PREPARATION AND PROPERTIES OF A MITOCHONDRIAL FRACTION FROM WHEAT STEM RUST UREDOSPORES

1964 ◽  
Vol 42 (5) ◽  
pp. 497-505 ◽  
Author(s):  
P. G. Williams ◽  
G. A. Ledingham
Keyword(s):  
Serum Albumin ◽  
Extraction Procedure ◽  
Krebs Cycle ◽  
Mitochondrial Fraction ◽  
Glass Beads ◽  
Endogenous Respiration ◽  
Wheat Stem Rust ◽  
Succinate Oxidation ◽  
Glass Homogenizer ◽  
Grinding Time

Ungerminated uredospores of Puccinia graminis f. sp. tritici Erikss. & Henn. race 15B were disrupted in a buffered sucrose serum albumin solution, using a "Teflon" pestle homogenizer and glass beads 80 μ in diameter. A particulate fraction was sedimented between 2000 and 30,000 × g and its oxidation of Krebs' cycle acids measured manometrically. Endogenous respiration of washed preparations was low or negligible. Oxygen uptake was observed with succinate (40 to 60 μl/hour mg protein), α-ketoglutarate, malate, citrate, and isocitrate (10 to 30 μl/hour mg protein) but not with fumarate or pyruvate. Succinate oxidation was sensitive to heat, cyanide, and malonate.Aspects of the extraction procedure were examined for effects on yield and activity. Serum albumin in the grinding medium, very low grinding speed, and short grinding time favored high activity per mg protein. Glass beads increased the yield with the "Teflon" pestle while an all-glass homogenizer gave preparations of low activity.Oxidatively active particle suspensions reduced Janus green B and were shown by electron microscopy to consist of vesicles and some mitochondria.

RADIOIMMUNOASSAYS OF 3,5,3′-TRIIODO-L-THYRONINE WITH AND WITHOUT A PRIOR EXTRACTION STEP

1975 ◽  
Vol 80 (1) ◽  
pp. 58-69 ◽  
Author(s):  
A. Burger ◽  
C. Sakoloff ◽  
V. Staeheli ◽  
M. B. Vallotton ◽  
S. H. Ingbar
Keyword(s):  
Human Serum Albumin ◽  
Serum Albumin ◽  
Human Serum ◽  
Normal Values ◽  
Covalent Binding ◽  
Extraction Procedure ◽  
Mixed Population ◽  
Coupling Agents ◽  
Extraction Step ◽  
The Mean

ABSTRACT Two radioimmunoassays for triiodothyronine (T3) are described, one of which includes an extraction step, while the other does not. To raise antibodies, two carrier proteins and different coupling agents were used, namely haemocyanin and diazotized benzidine or human serum albumin and carbodiimide. In the case of T3 coupled to haemocyanin by diazotized benzidine, evidence of covalent binding of the hapten to the protein was obtained. In the case of T3 coupled to human serum albumin, little evidence of covalent linkage was available. Nevertheless immunization was successful in both cases. The radioimmunoassay in unextracted serum was highly reproducible and precise (intra-assay variability 5.2% inter-assay variability 8.1%). Normal values were determined which clearly indicate a fall in the serum T3 concentration with increasing age. In men the fall occurs in the fifth decade. In women the T3 starts to fall only after 70 years of age. In 31 cases of hyperthyroidism the serum T3 concentration ranged from 2.26 to 10.4 ng T3/ml. In 10 cases of hypothyroidism the values ranged from 0 to 0.8 ng T3/ml. The radioimmunoassay using an extraction procedure was less extensively used since it was found to be less reproducible (intra-assay variability 7.5%, inter-assay 12.25%). The normal values were determined with a mixed population aged 20–50. The mean ± 2 sd was 0.9 ± 0.36 ng T3/ml (n=52). In 17 cases of hypothyroidism the values ranged from 0 to 0.6 ng T3/ml and in 22 cases of hyperthyroidism from 2 to 14.4 ng T3/ml.

scholarly journals The Effect of Narcotics on the Endogenous Respiration and Succinate Oxidation in Oyster Muscle

1948 ◽  
Vol 27 (2) ◽  
pp. 504-512
Author(s):  
George Frederick Humphrey
Keyword(s):  
Salicylic Acid ◽  
Oxygen Consumption ◽  
Benzoic Acid ◽  
Succinic Acid ◽  
Chloral Hydrate ◽  
Endogenous Respiration ◽  
Succinate Oxidation ◽  
Adductor Muscles

The action of the following compounds on homogenates of the adductor muscles of Saxostrea commercialis was studied: urethane, phenyl urethane, chloral hydrate, salicylic acid, morphine, caffeine, barbitone, benzoic acid, salicylamide and benzamide.It was found that, except for 0·05 M urethane, the endogenous oxygen consumption was reduced by all these compounds when present in concentrations ranging from o·001 to 0·05M. The oxidation of succinic acid was partially inhibited by these substances.These findings are discussed in relation to similar studies on other tissues.

scholarly journals Some aspects of carnitine metabolism in avocado (Persea americana) (Short Communication)

1973 ◽  
Vol 134 (2) ◽  
pp. 655-658 ◽  
Author(s):  
R. A. Panter ◽  
J. B. Mudd
Keyword(s):  
Short Communication ◽  
Higher Plants ◽  
Glyoxylate Cycle ◽  
Mitochondrial Fraction ◽  
Persea Americana ◽  
Succinate Oxidation ◽  
Two Systems ◽  
Carnitine Metabolism ◽  
The Glyoxylate Cycle

The fact that palmitoyl-l-carnitine is oxidized by avocado mitochondria at a rate comparable with that of succinate oxidation suggests that there are at least two systems for β-oxidation in higher plants. The carnitine-associated system is located in a mitochondrial fraction, whereas the glyoxylate-cycle-associated system is located in the glyoxysomes.

scholarly journals Alteration of Pseudomonas aeruginosa respiration by 4-(1-adamantyl)-phenol derivative

Biologija ◽  
2018 ◽  
Vol 64 (3) ◽  
Author(s):  
Daria M. Dudikova ◽  
Nina O. Vrynchanu ◽  
Valentyna I. Nosar
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Oxygen Consumption ◽  
Oxidative Phosphorylation ◽  
Respiratory Chain ◽  
Structural Integrity ◽  
Respiratory Control ◽  
Endogenous Respiration ◽  
Phenol Derivative ◽  
Succinate Oxidation ◽  
Intact Cells

Derivatives of 4-(1-adamantyl)-phenol are a promising class of antimicrobials affecting the structural integrity and functions of the bacterial cell membrane. The functioning of Pseudomonas aeruginosa respiratory chain and related system of oxidative phosphorylation was investigated before and after treatment with a derivative of 4-(1-adamantyl)-phenol (compound KVM-97). Oxygen consumption was measured polarographically with a Clark-type oxygen electrode. KVM-97 was tested at 0.5× and 1.0× MIC (minimum inhibitory concentration). Specific substrates of the respiratory chain (either 3.0 mM glutamate with 2.0 mM malonate or 3.0 mM succinate with 5.0 μM rotenone) were used. All reactions were stimulated by addition of ADP (0.2 mmol). It was found that at tested concentrations, KVM-97 inhibited the endogenous respiration and substrate oxidation in P. aeruginosa cells. The inhibiting effect was dose-dependent and more pronounced with succinate oxidation rather than glutamate oxidation. The respiratory control index value (RCI) in compound-treated cells was in average 1.5 times lower compared to the intact cells. The decrease in the RCI was related to changing the oxygen uptake rates in state 3 and state 4, which indicate the uncoupling of respiration and oxidative phosphorylation. The data obtained showed that 4-(1-adamantyl)-phenol derivative inhibits oxygen consumption and has uncoupling effects in P. aeruginosa cells.

scholarly journals Metabolism of free and esterified cholesterol by Leydig-cell tumour mitochondria

1973 ◽  
Vol 134 (2) ◽  
pp. 415-424 ◽  
Author(s):  
William R. Moyle ◽  
Robert L. Jungas ◽  
Roy O. Greep
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Leydig Cell ◽  
Bovine Serum ◽  
Incubation Temperature ◽  
Mitochondrial Fraction ◽  
Second Phase ◽  
Leydig Cell Tumour ◽  
Two Phases

1. Experiments were designed to localize intracellularly the enzymes and sterol substrates required for steroidogenesis in Leydig-cell tumours. Subcellular fractions were prepared by differential centrifugation of tumour homogenates. Both free and esterified cholesterol were associated primarily with the fractions sedimenting at 1400gav. and the lipid layer floating on the surface of the isolation tubes; they were not found in the mitochondria, where the conversion of cholesterol into pregnenolone occurred. 2. Hydrolysis of esterified cholesterol was required before it could be oxidized to pregnenolone. 3. An enzyme capable of hydrolysing cholesterol esters was located external to the mitochondria. 4. Mitochondria were subfractionated by allowing them to swell in 0.02m-phosphate buffer (pH7.2) and separating the inner and outer membranes by sedimentation in sucrose gradients. The outer membrane, identified by its content of monoamine oxidase, contained most of the cholesterol associated with the mitochondria. The inner membrane, identified by its content of succinate dehydrogenase, contained the cholesterol side-chain-cleaving enzyme and very little cholesterol. 5. Accumulation of sterols by the mitochondria was studied by incubating this fraction with labelled free and esterified cholesterol suspended in lipid-free bovine serum albumin. Two phases of cholesterol accumulation were observed. The first phase, requiring 10–15min, was independent of the incubation temperature, and was inhibited by the presence of bovine serum albumin in the incubation medium. The second phase of accumulation was independent of the serum albumin content of the medium but was inhibited by low incubation temperature. 6. Esterified cholesterol was not accumulated by the mitochondria after the initial rapid binding phase. 7. The findings suggest that cholesterol was not rapidly accumulated by the mitochondrial fraction in vitro and that mechanisms may be required to facilitate cholesterol transport into mitochondria in intact tumour cells during the periods in which steroidogenesis is stimulated maximally.

scholarly journals Technetium-99m-Labeled Autologous Serum Albumin: A Personal-Exclusive Source of Serum Component

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Yuh-Feng Wang ◽  
Yi-Chun Chen ◽  
Dian-Kun Li ◽  
Mei-Hua Chuang
Keyword(s):  
Nuclear Medicine ◽  
Serum Albumin ◽  
Safety Concern ◽  
Extraction Procedure ◽  
Autologous Serum ◽  
Transmitted Infection ◽  
Technetium 99M ◽  
Transfusion Transmitted Infection ◽  
Serum Component ◽  
Liver And Kidney

Technetium-99m human serum albumin (99mTc-HSA) is an important radiopharmaceutical required in nuclear medicine studies. However, the risk of transfusion-transmitted infection remains a major safety concern. Autopreparation of serum component acquired from patient provides a “personal-exclusive” source for radiolabeling. This paper is to evaluate the practicality of on-site elusion and subsequent radiolabeling efficacy for serum albumin. Results showed that the autologous elute contained more albumin fraction than serum without extraction procedure. Good radiochemical purity and stability were demonstrated after radiolabeling. Biodistribution study showed that labeled albumin accumulated immediately in the lung, liver, and kidney. It was cleared steadily and excreted in the urine. The biologic half-life was defined, and all samples passed the pyrogenicity and sterility tests. In conclusion, autoalbumin could be extracted and radiolabeled properly in a nuclear medicine setting. Moreover, the risk of transfusion-transmitted infection associated with nonautologous, multisource99mTc-HSA agents can be reduced.

scholarly journals Activity of liver mitochondrial NAD+-dependent dehydrogenases of the krebs cycle in rats with acetaminophen-induced hepatitis developed under conditions of alimentary protein deficiency

2016 ◽  
Vol 62 (2) ◽  
pp. 169-172 ◽  
Author(s):  
O.N. Voloshchuk ◽  
G.P. Kopylchuk
Keyword(s):  
Malate Dehydrogenase ◽  
Isocitrate Dehydrogenase ◽  
Control Level ◽  
Krebs Cycle ◽  
Fold Increase ◽  
Mitochondrial Fraction ◽  
Protein Deprivation ◽  
Drug Induced ◽  
Alimentary Protein Deficiency ◽  
Ketoglutarate Dehydrogenase

Activity of isocitrate dehydrogenase, a-ketoglutarate dehydrogenase, malate dehydrogenase, and the NAD+/NADН ratio were studied in the liver mitochondrial fraction of rats with toxic hepatitis induced by acetaminophen under conditions of alimentary protein deprivation. Acetaminophen-induced hepatitis was characterized by a decrease of isocitrate dehydrogenase, a-ketoglutarate dehydrogenase and malate dehydrogenase activities, while the mitochondrial NAD+/NADН ratio remained at the control level. Modeling of acetaminophen-induced hepatitis in rats with alimentary protein caused a more pronounced decrease in the activity of NAD+-dependent dehydrogenases studied and a 2.2-fold increase of the mitochondrial NAD+/NADН ratio. This suggests that alimentary protein deprivation potentiated drug-induced liver damage

Phosphatidic acid synthesis in the heart. 1. Effect of age and species difference in the mitochondrial and microsomal synthesis

1977 ◽  
Vol 55 (4) ◽  
pp. 308-314 ◽  
Author(s):  
K. J. Kako ◽  
G. Zaror-Behrens ◽  
S. D. Peckett
Keyword(s):  
Bovine Serum Albumin ◽  
Phosphatidic Acid ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Mitochondrial Fraction ◽  
Subcellular Fractions ◽  
Newborn Rats ◽  
Adult Rats ◽  
Rat Hearts ◽  
Weanling Rats

Rates of syntheses of monoacyl- and diacyl-glycerol 3-P (phosphate) were determined in the mitochondrial and microsomal fractions prepared from hearts of rats and rabbits, to compare characteristics of the acylation reactions by the two subcellular fractions. The assays were carried out with the subcellular fractions prepared from (i) hearts of hyperthyroid animals, and (ii) hearts of newborn and weanling rats. In addition, the effect of an addition of bovine serum albumin in the assay system was examined. (1) Administration of thyroid hormones increased the acyltransferase activity in rabbit hearts but not that in rat hearts. (2) Mitochondrial and microsomal fractions of hearts of newborn rats acylated glycerol 3-P at a rate 1.3–4 times greater than those of adult rats. The rate of acylation by the mitochondrial fraction of weanling rats was also high, but the rate of microsomal acylation was slightly lower than that of adult rats. By contrast, in newborn rats, diacylglycerol 3-P formation by the liver microsomes was not greater than that of the adults, although its formation by the newborn liver mitochondria was greater. (3) The accumulation of monoacylglycerol 3-P during the assay was accelerated by the addition of increasing amounts of bovine serum albumin. Therefore, the monoacylglycerol 3-P formation was less than 10% of the diacylglycerol 3-P formation in the assay containing no albumin and with the rat subcellular fractions, whereas nearly four times more monoacyl- than diacyl-glycerol 3-P was synthesized in the presence of 20 mg albumin. (4) The ratio of monoacyl- to diacyl-glycerol 3-P formation by the mitochondrial fraction was greater than that of microsomal fraction at any concentration of albumin in both rats and rabbits. At an equal albumin concentration in the assay, relatively more diacyl- than monoacyl-glycerol 3-P was formed in the mitochondrial fraction of newborn rat hearts as compared with adult hearts. (5) In conclusion, our data concerning the age and species differences in acyltransferase activities support a view that the mitochondrial fraction of both rat and rabbit hearts, in addition to the microsomal enzymes, is capable of catalyzing the de novo synthesis of phosphatidic acid.

Sign in / Sign up

Export Citation Format

Share Document