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J H Clark

Publications and source records attributed to J H Clark.

At least 19 recordsLinked to original sources

Radioligand binding and functional responses of ligands for human recombinant adenosine A(3) receptors.

The binding and functional properties of adenosine receptor ligands were compared in Chinese hamster ovary cells transfected with human adenosine A(3) receptors. Inhibition of [(125)I]-aminobenzyl-5'-N-methylcarboamidoadenosine ([(125)I]-AB-MECA) binding by adenosine receptor ligands was examined in membrane preparations. Inhibition of forskolin-induced cAMP accumulation by agonists was measured using a cAMP enzyme immunoassay. The rank order of agonist potency for both assays was N(6)-(3-iodobenzyl)-adenosine-5'-N-methyluronamide (IB-MECA) > 5'-N-ethylcarboxamidoadenosine (NECA) > (-)-N(6)-[(R)-phenylisopropyl] adenosine (R-PIA) > 4-aminobenzyl-5'-N-methylcarboxamidoadenosine (AB-MECA) > N(6)-cyclopentyl adenosine (CPA) > adenosine. The radioligand binding rank order of antagonist potency was N-[9-chloro-2-(2-furanyl)[1,2,4]-triazolo[1,5-c]quinazolin-5-benzeneacetamide (MRS1220) > 1,3-dipropyl-8-cyclopentylxanthine (DPCPX) > 8-phenyltheophylline (8-PT) > 8-(p-sulfophenyl)-theophylline (8-SPT). MRS1220 competitively inhibited the effect of IB-MECA on cAMP production, with a K(B) value of 0.35 nm. These data are characteristic of adenosine A(3) receptors. The absence of Mg(2+) and presence of guanosine 5'-(gamma-thio)triphosphate (GTPgammaS) significantly reduced agonist binding inhibition potency, indicating binding to high- and low-affinity states. The IB-MECA, NECA and R-PIA IC(50) values were greater for the cAMP assay than for radioligand binding, suggesting an efficient stimulus-response transduction pathway.

Adenosine↗

NutriDense corn grain and corn silage for dairy cows.

Twenty multiparous Holstein cows, 4 of them surgically fitted with ruminal cannulas, were used in a replicated 4 x 4 Latin square to compare the effects of whole-plant silage and grain produced from NutriDense (ND), leafy NutriDense (LND), or a conventional yellow dent (YD) hybrid on ruminal fermentation, total tract nutrient digestibility, and performance of lactating dairy cows. On a DM basis, diets contained 30.6% corn silage and 27.7% corn grain provided from the 3 hybrids according to the following combinations: 1) YD grain and YD silage, 2) YD grain and LND silage, 3) ND grain and YD silage, and 4) ND grain and LND silage. The average concentrations of crude protein, neutral and acid detergent fiber, and ether extract of LND silage and ND grain were higher, but the contents of nonfibrous carbohydrates and starch were lower than those of their YD counterparts. Although DM intake was similar among treatments, feeding ND grain, LND silage, or both reduced the intakes of nonfibrous carbohydrates and starch but increased the intake of ether extract. Apparent digestibility of starch in the total tract was highest for the diet that contained LND silage and YD grain, whereas the amount and percentage of ether extract that were apparently digested in the total tract was increased and tended to be increased, respectively, by the addition of ND grain, LND silage, or both to the diets. Ruminal fermentation parameters were unaffected by treatments except for the concentration of ammonia nitrogen in the ruminal fluid, which tended to be increased by the feeding of ND grain, LND silage, or both. Production of milk, crude and true protein, fat, lactose, and total solids did not differ among diets. Concentration of milk urea nitrogen increased when the ND grain, LND silage, or both were fed to the cows. Results indicate that ND grain and LND silage were similar to the conventional grain and silage for the feeding of lactating dairy cows.

Ammonia↗

Development of a system to predict feed protein flow to the small intestine of cattle.

A data set constructed from research trials published between 1979 and 1998 was used to derive equations to adjust published tabular values for the rumen-undegradable protein (RUP) content of feeds to better predict the passage of nonammonia nonmicrobial N (NANMN) to the small intestine of lactating dairy cows. Both linear and nonlinear forms of equations were considered for making adjustments. Iterative processes were used to estimate equation parameters. A logistic equation was developed and considered to be the most optimal for adjustment of published tabular RUP contents of feeds. The equation is a function of dietary dry matter intake (DMI) and includes terms for tabular RUP and nonprotein N contents of individual feeds. The equation has a standard error of prediction of 69.29 g of NANMN/ d per cow and a root mean square prediction error of 104.63 g of NANMN/d per cow. Independent evaluation of the equation indicated that the concept of variable RUP content for feeds based on DMI is correct. Further refinements may be needed as other data become available to quantify the effects of additional factors on the RUP value of feeds.

Animal Feed↗

Varying protein and starch in the diet of dairy cows. I. Effects on ruminal fermentation and intestinal supply of nutrients.

The main objective of this experiment was to examine the effects of the percentage and source of crude protein (CP) and the amount of starch in the diet of dairy cows on ruminal fermentation, nutrient passage to the small intestine, and nutrient digestibility. For this purpose, 6 multiparous Holstein cows fistulated in the rumen and duodenum that averaged 73 d in milk were used in a 6 x 6 Latin square design with a 2 x 3 factorial arrangement of treatments. Two sources of CP [solvent-extracted soybean meal (SBM) and a mixture of SBM and a blend of animal-marine protein supplements plus ruminally protected Met (AMB)] and 3 levels of dietary protein (about 14, 16, and 18%) were combined into 6 treatments. On a dry matter (DM) basis, diets contained 25% corn silage, 20% alfalfa silage, 10% cottonseed, 26.7 to 37% corn grain, and 4 to 13.5% protein supplement. Intakes and digestibilities in the rumen and total tract of DM, organic matter, acid and neutral detergent fiber were unaffected by treatments. Increasing dietary CP from 14 to 18% decreased the intake and apparent ruminal and total tract digestion of starch, but increased the proportion of starch consumed by the cows that was apparently digested in the small intestine. At 14% CP, starch intake and total tract digestion were higher for the AMB diet than for the SBM diet, but the opposite occurred at 16% CP. Across CP sources, increasing CP in the diet from 14 to 18% increased the intakes of N and amino acids (AA), and ruminal outflows of nonammonia N, nonammonia nonmicrobial N, each individual AA except Met, total essential AA, and total AA. Across CP percentages, replacing a portion of SBM with AMB increased the intake of Met and Val and decreased the concentration of ammonia N in the rumen, but did not affect the intake of other essential AA or the intestinal supply of any essential AA and starch. The ruminal outflow of microbial N, the proportional contribution of Lys and Met to total AA delivered to the duodenum, and milk yield were unaffected by treatments. Data suggest that the intake of N by high-producing dairy cows that consume sufficient energy and other nutrients to meet their requirements can be decreased to about 600 to 650 g daily without compromising the supply of metabolizable protein if the source and amount of dietary CP and carbohydrate are properly matched.

Animal Nutritional Physiological Phenomena↗

Varying protein and starch in the diet of dairy cows. II. Effects on performance and nitrogen utilization for milk production.

The main objective of this experiment was to examine the effects of the percentage and source of crude protein (CP) and the amount of starch in the diet of dairy cows on the lactational performance and use of N for milk production. Sixty multiparous Holstein cows were used in a 210-d lactational trial with a completely randomized design with a 2 x 3 factorial arrangement of treatments. Two sources of CP [solvent-extracted soybean meal (SBM) and a mixture of SBM and a blend of animal-marine protein supplements plus ruminally protected Met (AMB)] and 3 levels of dietary CP (means = 14.8, 16.8, and 18.7%) were combined into 6 treatments. On a dry matter (DM) basis, diets contained 25.0% corn silage, 20.0% alfalfa silage, 10.0% cottonseed, 26.7 to 37.0% corn grain, and 4.8 to 13.5% protein supplement, plus minerals and vitamins. Across the 210 d of lactation, the productive response of dairy cows to the source of supplemental CP depended on the concentration of CP in the diet. At 18.7% CP, cows fed SBM consumed more DM and produced more milk, 3.5% fat-corrected milk, fat, and true protein, but had lower efficiency of feed use and body condition score than cows fed AMB. At 16.8% CP, cows fed AMB produced more 3.5% fat-corrected milk, fat, and true protein than cows fed SBM. At 14.8% CP, cows fed SBM consumed more DM but produced less true protein and had lower feed efficiency than cows fed AMB. Across CP sources, cows fed 14.8% CP produced less fat-corrected milk and true protein than cows fed 16.8 and 18.7% CP. Across CP percentages, cows fed AMB produced more fat-corrected milk per kilogram of DM consumed than cows fed SBM. Despite these interactions, improvements in the gross efficiency of N use for milk production were achieved through reductions in the intake of N independently of the source of CP. Data suggest that the intake of N by high-producing dairy cows that consume sufficient energy and other nutrients to meet their requirements can be decreased to about 600 to 650 g daily if the source of RDP and RUP are properly matched with the source and amount of carbohydrate in the diet.

Ammonia↗

Rumen fermentation and intestinal supply of nutrients in dairy cows fed rumen-protected soy products.

Four multiparous lactating Holstein cows that were fistulated in the rumen and duodenum and that averaged 205 d in milk were used in a 4 x 4 Latin square design to evaluate the practical replacement of solvent-extracted soybean meal (SSBM) with soy protein products of reduced ruminal degradability. On a dry matter (DM) basis, diets contained 15% alfalfa silage, 25% corn silage, 34.3 to 36.9% corn grain, 19.4% soy products, 18.2% crude protein, 25.5% neutral detergent fiber, and 35.3% starch. In the experimental diets, SSBM was replaced with expeller soybean meal (ESBM); heated, xylose-treated soybean meal (NSBM); or whole roasted soybeans (WRSB) to supply 10.2% of the dietary DM. Intakes of DM (mean = 20.4 kg/d), organic matter, and starch were unaffected by the source of soy protein. Similarly, true ruminal fermentation of organic matter and apparent digestion of starch in the rumen and total tract were not altered by treatments. Intake of N ranged from 567 (WRSB) to 622 g/d (ESBM), but differences among soy protein supplements were not significant. Compared with SSBM, the ruminal outflow of nonammonia N was higher for NSBM, tended to be higher for ESBM, and was similar for WRSB. The intestinal supply of nonammonia nonmicrobial N was higher for NSBM and WRSB and tended to be higher for ESBM than for SSBM. However, no differences were detected among treatments when the flow to the duodenum of nonammonia nonmicrobial N was expressed as a percentage of N intake or nonammonia N flow. The ruminal outflow of microbial N, Met, and Lys was not altered by the source of soy protein. Data suggest that partially replacing SSBM with ESBM, NSBM, or WRSB may increase the quantity of feed protein that reaches the small intestines of dairy cows. However, significant improvements in the supply of previously reported limiting amino acids for milk production, particularly of Met, should not be expected.

Amino Acids↗

Impacts of the source and amount of crude protein on the intestinal supply of nitrogen fractions and performance of dairy cows.

The objective of this article was to review and summarize the significance of the amount and source of dietary crude protein supplements on the supply of nitrogen fractions passing to the small intestine and the performance of lactating dairy cows. A meta-analysis was used to evaluate 2 data sets, one for nitrogen flow to the small intestine and one for performance of cows. The response of dairy cows to rumen-undegradable protein supplements is variable. A portion of this variable response from research trials is explained by the source of crude protein in the control diet, the proportion and source of rumen-undegradable protein in the experimental diet, the effect of rumen-undegradable protein on microbial protein outflow from the rumen, the degradability and amino acid content of the rumen-undegradable protein, and the crude protein percentage of the diet. Compared with soybean meal, the mean milk production responses to feeding rumen-undegradable protein supplements ranged from -2.5 to +2.75%. Because of the large variation and small magnitude of response when rumen-undegradable protein supplements are fed compared with soybean meal, efficiency of nitrogen utilization and the cost to benefit ratio for these crude protein supplements may determine the source and amount of crude protein to feed to dairy cows in the future.

Amino Acids↗

Effect of feeding supplemental fibrolytic enzymes or soluble sugars with malic acid on milk production.

Two trials were conducted to evaluate effects of feeding supplemental fibrolytic enzymes or soluble sugars and malic acid on milk production. In trial 1, 257 cows at four sites were fed a basal diet consisting of no more than 60% of forage DM as corn silage and less than 40% as alfalfa hay. Cows were assigned randomly within site, parity, and two stages of lactation to: 1) control; 2) enzyme A; 3) enzyme B; and 4) soluble sugars and malic acid. There was a 14-d pretreatment and an 84-d treatment period. Enzyme solutions were sprayed on either the forage component or the TMR each day while mixing feed. Trial 2 was similar, except 122 cows at one site in the United Kingdom were fed diets containing forage that was 75% corn silage and 25% grass silage, and all cows began the study between 25 to 31 DIM. Mean milk productions for 233 cows that completed trial 1 were 32.9, 32.5, 32.4, and 32.9 kg/d for control, enzyme A, enzyme B, and soluble sugars and malic acid, respectively. Mean milk productions for 116 cows that completed trial 2 were 28.2, 27.9, 28.8, and 28.4 kg/d, respectively. In vitro analyses of the activities of enzyme solutions indicated that all major cellulose and hemicellulose degrading activities were present; however, the pH optima (approximate pH = 4 to 5) were more acidic, and the temperature optimum (approximately 50 degrees C) was greater than normal pH and temperature in the rumen. If fibrolytic activity in the rumen is a major mechanism of action of supplemental fibrolytic enzymes, it appears that considerable activity of these preparations was lost due to conditions in the rumen. In conclusion, feeding supplemental fibrolytic enzymes or malic acid with soluble sugars had no effect on milk production under the conditions used in this study.

Animal Nutritional Physiological Phenomena↗

Effects of tallow in diets based on corn silage or alfalfa silage on digestion and nutrient use by lactating dairy cows.

Six multiparous Holstein cows (average 31 days in milk; 36.3 kg/d of milk) fitted with ruminal cannulas were used in a 6 x 6 Latin square with 21-d periods to investigate the effects of diets that varied in forage source and amount of supplemental tallow. Isonitrogenous diets in a 2 x 3 factorial arrangement were based on either high corn silage (40:10 corn silage to alfalfa silage, % of dry matter) or high alfalfa silage (10:40 corn silage to alfalfa silage, % of dry matter) and contained 0, 2, or 4% tallow. Intakes of dry matter and total fatty acids were lower when cows were fed the high corn silage diet. Tallow supplementation linearly decreased dry matter intake. Milk yield was unaffected by diet; yields of milk fat and 3.5% fat-corrected milk were higher for the high alfalfa silage diet but were unaffected by tallow. Milk fat percentage was higher for the high alfalfa silage and tended to decrease when tallow was added to the high corn silage diet. Contents of trans-C18:1 isomers in milk fat were increased by high corn silage and tallow, and tended to be increased more when tallow was fed in the high corn silage diet. Ruminal pH and acetate:propionate were lower when high corn silage was fed. Ruminal acetate:propionate decreased linearly as tallow increased; the molar proportion of acetate was decreased more when tallow was added to the high corn silage diet. Ruminal liquid dilution rates were higher for the alfalfa silage diet; ruminal volume and solid passage rates were similar among diets. Total tract apparent digestibilities of dry matter, organic matter, crude protein, starch, energy, and total fatty acids were unaffected by diet. Digestibilities of neutral detergent fiber, acid detergent fiber, hemicellulose, and cellulose were lower when high corn silage was fed. The high alfalfa silage diet increased intakes of metabolizable energy and N, and increased milk energy and productive N. Tallow decreased the amount of N absorbed but had few other effects on utilization of energy or N. Tallow linearly increased concentrations of nonesterified fatty acids and cholesterol in plasma; cholesterol was increased by high alfalfa silage. Overall, forage source had more pronounced effects on production and metabolism than did tallow supplementation. Few interactions between forage source and tallow supplementation were detected except that ruminal fermentation and milk fat content were affected more negatively when tallow was fed in the high corn silage diet.

Animal Nutritional Physiological Phenomena↗

Soyhulls as an alternative feed for lactating dairy cows: a review.

Dairy producers use soyhulls, a byproduct of soybean processing, to replace either grain or forage in diets of lactating dairy cows. In view of the nutritional and economical value of soyhulls it is anticipated that this practice will continue to increase in popularity among nutritionists and producers of ruminant animals. This paper reviews information regarding the nutritional value of soyhulls and the effects of feeding this alternative feed on ruminal fermentation, nutrient digestion and utilization, and performance of dairy cows. Soyhulls can replace corn grain to supply about 30% of the dry matter (DM) in high-grain diets without negatively affecting either the fermentation or digestion of nutrients in the gastrointestinal tract or the performance of dairy cows. Additionally, data suggest that soyhulls might successfully replace forage to supply < or = 25% of the DM in diets of dairy cows when the supply of effective fiber, which includes a chemical and a physical component, remains adequate after including the hulls. However, caution should be exercised when data from different studies are extrapolated to practical situations because the response to feeding soyhulls appears to be largely affected by the type of carbohydrate being replaced by soyhulls; the amount, type, and physical form of the dietary forage; and the incidence of either negative or positive associative effects before and after the addition of soyhulls to the original diet. Unfortunately, the paucity of data from experiments in which soyhulls constituted more than 25 to 30% of the dietary DM restricts the ability to identify the maximum amount of soyhulls that can be used in diets of dairy cows. Information from studies in which > or = 25 to 30% of dietary DM supplied as either cereal grains or forages are replaced with soyhulls is needed to better understand and predict the production of dairy cows fed diets containing the hulls. This knowledge is essential for maximizing the use of soyhulls in diets for dairy cows.

Animal Feed↗

Performance of lactating dairy cows fed corn as whole plant silage and grain produced from a glyphosate-tolerant hybrid (event NK603).

Sixteen multiparous Holstein cows averaging 74 d in milk were used in a replicated 4 x 4 Latin square to compare the effects on animal performance of feeding whole plant silage and grain from a glyphosate-tolerant corn hybrid (event NK603), a nontransgenic control hybrid, and two commercial nontransgenic hybrids (DK647 and RX740). The grain and silage from the four corn hybrids were produced using the same procedures and under similar agronomic conditions at the University of Illinois. On a dry matter (DM) basis, diets contained 30% corn silage and 27.34% corn grain produced either from event NK603, a nontransgenic control, or commercial hybrids. Apart from the DM content of silages, the chemical composition of both grain and silage produced from the four corn hybrids were substantially equivalent. Feeding diets that contained event NK603 and DK647 hybrids tended to decrease DM intake (DMI) compared with the control nontransgenic and RX740. The intakes of crude protein (CP), acid and neutral detergent fiber, and nonfiber carbohydrates were not different for cows fed event NK603 and control diets. The RX740 diet resulted in the highest intakes of fiber and CP, whereas the DK647 diet resulted in the lowest intake of CP. These differences in nutrient intake arose from small variations in both the DMI and the chemical composition of feed ingredients and experimental diets. Production of milk and 3.5% fat-corrected milk; milk fat, CP, and true protein percentage and yield; milk urea N; milk total solids percentage and yield; and somatic cell count were not affected by treatments. These data indicate that the stable insertion of the gene that confers tolerance to glyphosate (event NK603) in the corn line used in this experiment does not affect its chemical composition and nutritional value for lactating dairy cows when compared with conventional corn.

Animals↗

Ruminal fermentation and nutrient digestion by dairy cows fed varying amounts of soyhulls as a replacement for corn grain.

Five multiparous Holstein cows cannulated in the rumen and duodenum that averaged 63 d in milk were used in a 5 x 5 Latin square design with 14-d periods to evaluate the incremental substitution of soyhulls for corn in the diet. Diets contained 23% alfalfa silage, 23% corn silage, and 54% concentrate on a dry matter (DM) basis. Pelleted soyhulls replaced corn in the concentrate to supply 0, 10, 20, 30, or 40% of the dietary DM. The intakes of DM and organic matter were unaffected by treatments. Intakes of acid detergent fiber and neutral detergent fiber increased linearly, but the intake of nonstructural carbohydrates decreased linearly as soyhulls increased from 0 to 40% of dietary DM. The amount of acid detergent fiber and neutral detergent fiber digested was increased whereas the amount of nonstructural carbohydrate digested was decreased in the rumen, in the lower digestive tract, and in the total digestive tract as soyhulls replaced corn in the diet. Passage to the duodenum of nonammonia N, microbial N, nonammonia nonmicrobial N, total essential amino acids, total nonessential amino acids, and total amino acids were not affected by treatments. Yield of milk (29.5 kg/d) was not affected by treatments in this experiment. In a companion experiment, cows fed the 40% SH diet produced 1.2 kg/day per cow less (P < 0.07) milk than cows fed the control diet which is similar to the 1.3 kg/day per cow less milk produced by cows fed the same 40% SH diet in this experiment. Differences in the source of energy (fiber vs. nonstructural carbohydrates), in the amount of fiber and nonstructural carbohydrates digested, and in the site of digestion in the gastrointestinal tract may cause a shortage of the source and/or amount of energy that is required for maximum milk production in high producing cows when more than 30% of the dietary DM that is supplied as corn is replaced with soyhullss.

Amino Acids↗

Performance of lactating dairy cows fed varying amounts of soyhulls as a replacement for corn grain.

Fifteen multiparous Holstein cows averaging 112 d in milk were used in a replicated 5 x 5 Latin square to evaluate the incremental substitution of soyhulls for corn in the diet. Diets contained 23% alfalfa silage, 23% corn silage, and 54% concentrate on a dry matter basis. Pelleted soyhulls replaced corn in the concentrate to supply 0, 10, 20, 30, or 40% of the dietary dry matter. Dry matter intake decreased linearly as soyhulls replaced corn in the diet, but the major decrease in dry matter intake occurred when soyhulls provided 30 and 40% of the dietary dry matter. Intakes of both acid and neutral detergent fiber increased linearly as soyhulls increased from 0 to 40% of dietary dry matter. Production of milk tended to decrease when soyhulls supplied 40% of the dietary dry matter. Production of 3.5% fat-corrected milk, milk crude protein percentage and yield, milk urea N, and total solids yield were not affected by treatments. Production of true protein, but not percentage, tended to decrease by about 5% when soyhulls supplied 40% of the dietary dry matter. Increasing the percentage of soyhulls in the dietary dry matter increased linearly milk fat content and yield, and total solids content in milk. These data suggest that soyhulls can successfully supply up to about 30% of the dry matter intake of midlactation cows without depressing animal performance. Furthermore, replacing part of the corn with soyhulls in high grain diets may be viable when milk fat has a high monetary value or when soyhulls can be purchased at a more competitive price than grains on a nutrient content basis.

Animal Feed↗

Fatty acid metabolism in liver of dairy cows fed supplemental fat and nicotinic acid during an entire lactation.

Liver biopsies from 38 multiparous Holstein cows were used to determine rates of peroxisomal beta-oxidation and total beta-oxidation of palmitate in liver homogenates and contents of total lipid, triglyceride, and glycogen during the lactation cycle. Cows were assigned to one of four diets from wk 4 through wk 42 of lactation: control, control plus nicotinic acid (12 g/d), supplemental fat, or supplemental fat plus nicotinic acid. Liver biopsies were obtained at wk 3 (covariate), 6, 12, 24, and 42 of lactation. Neither supplemental fat nor nicotinic acid affected palmitate oxidation in liver homogenates or liver composition. Peroxisomal beta-oxidation capacity and the ratio of peroxisomal to total beta-oxidation decreased from wk 3 to 12 and then increased at wk 42. Contents of total lipid and triglyceride decreased, and content of glycogen increased, from wk 3 to 12. Total oxidation capacity in liver homogenates was correlated negatively with total lipid and triglyceride in liver, yields of milk and solids-corrected milk (SCM), and plasma nonesterified fatty acids (NEFA), and was correlated positively with liver glycogen, dry matter intake (DMI), energy balance, and plasma glucose. Peroxisomal beta-oxidation was correlated negatively with yields of milk and SCM. The ratio of peroxisomal to total beta-oxidation was correlated positively with liver total lipid, liver TG, and plasma NEFA and negatively with DMI and energy balance. When only data from wk 3 postpartum were considered, both total and peroxisomal beta-oxidation were correlated negatively with hepatic concentrations of total lipid and TG. Peroxisomal beta-oxidation in liver of dairy cows is not affected by feeding supplemental fat or nicotinic acid during wk 4 to 42 of lactation but may be a part of the hepatic adaptations to negative energy balance.

Animals↗

Accuracy and precision of computer models to predict passage of crude protein and amino acids to the duodenum of lactating cows.

To evaluate the ability of several models to accurately and precisely predict the passage of crude protein (CP) and amino acids to the duodenum of lactating cows, we simulated data from six published studies using the 1989 National Research Council equations, the Mepron Dairy Ration Evaluator (version 1.1), the University of Pennsylvania release of the Net Carbohydrate and Protein System (version 2.12p), the Cornell Net Carbohydrate and Protein System (version 3), and CPM Dairy (version 1.0). Models overestimated the passage of CP from microbes by an average of 323 g/d, and underestimated the passage of CP from feed by an average of 874 g/d. These two errors were partially canceled when CP from microbes and feed were summed to estimate passage of total CP to the duodenum. Many dietary composition variables appeared to bias the predictions; however, the influence of any one variable was small. The efficiency of modeling was high for most predictions but was variable for predicting passage of specific individual amino acids to the small intestine depending on the model selected. These simulations indicated no obvious advantage for any model over the others tested. The models responded to changes in diets by altering the amount of protein from microbes and feed that reached the duodenum, resulting in improved accuracy of predictions of duodenal CP passage compared with simply assuming a constant value for passage of CP to the duodenum.

Amino Acids↗

Prediction of crude protein and amino acid passage to the duodenum of lactating cows by models compared with in vivo data.

To determine whether statistical inferences obtained from predictions by models were similar to those of measured data from individual cows, data from six research trials published between 1989 and 1997 were simulated using the 1989 National Research Council Model, the Mepron Dairy Ration Evaluator (version 1.1), The University of Pennsylvania release of the Net Carbohydrate and Protein System (version 2.12p), The Cornell Net Carbohydrate and Protein System (version 3), and the CPM Dairy (version 1.0). Both predicted and measured protein fractions were analyzed by ANOVA and compared to determine whether statistical inferences among treatments from predictions by the models were similar to those from the measured data. The interpretations and statistical inferences of measured data did not always agree with those for predicted data. All models responded to changes in diet composition and often predicted that dietary changes would result in statistically different amounts of protein and amino acids passing to the duodenum than were observed in the measured data. The direction of predicted change among treatments for passage of nitrogen fractions to the duodenum also did not agree with the measured data a large percentage of the time. Discrepancies in ANOVA and interpretations between predicted and measured data may be due to the reduction in variation associated with modeling biological systems, associative effects of feeds not accounted for by models, inadequate equations in the models, inadequate description of feeds, or experimental error in measured data. Before model simulations of duodenal flow of crude protein and amino acids can be substituted for experimental measurements, better descriptors of main dietary effects, microbial protein production, ruminal protein degradation, and interactions among dietary factors must be developed.

Amino Acids↗

Initial results of solitary pancreas transplants performed without regard to donor/recipient HLA mismatching.

BACKGROUND: We hypothesized that solitary pancreas transplants could be performed successfully even in the presence of poor HLA matching if an aggressive approach were taken with regard to immunosuppressive protocol and the performance of allograft biopsy. METHODS: Seven pancreas-after-kidney transplants and seven pancreas transplants alone were performed without consideration given to the degree of HLA mismatching (MM) using tacrolimus (FK506)/mycophenolate mofetil (MMF)/prednisone maintenance therapy. Mean (+/-SD) total HLA MM was 4.8+/-1.2. All patients were followed for at least 6 months. The first four cases were induced with ATGAM for 7 to 10 days. In the remaining 10 cases, an ultrasound-guided percutaneous needle biopsy was attempted on a protocol basis 10 days after completing induction with OKT3 for 7 (n=2) or 14 (n=8) days. RESULTS: Overall patient survival, graft survival, and incidence of acute rejection requiring treatment were 86, 79, and 50%, respectively. Two patients receiving ATGAM developed grade III-IV rejection at 3 weeks. Both patients receiving OKT3 for 7 days developed early grade III rejection. However, only three of eight patients receiving OKT3 for 14 days developed rejection requiring treatment. Protocol biopsy was successfully performed in six of seven patients and uncovered three cases of otherwise undetectable grade III-IV rejection. CONCLUSIONS: Although based on a small number of cases, our results suggest that solitary pancreas transplants with a poor HLA match can be performed with an acceptable rejection incidence and graft survival rate using an OKT3/FK506/MMF/prednisone regimen with protocol biopsy.

Adolescent↗

Epidermal growth factor activates reproductive behavior independent of ovarian steroids in female rodents.

Sex steroids exert profound influence on neural development and function through activation of intranuclear receptors. However, during sexual differentiation and at onset of puberty, intracerebral estrogen (E) availability is subsequent to these effects. The potent mitogen epidermal growth factor (EGF) activates estrogen receptor (ER)-dependent transcription in cultured cells in the absence of exogenous E. Since reproductive behavior in female rodents is the result of E-dependent transcriptional activity and protein synthesis, lordosis serves as a well established in vivo model for probing cellular and molecular mechanisms of steroid receptor-dependent behavior. Here we demonstrate that EGF can signal through the classical E receptor (ERalpha) to alter in vivo function in rodent central nervous system. EGF and EGF receptor ligands induced lordosis in a dose- and time-dependent manner in the absence of steroid treatment in ovariectomized rats and mice. Using antisense oligonucleotides, pharmacological and antibody blockade, and mutant mice, we also report that this behavioral responsiveness is mediated through ERalpha by specific stimulation of membrane-bound EGF receptors and EGF receptor-specific tyrosine kinase rather than by direct ligand activation of the ERalpha. Of biological significance, delayed onset of puberty and the absence of synchronization between reproductive behavior and ovulation was detected in intact mutant Wa-2 mice that express a naturally occurring point mutation in the EGF receptor. To our surprise, EGF-mediated behavior was independent of progesterone (P) and progesterone receptor (PR) since antiprogestins, PR antisense oligonucleotides, and targeted disruption of PR in ovariectomized transgenic mice failed to impede the display of lordosis after EGF. Finally, we also found that another growth factor, insulin-like growth factor-1, which provokes ER-dependent transcription in vitro, activates mating behavior in a similar E-independent manner. Thus, growth factor mediation of ER-targeted function may be a universal feature in the rodent central nervous system, raising critical questions about the role of growth factors in mediating ER-dependent processes in development and reproduction.

Aging↗