Effect of a thyroid hormone treatment on brain protein synthesis in rats

The effect of the thyroid hormone on the rate of brain protein synthesis in rats was studied. Experiments were conducted on three groups of rats given 6-propyl-2-thiouracil (PTU, a thyroid inhibitor) without a triiodothyronine (T3) treatment, those treated with PTU + T3, and those treated with neither PTU nor T3 (control). The fractional rates of protein synthesis in the brain, liver, and kidney of rats given PTU + T3 were significantly greater than those in rats given PTU alone. In the brain and kidney, the RNA activity [g of protein synthesized/(g of RNA.d)] were significantly correlated with the fractional rates of protein synthesis. In the liver and kidney, the RNA concentration (mg of RNA/g of protein) was related to the fractional rate of protein synthesis. These results suggest that the thyroid hormone treatment would be likely to increase the rate of protein synthesis in the brain of rats, and that the RNA activity is, at least partly, related to the fractional rate of brain protein synthesis.     

Deprivation of dietary nucleotides decreases protein synthesis in the liver and small intestine in rats

BACKGROUND & AIMS: Dietary nucleotides are reported to influence the growth and functioning of the liver and small intestine. The aim of this study was to examine the mechanism by which nucleotides exert their effects in these tissues by assessing protein synthesis activity and related parameters in the presence or absence of dietary nucleotides. METHODS: Rats were fed a purified diet with or without nucleotides for 10 days. Fractional protein synthesis rate, RNA and DNA concentrations, polysome size distribution, and number of ribosomes were assessed. RESULTS: Fractional protein synthesis rates of the liver and small intestine were lower in the nucleotide-deprived group than in the control group. In the liver, RNA concentration was also lower in the nucleotide-deprived group, but values in the small intestine were similar in the two groups. In the liver, deprivation of nucleotides resulted in a reduction in the number of ribosomes and in polysome breakdown. Protein and DNA concentrations did not vary in the liver; however, the concentration of DNA was lower in the small intestine of the nucleotide-deprived group than in the control group. CONCLUSIONS: Dietary nucleotides can modulate protein synthesis in the liver and small intestine as a result of tissue-specific nucleic acid changes.     

Effect of varying amino acid levels on protein metabolism in nephrotic rats during total parenteral nutrition

In an attempt to determine appropriate diet in nephrotic syndrome, nephrotic rats, induced by puromycin aminonucleoside, were nourished by total parenteral nutrition fluid containing the same energy, but three different levels (1.65, 3.3, and 6.6%) of amino acids for 7 d. The fractional rate of total protein synthesis in the liver was determined by injecting a flooding dose of [3H]phenylalanine. The proportion of newly synthesized proteins retained and exported by the liver was estimated by injecting a tracer dose of [14C]leucine and then measuring the protein radioactivity remaining in the liver and present in the plasma after secretion was completed. Nephrotic animals synthesized more protein than control animals. Although the absolute synthesis rates of total protein in liver were increased with increasing amino acid administration, the absolute rates of synthesis of albumin were higher in the 3.3% group than in the other groups in nephrotic rats. However, kidney protein synthesis in nephrotic rats was higher in the 1.65% group than in the 3.3% group. Interestingly, the 3.3% group revealed the smallest urinary excretion of total protein and albumin. In addition, in the 3.3% group, plasma concentrations of total protein and albumin were higher, and plasma concentrations of total cholesterol and triglyceride were lower than in other groups. It was concluded that the 3.3% group, corresponding to a normal protein diet, has the greatest salutary effect on urinary protein excretion, followed by protein and lipid metabolism, in nephrotic rats. Not only protein intake but also the energy:protein ratio are important for diet therapy in nephrotic animals. The technique of total parenteral nutrition may be useful in defining the factors involved in glomerular permeability or permselectivity and intracellular protein metabolism.     

Effects of zinc deficiency on protein synthesis and expression of specific mRNAs in rat liver

The effects of zinc deficiency on protein synthesis and expression of specific mRNAs were assessed in rat liver. Zinc deficiency had no apparent effect on liver weight, protein content, or RNA content when these properties were compared with values obtained using pair-fed rats. However, zinc deficiency resulted in a lower rate of hepatic protein synthesis. The decreased rate of protein synthesis was due to a decrease in the rate of synthesis of proteins retained in the liver, with no apparent change in the synthesis of secreted proteins. Analysis of expression of specific gene products, as assessed by in vitro translation of total RNA followed by two-dimensional gel analysis, showed that the expression of only a few mRNAs was altered by zinc deficiency. The patterns of change in gene expression resulting from zinc deficiency varied from almost complete repression to full expression. In additional studies, cDNA clones to serum retinol-binding protein and transthyretin were used to examine the effect of zinc deficiency on the relative abundance of mRNA for these two proteins. The relative abundance of mRNA for transthyretin was specifically elevated as a result of zinc deficiency. In contrast, the relative abundance of mRNA for hepatic serum retinol-binding protein was increased in both zinc-deficient and pair-fed rats. Therefore, the observed change in mRNA for serum retinol-binding protein was apparently at least in part due to the inanition that accompanies zinc deficiency. Overall, the results suggest that zinc can regulate the synthesis of specific proteins in rat liver through changes in the relative abundance of specific mRNAs.     

Measurement of fetal tissue protein synthetic rate in the lamb in utero

The fractional synthesis rate of the mixed proteins in the organs of fetal lambs at 123 to 130 days' gestation were calculated from measurements of the rate of uptake of L14Clysine by the protein in the steady state in utero. The fractional synthesis rates were 82, 121 and 99 per cent per day for the brain, liver and cardiac muscle respectively; the average values of 40 per cent for skeletal muscle was significantly smaller than for the other tissues. The fetal fractional protein synthetic rates measured were similar to those of the newborn lamb, but faster than in the adult sheep by a factor of 10. Similar differences in synthesis rates have been observed between the young and adult rat and the possible reasons for the change in fractional rate during growth are discussed.     

Soybean protein suppresses hepatic lipogenic enzyme gene expression in Wistar fatty rats

The effects of dietary soybean protein on lipogenic enzyme gene expression in livers of genetically fatty rats (Wistar fatty) have been investigated. When Wistar fatty rats and their lean littermates (7-8-wk old) were fed a casein or soybean protein isolate diet containing hydrogenated fat (4% hydrogenated fat plus 1% corn oil) or corn oil (5%) for 3 wk, the hepatic messenger RNA concentrations and activities of lipogenic enzymes were significantly lower in rats fed soybean protein than in those fed casein, regardless of genotype or dietary fat. The conversion rates of thyroxine to triiodothyronine by liver microsomes and plasma triiodothyronine concentrations were lower in the fatty rats than in the lean rats and were significantly greater in rats fed soybean protein than in those fed casein. Conversely, plasma and liver triacylglycerol concentrations were lower in soybean protein-fed fatty and lean rats than in those fed casein. The body weight was less in the fatty rats fed soybean protein than in those fed casein after 3 wk of feeding. Moreover, dietary polyunsaturated fatty acids suppressed lipogenic enzyme gene expression in the lean rats but did not in the fatty rats. Dietary soybean protein appeared to be useful for the reduction of obesity.     

Metallothionein and its relationship to the metabolism of dietary zinc in rats

Metallothionein (MTN), a low molecular weight cytoplasmic metalloprotein has previously been implicated in the metabolism of Zn. Experiments were conducted with rats to determine the effect of feeding varying levels of dietary Zn for short periods on the increase or decrease in MTN-bound Zn. The results indicated that MTN-Zn in both liver and intestinal mucosal cytosol responded rapidly to an altered dietary Zn level and that serum Zn was directly related to the appearance of MTN. 65Zn absorption also responded rapidly to a change in dietary Zn and was inversely correlated with intestinal mucosal MTN-Zn. Hepatic 65Zn uptake appeared to be directly correlated with liver MTN-Zn. A 24 hour fast increased MTN-Zn in rats fed two different levels of Zn except in those rats that were previously treated with actinomycin D. The antibiotic, which blocks DNA-dependent RNA synthesis, appeared to block Zn uptake from blood to the liver. These findings offer support for the involvement of MTN in zinc metabolism.     

Rate of glutamate synthesis from leucine in rat brain measured in vivo by 15N NMR

The rate of glutamate synthesis from leucine by the branched-chain aminotransferase was measured in rat brain in vivo at steady state. The rats were fed exclusively by intravenous infusion of a nutrient solution containing [15N]leucine. The rate of glutamate synthesis from leucine, determined from the rate of increase of brain [15N]glutamate measured by 15N NMR and the 15N enrichments of brain and blood leucine analyzed by gas chromatography-mass spectrometry, was 0.7-1.8 micromol/g/h at a steady-state brain leucine concentration of 0.25 micromol/g. A comparison of the observed fractional 15N enrichments of brain leucine (0.42 +/- 0.03) and glutamate (0.21 +/- 0.015) showed that leucine provides approximately 50% of glutamate nitrogen under our experimental condition. From the observed rate (0.7-1.8 micromol/g) and the known Km of the branched-chain aminotransferase for leucine (1.2 mM), the rate of glutamate synthesis from leucine at physiological brain leucine concentration (0.11 micromol/g) was estimated to be 0.35-0.9 micromol/g/h, with leucine providing approximately 25% of glutamate nitrogen. The results strongly suggest that plasma leucine from dietary source, transported into the brain, is an important external source of nitrogen for replenishment of brain glutamate in vivo. Implications of the results for treatment of maple-syrup urine disease patients with leucine-restricted diet are discussed.     

Effects of iron deficiency and iron overload on manganese uptake and deposition in the brain and other organs of the rat

Manganese (Mn) is an essential trace element at low concentrations, but at higher concentrations is neurotoxic. It has several chemical and biochemical properties similar to iron (Fe), and there is evidence of metabolic interaction between the two metals, particularly at the level of absorption from the intestine. The aim of this investigation was to determine whether Mn and Fe interact during the processes involved in uptake from the plasma by the brain and other organs of the rat. Dams were fed control (70 mg Fe/kg), Fe-deficient (5-10 mg Fe/kg), or Fe-loaded (20 g carbonyl Fe/kg) diets, with or without Mn-loaded drinking water (2 g Mn/L), from day 18-19 of pregnancy, and, after weaning the young rats, were continued on the same dietary regimens. Measurements of brain, liver, and kidney Mn and nonheme Fe levels, and the uptake of 54Mn and 59Fe from the plasma by these organs and the femurs, were made when the rats were aged 15 and 63 d. Organ nonheme Fe levels were much higher than Mn levels, and in the liver and kidney increased much more with Fe loading than did Mn levels with Mn loading. However, in the brain the increases were greater for Mn. Both Fe depletion and loading led to increased brain Mn concentrations in the 15-d/rats, while Fe loading also had this effect at 63 d. Mn loading did not have significant effects on the nonheme Fe concentrations. 54Mn, injected as MnCl2 mixed with serum, was cleared more rapidly from the circulation than was 59Fe, injected in the form of diferric transferrin. In the 15-d-rats, the uptake of 54Mn by brain, liver, kidneys, and femurs was increased by Fe loading, but this was not seen in the 63-d rats. Mn supplementation led to increased 59Fe uptake by the brain, liver, and kidneys of the rats fed the control and Fe-deficient diets, but not in the Fe-loaded rats. It is concluded that Mn and Fe interact during transfer from the plasma to the brain and other organs and that this interaction is synergistic rather than competitive in nature. Hence, excessive intake of Fe plus Mn may accentuate the risk of tissue damage caused by one metal alone, particularly in the brain.     

Estimation of tissue protein synthesis in rats fed diets labeled with (U-14C)tyrosine

The rate of liver and muscle protein synthesis has been measured in 27 rats after feeding L-[U-14C]tyrosine in L-amino acid diets prepared as agar gels. Constant specific activity of the free tyrosine pool, as indicated by constant excretion of 14CO2, was reached within 2 h of feeding and was maintained for the remaining 6 h of the 8-h experiment. Muscle protein synthesis was decreased (P less than 0.05) in rats fed a 0.3% methionine diet compared with rats fed this diet supplemented with 0.51% cystine (fractional rate of synthesis, ks: 0.098 vs. 0.121). No effect (P greater than 0.05) of these diets on liver protein synthesis was observed (ks: 0.603 vs. 0.532). Protein synthetic rate was also determined by the constant-intravenous infusion technique in 17 rats fed unlabeled diets. The two techniques gave similar estimates. Restraint of the rats or the infusion of saline had no measurable effect on the rate of protein synthesis in rats fed labeled diets. This feeding technique is essentially equivalent to the constant-infusion technique and offers an easier, more physiological approach to achieving a steady state.     

Changes in the expression of protein kinase C (PKC), phospholipases C (PLC) and D (PLD) isoforms in spleen, brain and kidney of the aged rat: RT-PCR and Western blot analysis

The age-dependent changes of expression of protein kinase C (PKC), phospholipase C (PLC) and phospholipase D (PLD) isozymes were analyzed in spleen, brain and kidney of young-adult (12-16 week-old) and aged (82-88 week-old) rats. The activities of spleen cPKC and nPKC were significantly decreased by nearly 35 and 30% in aged rats compared to those of young adults, respectively (P < 0.05). The level of PKC beta1 was significantly decreased in aged rats as assessed by RT-PCR and Western blot analyses. In aged rat brain where the activity of cPKC was significantly decreased by nearly 25% (P < 0.05), PKC alpha and beta1 isozymes were significantly down-regulated. In kidney, the level of PKC beta2 was decreased. In spleen the both mRNA and protein levels of PLC beta2 and gamma2 were significantly down-regulated in aged rat (P < 0.05). PLC beta1 was also significantly lower in aged rat brain (P < 0.05) as assessed by RT-PCR and Western blotting. Moreover, PLC beta1 was significantly down-regulated in both mRNA and protein levels in aged rat kidney (P < 0.05). In contrast, the tissues examined, the expressions of PLD isozymes (PLD1a, 1b and 2) were rather stable in the course of aging. These results indicate that mRNAs of PLD isozymes were rather stable but that particular PKC and PLC isozymes were down-regulated in different tissues during aging, suggesting age-dependent decline of specific PKC and PLC isozymes in organs which may, at least in part, be implicated in tissue dysfunction with aging.     

Brain iron, transferrin and ferritin concentrations are altered in developing iron-deficient rats

To study the iron, transferrin, and ferritin distribution at subcellular levels in response to acute dietary iron deficiency, we tested the hypothesis that early post-weaning iron deficiency can change iron and iron regulatory protein concentrations in rat brain. Male Sprague-Dawley rats were fed diets containing either 2 or 35 micrograms iron/g for 2, 3 or 4 wk starting at 21 d of age. Brain iron, transferrin and ferritin concentrations in cytosolic and microsomal fractions of either whole brain or pons and cerebellum were then determined. After 14 d of dietary iron restriction, brain iron concentrations were 50% lower in the microsomal fraction and 30% lower in cytosol compared with controls. Brain cytosolic transferrin concentration almost doubled in the same animals. Brain ferritin concentration in fractions from rats fed the iron-deficient diet for 14 d was lower than in controls, but then remained fairly constant. Absolute brain weight and total brain protein contents were unaffected by iron restriction. This study extends previous research by demonstrating that the brain responds to changes in body iron status with a change in transferrin concentration. If the dietary restriction is quite severe, this adaptation is insufficient. This study also notes that brain ferritin decreases with decreasing body iron status, though it was less responsive than nonheme iron in liver. The concept that iron enters the brain through a highly regulated endocytotic process at the blood brain barrier, that undoubtedly involves the regulation of transferrin receptors in capillary endothelial cell, is supported by our observation of elevated transferrin concentrations in brain of iron-deficient rats.     

Rates of lysine catabolism are inversely related to rates of protein synthesis when measured concurrently in adult female rats induced to grow at different rates

To test the effect of changes in the rate of protein synthesis on amino acid oxidation, both were studied concurrently in individual 200-g female Sprague-Dawley rats. In a growth trial (Experiment 1), recombinant bovine somatotropin (rbST) was injected subcutaneously (0, 2 or 12 mg/d) over 6 d (n = 4 rats per rbST level). Weight gain increased with rbST level (P < 0.01); 1.96 +/- 0.8, 4.24 +/- 0.8 and 8.67 +/- 0.8 g/d, respectively. After treatment with rbST (0 or 12 mg/d) for 4 d (Experiment 2), rats were injected via a tail vein catheter with valine (400 mmol, 4.07 mBq L-[3,4(n)-3H]valine) at 0, 4, 10, 13 or 16 h after the daily rbST injection and killed 20 min later. This flooding dose was 5 to 6 times, not 10 times, the free pool as hoped. Protein synthesis in rbST-treated rats increased 46% in muscle (P < 0.001) and 36% in liver (P < 0.01). The ks was unaltered with time after rbST injection (0-16 h, P > 0.05). When 600 mmol valine (4.4 mBq L-[3,4(n)-3H]valine) was used in Experiment 3, specific activity (SA) of free valine was constant over 20 min and was 94 +/- 4% of that injected. Finally, in Experiment 4, protein synthesis and amino acid oxidation rates measured in the same rat revealed a 35% increase (P < 0.01) in protein synthesis in hind leg muscle and a 29% increase in liver (P < 0.05) from rbST-injected (12 mg/d) rats (n = 6). Lysine oxidation was estimated by continuous (12 h) infusion of L-[1-14C]lysine via the opposite tail vein catheter. Expired CO2 was collected over 20-min intervals and SA at plateau was estimated by fitting an exponential model. Lysine oxidation was reduced (P < 0.05) by 44% in rbST-treated rats. The idea that an increase in protein synthesis results in decreased amino acid oxidation remains tenable.     

Changes in brain protease activity in aging

We measured changes in protease activity with aging, conducting assays of cathepsin D and calpain II activities and the rate of degradation of cytoskeletal proteins, preparing the enzymes and substrates from young and aged brains. Calpain preparations added to the young and to the aged substrates were standardized with casein as substrate so that age-related changes in calpain specificity and substrate susceptibility were measured. Several age-related differences were observed in substrate susceptibility and in enzyme activity. With respect to substrate, the neurofilament protein from young animals was somewhat more susceptible to calpain action than that from older animals. With respect to enzyme activity, calpain from aged brain cleaved neurofilament protein at a faster rate than did calpain from young. With neurofilaments, the most rapid breakdown usually occurred when enzyme from aged tissue was incubated with substrate from young. Kidney enzyme of aged rats incubated with neurofilament substrate of aged rats resulted in a more rapid breakdown than enzyme of young kidney incubated with substrate of young. The age dependence of tubulin breakdown was somewhat different from that of neurofilament breakdown. The most rapid breakdown usually occurred when using enzyme from young with tubulin from young. Incubation of neurofilament protein or tubulin with cathepsin D did not reveal any differences with aging. These studies suggest that an increase in enzyme activity observed previously during aging may also include changes in the properties of the enzyme (substrate specificity) and/or in the properties of their endogenous substrates (susceptibility to breakdown).     

Effect of glycerol on lipogenic enzyme activities and on fatty acid synthesis in the rat and chicken

The influence of glycerol on the rates of fatty acid snythesis in liver slices from rats and chickens in pieces of adipose tissue from rats was first studied. Then the effect of dietary glycerol on lipid metabolism in rats and cheickens was examined. Media containing 3 or 10 mM glycerol depressed the rate of glucose conversion to fatty acids in rat liver slices. However, media containing up to 25 mM glycerol did not influence the rate of fatty acid synthesis in chick liver slices. The inhibitory action of glycerol in rat liver slices might occur at the level of glucose (or glycogen) conversion to pyruvate because glycerol did not inhibit pyruvate or acetate conversion to fatty acids. Rats and chickens were fed glycerol containing diets for either 3 days or 3 weeks. Feeding diets containing 20.5 parts glycerol (22% of dietary energy) to rats or chickens did not influence the growth rate of the animals. However, substitution of 42.2 parts glycerol (43% of dietary energy) for glucose in the diet significantly depressed food intake and growth rate in both rats and chickens. The activities of citrate cleavage enzyme, fatty acid synthetase and malic enzyme in livers of rats fed the glycerol-containing diets were dramatically increased. However, this stimulation of enzyme activity occurred without a concomitant increase in the in vivo rate of fatty acid synthesis in the rat liver. In the chicken, unlike the rat, dietary glycerol did not stimulate but instead decreased hepatic malic enzyme and fatty acid synthetase activities. No significant differences in adipose tissue lipogenic enzyme activities or in the rates of fatty acid synthesis were observed in rats fed glycerol-containing diets. The lipogenic response to glycerol feeding depends on the species as well as the organ.     

Molecular cloning and biochemical characterization of bovine spleen myristoyl CoA:protein N-myristoyltransferase

Myristoyl-CoA:protein N-myristoyltransferase (NMT) is an essential eukaryotic enzyme that catalyzes the cotranslational transfer of myristate to the NH2-terminal glycine residue of a number of important proteins of diverse function. We have isolated full-length cDNA encoding bovine spleen NMT (sNMT). The single long open reading frame of 1248 bp of sNMT specifies a protein of 416 amino acids with a predicted mass of 46,686 Da. The protein coding sequence was expressed in Escherichia coli resulting in the production of functionally active 50-kDa NMT. Deletion mutagenesis showed that the C-terminus is essential for activity whereas up to 52 amino acids can be deleted from the N-terminus without affecting the function. One of the N-terminal deletions resulted in threefold higher NMT activity. Genomic Southern analysis indicated the presence of two strong hybridizing bands with three different restriction enzyme digests suggesting the possibility of two copies of the NMT gene in the bovine genome. RNA blot hybridization analysis of total cellular RNA prepared from bovine brain, heart, spleen, lung, liver, kidney, and skeletal muscle probed with bovine sNMT cDNA revealed a single 1.7-kb mRNA. Western blot analysis of various bovine tissues with human NMT peptide antibody indicated a common prominent immunoreactive band with an apparent molecular mass of 48.5-50 kDa in all tissues. Additional immunoreactive bands were observed in brain (84 and 50 kDa), lung (58 kDa), and skeletal muscle (58 kDa). Activity measurements demonstrated that brain contained the highest NMT activity followed by spleen, lung, kidney, heart, skeletal muscle, pancreas, and liver. It appears therefore that mRNA and protein expression do not correlate with NMT activity, suggesting the presence of regulators of the enzyme activity.     

Inhibitory effect of crocetin on intracellular nucleic acid and protein synthesis in malignant cells

The possibility that dietary intake of diverse naturally occurring compounds may influence the occurrence of cancer is receiving considerable scientific attention. Previously, it was reported that an extract (Crocus sativus), which contains carotenoids, had an antitumor effect and inhibited colony formation and nucleic acid synthesis by malignant human cells. Epidemiological and experimental research has indicated that carotenoids might act as antitumor agents. We have studied crocetin, a carotenoid isolated from saffron, which has been shown to have biological activity. In our experiments we utilized three malignant human cell lines: HeLa (cervical epitheloid carcinoma), A549 (lung adenocarcinoma) and VA13 (SV-40 transformed fetal lung fibroblast) cells. The effect of crocetin on colony formation and cellular DNA, RNA and protein synthesis in these cells has been examined. Incubation of these cells with crocetin for 3 h caused a dose-dependent inhibition of nucleic acid and protein synthesis. Crocetin also had a dose-dependent inhibitory effect on DNA and RNA synthesis in isolated nuclei and suppressed the activity of purified RNA polymerase II.     

Influence of environmental temperature on in vivo energy expenditure in vitro ouabain-sensitive respiration in duodenal mucosa and liver in rats fed different levels of dietary fiber or protein

Seventy two Wistar rats were used in two repeat studies to investigate the effect of environmental temperature (18 degrees C or 28 degrees C) and increasing levels of dietary fibre (low, 68 g/kg DM; medium 110 g/kg DM; high, 157 g/kg DM) or protein (low, 91 g/kg DM; medium, 171 g/kg DM; high, 262 g/kg DM) on digestive tract, visceral organ size, energy metabolism, and respiration attributable to Na+,K(+)-ATPase activity in duodenal mucosa and liver. Total and ouabain-sensitive (a measure of Na+,K(+)-ATPase activity) O2 consumption in vitro of tissues were measured polarographically using a Clark-style YSI biological O2 monitor. Whole body heat production (in vivo) was measured using open-circuit respiration chambers. The weight of the visceral organs was higher in rats housed at 18 degrees C than at 28 degrees C. The empty weight of the small intestine, caecum, and colon increased as the level of dietary fibre increased (P 0.05). Heat production as a proportion of metabolizable energy was higher (P < 0.05) at 18 degrees C than at 28 degrees C in the first experiment but this difference was significant in the second experiment. Rats fed the low protein diet had significantly higher (P > 0.05) heat production than those fed medium or high protein diets. Compared to 28 degrees C, environmental temperature of 18 degrees C caused an increased total and ouabain-sensitive O2 consumption in duodenal mucosa. There was no significant effect of environmental temperature on total and ouabain-sensitive O2 consumption in the liver. However, ouabain-sensitive O2 consumption in liver was significantly higher (P 0.05) when rats were fed a low protein diet compared to the medium or high protein diet. Total and ouabain-sensitive O2 consumption increased in duodenal mucosa of rats fed low level of dietary fibre compared to the medium or high dietary fibre diets. The in vitro results corresponded with the whole animal energy expenditure and O2 consumption in vivo.     

Studies on transmembrane and paracellular phenomena in the foot of the slug agriolimax reticulatus (Mü)

Employing livers from rats fed on a protein-free diet for two weeks, the effects of protein deficiency on both biosynthesis and degradation of rRNA were investigated and the following results were obtained. 1. Protein deficiency led to a decrease of total liver RNA content per DNA to about 80% of that in normal rat liver. 2. From the kinetics of rRNA labelling with [14C]orotic acid in vivo, the half-lives of cytoplasmic rRNA's of normal and protein-deficient rat livers were determined to be 6.2 and 5.1 days, respectively. Furthermore, considering the pool size of rRNA in rat liver, the turnover rate of cytoplasmic rRNA was calculated to be 0.212 pmole/min/mg of nuclear DNA in normal rats and 0.240 pmole/min/mg of nuclear DNA in protein-deficient rats. 3. From the electrophoretic patterns of nucleolar RNA's of both groups of rat livers labeled with [14C]orotic acid, the time courses of the specific activities of nucleolar 45S, 32S, and 28S rRNA's were analysed and the half-life of each nucleolar RNA in both groups of rat livers was determined. Nucleolar 45S, 32S, and 28S RNA's had half-lives of 6.0, 15.9, and 26.5 min in normal rats, respectively, and 5.5, 19.4, and 22.9 min in protein-deficient rats, respectively Considering the pool size of each nucleolar RNA obtained from the leectrophoretic pattern, the turnover rates of 45S, 32S, and 28S RNA's were calculated to be the same, i.e., o.189 pmoles/min/mg of nuclear DNA, in normal rat liver and 0.372, 0.372, and 0.358 pmoles/min/mg of nuclear DNA in protein-deficient rat liver, respectively. 4. These results indicate that protein deficiency increased both the rate of degradation of cytoplasmic rRNA and that of nucleolar rRNA synthesis in rat liver. While in normal rat liver the rates of rRNA synthesis and degradation were rather similar, the rate of rRNA synthesis in protein-deficient rats was about 1.5 times higher than that of its degradation. Therefore, the decrease of total liver RNA content in protein deficiency might be accounted for by stimulated degradation of rRNA in the nucleus. 5. The activities of RNase in nuclear fractions of both groups of rat livers were compared. Both activities of nuclear acid RNase and especially that of the free form of alkaline RNase in protein-deficient rat liver were higher than those in normal rat liver.