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Orthotopic liver transplantation is the most effective treatment for fulminant hepatic failure. As an alternative treatment, an efficient extracorporeal bioartificial liver should contain a large yield of functional hepatocytes with an immunoprotective barrier, for providing temporary adequate metabolic support to allow spontaneous liver regeneration or for acting as a bridge toward transplantation. Survival, proliferation, and functions of porcine hepatocytes were evaluated in primary cultures and after embedding in alginate beads, which were subsequently coated with a membrane made by a transacylation reaction between propylene glycol alginate and human serum albumin. Disruption of total pig livers by collagenase perfusion/recirculation allowed the obtention of up to 10(11) hepatocytes with a viability greater than 95%. Hepatocytes in conventional cultures or embedded in coated alginate beads survived for about 10 days, secreted proteins, particularly albumin, and maintained several phase I and II enzymatic activities, namely ethoxyresorufin-O-deethylase, oxidation of nifedipine to pyridine, phenacetin deethylation to paracetamol, glucuroconjugation of paracetamol, and N-acetylation of procainamide. Typical features of mitosis and [3H]thymidine incorporation indicated that porcine hepatocytes proliferated in both conventional cultures and alginate beads. The efficacy of the membrane surrounding alginate beads for protecting cells from immunoglobulins was tested by embedding HLA-typed human lymphocytes, which were subsequently incubated with specific anti-HLA immunoglobulin G and complement. These data show that large yields of porcine hepatocytes that are embedded in coated alginate beads remain functional and are isolated from large molecular weight molecules, such as immunoglobulins. This system represents a promising tool for the design of an extracorporeal bioartificial liver, containing xenogeneic hepatocytes, to treat acute liver disease in humans.

Two experiments were carried out with piglets, 3-5 kg live weight, to evaluate the effects of feeding a tryptophan (TRP)-deficient diet for 2 weeks on protein synthesis rates measured in vivo 2 h after a meal.In the first experiment on twenty piglets fed on 250 g protein/kg diets, T R P deficiency (0.77 g/16 g nitrogen) as compared with adequacy (1.17 g/16 g N) significantly decreased feed intake, growth performance and fractional protein synthesis rates (FSR), without variation of RNA in longissirnus dorsi (LD) and with parallel increases in RNA in semitendinosus (ST) muscle and liver. In the second experiment thirty-two piglets were tube-fed deficient and adequate diets at the two feeding levels (LF) previously achieved. Both T R P and LF significantly increased growth performance and FSR, but not RNA, in LD and ST muscle, with a trend to a synergy between the two factors (TRP x LF interaction). In another muscle, trapezius (TR), the same interaction was only apparent in RNA content. Among the three muscles it was in LD that FSR was the most responsive to dietary TRP (significant muscle x T R P interaction). In the liver the TRP x LF interaction on FSR and not RNA was the major significant effect, indicating that higher T R P and higher LF were both required to get the maximum protein synthesis rate. At 30 min after a meal the same significant interaction effect was shown on plasma glucose, whilst the higher LF increased plasma insulin with both diets. After a further 30 min the appearance of a similar significant effect of the T R P x L F interaction on plasma insulin resulted from its abatement when the deficient diet had been fed at high LF. These results suggest that dietary TRP deficiency decreased muscle and liver protein synthesis rates in relation to a decrease in the post-prandial release of insulin following a decreased rate of nutrient absorption.

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Two replicates of three multiparous crossbred Large White x Landrace lactating sows were used to develop a technique for the continuous direct measurement of the blood flow through the mammary gland using transit time ultrasound. Four to six days after farrowing, an ultrasonic transit time flow probe was implanted around the right external pudic artery in order to measure the short-term variations of mammary blood flow through this vessel in response to postural change (standing vs lying), meal distribution, hand-milking, and weaning. After surgery, all sows were fed 3.8 kg/d of a lactation diet and housed either at 20 or 28 degrees C. The implantation of the ultrasonic blood flow probe was successful in all six operated sows. Postmortem examination did not indicate the presence of infection, any collateral bypassing the flow probe, or a reduction of artery diameter. The right pudic artery mammary blood flow (PMBF) was measured for 8.5 h over two periods of three days (d 11 to d 13 and d 18 to d 20 of lactation). The PMBF averaged 910 +/- 238 mL/min but was variable within 1 d. Compared with the lying position, PMBF was decreased (- 6%, P < 0.05) when sows were standing. Between 0 to 15 and 16 to 30 min after oxytocin injection (t = 0) and hand-milking, PMBF remained constant (P = 0.05; 801 vs 767 mL/ min) and increased (P = 0.02), respectively, in comparison with the mean calculated over the preceding 30-min period (982 vs 784 mL/min). The PMBF increased (P < 0.05) after meal distribution and reached a peak 65 min later (i.e., 980 mL/min). The PMBF decreased regularly after separation of piglets at weaning; at 8 and 16 h after weaning, PMBF was 60 and 40% of the value recorded before weaning, respectively. Assuming that PMBF drains one-quarter of the whole mammary gland, it can be calculated that blood flow through the entire mammary gland averages 3.6 L/min and that about 470 L of blood are required to produce 1 kg of milk. The proposed methodology constitutes a new technique to measure direct mammary blood flow and its short-term factors of variation.