly allocated to 3 dietary treatments: no supplemental yeast culture (Control), 1 L/d of CY (33.1 g yeast) per cow, and 2 L/d of GY (153.2 g glycerol and 31.6 g yeast) per cow. During the 60-d experiment, values of air temperature and relative humidity inside the barn were recorded hourly every 3 d to calculate temperature-humidity index (THI). Weekly rectal temperatures (RT) and respiration rates and daily DMI and milk yield were recorded for all cows. Milk and blood samples were taken twice monthly, and BW and BCS were obtained on d 0 and 60. In this experiment, THI values indicated cows experienced a moderate heat stress. Cows supplemented with CY and GY had greater yields of milk, energy-corrected milk and milk fat, and milk fat percent but lower HSP70 mRNA expression in peripheral blood lymphocytes than Control cows (P < 0.05). Supplementing CY and GY tended (P < 0.15) to decrease RT at 1400 h, increase milk protein yield and erythrocyte glutathione, and reduce plasma urea nitrogen compared with Control. Lower plasma NEFA concentration and HSP70 mRNA expression in peripheral blood lymphocytes (P < 0.05) and tendencies towards greater plasma glucose concentration (P = 0.11) but less BW loss (P = 0.14) were observed in GY relative to CY cows. In conclusion, either CY or GY supplementation partially mitigated the negative effects of heat stress on performance and HSP70 mRNA expression of lactating cows, and GY supplementation provided additional improvements in energy status and HSP70 gene expression of lactating cows.

Bovine neuronal vesicular glutamate transporter activity is inhibited by ergovaline and other ergopeptines.

l-Glutamate (Glu) is a major excitatory neurotransmitter responsible for neurotransmission in the vertebrate central nervous system. Vesicular Glu transporters VGLUT1 and VGLUT2 concentrate (50mM) Glu [Michaelis constant (measuring affinity), or K(m),=1 to 4mM] into synaptic vesicles (SV) for subsequent release into the synaptic cleft of glutamatergic neurons. Vesicular Glu transporter activity is dependent on vacuolar H(+)-ATPase function. Previous research has shown that ergopeptines contained in endophyte-infected tall fescue interact with dopaminergic and serotoninergic receptors, thereby affecting physiology regulated by these neuron types. To test the hypothesis that ergopeptine alkaloids inhibit VGLUT activity of bovine cerebral SV, SV were isolated from cerebral tissue of Angus-cross steers that were naive to ergot alkaloids. Immunoblot analysis validated the enrichment of VGLUT1, VGLUT2, synaptophysin 1, and vacuolar H(+)-ATPase in purified SV. Glutamate uptake assays demonstrated the dependence of SV VGLUT-like activity on the presence of ATP, H(+)-gradients, and H(+)-ATPase function. The effect of ergopeptines on VGLUT activity was evaluated by ANOVA. Inhibitory competition (IC(50)) experiments revealed that VGLUT-mediated Glu uptake (n=9) was inhibited by ergopeptine alkaloids: bromocriptine (2.83±0.59µM)

A cross-ethnic comparison on incidence of suicide.

BACKGROUND: Suicide rates vary widely across nations and ethnic groups. This study aims to explore potential factors contributing to inter-ethnic differences in suicide rates. METHOD: Study subjects came from a case-control psychological autopsy study conducted in Taiwan, including 116 consecutive suicides from two aboriginal groups and Taiwanese Han; 113 of them each matched with two living controls. Gender-, age- and method-specific suicide rates, population attributable fraction (PAF) of suicide for five major risk factors, help-seeking before suicide and emergency medical aid after suicide were compared between the three ethnic groups. RESULTS: One aboriginal group (the Atayal) had significantly higher adjusted rate ratios (RR) of suicide than the other aboriginal group (the Ami) [RR 0.20, 95% confidence intervals (CI) 0.12-0.34] and the Han (RR 0.26, 95% CI 0.16-0.40). Such differences can be explained by higher PAFs of suicide for three major risk factors (substance dependence, PAF 47.6%, 95% CI 25.5-64.2; emotionally unstable personality disorder, PAF 52.7%, 95% CI 32.8-69.0; family history of suicidal behaviour, PAF 43.5%, 95% CI 23.2-60.2) in this group than in the other two groups. This higher suicide rate was substantially reduced from 68.2/100 000 per year to 9.1/100 000 per year, comparable with the other two groups, after stepwise removal of the effects of these three risk factors. Suicide rates by self-poisoning were also significantly higher in this group than in the other two groups. CONCLUSIONS: Higher rates of specific risk factors and use of highly lethal pesticides for suicide contributed to the higher suicide rate in one ethnic group in Taiwan. These findings have implications for developing ethnicity-relevant suicide prevention strategies.

Soft tissue infections related to peripheral intravenous catheters in hospitalised patients: a case-control study.

Peripheral intravenous (i.v.) catheter-related soft tissue infections begin with local skin and soft tissue inflammation, which can progress to cellulitis or even tissue necrosis requiring aggressive surgical treatment. We conducted a matched case-control study to investigate risk factors for peripheral i.v. catheter-related soft tissue infections in hospitalised patients. We retrospectively identified 46 cases that occurred during 2006-2008 in two teaching hospitals. Each case was randomly matched with four control subjects from the same ward and on the same day that the soft tissue infections arose. Risk factors were analysed using conditional logistic regression. Multiple regression analysis identified the following independent risk factors: >24h of continuous i.v. fluid infusion (odds ratio: 5.2, P=0.001), insertion site in lower extremity (8.5, P=0.003), use of an infusion pump (4.6, P=0.023), and hospitalisation due to a neurological or neurosurgical condition (3.6, P=0.018). The population-attributable fractions (the percentage of cases in the study population that could be prevented if the exposure were removed) were 40%, 19%, 24% and 25%, respectively. Minimising unnecessarily prolonged i.v. fluid infusion and avoidance of insertion in the lower extremity may significantly reduce the incidence of peripheral i.v. catheter-related soft tissue infection in the study hospitals.

Metabolic acidosis in sheep alters expression of renal and skeletal muscle amino acid enzymes and transporters.

To determine the effect of metabolic acidosis on expression of L-Gln, L-Glu, and L-Asp metabolizing enzymes and transporters, the relative content of mRNA, protein, or mRNA and protein, of 6 enzymes and 5 transporters was determined by real-time reverse transcription-PCR and immunoblot analyses in homogenates of kidney, skeletal muscle, and liver of growing lambs fed a common diet supplemented with canola meal (control; n = 5) or HCl-treated canola meal (acidosis; n = 5). Acidotic sheep had a 790% greater (P = 0.050) expression of renal Na(+)-coupled neutral AA transporter 3 mRNA and a decreased expression of renal glutamine synthetase mRNA (47% reduction, P = 0.037) and protein (57% reduction, P = 0.015) than control sheep. No change in renal cytosolic phosphoenolpyruvate carboxykinase (protein and mRNA), glutaminase (mRNA), or L-Glu dehydrogenase (protein) was found. In skeletal muscle, acidotic sheep had 101% more (P = 0.026) aspartate transaminase protein than did control sheep, whereas no change in the content of 3 Na(+)-coupled neutral AA transporters (mRNA) or 2 high-affinity L-Glu transporter proteins was found. In liver, no change in the content of any assessed enzyme or transporter was found. Collectively, these findings suggest that tissue-level responses of sheep to metabolic acidosis are different than for nonruminants. More specifically, these results indicate the potential capacity for metabolism of L-Asp and L-Glu by skeletal muscle, and L-Gln absorption by kidneys, but no change in hepatic expression of L-Gln metabolism, elaborates previous metabolic studies by revealing molecular-level responses to metabolic acidosis in sheep. The reader is cautioned that the metabolic acidosis model employed in this study differs from the increased plasma lactate-induced metabolic acidosis commonly observed in ruminants fed a highly fermentable grain diet.

The small intestinal epithelia of beef steers differentially express sugar transporter messenger ribonucleic acid in response to abomasal versus ruminal infusion of starch hydrolysate.

In mammals, the absorption of monosaccharides from small intestinal lumen involves at least 3 sugar transporters (SugT): sodium-dependent glucose transporter 1 (SGLT1; gene SLC5A1) transports glucose and galactose, whereas glucose transporter (GLUT) 5 (GLUT5; gene SLC2A5) transports fructose, across the apical membrane of enterocytes. In contrast, GLUT2 (gene SLC2A2) transports all of these sugars across basolateral and apical membranes. To compare the distribution patterns and sensitivity with nutritional regulation of these 3 SugT mRNA in beef cattle small intestinal tissue, 18 ruminally and abomasally catheterized Angus steers (BW approximately 260 kg) were assigned to water (control), ruminal cornstarch (partially hydrolyzed by alpha-amylase; SH), or abomasal SH infusion treatments (n = 6) and fed an alfalfa-cube-based diet at 1.3 x NE(m) requirement. The SH infusions amounted to 20% of ME intake. After 14- or 16-d of infusion, steers were killed; duodenal, jejunal, and ileal epithelia harvested; and total RNA extracted. The relative amount of SugT mRNA in epithelia was determined using real-time reverse transcription-PCR quantification methods. Basal expression of GLUT2 and SGLT1 mRNA was greater (P < 0.09) by jejunal than by duodenal or ileal epithelia, whereas basal content of GLUT5 mRNA was greater (P < or = 0.02) by jejunal and duodenal than by ileal epithelia. The content of GLUT5 mRNA in small intestinal epithelia was not affected (P > or = 0.16) by either SH infusion treatment. In contrast, GLUT2 and SGLT1 mRNA content in the ileal epithelium was increased (P < or = 0.05) by 6.5- and 1.3-fold, respectively, after abomasal SH infusion. Duodenal SGLT1 mRNA content also was increased (P = 0.07) by 64% after ruminal SH infusion. These results demonstrate that the ileum of beef cattle small intestine adapts to an increased luminal supply of glucose by increasing SGLT1 and GLUT2 mRNA content, whereas increased ruminal SH supply results in duodenal upregulation of SGLT1 mRNA content. These adaptive responses of GLUT2 and SGLT1 mRNA to abomasal or ruminal SH infusion suggest that beef cattle can adapt to increase their carbohydrate assimilation through small intestinal epithelia, assuming that altered SugT mRNA contents reflect the altered transport functional capacities.

Effect of dietary glycemic index on substrate transporter gene expression in human skeletal muscle after exercise.

BACKGROUND/OBJECTIVES: Skeletal muscle plays important role in the regulation of whole-body metabolism. In skeletal muscle, uptakes of glucose and fatty acid from circulation are facilitated by transmembrane substrate transporters GLUT4 and FAT/CD36, respectively. The aim of this study was to determine the effect of dietary glycemic index (GI) on GLUT4 and FAT/CD36 gene expressions in human skeletal muscle after a single bout of exercise. SUBJECTS/METHODS: Eight male subjects completed a 60-min cycling exercise at 75% maximal oxygen consumption (VO(2 max)), and were immediately fed an isocaloric meal containing either high-GI (HGI) or low-GI (LGI) diets, with similar proportions of carbohydrate, fat and protein in a crossover design. Muscle samples from deep vastus lateralis were taken by needle biopsy immediately after exercise and 3 h after exercise. RESULTS: After exercise, the HGI diet produced significantly greater glucose and insulin responses compared with the LGI diet, as indicated by the greater area under the curves. Both diets resulted in rapid reductions in plasma fatty acid and glycerol below fasting level. GLUT4 mRNA was downregulated by both HGI and LGI diets to a comparable extent, whereas GLUT4 protein levels were not changed during this short period. FAT/CD36 mRNA and protein levels were substantially decreased with the HGI diet below baseline, but not with the LGI diet. CONCLUSION: This study found a significant dietary GI effect on post-exercise FAT/CD36 gene expression in human skeletal muscle. This result implicates that the differences in dietary GI are sufficient to alter fat metabolism.

Ruminal and abomasal starch hydrolysate infusions selectively decrease the expression of cationic amino acid transporter mRNA by small intestinal epithelia of forage-fed beef steers.

Although cationic amino acids (CAA) are considered essential to maximize optimal growth of cattle, transporters responsible for CAA absorption by bovine small intestinal epithelia have not been described. This study was conducted to test 2 hypotheses: 1) the duodenal, jejunal, and ileal epithelia of beef cattle differentially express 7 mRNA associated with 4 mammalian amino acid (AA) transport activities: y(+) (CAT1), B(0,+) (ATB(0,+)), b(0,+) (b(0,+)AT and rBAT), and y(+)L (y(+)LAT1, y(+)LAT2, and 4F2hc), and 2) the expression of these mRNA is responsive to small intestinal luminal supply of AA substrates (derived from ruminal microbes) or glucose-derived energy (from starch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (body weight = 260 +/- 17 kg) fed an alfalfa cube-based diet at 1.33 x net energy for maintenance requirement were assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control); ruminal SH and abomasal water infusion; and ruminal water and abomasal SH infusion. The dosage of SH infusion amounted to 20% of metabolizable energy intake. After 14 or 16 d of infusion, steers were slaughtered, duodenal, jejunal, and ileal epithelia were harvested, and total RNA was extracted. The relative amounts of mRNA expressed by epithelia were quantified using real-time reverse transcription-PCR. All 7 mRNA species were expressed by the epithelium from each region, but their abundance differed among the regions. Specifically, duodenal expression of CAT1 and ATB(0,+) mRNA was greater than jejunal or ileal expression; ileal expression of b(0,+)AT, rBAT, and y(+)LAT1 mRNA was greater than jejunal or duodenal expression, whereas the expression of y(+)LAT2 and 4F2hc mRNA did not differ among the 3 epithelia. With regard to SH infusion effect, ruminal infusion down-regulated or tended to down-regulate the jejunal expression of CAT1, rBAT, y(+)LAT2, and 4F2hc mRNA. Abomasal infusion down-regulated the jejunal expression of y(+)LAT2 mRNA and tended to down-regulate the jejunal expression of 4F2hc mRNA. This study characterized the pattern of CAA transporter mRNA expressed by growing beef cattle fed an alfalfa-based diet. Moreover, this study demonstrated that increasing the luminal supply of microbe-derived AA (by ruminal supplementation of SH) results in a reduced capacity of apical and basolateral membrane to transport of CAA, whereas increasing luminal glucose supply (by abomasal supplementation of SH) reduces only the basolateral transport capacity, assuming that CAA transporter mRNA content represents functional capacity.

Basal expression of nucleoside transporter mRNA differs among small intestinal epithelia of beef steers and is differentially altered by ruminal or abomasal infusion of starch hydrolysate.

In ruminants, microbial-derived nucleic acids are a major source of N and are absorbed as nucleosides by small intestinal epithelia. Although the biochemical activities of 2 nucleoside transport systems have been described for cattle, little is known regarding the regulation of their gene expression. This study was conducted to test 2 hypotheses: (1) the small intestinal epithelia of beef cattle differentially express mRNA for 3 concentrative (CNT1, 2, 3) and 2 equilibrative (ENT1, 2) nucleoside transporters (NT), and (2) expression of these NT is responsive to small intestine luminal supply of rumen-derived microbes (hence, nucleosides), energy (cornstarch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (260 +/- 17 kg of BW) were fed an alfalfa cube-based diet at 1.33x NE(m) requirement. Six steers in each of 3 periods were blocked by BW (heavy vs. light). Within each block, 3 steers were randomly assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control), ruminal SH infusion/abomasal water infusion, or ruminal water infusion/abomasal SH infusion. The dosage of SH infusion amounted to 20% of ME intake. After a 14-or 16-d infusion period, steers were slaughtered, and duodenal, jejunal, and ileal epithelia were harvested for total RNA extraction and the relative amounts of mRNA expressed were determined using real-time RT-PCR quantification methodologies. All 5 NT mRNA were found expressed by each epithelium, but their abundance differed among epithelia. Specifically, jejunal expression of all 5 NT mRNA was higher than that by the ileum, whereas jejunal expression of CNT1, CNT3, and ENT1 mRNA was higher, or tended to be higher, than duodenal expression. Duodenal expression of CNT2, CNT3, and ENT2 mRNA was higher than ileal expression. With regard to SH infusion treatments, ruminal infusion increased duodenal expression of CNT3 (67%), ENT1 (51%), and ENT2 (39%) mRNA and ileal expression of CNT3 (210%) and ENT2 (65%) mRNA. Abomasal infusion increased (54%) ileal expression of ENT2 mRNA and tended to increase (50%) jejunal ENT2 mRNA expression. This study has uniquely characterized the pattern of NT mRNA expression by growing beef cattle and found that the mRNA abundance for CNT3, ENT1, and ENT2 in small intestinal epithelia can be increased by increasing the luminal supply of nucleotides (CNT3, ENT1, ENT2) or glucose (ENT2).

Identification and expression pattern of cationic amino acid transporter-1 mRNA in small intestinal epithelia of Angus steers at four production stages.

Although dietary supplementation of cationic AA (CAA), especially l-Lys, is known to be essential for optimal growth of beef cattle, the proteins responsible for absorption of CAA by bovine intestinal epithelia have not been described. Cationic AA transporter-1 (CAT-1) is a major intestinal CAA transporter, demonstrating a high-affinity (muM) transport activity for l-Lys in other mammals, and is widely expressed by small intestinal epithelia of nonruminants, but neither sequence nor expression pattern data exist for CAT-1 in cattle. Therefore, the goal of this research was to compare the relative expression (putative) of CAT-1 mRNA by duodenal, jejunal, or ileal small intestinal epithelia across and within commercially relevant beef cattle production and development stages. Twenty-four Angus steers were assigned randomly (n = 6) to 1 of 4 treatments (suckling, weanling, growing, and finishing) after all steers were born. Duodenal, jejunal, and ileal epithelia were scraped, and total RNA was extracted after the steers were killed at 32, 184, 248, or 423 d of age. Average daily gains of the steers did not differ (1.09 +/- 0.05 kg/d) among stages, whereas the small intestinal length relative to BW decreased (P < 0.01) with steer development. Using standard reverse transcription-PCR cloning techniques, we generated a partial-length bovine CAT-1 complementary DNA (695 bp; GenBank accession no. DQ399522) from jejunal mRNA samples, which possessed 89 and 87% identities to pig and human CAT-1 orthologs, respectively. On the basis of this bovine-specific genetic data, a real-time PCR-based assay of reverse-transcribed mRNA was developed and used to measure relative changes in bovine CAT-1 mRNA abundance in intestinal epithelia as steers developed. The CAT-1 mRNA was expressed by the duodenum, jejunum, and ileum of all 4 production stages. In contrast to expression by duodenal or ileal epithelium, jejunal expression of CAT-1 mRNA by growing steers was greater (P = 0.005) than that by suckling, weanling, or finishing steers. In terms of the expression of CAT-1 mRNA within production stage, jejunal expression was greater (P = 0.002) than that by duodenum or ileum for growing steers. In contrast, no intestinal site difference was found for suckling, weanling, or finishing steers. These data indicate that previously reported Na(+)-independent uptake of Lys by jejunal and ileal epithelia likely occurred by CAT-1, and that the potential capacity for CAT-1-mediated uptake of CAA for beef steers may be greatest for the "growing" phenotype.

The effect of feeding low-phytate barley-soybean meal diets differing in protein content to growing pigs on the excretion of phosphorus and nitrogen.

An experiment was conducted with growing pigs to determine the excretion of P and N in 4 barley-based diets formulated to contain 18 or 15% CP by using a normal barley (NB) or a low-phytate barley (LPB). The NB contained 0.31% total P and 0.19% phytate P; the LPB contained 0.32% total P and 0.01% phytate P. The diets were supplemented, when so required, with lysine, methionine, threonine, and tryptophan to meet their apparent ileal digestible supplies according to the NRC (1998). The diets containing NB were supplemented with inorganic P to meet the NRC (1998) recommendation for available P (0.23%). The diets containing LPB were not supplemented with inorganic P because these contained sufficient available P (0.27%). Eight barrows with an average BW of 20.9 kg were assigned to the 4 dietary treatments according to a repeated 4 x 4 Latin square design. The diets were fed at a rate of 2.5 times the ME requirement for maintenance. The barrows were fed twice daily, at 0800 and 1500, equal amounts each meal. Water was added to the feed at a ratio of 2.5:1. Each experimental period consisted of a 7-d adaptation period followed by a 5-d collection of feces and urine. The substitution of NB with LPB decreased (P < 0.001) the total P excretion by 38 and 43% for the 18 and 15% CP diets, respectively. Reducing the CP content from 18 to 15% decreased (P < 0.001) the N excretion by 29 and 32% for the NB and LPB diets, respectively. With the reduction in CP content, there was a decrease (P < 0.001) in the amount of N retained. The N:P ratio in manure of pigs fed the LPB diets was greater (P < 0.001) than from pigs fed the NB diets. These data indicate that P and N excretion can be greatly reduced by substitution of NB by LPB, and also by the reduction of the CP content, in diets for growing pigs.

Effect of phytase supplementation to a low- and a high-phytate diet for growing pigs on the digestibilities of crude protein, amino acids, and energy.

Supplementation of microbial phytase usually improves the digestibility and utilization of phosphorus in feedstuffs of plant origin. The effect of phytase supplementation on the digestibilities of AA also has been examined, but the results have been inconsistent. This study was carried out to determine the effect of phytase (Natuphos) supplementation, at a rate of 2,000 phytase units/kg, to two basal diets on the apparent ileal digestibilities (AID) of GE, CP, and AA, and on the apparent total-tract digestibilities (ATTD) of CP and GE. The basal diets contained 18% CP and were formulated (as-fed basis) to contain either a low (0.22%) or high content (0.48%) of phytate P. The high-phytate diet contained 20% rice bran, which is a rich source of phytate and has low intrinsic phytase activity. Eight barrows (average initial BW = 40.6 kg), fitted with a simple T-cannula at the distal ileum, were fed the four diets according to a replicated 4 x 4 Latin square design. The pigs were fed twice daily at 0800 and 2000, equal amounts each meal, at a rate of 2.4 times the daily maintenance requirement for ME. Each experimental period comprised 14 d. Ileal digesta were collected from 0800 to 2000 on d 12, 13, and 14. Feces were collected from 0800 on d 8 until 0800 on d 12. Chromic oxide was used as the digestibility marker. The AID of GE, CP, and AA and the ATTD of CP and GE were less in the high- than in the low-phytate diet (P < 0.01). With the exception of glutamic acid, phytase supplementation did not affect (P > 0.10) the AID of CP and AA. There was no effect (P > 0.05) of phytase on the ATTD of CP and GE. These results show that if a response occurs to phytase supplementation, it is independent of the dietary phytate content.

Effect of phytase supplementation to diets for weanling pigs on the digestibilities of crude protein, amino acids, and energy.

Four experiments were conducted with weanling pigs fitted with a simple T-cannula at the distal ileum, to determine the effect of phytase supplementation to four diets on the apparent ileal digestibilities (AID) of CP and AA, and the apparent total-tract digestibilities (ATTD) of CP and DE. Phytase (Natuphos, DSM Food Specialties, Delft, The Netherlands) was supplemented at rates of 0, 500 or 1,000 FTU/kg to the four diets. A 20% CP (as-fed basis) corn-soybean meal diet was used in Exp. 1; a 20% CP wheat-soybean meal diet in Exp. 2; a 20% CP wheat-soybean meal-canola meal diet in Exp. 3; and a 19% CP barley-peas-canola meal diet in Exp. 4. In each experiment, six barrows, fitted with a simple T-cannula at the distal ileum, were fed the basal plus phytase-supplemented diets according to a repeated 3 x 3 Latin square design. Each experimental period comprised 14 d. The piglets were at fed 0800 and 2000 daily, equal amounts for each meal, at a daily rate of at least 2.4 times the maintenance requirement for ME. Feces were collected from 0800 on d 8 until 0800 on d 12 of each experimental period. Ileal digesta were collected from 0800 to 2000 on d 12, 13, and 14. Chromic oxide was used as the digestibility marker. The average initial and final BW (average of all experiments) were 7.9 and 16.5 kg, respectively. Phytase supplementation did not improve the AID of CP and AA in Exp. 1, 2, and 4; however, there were improvements (P < 0.05) or tendencies (P < 0.10) toward improvements in the AID of CP and AA or the ATTD of CP and the content of DE with phytase supplementation in Exp. 3. These results suggest that the AA response factor to microbial phytase supplementation depends on diet composition.

Temporal differences in relative phasing of gait initiation and first step length in patients with cervical and lumbosacral spinal cord injuries.

STUDY DESIGN: Comparison group design. OBJECTIVE: To compare the temporal distance factors during gait initiation between patients with incomplete cervical spinal cord injury, incomplete lumbosacral spinal lesion, and unimpaired control adults. SETTING: Human performance and movement analysis laboratory, Taiwan. PARTICIPANTS: Five patients with an incomplete cervical spinal cord injury (Group 1), five patients with an incomplete lumbosacral spinal lesion (Group 2) and nine unimpaired control adults (Group 3). METHODS: Subjects underwent a three-dimensional gait analysis. The total gait initiation period, reaction time, each relative phasing of gait initiation and the length of the first step were identified by using the kinematic measurement system. MAIN OUTCOME MEASURES: The total gait initiation period (start of the auditory cue for gait initiation to heel-strike of the first swing leg); each relative phasing of gait initiation indicated that the duration of the preparatory phase (start of auditory cue for gait initiation to heel-off of the first swing leg), the duration of the push-up phase (heel-off to toe-off of the first swing leg), and the duration of the single-stance phase (toe-off to heel-strike of the first swing leg) established by the total gait initiation period; and the length of the first step. RESULTS: The gait initiation period was greater in Groups 1 and 2 than that of Group 3 (P<0.05). Each relative phasing including the duration of the preparatory phase, the push-up phase, and the swing phase relative to the total gait initiation period, did not differ among Groups 1-3 (P>0.05). The length of the first step, measured while the nonpreferred leg stepped first in Groups 1 and 2, was shorter than that of Group 3 (P<0.05). CONCLUSIONS: Patients with incomplete cervical spinal cord injuries or lumbosacral spinal lesions took more time in gait initiation than unimpaired control adults. The first step length also reduced in these patients while the nonpreferred leg stepped first, as compared to unimpaired control adults. The data indicated that centrally programmed gait initiation might be preserved in ASIA-D spinal patients who, in this study, executed gait initiation with varying temporal distance strategies to compensate for peripheral impairments, as compared to unimpaired control adults.