Regulation of oxidative phosphorylation by adrenaline and insulin, effect of the insulin-dependent cytoplasmic factor

The insulin-dependent cytoplasmic factor-regulator (IDR) was shown to inhibit pyruvate and succinate oxidation by the rat liver mitochondria. In view of the fact that insulin increases and adrenaline decreases IDR activity in the liver cytoplasm it is suggested that oxidation of the substrate in mitochondria in vivo is regulated by changes in IDR content in cytoplasm. It was shown that adrenaline-activated oxidation apart from decreased activity of IDR in cytoplasm is induced by a different mechanism not related to IDR content.     

Factors regulating gluconeogenesis in chick embryo liver]

The ratio NAD+/NADH in cytoplasm and mitochondria of chicken embryo liver does not change up to the stage of hatching. After the hatching this ratio decreases 2-fold in both cytoplasm and mitochondria. The hatching is also accompanied by the decrease of total and mitochondrial contents of oxaloacetate and of oxaloacetate/malate ratio, the activity of citrate synthase and the ratio acetyl-CoA/CoA being unchanged.     

Ratio of concentration of ions of uni- and bivalent metals in the tissues of developing chick embryos and 1-day-old chickens]

The ratio of concentrations Na+/K+ decreases in the brain and liver and increases in the cardiac muscle during the chick embryogenesis. The maximum concentration of Ca2+ was noted in the tissues under study on 14th day of chick embryonic development and its content decreased reliably at the subsequent stages. The concentration of Mg2+ in the tissues under study decreased during the embryogenesis as well.     

The effects of surgical stress and short-term fasting on protein synthesis in vivo in diverse tissues of the mature rat.

1. We measured fractional rates of protein synthesis, capacities for protein synthesis (i.e. RNA/protein ratio) and efficiencies of protein synthesis (i.e. protein-synthesis rate relative to RNA content) in fasted (24 or 48 h) or fasted/surgically stressed female adult rats. 2. Of the 15 tissues studied, fasting caused decreases in protein content in the liver, gastrointestinal tract, heart, spleen and tibia. There was no detectable decrease in the protein content of the skeletal muscles studied. 3. Fractional rates of synthesis were not uniformly decreased by fasting. Rates in striated muscles, uterus, liver, spleen and tibia were consistently decreased, but decreases in other tissues (lung, gastrointestinal tract, kidney or brain) were inconsistent or not detectable, suggesting that, in many tissues in the mature rat, protein synthesis was not especially sensitive to fasting. 4. In fasting, the decreases in fractional synthesis rate resulted from changes in efficiency (liver and tibia) or from changes in efficiency and capacity (heart, diaphragm, plantaris and gastrocnemius). In the soleus, the main change was a decrease in capacity. 5. Surgical stress increased fractional rates of protein synthesis in diaphragm (where there were increases in both efficiency and capacity) by about 50%, in liver by about 20%, in spleen by about 40%, and possibly also in the heart. In liver and spleen, capacities were increased. In other tissues (including the skeletal muscles), the fractional rates of protein synthesis were unaffected by surgical stress.     

基于核酸定量的核质分离质控方案

目的:拟建立一种方便快捷、经济有效的细胞核质分离鉴定方法。方法:本研究从DNA水平进行核质分离鉴定,选择GAPDH、ND1分别作为细胞核和细胞质标志基因,并根据GAPDH及ND1序列保守区设计引物,基于荧光定量PCR方法定性检测核质分离的效果。随后将本鉴定方法应用于其他种类细胞(BEAS-2B细胞、GT1-7细胞)及组织(小鼠心脏、肝脏、大脑)的核质分离鉴定。结果:在293T细胞应用该鉴定方法鉴定核质分离效果,结果显示:GAPDH、ND1在核组、质组中的含量存在明显差异,其中核标志基因GAPDH在细胞核中的比例达到了95%以上,质标志基因ND1在细胞质中的比例也达到了90%左右,这与从RNA水平及蛋白水平鉴定核质分离的结果一致。在其他种类的细胞(BEAS-2B细胞、GT1-7细胞)及组织(小鼠心脏、肝脏、大脑)应用该方法结果显示:细胞核组分中质标志基因ND1含量比293T细胞的有所增加,但仍可以实现核质分离鉴定。结论:本研究所建立的核质分离质控方法可以实现从DNA水平进行核质分离的鉴定,该方法更加经济、快捷。     

Glycogen content and glucose-6-phosphatase activity in the tissues of the adult frog and tadpole

Studies have been made on the content of glycogen and the activity of glucose-6-phosphatase in tissues of adult frog Rana temporaria (liver, brain, pia mater, n. ischiadicus, fast and slow muscles) and tadpoles (liver, tail muscles). It was found that the enzymic activity is somewhat higher in the liver of tadpoles than that in the liver of adult frogs, being low in the tail muscles. Pia mater and n. ischiadicus exhibit higher activity of glucose-6-phosphatase as compared to the liver. In tadpoles, glycogen content of the liver increases from the 40th stage of metamorphosis and decreases in the tail muscles to the 42nd-50th stages. Glycogen content in tissues other than liver of adult frogs is higher than in similar tissues of mammals.     

The characteristics of nitrogen metabolism in the tissues of rabbits with ammonium toxicosis]

Disturbances of nitrogen metabolism under acute ammonium toxicosis have been studied in tissues of rabbit. A sharp increase of the ammonium content in the blood and tissues of the liver and kidneys is accompanied by an increase in the glutamine and glutamate level in all tissues. The level of urea nitrogen in the blood of rabbits increases. The activity of phosphate-independent and phosphate-activated glutaminase also increases in tissues of the liver and kidneys, while arginase activity decreases as compared with the control, which is connected with fall of the ATP level under hyperammonemia. A nomograph method of representation of the redox state has been used.     

Peculiarities of urea distribution in tissues of rats of different age

The content of urea was studied in protein-free filtrates of the liver kidneys, skeletal muscles, myocardium, spleen, brain tissues and blood serum as well as in urine of 1, 3, 6, 12 and 24-month rats. It is shown that at the age of 6 months the content of urine in most tissues under study is significantly decreased (by 42-73%), at the age of 12 months in the spleen and at the age of 24 months in the brain tissues as compared to the one-month animals. The level of urine decrease in the liver and brain tissues of 24-month animals is less pronounced than in other tissues, that corresponds to age peculiarities of their protein metabolism. A decrease of blood consumption per weight unit and a relative increase in the amount of nitrogen excreted with urine are observed with ageing. The arginase activity in the liver decreases essentially only in 3-month animals. A conclusion is drawn that peculiarities of food consumption and the character of changes in the urea content in tissues and urine are adaptation manifestation of an age decrease in the intensity of nitrogen metabolism and protein demand of the organism.     

Changes in subfraction composition of chicken lysine-rich histone during ontogenesis]

Lysine-rich histone isolated from different chicken tissues was separated electrophoretically into 4-5 subfractions. The subfrations reffered to as 1, 2, 3, and 4, occur in each the tissue studied, erythrocyte lysine-rich histone containing an additional subfraction 1a. F1 histone from mitotically active tissues (intestinal mucosa, thymus, testes) has a higher content of subfraction 2, while the same histones from mototically inactive tissues (liver, heart, brain) contain an elevated amount of subfraction 3. F1 histone isolated from liver, brain and heart of 21-day embryo has much more of subfraction 2, than the same histone of adult animal. During the chicken development from hatching till maturation the content of subfraction 2 in these organs decreases, and the content of subfraction 3 increases. The rate of this change in liver corresponds to the rate of DNA synthesis. In F1 histone of erythrocytes the content of subfraction 4 falls down during the post hatching ontogenesis.     

Activation of carboxylation as a factor in correcting the metabolic disorder in diabetes mellitus

The paper deals with studies of the effect of the 10-day complex therapy with carbostimulin application on the content of the tricarboxylic cycle and glycolysis metabolites, total lipids, cholesterol, triglycerides in blood and potassium and sodium in blood serum of patients with diabetes mellitus. It is established that under the effect of carbostimulin the content of oxaloacetate, lactate and alpha-glycerophosphate becomes normal, the lactate/pyruvate ratio elevated in diabetes decreases, which evidences for intensification of reduction processes in the cytoplasm.     

The time-course of the effects of ethanol on the redox and phosphorylation states of rat liver

1. The time-course of the effects of ethanol administration on the metabolite concentrations, redox states and phosphorylation state was studied in the freeze-clamped liver of starved rats. The response was found to vary with the time after ethanol administration. 2. Administration of ethanol caused an immediate decrease in the [NAD(+)]/[NADH] ratio of both cytoplasm and mitochondria, which persisted over the 30min studied. 3. The free cytoplasmic [NADP(+)]/[NADPH] ratio in liver decreases immediately after ethanol administration but returns nearly to control values after 15min. 4. The cytoplasmic [ATP]/[ADP][HPO(4) (2-)] ratio is elevated 15min after ethanol administration in the starved rat. 5. The rapid and large changes in most metabolite concentrations measured appeared to result from the maintenance of near-equilibrium in a wide interlinked network. 6. Differences between fed and starved rats 15min after ethanol administration were slight.     

The redox state of free nicotinamide-adenine dinucleotide in the cytoplasm and mitochondria of rat liver

1. The concentrations of the oxidized and reduced substrates of the lactate-, beta-hydroxybutyrate- and glutamate-dehydrogenase systems were measured in rat livers freeze-clamped as soon as possible after death. The substrates of these dehydrogenases are likely to be in equilibrium with free NAD(+) and NADH, and the ratio of the free dinucleotides can be calculated from the measured concentrations of the substrates and the equilibrium constants (Holzer, Schultz & Lynen, 1956; Bücher & Klingenberg, 1958). The lactate-dehydrogenase system reflects the [NAD(+)]/[NADH] ratio in the cytoplasm, the beta-hydroxybutyrate dehydrogenase that in the mitochondrial cristae and the glutamate dehydrogenase that in the mitochondrial matrix. 2. The equilibrium constants of lactate dehydrogenase (EC 1.1.1.27), beta-hydroxybutyrate dehydrogenase (EC 1.1.1.30) and malate dehydrogenase (EC 1.1.1.37) were redetermined for near-physiological conditions (38 degrees ; I0.25). 3. The mean [NAD(+)]/[NADH] ratio of rat-liver cytoplasm was calculated as 725 (pH7.0) in well-fed rats, 528 in starved rats and 208 in alloxan-diabetic rats. 4. The [NAD(+)]/[NADH] ratio for the mitochondrial matrix and cristae gave virtually identical values in the same metabolic state. This indicates that beta-hydroxybutyrate dehydrogenase and glutamate dehydrogenase share a common pool of dinucleotide. 5. The mean [NAD(+)]/[NADH] ratio within the liver mitochondria of well-fed rats was about 8. It fell to about 5 in starvation and rose to about 10 in alloxan-diabetes. 6. The [NAD(+)]/[NADH] ratios of cytoplasm and mitochondria are thus greatly different and do not necessarily move in parallel when the metabolic state of the liver changes. 7. The ratios found for the free dinucleotides differ greatly from those recorded for the total dinucleotides because much more NADH than NAD(+) is protein-bound. 8. The bearing of these findings on various problems, including the following, is discussed: the number of NAD(+)-NADH pools in liver cells; the applicability of the method to tissues other than liver; the transhydrogenase activity of glutamate dehydrogenase; the physiological significance of the difference of the redox states of mitochondria and cytoplasm; aspects of the regulation of the redox state of cell compartments; the steady-state concentration of mitochondrial oxaloacetate; the relations between the redox state of cell compartments and ketosis.     

A morphologic and morphometric study of the mitochondria in several hepatoma cell lines and in isolated hepatocytes.

Some characteristics of the mitochondria of hepatocytes and of three hepatoma cell lines have been compared. By means of stereologic analysis of electron micrographs of cross-sections through cells the volume of mitochondria per unit volume of cell cytoplasm and the surface areas of the mitochondrial envelope and cristae membranes have been measured. The relative mitochondrial volume in the cytoplasm decreases with increasing growth rate but the surface area of outer and cristae membranes per unit volume of mitochondria is not altered. The internal organization of hepatoma mitochondria, however, differs distinctly from that of normal liver mitochondria as evident from electron micrographs; the hepatoma cells contain mitochondria in which parallel cristae appear to cross the whole mitochondrial profile unlike the irregular, short cristae seen in normal liver mitochondria. Furthermore, in the fast-growing hepatoma cells the mitochondrial matrix appears less dense than in the hepatocyte. Hepatoma cells contain less organized rough endoplasmic reticulum than normal liver cells and the spatial relationship of the mitochondria to the rough cisternae, seen in the hepatocyte, is absent in the fast-growing hepatoma cell lines. It is concluded that hepatoma cells have fewer mitochondria than normal liver cells, but that the organelles have a normal content of inner membranes.     

Prothymosin alpha and parathymosin: mRNA and polypeptide levels in rodent tissues

Blot hybridization analyses have established the presence of mRNAs for prothymosin alpha (ProT alpha) and for parathymosin (ParaT) in rat and mouse lung, liver, kidney, and brain, confirming the biosynthesis of these peptides in nonlymphoid tissues. In these tissues the levels of mRNAs paralleled the content of the polypeptides, determined with specific radioimmunoassays. The mRNA levels also confirmed the reciprocal relation between the two polypeptides; ProT alpha and its mRNA were found in highest concentrations in spleen and thymus, followed by lung, kidney, and brain, with lowest concentrations in liver. On the other hand, liver contained highest concentrations of ParaT and the mRNA for ParaT, with lowest levels present in spleen and thymus. In comparison to tissues from young (6-8 week) mice, older (18 month) mice contained lower concentrations (20-40%) of both polypeptides, with qualitatively similar decreases in mRNA content.     

Radiation-induced lipid peroxidation, a fish diet and modulation of the effects by vitamin E

Data are presented in this paper on the effect of vitamin E on rats given a fish diet after whole-body gamma-irradiation. The content of lipid peroxidation products in rat plasma, brain and liver and also the content of vitamin E have been investigated. Irradiation increases lipid peroxidation in the studied tissues and decreases vitamin E content. This process is aggravated by the fish diet. Vitamin E given in addition to fish diet helps the organism to stabilize the antioxidant homeostasis at a qualitatively different level.     

Effect of cestodes on the tissue lipid composition of the burbot and stickleback

The effect of cestodes of the genus Diphyllobothrium on the lipoid contents of tissues of freshwater fishes has been studied. The level of general lipids in the liver of burbot and stickleback infected with plerocercoids of D. latum and D. vogeli, respectively, decreases. It occurs in general on account of decrease in contents of structural lipids, phospholipids, while the contents of triacylglycerines, cholesterol and its ethers change but negligibly. Simultaneous parasitism of plerocercoids of D. latum and Triaenophorus nodulosus in the liver of burbot causes especially great changes in the lipoid metabolism. Certain changes in the ratio of some phospholipid fractions take place in infected tissues: the decrease in the level of phosphatidylcholine and the parallel rise in the fraction of lysophosphatidylcholine. The ratio between other phospholipids virtually does not change.     

Changes in levels of cyclic AMP and cyclic GMP in various tissues of the rat induced by cold acclimation

In the rat, the effects of cold acclimation on the content of cyclic AMP and cyclic GMP were studied in various tissues concerned with increased heat production: brown and white adipose tissue, liver, heart, diaphragm, lungs, adrenals, thyroid. Significant cold-induced variations were observed only in those tissues in which the lipid metabolism is enhanced by cold (adipose tissues and liver). In these tissues, decrease in the cAMP/cGMP ratio indicates a role of cGMP in the regulation of the increased lipid metabolism.     

Taurine distribution in rat tissues during development.

1. Taurine levels have been determined in eight rat organs. 2. During postnatal growth the taurine content in retina, heart, small intestine, spleen and lung increases with advancing age, although adult values are not reached at the same time. 3. In contrast the taurine content decreases with age in brain cortex, liver and kidney. 4. The taurine in subcellular fractions of adult, 20-day-old and 5-day-old rat tissues exists predominantly in the cytosol of the cell. Taurine content in particulate fractions shows marked variations during development in the different organs. 5. Taurine distribution in the subcellular fractions suggests that some of the cellular taurine in the tissues is not freely mobile in cytosol.     

Measurements of protein carbonyls, ortho- and meta-tyrosine and oxidative phosphorylation complex activity in mitochondria from young and old rats.

Mitochondrial bioenergetic function is often reported to decline with age and the accumulation of oxidative damage is thought to contribute. However, there are considerable uncertainties about the amount and significance of mitochondrial oxidative damage in aging. We hypothesized that, as radical production in mitochondria is greater than the rest of the cell, protein oxidative damage should accumulate more in mitochondria than the cytoplasm, and that this relative accumulation should increase with age. To test these hypotheses we measured the accumulation of three markers of protein oxidative damage in liver, brain, and heart from young and old rats. Ortho- and meta-tyrosine levels in protein hydrolysates were measured by a gas chromatography/mass spectrometry assay, and protein carbonyl content was determined by ELISA. Using these assays we found no evidence for increased protein oxidative damage in mitochondria relative to the cytosol. Most increases found in protein oxidative damage on aging were modest for all three tissues and there was no consistent pattern of increased oxidative damage in mitochondrial proteins on aging. Mitochondrial oxidative phosphorylation complex activities were also assessed revealing 39-42% decreases in F0F1--ATP synthase activity in liver and heart on aging, but not in other oxidative phosphorylation complexes. These findings have implications for the contribution of mitochondrial oxidative damage and dysfunction to aging.