Münchmeyer R

Untersuchungen zur Sezernierung endogener Aminosäuren in den Verdauungstrakt und zur Aminosäurenresorption beim Schwein

A trial was performed with 2 fistula pigs (each with 2 fistulas, one located about 30 cm below the pyloric orifice and the other at the end of the small intestine). Animal A received a casein diet containing 14% crude protein for a period of 2 weeks before the tracer amino acid was administered. Animal B received the same diet for a period of 10 days and was then fed a diet (at the same protein level) containing gluten as sole protein source. The two tracer amino acids, 14C-U-L-leucine and 3H-4,5-(N)-L-lysine, were injected intravenously. The passage rates for dry matter, organic matter and N measured at the beginning of the small intestine were higher than the rate of intake. The rate of passage of amino acids was also found to be increased relative to the rate of intake. In general, this increase involved the non-essential amino acids to a much larger extent. A considerable proportion of the amino acids passing into the large intestine is not excreted with the faeces but is probably converted in catabolic processes. It is for this reason that any values for the efficiency of amino acid absorption calculated on the basis of data on the faecal excretion of amino acids will not provide conclusive evidence for the availability of dietary amino acids in processes of the intermediate metabolism. The rate of secretion of 3H and 14C radioactivity into the digesta of the small intestine was found to increase rapidly within 1-2 hrs after administration of the tracer amino acids. The 14C radioactivity detected was found to be almost exclusively derived from 14C leucine while only about 60% of the 3H activity found in the digesta of fistula I were shown to be bound to lysine. Labelled lysine and leucine (of endogenic origin) are absorbed into the small intestine at a slower rate (i.e. endogenic proteins are less efficiently digested) than the non-radioactive amino acids (of exogenic origin) so that a process of concentration of endogenic amino acids is observed towards the end of the small intestine.

Studies on the secretion of amino acids and of urea into the gastro intestinal tract of pigs

Three pigs, of 34 kg live weight, were each fitted with re-entrant cannulas both in the duodenum and terminal ileum and catheters in the jugular vein and in the carotid artery. Pigs received a diet based on wheat and dried skimmed milk in equal amounts at 12 h intervals. During the preliminary period the digesta flowing from both duodenal and ileal cannulas were collected over 12 h after feeding on two consecutive days and half of them were reintroduced into the gut and half were stored at -20 degrees C. During the experimental period 15N-urea was infused into the jugular vein for 12 hours starting with the morning meal. Total amount of urea infused was 5 g containing 1.22 g 15N-excess. The digesta from both proximal duodenal and ileal cannulas were collected and stored, while the digesta from the preliminary period were reintroduced into the respective distal cannulas. Blood samples were taken at different time of infusion. At the end of infusion period the animals were sacrificed and samples of the contents of the digestive tract and tissues were taken. Urea flux calculated according to atom-% 15N-excess of urea N in plasma was 1.23 to 2.37 g/kg body weight/day. In the duodenal digesta 94.5 +/- 0.2 and in ileal digesta 57.1 +/- 7.39 per cent of 15N were in the TCA soluble fraction. The total amount of 15N in the duodenal digesta was 1.7 to 6.3 times greater than in the ileal digesta. Only small amount of 15N was found in the caecum and almost none in the contents of colon and rectum. It is concluded that urea is secreted into all parts of the digestive tract, the main sites of urea secretion being pancreatic juice and/or bile as well as the small intestine. The total amount of urea secreted is assumed to be similar to the daily urea excretion.Three pigs of 34 kg live weight were fitted with a re-entrant cannula in the duodenum, and with two catheters placed in the jugular vein and carotid artery. They were fed 1.2 kg/d of wheat-dried-skimmed milk diet. Digesta from the proximal duodenal cannula were collected for 12 h on 2 consecutive days; 50% were reintroduced into the respective distal cannula and 50% were stored at -20 degrees C. Three days later 14C-leucine was infused into the jugular vein for 12 h, starting with the morning meal. During this period the digesta from the proximal cannula were collected and stored for analysis while the digesta collected previously were reintroduced into the distal cannula. Blood samples were taken from the carotid artery. The total flow of duodenal digesta in 12 h was 5760 +/- 530 g. On average 70 percent of the radioactivity in digesta was associated with the TCA-precipitable fraction. During hours 0-3 and 11-12 of infusion 60-70% and 96-98% of the radioactivity in the TCA precipitable fractions was in leucine. In the TCA soluble fraction only 30-40 per cent of the radioactivity was associated with leucine. At the plateau 2.1 and 3.3% of infused 14C-leucine and of radioactivity were recovered in the duodenal digesta. The calculated amount of endogenous protein passing the duodenum was 20.4 g/d/pig. The results are discussed in relation to previous studies on protein synthesis and secretion in which 14C- and 15N-amino acids were used.

Flow of endogenous and exogenous amino acids along the gut of pigs

Digesta were collected from 5 pigs of 33 kg live weight fitted with re-entrant cannulas in the duodenum (within 20-30 cm of the pylorus) and terminal ileum. The pigs received a diet of barley, soya bean oilmeal and a vitamin and mineral mixture. The flow rates of digesta, total nitrogen and the individual amino acids were measured at different time after feeding and during two 24 h periods. A marked increase in the flow of digesta, nitrogen and amino acids was seen in the duodenum after feeding. Total flow during 24 h of nitrogen and amino acids except His, Val, Leu, Phe and Met exceeded intake. Output of nitrogen and amino acids from the duodenal cannula was 117 and 108% of intake, respectively. A method to calculate the ratio of endogenous amino acids in digesta based on the amino acid composition of digesta, diet and endogenous secretions was developed. The calculated amounts of endogenous amino acids passing the proximal duodenum and terminal ileum were 32.2 and 21.9 g per 24 h, respectively. The greatest amount of endogenous amino acids passed through the duodenal cannula in the first two hours after feeding (2-3 g/h) and then gradually decreased to 1 g per hour. The results are discussed in relation to other studies on the secretion of endogenous protein and its amino acid composition.

Untersuchungen zur Wirkung von Proteinaseinhibitoren bei Ratten

: Male Wistar rats (initial body weight 90 g) were fed ad libitum a whole-egg diet containing 10,5% crude protein. The animals of the experimental group received in each case of 1 mg leupeptin per 100 g of body weight in 12 hrs-intervals by i. p.-injection (3 days of treatment). Control animals got a leupeptin free solution. In addition, lysine dihydrochloride-alpha-15N was applied during the first three days of experiment to all animals and the nitrogen balance was determined. Urine from the N-Balance collection was analysed for 3-methyl-histidine excretion in order to calculate the degradation rate of myofibrillar proteins. On the fourth day the fractional rate of protein synthesis in several organs was estimated using the continuous infusion technique with 14C-leucine and 14C-lysine. The apparent biological half-lives of tissue protein were determined by a triple labelling technique, with (14C)-guanidino-L-arginine, L-5-3H-arginine and 15N-Lysine. The short-term treatment 3 days) with leupeptin did not affect the weight gain, the apparent digestibility of nitrogen and the N-balance. The fractional rate of protein synthesis was highest in the small intestine followed by the large intestine, liver and skeletal muscle and no influence of leupeptin treatment was observed. Furthermore no differences in the degradation rates of myofibrillar proteins between treated and untreated animals were found. The 3-methyl-histidine excretion via urine was 1.44 mg . kg-1 day-1 in both groups corresponding to a fractional rate of degradation of myofibrillar proteins of 2,5% per day. Apparent half-lives of tissue proteins in the small intestine, large intestine and liver, respectively, were shortest when estimated from the decay curves for the 14C-label and longest from the curves for the 15N-label. Leupeptin treatment resulted in prolonged apparent half-lives of the proteins in the large intestine and of the slowly turning over proteins in the liver. However, this effect seems to be caused rather by an increased reutilization of labelled amino acids than by a decreased protein degradation. Before continuing this kind of work the rate of uptake of injected leupeptine into tissues has to be investigated. Studies dealing with the in vivo action of proteinase inhibitors on protein metabolism have to include estimations of N-balance, protein synthesis rate, intracellular degradation rate of proteins as well as amino acid reutilization.

Untersuchungen zur Charakterisierung der Radioaktivitäts-verteilung im Organismus und zum Radioaktivitätseinbau in Gewebeproteine bei monogastrischen Tieren nach i. v.-Injektion von Traceraminosäuren

The studies were carried out with pigs and rats. The radioactive animo acids (14C leucine and 3H lysine) were administered to the pigs by way of a catheter tube into the jugular vein. Subsequently, the time pattern of the distribution of the specific amino acid radioactivity was followed in the TCE soluble and Tce precipitable fractions of the blood plasma (TCE= trichloro-acetic acid). The radioactive labelling in rats was carried out by injecting 14C leucine into the portal vein. The animals were killed after incorporation periods from 2 to 60 mins, and the levels of specific radioactivity were estimated in the TCE soluble and TCE precipitable fractions of the blood plasma, in the liver and in the skeletal muscles. The experimental results clearly indicated that the specific radioactivity of the tracer amino acids and the rate of incorporation of radioactivity into tissue proteins were greatly influenced by the size of the free amino acid pool within the range of distribution of the tracer. An estimation of the magnitude of the pool of free amino acids within the distribution range of the tracer can be obtained from the curve pattern for the decline of specific radioactivity of the corresponding free amino acid in the blood plasma. This pool exhibits a high rate of turnover. In all studies made to evaluate in vivo processes of protein synthesis by use of radioactive tracer amino acids it will be particularly important that consideration should be given to the specific radioactivity of the amino acid in the precursor pool for protein synthesis.

[Action of proteinase inhibitors. 4. Effect of short-term application of chymostatin on nitrogen metabolism in the digestive tract and protein metabolism in tissue].

Chymostatin is an effective inhibitor of intracellular proteinases in vitro. In the present experiment male rats were injected intraperitonealy during a 3 days period twice daily with a solution containing 0,9 mg Chymostatin per 100 g live weight. Reference animals received a control injection containing the same solvents but no chymostatin. During this period a daily nitrogen balance was made and metabolic faecal nitrogen and true digestibility of nitrogen were estimated using 15N-labelled animals. Furthermore, apparent biological half lives of proteins in liver and intestinal tissues were determined following the decay curves for radioactivity in proteins 48 hours after injection of L-[5-3H]-arginine und L-[guanido-14C]-arginine. The fractional rate of protein synthesis in tissues was measured by a 6 hours continuous infusion technique with L-[U-14C]-tyrosine and L-[U-14C]-leucine. Among the parameters estimated only the apparent biological half lives of proteins in liver and intestinal tissues were influenced by chymostatin. However, the prolonged half lives seemed to be rather an effect of an increased reutilisation of amino acids resulting from the intracellular protein breakdown than a decreased rate of protein degradation. The in vivo effect of the proteinase inhibitor was by far inferior compared with the action in vitro. Factors like distribution, degradation and excretion of the inhibitor could be responsible for the moderate in vivo action of chymostatin.