The incorporation of acetate,stearate and d(?)-β-hydroxybutyrate into milk fat by the isolated perfused mammary gland of the goat

1. Mammary glands of lactating goats were perfused for 12.5-15hr. with heparinized whole blood and infused with a substrate mixture of glucose, acetate and amino acids (and sometimes chylomicra) containing either [1-(14)C]acetate, d(-)-beta-hydroxy[1-(14)C]butyrate or [U-(14)C]stearate. 2. There was a substantial net uptake of acetate by the glands and transfer of radioactivity into milk fat. Acetate was extensively utilized for the synthesis of milk fatty acids of chain length up to C(14) and to a smaller extent for the synthesis of palmitate. 3. There was a small and variable net uptake of stearate and beta-hydroxybutyrate and negligible oxidation of these substrates. However, tissue uptake was demonstrated by a substantial fall in specific radioactivity across the glands and an extensive transfer of radioactivity into milk fatty acids. 4. With beta-hydroxybutyrate the labelling of milk fat was very similar to that with acetate, but the distribution of radioactivity suggested a cleavage into C(2) fragments of about 40%. 5. Labelled stearate gave rise to highly labelled stearate and oleate in the milk fat. Small amounts of radioactivity were detected in stearate of plasma triglycerides and oleate of plasma free fatty acids. 6. In experiments where there was a decline in milk-fat secretion late in perfusion, the milk fatty acids showed a marked decline in the proportion of stearate and oleate and a rise in the proportion of myristate and palmitate. This did not occur in experiments where milk-fat secretion was maintained at a higher level. 7. The present results confirm that there is a large pool of long-chain fatty acids in mammary tissue that can act as an endogenous source of these substrates.     

Empirical prediction of net splanchnic release of ketogenic nutrients,acetate, butyrate and β-hydroxybutyrate in ruminants: a meta-analysis

For energy feeding systems for ruminants to evolve towards a nutrient-based system, dietary energy supply has to be determined in terms of amount and nature of nutrients. The objective of this study was to establish response equations of the net hepatic flux and net splanchnic release of acetate, butyrate and β-hydroxybutyrate to changes in diet and animal profiles. A meta-analysis was applied on published data compiled from the FLuxes of nutrients across Organs and tissues in Ruminant Animals database, which pools the results from international publications on net splanchnic nutrient fluxes measured in multi-catheterized ruminants. Prediction variables were identified from current knowledge on digestion, hepatic and other tissue metabolism. Subsequently, physiological and other, more integrative, predictors were obtained. Models were established for intakes up to 41 g dry matter per kg BW per day and diets containing up to 70 g concentrate per 100 g dry matter. Models predicted the net hepatic fluxes or net splanchnic release of each nutrient from its net portal appearance and the animal profile. Corrections were applied to account for incomplete hepatic recovery of the blood flow marker, para-aminohippuric acid. Changes in net splanchnic release (mmol/kg BW per hour) could then be predicted by combining the previously published net portal appearance models and the present net hepatic fluxes models. The net splanchnic release of acetate and butyrate were thus predicted from the intake of ruminally fermented organic matter (RfOM) and the nature of RfOM (acetate: residual mean square error (RMSE)=0.18; butyrate: RMSE=0.01). The net splanchnic release of β-hydroxybutyrate was predicted from RfOM intake and the energy balance of the animals (RMSE=0.035), or from the net portal appearance of butyrate and the energy balance of the animals (RMSE=0.050). Models obtained were independent of ruminant species, and presented low interfering factors on the residuals, least square means or individual slopes. The model equations highlighted the importance of considering the physiological state of animals when predicting splanchnic metabolism. This work showed that it is possible to use simple predictors to accurately predict the amount and nature of ketogenic nutrients released towards peripheral tissues in both sheep and cattle at different physiological status. These results provide deeper insight into biological processes and will contribute to the development of improved tools for dietary formulation.     

A comprehensive ex vivo functional analysis of human NKT cells reveals production of MIP1-α and MIP1-β, a lack of IL-17, and a Th1-bias in males

NKT cells contribute to the modulation of immune responses and are believed to be important in the pathogenesis of autoimmune and infectious diseases, as well as cancer. Variations in the composite NKT cytokine response may determine individual disease susceptibility or severity. Due to low frequencies in peripheral blood, knowledge of the breadth of ex vivo human NKT cell functions has been limited. To bridge this gap, we studied highly purified NKT cells from PBMC of healthy donors and assessed the production of 27 effector functions using sensitive Elispot and multiplex bead assays. We found the ex vivo human NKT cell response is predominantly comprised of the chemokines MIP1-α, and MIP1-β as well as the Th1 cytokines IFN-γ and TNF-α. Although lower in magnitude, there was also significant production of IL-2, IL-4, and perforin after mitogen stimulation. Surprisingly, little/no IL-5, IL-6, IL-10, or IL-13 was detected, and no subjects' NKT cells produced IL-17. Comparison of the NKT functional profiles between age-matched male and female subjects revealed similar IL-4 responses, but higher frequencies of cells producing IFN-γ and MIP1-α, from males. There were no gender differences in the circulating NKT subset distribution. These findings implicate chemokines as a major mechanism by which NKT cells control responses in humans. In addition, the panoply of Th2 and Th17 cytokine secretion by NKT cells from healthy donors may not be as pronounced as previously believed. NKT cells may therefore contribute to the gender bias found in many diseases.     

The diuretic effects of CL 115,129 (d,1-15-deoxy-16-hydroxy-16(α/β)-vinyl-prostaglandin E2) and ℓ-prostaglandin E2 in dogs

CL 115,129, the corresponding carboxylic acid and major metabolite of CL 115,347 (d,1-15-deoxy-16-hydroxy-16(α/β)-vinyl-prostaglandin E₂ methyl ester), a potent orally and transdermally long acting antihypertensive agent, infused at 0.1 μg/kg/min into the left renal artery of sodium pentobarbital anesthetized beagle dogs increased urinary volume, sodium (Na⁺), potassium (K⁺) and chloride (Cl⁻) excretion of the left kidney 289, 201, 101 and 229%, respectively, over the 30 min vehicle-treated control periods. At 0.3 μg/kg/min CL 115,129 caused a 475 and 336% increase in urinary volume and Na , respectively. ℓ-Prostaglandin E₂ (ℓ-PGE₂) infused at 0.1 μg/kg/min into the left renal artery increased urinary volume, Na⁺, K⁺ and Cl⁻ excretion of the left kidney of anesthetized beagle dogs 416, 234, 112 and 255%, respectively, over the control. Both CL 115,129 and ℓ-PGE₂ did not affect the systemic arterial blood pressure or the electrolyte excretion of the contralateral kidney. It is concluded that in contrast to other conventional vasodilators, which may cause severe water and electrolyte retention, CL 115,347, via its metabolite CL 115,129, may cause diuresis and natriuresis in many clinical settings when used as an antihypertensive.