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[Comparison of estradiol benzoate and dihydrotestosterone propionate with testosterone propionate on the induction of male-like sexual behavior in female sheep ovariectomized as adults].

Daily I/M injections of 200 microgram of oestradiol benzoate induce in female Sheep, ovariectomized as adults, male-like sexual reactions similar to what is obtained with 10 mg testosterone propionate per day. Simultaneously, a permanent female sexual receptivity is observed. Oestradiol benzoate at the dose of 20 microgram/day, and dihydrotestosterone propionate (10 mg/day) are both totally ineffective. The result support the hypothesis of the necessity of the aromatization of testosterone for the action on sexual behaviour.

Animals

Gluconeogenesis in ruminants: propionic acid production from a high-grain diet fed to cattle.

Effects of amount of feed intake on in vivo remen propionate production and the reproducibility of measurements of propionate production rates were investigated in five dairy steers weighing 142 to 228 kg. Each steer received 275 g of an 80%-grain diet every 2 hours, and three of the five steers later received 500 g/2 hours. Rumen propionate pool sizes and production rates were determined at both intakes by administering a single dose of [1-14C] propionate via rumen fistula and observing decreasing specific activity of rumen propionate for 4 hours. A linear regression equation was calculated from the natural logs of decreasing specific activity. Propionate pool sizes averaged 30.6 and 47.4 g, and production rates averaged 579 and 1032 g/day at feed intakes of 275 and 500 g/2 hours. Thus, propionate production changed in almost direct proportion to feed intake. Acetate to propionate ratios were about 3:1 in rumen fluid. Considerable variability in estimates of propionate pool sizes and production rates was found both within and among steers. The results indicate propionate production is nearly equivalent to blood glucose turnover determined previously; all propionate produced in the rumen, however, is not used for glucose synthesis.

Animals

Factors determining the sequence of oxidative decarboxylation of the 2- and 4-propionate substituents of coproporphyrinogen III by coproporphyrinogen oxidase in rat liver.

Coproporphyrinogen oxidase (EC 1.3.3.3) catalyses the oxidative decarboxylation of the 2- and 4-propionate substituents of coproporphyrinogen III to form protoporphyrinogen IX. A 4-propionate-substituted porphyrinogen, harderoporphyrinogen, which is also a substrate for coproporphyrinogen oxidase, is formed during the reaction. Synthetic [(14)C]coproporphyrinogens III, specifically labelled in the carboxyl carbon atoms of either the 2- or 4-propionate substituents, were used to measure the rate of decarboxylation of each substituent by rat liver coproporphyrinogen oxidase. The experimental results, together with the recognition that in all known substrates of coproporphyrinogen oxidase only those propionate groups flanked by a specific arrangement of substituents are decarboxylated, indicate that the 4-propionate group of coproporphyrinogen III cannot be attacked until the 2-propionate group has been decarboxylated. Production of (14)CO(2) from the substrate labelled in the 2-propionate group therefore measures the formation of harderoporphyrinogen, whereas (14)CO(2) from the 4-propionate-labelled substrate measures protoporphyrinogen IX formation. The rate of harderoporphyrinogen formation is about twice that of protoporphyrinogen, and this ratio is unchanged by varying the concentration of coproporphyrinogen III or by competitive inhibition of the enzyme. When coproporphyrinogen III is present in an excess, two fractions of harderoporphyrinogen can be distinguished. One accumulates during the reaction, and the other, which is destined to become protoporphyrinogen IX, does not equilibrate with added harderoporphyrinogen. It is suggested that both decarboxylations take place at the same active centre, which becomes temporarily inaccessible to coproporphyrinogen III and added harderoporphyrinogen, and that the molecule rotates after the first decarboxylation to allow the second to take place.

Animals

Rumen propionate and blood glucose kinetics in growing cattle fed isoenergetic diets.

The relationship between rumen propionate production and blood glucose kinetics was examined in four rumen-fistulated Holstein steers fed isoenergetic amounts of 80/20 (G) and 30/70 (R) grain/chopped alfalfa hay diets at 2-hour intervals. Single-injection rumen propionate and blood glucose kinetics were determined in consecutive 4-hour periods by using [6-3H]glucose intravenously then [1-14C]propionate intraruminally. Rumen propionate specific activity was determined after isolation and quantitation by high-pressure liquid chromatography. Average rumen propionate production rates and pool sizes were 441 g/day and 32.0 g when diet R was fed but increased to 510 g/day (P less than 0.05) and 36.5 g (P less than 0.10), respectively, when diet G was fed. Propionate production as related to digestible energy (DE) intake, averaged 0.56 mole/Mcal DE for R and 0.64 mole/Mcal DE for G. Rumen propionate turnover times were similar for both diets. Despite differences in propionate availability, there were no significant dietary differences in glucose kinetic parameters. Average glucose pool sizes and irreversible losses were 27.0 g and 585 g/day for R and 27.0 g and 582 g/day for G.

Animals

[Role of propionic acid, yeasts and ethyl alcohol in regulating the activity of H-factor in Drosophila simulans].

Effects of yeast, propionic acid and ethanol on the activity of H-factor, which sharply increases the frequency of somatic recombination in X-chromosomes of dorsal prothoracal disc cells in Drosophila simulans are studied. The frequency of yellow and singed mosaic spots in heterozygous yw ++/++sn1vHD. melanogaster females, inherited H-factor from the father (the stock sn1v) is estimated. The results of the varience analysis have shown that yeast and propionic acid regulate the activity of H-factor in cells of dorsal prothoracal disc, the interaction of yeast and propionic acid being also observed. Yeasts (or some unknown product of their metabolism) are the activator of H-factor; thus, when larvae eat much yeast, the frequency of yellow and singed mosaic spots in humeral region becomes high. A decrease of mosaic spot frequency under the increase of propionic acid content in nutrition medium is a result of the inhibitory effect of propionic acid on the yeast growth, but not of the direct repression of H-factor activity. So, propionic acid may be considered as a regulator of the second order. Ethanol does not activate H-factor. Changes in the content of yeast and propionic acid in nutrition medium do not affect the frequency of yellow and singed mosaic spots in other regions of D. simulans body, except humeral.

Animals

Applicability of an enzymatic quantitation of methylmalonic, propionic, and acetic acids in normal and megaloblastic states.

A rapid sensitive spectrophotometric assay for the measurement of methylmalonic and propionic acids in urine is described. The assay is based upon the quantitation of propionic acid using acetyl coenzyme A synthetase isolated from baker's yeast. This enzyme is highly specific for acetate and propionate, and acetate interference is eliminated by conversion to citrate. Methylmalonic acid was assayed by converting it to propionate by heat decarboxylation and then measuring the propionate increment over the endogenous amount in the noncarboxylated sample. Studies of urine obtained from normal subjects (by isolation, partial purification, and then assay by the isotope dilution technique) demonstrated urinary excretion of less than 1 mg of propionic acid and 1-5 mg of methylmalonic acid per day. In 22 consecutive patients with documented vitamin B12 deficiency, methylmalonic acid excretion in excess of 30 mg/24 hr was found. In four other patients, with only neurologic involvement methylmalonic aciduria aided in identifying B12 deficiency as an etiologic factor. Methylmalonic acid excretion was measured by direct assay of an aliquot of urine, requiring neither a valine load nor special extraction procedures. Propionic aciduria was variably increased in B12 deficiency and did not correlate either with the severity of the deficit or degree of methylmalonic aciduria. The assay was performed on urine, but it is potentially applicable to tissue extracts. In addition, this assay method can be utilized for the quantification of urine acetate levels as well.

Acetates

Participation of propionate in cholesterol biosynthesis by rat liver.

Incorporation of (214C) propionate into cholesterol was demonstrated using rat liver slices and homogenates. The incorporation of (214C) propionate was greater than that of (214C) acetate. Using the same liver homogenate preparation (214C) succinate and (214C) pyruvate were incorporated into cholesterol to a lesser extent than (214C) acetate and (214C) propionate. Addition of unlabeled acetate failed to dilute the incorporation of (214C) propionate. Incorporation of the 2 and 3 carbon atoms pf propionate were equal; little incorporation of the 1 carbon atom was demonstrable. These results indicate that propionate is an excellent source of 2 carbon units for isoprenoid biosynthesis; the intermediary pathway does not involve a common acetate pool nor can these results be satisfactorily explained by citric acid intermediary metabolism.

Acetates

A relationship between the molar proportion of propionic acid and the clearance rate of the liquid phase in the rumen of the sheep.

1. Four rumen-cannulated sheep were given a forage mixture (F) of chopped hay-ground, pelleted, dried grass (92:8, w/w) and two concentrate mixtures (C and S) of ground barley-ground hay-flaked maize (46:24:30 and 56:24:20, by wt respectively) in twenty-four hourly meals each day. Each of the diets was offered in successive periods of 16 d to give a feeding sequence F-S-C-S for one pair of sheep and C-S-F-S for the other pair. 2. The average composition (mol/100 mol) of the mixture of short-chain fatty acids, acetic, propionic and butyric, in the rumen was respectively 70-1, 18-5 and 7-5 with diet F, and 55-8, 24-8 and 13-6 with diet C. With diet S, the pattern of fermentation varied both between animals and in the same animal for different periods having either 'high' (28-39 mol/100 mol) or 'low' (16-21 mol/100 mol) proportions of propionic acid. On average when diet S followed diet F there was less propionic acid in the fermentation mixture than when diet S followed diet C (59-3 acetic, 22-2 propionic and 14-1 butyric as compared with 52-7, 29-4 and 13-1 respectively) but this trend was not significant and there was evidence of interactions between the feeding sequences and the individual sheep. 3. The mean concentrations of ammonia, sodium, potassium and chloride were similar for all diets but the pH and concentrations of calcium, magnesium and phosphorus tended to be higher and the buffering capacity lower for diet F than for diets C or S. In animals receiving diet S there was no relationship between the concentrations of minerals, the pH or buffering capacity and the pattern of fermentation except for ammonia, the concentration of which was high when the molar proportion of propionic acid was low. 4. Rumen volume, outflow rate and clearance rate, determined using polyethylene glycol, were higher for diet F than for diets C and S but within each diet, particularly for diet S, values varied considerably between sheep and between periods. 5. There was evidence of an interrelationship between the molar proportion of propionic acid in the fermentation products and the clearance rate, which indicated that the clearance rate may be an important factor influencing the pattern of fermentation in the rumen.

Acetates

Biotransformation of 3-(2',4',5'-triethoxybenzoyl) propionic acid, a new biliary smooth muscle relaxant with choleretic activity, in rats.

1. By the combined use of deuterium labelling and gas chromatographic-mass spectrometric analysis, the metabolite pattern of 3-[2',4',5'-triethoxybenzoyl) propionic acid (triethoxybenzoylpropionic acid), a new biliary smooth muscle relaxant with choleretic activity, has been determined in the rat. 2. The metabolites excreted in urine and/or bile were isolated and characterized as follows: the parent drug, 3-(2',5'-diethoxy-4'-hydroxybenzoyl)propionic acid (metabolite II),3-(2'-ethoxy-4'-hydroxy-5'-methoxybenzoyl)propionic acid or 3-(2'-ethoxy-5'-hydroxy-4'-methoxybenzoyl)propionic acid (III), 3-(2',4'-diethoxy-5'-hydroxybenzoyl)propionic acid (V) and 3-[2',4'-diethoxy-5'-(2-hydroxyethoxy)benzoyl]propionic acid (VI). 3. The primary route of biotransformation of triethoxybenzoylpropionic acid in the rat was either O-de-ethylation at the C-4' and C-5' positions or beta-hydroxylation of the ethoxy group at the C-5' position in the parent molecule. 4. All these excreted products except metabolite VI were eliminated both unconjugated and in conjugates (probably glucuronides and/or sulphates). 5. The rat excreted mainly metabolites V and VI in the urine and parent drug, metabolites V and VI in the bile.

Animals

Factors influencing rumen fermentation: effect of hydrogen on formation of propionate.

The effect of hydrogen on fermentation of lactate, pyruvate, fumarate, and succinate by resting rumen microorganisms has been investigated. Under an atmosphere of nitrogen, lactate was fermented to yield acetate as the major product (85 to 100 mole %) and propionate (0 to 17 mole %) and butyrate (0 to 3%) as secondary products. Under hydrogen, there was increased formation of both propionate and total volatile fatty acids. The amount of propionate increased 4 to 8 times and total volatile fatty acids 2.5 to 3.2 times. Propionate formation was proportional to the hydrogen concentration and reached a maximum at a partial pressure of hydrogen of .2 N/m2. With [2-carbon-14] lactate, propionate was formed via the dicarboxylic acid pathway under both nitrogen or hydrogen. Hydrogen did not affect significantly the fermentation of pyruvate or succinate. With fumarate under hydrogen, propionate and total volatile fatty acids increased 6.8 and 2 times while acetate was unchanged. The mechanism by which hydrogen exerts these effects is discussed in relation to the role of methanogenesis in the rumen.

Animals

On the effects of propionate and other short-chain fatty acids on sodium transport by the toad bladder.

1. Propionate and other unbranched short-chain fatty acids, butyrate, pentanoate, hexanoate and octanoate were found to both stimulate and inhibit active sodium transport by the toad bladder, as measured by the short-circuit current (s.c.c.). 2. Stimulation alone followed addition of low concentrations of fatty acids (0.1-1.0 mM) to either the serosal or mucosal bathing medium; stimulation was also seen after an initial period of inhibition in response to higher concentrations (approx. 5 mM) of some compounds. 3. Inhibition alone followed addition of high concentrations (5-20 mM) of these compounds. The duration and magnitude of the inhibition varied with increasing concentration and chain length of the fatty acid, and was greater following mucosal addition than serosal addition. 4. The inhibitory effect of mucosal propionate increased with decreasing pH of the mucosal bathing medium. 5. Inhibition by the fatty acids was completely reversed upon removing the compound from the bathing medium, and stimulation characteristically followed. 6. In studies designed to evaluate the role of metabolism of the fatty acids in their mucosal inhibitory effects it was found that 14-c-labelled propionate, when added to the mucosal surface of the bladder, was converted to 14-CO2, and mucosal succinate and alpha-oxoglutaric acid at 20 mM inhibited the s.c.c. slightly. However, malonate did not interfere with inhibition by mucosal propionate and two non-metabolizable acids, dimethylpropionate and benzoate, induced inhibition (and no stimulation) of the s.c.c. 7. In the presence of an inhibitory concentration of fatty acid, the ability of the bladder to respond to added pyruvate was reduced in proportion to the reduction in the level of the s.c.c., whereas the natriferic response to vasopressin was largely intact. 8. We conclude that stimulation of sodium transport by propionate and other short-chain fatty acids is due to metabolism of the compounds and provision of energy to the sodium transport mechanism. The basis of the inhibition appears complex. It may in part depend on metabolism of the fatty acids and/or uncoupling of oxidative phosphorylation, with resultant reduction in net ATP production for the sodium transport mechanism. However, the inhibition may also be caused in part by a direct effect on the mucosal entry of sodium into the transporting epithelial cells.

Animals

Effects of monensin on in vivo rumen propionate production and blood glucose kinetics in cattle.

Four rumen-fistulated steers (154 to 253 kg), fed two different diets in succession, were used to determine effects of monensin on rumen propionate production rates and blood glucose kinetics as determined by single-injection isotope-dilution techniques. A high-roughage and a high-grain diet, with and without 150 mg of monensin daily, were fed isoenergetically at 2-hour intervals. Monensin increased rumen propionate pool sizes from 32 to 57 g for the high-roughage diet and from 37 to 66 g for the high-grain diet and increased rumen propionate production rates from 441 to 659 g/day for the high-roughage diet and from 510 to 899 g/day for the high-grain diet. Molar percentages of rumen propionate were increased significantly by monensin in the high-grain diet. Blood glucose pool sizes were not changed significantly by either monensin or isoenergetic diets. Monensin increased irreversible losses of glucose from 582 to 677 g/day for the high-grain diet. Monensin tended to increase glucose total entry rates for both diets and to increase irreversible loss of glucose for the high-roughage diet but the differences were not significant. Thus, increases in glucose kinetics are minor in contrast to major increases of rumen propionate production caused by monensin.

Animals

Direct anabolic metabolism of three-carbon propionate to a six-carbon metabolite occurs in vivo across tissues and species.

Anabolic metabolism of carbon in mammals is mediated via the one- and two-carbon carriers S-adenosyl methionine and acetyl-coenzyme A. In contrast, anabolic metabolism of three-carbon units via propionate has not been shown to extensively occur. Mammals are primarily thought to oxidize the three-carbon short chain fatty acid propionate by shunting propionyl-CoA to succinyl-CoA for entry into the TCA cycle. Here, we found that this may not be absolute as, in mammals, one nonoxidative fate of propionyl-CoA is to condense to two three-carbon units into a six-carbon trans-2-methyl-2-pentenoyl-CoA (2M2PE-CoA). We confirmed this reaction pathway using purified protein extracts provided limited substrates and verified the product via LC-MS using a synthetic standard. In whole-body in vivo stable isotope tracing following infusion of 13C-labeled valine at steady state, 2M2PE-CoA was found to form via propionyl-CoA in multiple murine tissues, including heart, kidney, and to a lesser degree, in brown adipose tissue, liver, and tibialis anterior muscle. Using ex vivo isotope tracing, we found that 2M2PE-CoA also formed in human myocardial tissue incubated with propionate to a limited extent. While the complete enzymology of this pathway remains to be elucidated, these results confirm the in vivo existence of at least one anabolic three- to six-carbon reaction conserved in humans and mice that utilizes propionate.

Acetyl Coenzyme A

Inhibition of acetate and propionate formation upon aeration of resting cells of the anaerobic Propionibacterium shermanii: evidence of the Pasteur reaction.

When resting cell suspensions of the anaerobic P. shermanii were brought to an oxygen concentration of 0.64 mumoles/ml, acid formation was completely inhibited. The cells started to respire on the propionic acid previously accumulated furing anaerobiosis. Glucose consumption was concomitantly decreased to about 60 percent of the rate during anaerobiosis. As the viability of the cells was not affected by the transition to aerobic conditions, the changes observed upon aeration were ascribed to the regulatory properties of the Pasteur reaction. Damage inflicted by oxygen was encountered in the rapid inactivation of propionate respiration. This damage outlived the time of oxygenation, and was manifested during the subsequent anaerobiosis in the decreased activity to form propionate. This indicates that oxygen may inactivate one (or more) enzyme(s) involved in the metabolism of propionate. The viability of cells in buffer, and glucose-containing buffer, was found to be only insignificantly decreased by oxygen in the range from 0 to 500 mumoles of oxygen per g of wet cells.

Acetates

The utilization of diets containing acetate, propionate or butyrate salts by growing lambs.

1. In a comparative slaughter experiment growing lambs were given concentrate diets in which 7, 15 or 22% of the metabolizable energy (ME) provided by barley was replaced by sodium and calcium salts of acetic acid, or 22% of ME was replaced by Na and Ca salts of propionic or butyric acids. 2. The efficiency of utilization for fattening (kf) of the diets containing 0, 7, 15 or 22% of ME as acetate was 57.2, 59.6, 54.1 and 48.8 (SE +/- 1.8) respectively, the last value being significantly lower (P less than 0.001) than the first. The kf for successive increments of acetate was 90, 37 and 19% (SE +/ 3), the decrease being significant (P less than 0.001). 3. The kf value of the diets containing 22% of ME as propionate or butyrate respectively were 48.7 and 50.6 (SE +/- 1.8), both values being significantly lower than the control (P less than 0.01). The partial kf of propionate was 19 +/- 13, and of butyrate 28 +/- 13%. 4. It is concluded that the experiment provided evidence that the efficiency with which acetateis utilized for energy retentionis not constant, but varies with its contribution to ME. The experiment also provided some evidence that large amounts of propionate and butyrate may be inefficiently utilized by growing lambs, although poor utilization of high levels of volatile fatty acid (VFA) salts per se cannot be entirely excluded.

Acetates

Pathway of succinate and propionate formation in Bacteroides fragilis.

Cell suspensions of Bacteroides fragilis were allowed to ferment glucose and lactate labeled with (14)C in different positions. The fermentation products, propionate and acetate, were isolated, and the distribution of radioactivity was determined. An analysis of key enzymes of possible pathways was also made. The results of the labeling experiments showed that: (i) B. fragilis ferments glucose via the Embden-Meyerhof pathway; and (ii) there was a randomization of carbons 1, 2, and 6 of glucose during conversion to propionate, which is in accordance with propionate formation via fumarate and succinate. The enzymes 6-phosphofrucktokinase (pyrophosphate-dependent), fructose-1,6-diphosphate aldolase, phosphoenolpyruvate carboxykinase, malate dehydrogenase, fumarate reductase, and methylmalonyl-coenzyme A mutase could be demonstrated in cell extracts. Their presence supported the labeling results and suggested that propionate is formed from succinate via succinyl-, methylmalonyl-, and propionyl-coenzyme A. From the results it also is clear that CO(2) is necessary for growth because it is needed for the formation of C4 acids. There was also a randomization of carbons 1, 2, and 6 of glucose during conversion to acetate, which indicated that pyruvate kinase played a minor role in pyruvate formation from phosphoenolpyruvate. Phosphoenolpyruvate carboxykinase, oxaloacetate decarboxylase, and malic enzyme (nicotinamide adenine dinucleotide phosphate-dependent) were present in cell extracts of B. fragilis, and the results of the labeling experiments agreed with pyruvate synthesis via oxaloacetate and malate if these acids are in equilibrium with fumarate. The conversion of [2-(14)C]- and [3-(14)C]lactate to acetate was not associated with a randomization of radioactivity.

Acetates