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Alternative models for analyses of liver and mammary transorgan data were formulated and fitted to liver and mammary data sets respectively. The models considered metabolite inputs to and effluxes from an extracellular pool. In general, fits were greatly improved over previous efforts using other models (Miller et al. 1991a; Hanigan et al. 1992; Wray-Cahen et al. 1997). Errors of prediction were generally less than 15% for liver and less than 20% for mammary glands. With the possible exception of glutamine for the udder, all metabolites exhibited linear responses to extracellular concentrations within the observed ranges of inputs. However, prediction biases were evident for beta-hydroxybutyrate, acetate, and propionate by liver and for arginine, histidine, citrulline and glycerol by mammary tissue. These biases were hypothesized to be caused by the existence of additional regulatory complexity. With the exception of histidine, parameter estimates for essential amino acid removal by liver were 2-3-fold lower than for mammary gland. Infusion of an amino acid mixture into the mesenteric vein did not alter parameter estimates for removal of amino acids by the liver. Treatment of cows with bovine somatotropin resulted in changes in mammary parameter estimates for aspartate, glutamate, leucine, phenylalanine, glucose, and glycerol.

Mammary gland tissues from three lactating Holstein cows were sampled and slices incubated with various concentrations of glucose and in combinations with other substrates in Krebs-Ringer bicarbonate-based media. Conversions of 1-and 2-carbon-14 glucose to carbon dioxide, lactose, glyceride-glycerol, citrate, and fatty acids were determined. Glucose utilization increased asymptotically with glucose availability. Carbon dioxide, lactose, and glyceride glycerol production accounted for nearly 90% of glucose recovered. Only trace quantities of glucose were recovered as fatty acids. Estimates of apparent affinities for glucose use were in a physiological concentration range of glucose. Oxidation of glucose was positively correlated with availability of acetate. Acetate availability increased the ratio of 1-carbon-14 glucose to 2-carbon-14 glucose oxidation and decreased glucose conversion to fatty acids. Glucose conversion to glyceride-glycerol was not affected by acetate availability. Primary uses of glucose are in lactose and glyceride glycerol synthesis and formation of reduced nicotinamide adenine dinucleotide phosphate for lipogenesis. Extensive use of glucose for these purposes may limit amounts available for fatty acid synthesis.

Mammary gland tissue slices from three lactating Holstein cows were incubated in Krebs-Ringer bicarbonate-based media with varying concentrations of lactate and other substrates. Conversions of 1- and 2-carbon-14 of lactate to carbon dioxide, fatty acids, citrate, glyceride glycerol, and lactose were determined. Effects of acetate, glucose, beta-hydroxybutyrate, stearate, and pyruvate on lactate metabolism were evaluated. Oxidation of lactate increased asymptotically with lactate concentration. Low acetate concentration stimulated oxidation of carbon 2 of lactate slightly while higher acetate availability inhibited lactate oxidation. Conversion of lactate to fatty acids increased linearly with lactate concentration. This conversion of lactate was inhibited strongly by acetate. Significant conversion of carbon 2 of lactate to glyceride-glycerol but not lactose was detected. Bovine mammary glands have the capacity of utilize sizeable quantities of lactate for oxidation and lipogenesis. Both phenomena are highly dependent on availability of acetate. Pyruvate dehydrogenase and citrate lyase could represent important regulatory sites in vivo for determination of tissue preference for acetate over lactate. Results indicate factors other than an inactive malate transhydrogenation cycle likely limit glucose conversion to fatty acids.

Slices from mammary glands from six lactating Holstein cows were incubated in Krebs-Ringer bicarbonate-based media in the presence of varying concentrations of acetate, glucose, and lactate. Conversions of acetate to carbon dioxide, fatty acids, citrate, glyceride-glycerol, and lactose were determined. Specific effects of beta-hydroxybutyrate and stearate availability also were evaluated. Utilization of acetate for all functions usually followed Michaelis-Menten kinetics with apparent affinities in the physiological concentration range of acetate. Availability of glucose had a large "permissive" effect on acetate to conversion to fatty acids. However, acetate metabolism was independent of the availability of lactate. Conversion of acetate to glyceride-glycerol was significant. This indicates the possibility of bidirectional glycolytic flux, which could limit glucose conversion to fatty acids. Addition of stearate caused general increases of acetate metabolism. Conversion of acetate to fatty acids, carbon dioxide, and citrate was elevated in the absence of beta-hydroxybutyrate. The latter observation indicates beta-hydroxybutyrate may not be a required initiator of fatty acid synthesis in mammary tissue.