Biomedical subjects
Publications and source records attributed to O Simon.
Secretion into the Gastro-Intestinal Tract of Amino Acids Labelled with Radioactive Isotopes. (4) 14C-Labelling and 14C-Leucine in the Gastro-Intestinal Tract after Subcutaneous Injection of 14C-Leucine to Rats with Tied-off Pancreatic Gland.
Male Wistar rats (of 60 g live weight) allotted in 10 groups were fed diets with gradually increasing lysine levels ranging from 1.4 to 7.4 g lysine/16 g N. Feed intake was restricted so much that the experimental animals did not change their live weights during the last 3 days of the 8-day experiment period. On the 7th experimental day, 4 animals of each group were injected i.p. 14-C-L-lysine, the 14CO2-excretion being subsequently measured over a period of 2 hours. On the next day, 6 animals of each group were applied an i.p. injected of 15N-L-lysine, the urine being collected over the following 24-hour period to measure the 15N-frequency. Applying both labelling methods, an increased catabolisation of the amino acid was observed after the metabolically necessary lysine requirement had been covered. The methods are very sensitive and revealed, under the experimental conditions chosed, a lysine requirement coverage of about 3 g lysine/16 g N. The possibility of using also 15N-labelled compounds in the metabolism-oriented amino acid requirement determination is likely to facilitate the transfer of the methodology to farm animals and would thus allow to study the amino acid requirement of man. The metabolism-oriented amino acid requirement determination will likewise allow to estimate exact amino acid requirement data under conditions that cannot be rated on the basis of productive yields.
During a 15-day period, growing Wistar rats (75 g) were fed ad lib. a whole-egg diet containing 10% crude protein (control group) and the same diet + a daily subcutaneous injection of 1 mg of leupeptin per animal (experimental group), respectively. To investigate the influence of the peptide aldehyde leupeptin on the M-metabolism, N-balance trials were carried out between the 1st and the 6th, and the 10th and the 15th experimental days. Leupeptine was not found to influence the true N-digestibility. However, the intermediary N-utilization, characterized by the biological value of the dietary protein, deteriorated in comparison with the control group. The peptide aldehyde under study resulted in an extension of the biological half-life of the proteins in the tissue of the small and large intestine. It was measured via the decline of radioactivity in protein following application of L-guanidino-14 C-arginine and L-(4,5-3H)-leucine. The amino acid utilization appears to have been increased as well. In connection with previous studies (SIMON et al., 1976) i, which, under somewhat different experimental conditions, leupeptine resulted in a higher intermediary N-utilization, the mentioned agent is supposed to be able to influence N-utilization. However, additional studies are required to make statements regarding the influence of dosis and metabolic situation.
A daily s.c. injection of 0.5 mg leupeptin for 7 days does not influence the digestibility of food-N but seems to improve the N-retention. A daily injection of 1.0 mg leupeptin for 14 days is without influence on the digestibility of N, too, but the N-retention was decreased. At this leupeptin level the half-lives of intestinal proteins were prolonged. The data indicate the possibility to influence the N-retention using leupeptin.
Open- and closed-eyed EEG records were made in frontal, temporal and occipital regions, in order to examine the time relationships between various of their characteristics, in connection with the appearance of subclinical seizure patterns. 1. In the range of alpha frequency, both with open and closed eyes, the standard deviation of frequency over all brain areas decreased both before and after subclinical seizure patterns as compared to periods without pattern, and the number of temporal waves was reduced. Frontally, the mean amplitude was increased only in open-eyed records. The time relationship of the appearance of alpha maxima differed over all regions, but was closest in frontal records in both open- and closed-eyed records, and independent of the pathological pattern. In closed-eyed frontal records, a changeover in the time lead between the hemispheres could be observed, with one side leading during the period preceding the pattern, and the other after the pattern. 2. In the range of theta frequency, occipital open-eyed records showed slowing of the frequency both before and after the subclinical seizure pattern, with an increase in the standard deviation. In closed-eyed records made in the period preceding the pattern, an increase of the mean amplitude was observed frontally and occipitally, as well as an increase of the standard deviation of the amplitude in all regions. Slowing of the theta frequency was encountered in the frontal regions. Differences in the periods preceding and following subclinical seizures were to be seen only in the theta range.
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.
Short-term trials with a triazin derivative and a peptide aldehyde were carried out to investigate the in-vivo administration of proteinase inhibitors in rats for improving the N balance of the animals. The results of N balances suggest that the peptide aldehyde may be regarded as a promising substance that may be used to increase the rate of utilization of the absorbed N in the intermediary metabolism. The short-term use of both active compounds did not affect the true N digestibility. The two compounds did not influence the concentration of free amino acids in the liver and muscle tissue under the experimental feeding conditions used. Similarly, tests of the biological half-life of proteins taken from the small and large intestine did not reveal any changes due to the presence of the inhibitors. Animals receiving a protein-free diet exhibited longer half-life periods for proteins from the small and large intestine compared with animals of the other groups. A protein-free nutrition of the animals caused a general increase in the levels of free non-essential amino acids in the liver. This was accompanied by a simultaneous decrease in essential amino acids. The same type of nutrition led to a general decline in the concentration of free amino acids in muscular tissue, particularly of the essential amino acids. High-protein nutrition led especially to an increase in the concentration of branched-chain amino acids in both types of tissues investigated.
The paper first presents a survey of the existing literature where studies on the effects of isobutylidene diurea (IBDH) as NPN source for ruminants are discussed. Following this the authors discuss their own experiments on IBDH turnover. IBDH showed no toxi cation when tested in toxicity studies with rats which were carried out under standardized conditions. Within the framework of metabolism trial 4 growing male sheep (average liveweight: 30 kg) were fed a semisynthetic diet containing IBDH as sole N source. Each animal received 60 g IBDH per day. The average value for the apparent N digestibility was found to be 77.6%, the percentage for the average N ballance was 27.6%. The blood of the sheep was investigated, on a comparative basis, for the behaviour of free amino acids after the administration of an experimental semisynthetic diet containing IBDH or extracted soya bean meal as sole N source. The use of IBDH produced a higher concentration of free amino acids and a five-fold increase if urea concentrations in the blood plasma but reduced the Asp, Cys and Tyr levels when compared with the feeding of extracted soya bean meal. Similarly, IBDH supplementation increased the relative proportions of Gly, Lys, and His in blood plasma but reduced those of Asp, Val and Tyr. A positive relationship between the rate of IBDH hydrolysis and the decrease in pH was found in experiments studying the hydrolysis behaviour of IBDH at pH values from 1-7.
Male experimental rats (100 gm liveweight) were distributed into 10 groups of 8 animals each and received balanced diets, with the exception of lysine which was added to the diets in graded amounts in such a way that the lysine content of the diets ranged from 2.44 to 5.92 gm/16 gm N. After a feeding period of 7 days the animals received 3H- and 14C lysine injected intraperitoneally, 4 animals of each group were investigated for the total CO2 excretion and 14CO2 excretion during the first 2 hrs after the injection and for the urinary excretion of radioactivity (48 hours). The remaining animals in each group were used for determining the plasma amino acids and for establishing the specific radioactivity of free lysine in the liver and muscles after an 1-hour incorporation period. Total CO2 excretion was not found to be influenced by the lysine contents while the level of excretion of 14C activity through CO2 and that of specific 14C activity of CO2 increased with increasing lysine concentrations. This produced a broken curve pattern, showing an increased release of 14CO2 (under maintenance conditions) if the diet contained 4 gm lysine/16 gm N and more. Investigations for the specific 14C activity of free lysine in the liver, the main site of lysine oxidation, showed that the increase in 14CO2 release was due to an enhanced rate of lysine catabolism and was not brought about by changes in the pool volume or in specific radioactivity. The levels of urinary 14C excretion were not found to be related to the lysine content of the diets, whereas the curve pattern of 3H excretion observed 5 to 8 hrs after injection was similar to that of 14CO2 excretion. The lysine content of blood plasma and the content of free lysine in the liver increased continuously with increasing levels of dietary lysine. The methodological studies made in the present paper showed that in scientific research a determination of amino acid requirements on the basis of CO2 oxidation data may be a very exact and sensitive method. It will also yield values for maintenance requirements.
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.
4 different groups of experimental animals (rats) were injected 14C leucine at 4 different levels of specific 14C activity. Groups A and B received a dosis of leucine corresponding to the daily leucine requirements of the animals. The specific 14C activity was high in group A and low in group B. Groups C and D were injected a leucine dosis at a level of 10% from the groups A and B. the specific 14C activity being again high (group C) or low (group D). The results showed that a true evalution of the experiment could lnly be achieved if the values were related to the true rate of labeling (DPM/mg N or specific 14C leucine activity). The "percentage of the injected dosis" proved to be unsuitable for the analysis of the experimental results when taken as reference frame. The levels of specific 14leucine activity in free plasma leucine were in the same order as the specific activities of the injected solutions (C greater than A greater than D greater than B) whereas the order of specific 14C leucine activities in the TCE-soluble fraction of the pancreatic gland (TCE-trichloroacetic acid) was found to be A greater than C greater than B greater than D. This was due to the higher rate of 14C-leucine transfer into the protein fraction in the animals of group C and D receiving a normal supply of leucine. In these groups an equilibrium in specific 14C-leucine activity between the TCE-soluble and TCE-precipitable fractions was achieved 20-30 mins after injection. A relatively high degree of labelling of the pancreatic protein was noted as early as about 2 minutes after the injection of 14C-leucine. This indicates that in studies on amino acid absorption the secretion of those amino acids that are first absorbed and then resecreted via the pancreatic gland should always be taken into account.
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Growing rats received either a protein-free experimental diet or a diet containing 15% crude protein (whole egg). The crude fibre content of the diet was adjusted to 5%, 10%, or 20% by adding various proportions of what straw meal. Following a preliminary 7-day period of feeding the animals received L-14C-Leucine administered by stomach tube or by subcutaneous injection. In a 72-hour post-experimental period analyses were made to investigate the urinary and faecal excretion of radioactivity. About 50% of the amount of radioactivity excreted with the feaces were of endogenic origin. In general, higher percentages of straw meal in the diet produced higher levels of faecal 14C excretion. It was only in the animals placed on the whole egg diet that the TCA soluble portion of radioactivity was found to rise with the increasing straw content of the diet. The experimental results substantiate the assumption that native crude fibre influences the process of intestinal sloughing and, additionally, it is capable of abosrbing or adsorbing amino acids, peptides or protein, due to its spatial configuration. Similarly, it affects the digestion and resorption of proteins.
Male Albino rats (90-100 g) were fed ad libitum (with limited periods of feeding) for 14 days. The diets were adjusted to a crude protein content of 10%. Powdered whole egg, fish meal, yeast and gelatine were used as protein sources. Additionally, one group of rats was fed a protein-free diet. On the 15th day of experiment the rats were fed a test diet at a level of 2 g per 100 g of body weight. 2 hrs after that the rats received 25 muCi of 3H glycine and 5 muCi of 14C-L-Leucine per 100 g of body weight administered by way of intragastric infusion. It was found that a large proportion of the radioactive amino acids were absorbed as early as after 0.5 hr. The highest rate of absorption was observed in animals fed dietary proteins of poor quality or a protein free diet, so that in animals receiving a gelatine diet or a protein-free diet only 68.4% or 56.4% of the administered amount of 14C activity were detected inside the gastro intestinal tract after 0.5 hr. Analogous data for the 3H activity were 52.4% and 25.3%. Maximum absorption occurred after 3-7 hrs. Following this the level of radioactivity in the intestinal contents again increased reaching a peak value after 14-24 hrs; in the case of 14C activity this peak value amounted to 25.4% of the administered dose in animals fed the gelatine diet and 32.8% in the group receiving the protein-free diet. It was established that the major proportion of the resecreted amount of 14C activity was present in leucine. Until 72 hrs after the intake of 14C activity the level of radioactivity was again found to decline, a processes which was induced by processes occurring in the large intestines. Moreover, evidence was obtained in confirmation of previous findings, indicating that the composition of faecal amino acids was constant and unaffected by dietary proteins.
Male Albino rats (weighing 90-100 gms) were fed ad libitum for 14 days with limited periods of access to food. Powdered whole egg (V), fish meal (F), yeats (H), and gelatine (10% protein in dry matter) used as protein sources. Additionally, one group of rats received a protein-free (e). Radioactive tracers were administered by intragastric infusion of 25 mu Ci 3H glycine and 5 mu Ci 14C L-leucine per 100 gm of body weight 2 hrs after the feeding of 2 gm of the experimental diet per 100 gms of body weight. The level of uptake of radioactive tracers from the different sections of the gastro-intestinal tract was measurels of 3H and 14C labelling in intestinal tissues were observed 3 hrs and 7 hrs after infusion. The level of 14C labelling was found to be negatively correlated and the level of 3H labelling was positively correlated with the biological value of the diet. Intestinal tissues are capable of storing considerable amounts of 14C radioactivity. So, 72 hrs after infusion, the following levels of 14C radioactivity (expressed as percentage of the total dose of radioactivity) were found in tissues of the gastro-intestinal tract: whole egg: 8.4%, fish meal: 9.6%, yeast: 13.1%, gelatine: 14.9%; protein-free diet; 14.2%. The quotients correlating the levels of radioactivity from the intestinal contents with that found in the intestinal wall suggest that the walls of the small intestine possess a high capacity for absorption. At all times of radioactive measurements the walls of the small intestine were found to contain higher levels of both 14C and 3H radioactivity than the contents of small intestine.
Growing male rats received diets of varying biological value (protein sources: powdered whole egg (V); fish meal (F); yeast (H); gelatine (G); protein-free diet (e)) for a 14-day feeding period. Subsequently, 14C leucine and 3H glycine were administered intragastrically. The level of uptake of 14C and 3H radioactivity into blood plasma, liver and muscular tissue and the rate of incorporation of the radioactive tracers into the proteins of these tissues was examined. A negative correlation was found to exist between the incorporation of radioactivity into liver proteins and the biological value of dietary proteins, the former being mainly dependent on the level of incorporation into the liver. For muscular proteins the rate of incorporation decreases with the decreasing biological value of the dietary proteins. This may be attributed to the fact that with poor protein nutrition the rate of protein synthesis in the skeletal muscles is also reduced. Comparative studies on the specific 14C radioactivity from free leucine made in the group on the protein-free diet and in the group receiving the whole egg diet showed that the leucine pool of the skeletal muscles was markedly redueced in animals fed a protein-deficient diet while the leucine pool in the liver remained comparatively constant.
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