L-carnitine Concentrations Research Articles

Influence of intravenous L-carnitine administration in sheep preceding an oral urea drench.

Two experiments were conducted to investigate the effect of i.v. administration of L-carnitine on selected metabolites in sheep and to determine the feasibility of using L-carnitine to ameliorate the deleterious effects of hyperammonemia in sheep. In Exp. 1, i.v. L-carnitine solutions were administered at three levels in a replicated Latin square: 0 (CONT), 6.36 (CAR 1), and 12.72 (CAR 2) mmol L-carnitine/kg x (75) BW using Suffolk ewes (n = 6; average BW 75+/-3 kg). Plasma L-carnitine concentration was increased (P<.05) by treatment (51.9 vs 102.3, and 96.4 micromol/L in CONT, CAR 1, and CAR 2, respectively). Plasma glucose concentration was elevated (P<.05) in CAR 2 and CAR 1. Plasma NEFA concentration was highest (P<.05) in CAR 2. Area under the response curve for glucose was greater (P<.02) in CAR 2. In Exp. 2, Suffolk ewes (n = 16; average BW 48+/-2 kg) were used in a randomized complete block design with a 2x2 factorial treatment arrangement to determine the effects of i.v. L-carnitine administration during an oral urea load test (OULT). L-Carnitine (0 and 6.36 mmol/kg x (75) BW) was administered i.v. at 30 min, and an oral urea drench (50% wt/vol; 0 and 300 mg/kg BW) was administered at 60 min. Plasma L-carnitine was increased (P<.0001) by i.v. L-carnitine. Plasma ammonia N was highest (P<.0001) in the UREA treatment compared with the CONT, CARN, and CARN + UREA treatments (148 vs 95, 101, and 108 micromol/L, respectively). Intravenous L-carnitine administration influenced plasma glucose and NEFA concentrations in sheep and, when administered 30 min preceding an OULT, prevented the development of subclinical hyperammonemia in sheep.

Secondary abnormality of carnitine biosynthesis results from carnitine reabsorptional system defect in juvenile visceral steatosis mice

We characterized the l-carnitine transport system which is defective in the kidney of juvenile visceral steatosis (JVS) mice by using kidney slices and carnitine-related compounds, and evaluated the influence of the transport defect on the biosynthetic pathway of carnitine. The JVS mouse transport system defect, calculated as the difference in the transport activity between control and JVS mice, was simulated in control by γ-butyrobetaine ( γ-BB) and acetyl l-carnitine. γ-BB hydroxylase activity in the liver of JVS mice was double that of control mice, but the hepatic level of γ-BB in JVS mice was lower than in control mice, suggesting that the conversion of γ-BB to carnitine is not activated in the liver of JVS mice. JVS mice showed higher fractional excretions not only of l-carnitine but also of γ-BB and acetyl l-carnitine than control mice, indicating disturbed reabsorption of γ-BB and acetyl l-carnitine. The disturbed reabsorption of γ-BB in JVS mice is consistent with the fact that the amount of urinary γ-BB in JVS mice was four times that of control. The sum of the concentrations of l-carnitine, acetyl l-carnitine and γ-BB in the urine of JVS mice was not significantly different from that of the control, suggesting no remarkable increase of biosynthesis of γ-BB and carnitine in JVS mice. All these findings suggest that the carnitine transport system plays a role in the transport of γ-BB and that carnitine deficiency is aggravated by the disturbed reabsorption of γ-BB in the kidney.

The application of the differential pH method to the biochemical evaluation of seminal plasma

Objective: To apply the differential pH method to the biochemical evaluation of seminal plasma. Design and Methods: Fructose, citric acid and free L-carnitine concentration in human seminal plasma were measured by the differential pH method. These are routinely taken as markers of seminal vesicle, prostate and epididymal function, respectively. The pH differential method was then compared with standard spectrophotometry. Results: The linearity, recovery and imprecision calculation for each assay were found to be reliable and the method correlated well with the reference spectrophotometric technique. In addition, the pH differential method showed certain advantages compared to spectrophotometry: a) speed of procedure (40 to 120 sec/assay); b) ability to measure the analytes even in small volumes of seminal plasma (10 μL to 50 μL) without deproteinization or dilution, thus eliminating manual operations. Conclusions: The differential pH method is an innovative approach to the biochemical assessment of seminal plasma.

Effect of dehydroepiandrosterone sulfate on carnitine acetyl transferase activity and l-carnitine levels in oophorectomized rats

Alteration in energy metabolism of postmenopausal women might be related to the reduction of dehydroepiandrosterone sulfate (DHEAS). DHEA and DHEAS decline with age, leveling at their nadir near menopause. DHEA and DHEAS modulate fatty acid metabolism by regulating carnitine acyltransferases and CoA. The purpose of this study was to determine whether dietary supplementation with DHEAS would also increase tissue l-carnitine levels, carnitine acetyltransferase (CAT) activity and mitochondrial respiration in oophorectomized rats. Plasma l-carnitine levels rose following oophorectomy in all groups ( P<0.0001). Supplementation with DHEAS was not associated with further elevation of plasma l-carnitine levels, but with increased hepatic total and free l-carnitine ( P=0.021 and P<0.0001, respectively) and cardiac total l-carnitine concentrations ( P=0.045). In addition, DHEAS supplementation increased both hepatic and cardiac CAT activities ( P<0.0001 and P=0.05 respectively). CAT activity positively correlated with the total and free carnitine levels in both liver and heart ( r=0.764, r=0.785 and r=0.700, r=0.519, respectively). Liver mitochondrial respiratory control ratio, ADP:O ratio and oxygen uptake were similar in both control and supplemented groups. These results demonstrate that in oophorectomized rats, dietary DHEAS supplementation increases the liver and heart l-carnitine levels and CAT activities. In conclusion, DHEAS may modulate l-carnitine level and CAT activity in estrogen deficient rats. The potential role of DHEAS in the regulation of fatty acid oxidation in postmenopausal women is worthy of investigation.

Effects of pivalic acid and sodium pivalate on L-carnitine concentrations in the cauda epididymidis and on male fertility in the hamster.

In this study, administration of pivalic acid or its sodium salt was found to decrease the L-carnitine concentration in the epididymal lumen of the hamster; it also tested whether this decrease affected sperm cell motility, chromatin structure, or fertilizing capacity. Provision of pivalic acid or its sodium salt (20 mM or 40 mM) in the drinking water of mature male golden hamsters for 30 days reduced (by 72%, 75%, and 83% in three experiments) the L-carnitine concentration of the cauda epididymidis but did not inhibit sperm chromatin condensation, as assessed by flow cytometry. The treatments did not alter the location of motile sperm in the epididymidis nor did they appreciably affect the motility of sperm obtained from the distal cauda epididymidis. The numbers and percentage of ova that reached the 2-cell stage 36-40 h after uterine insemination with spermatozoa from control and treated hamsters served as a measure of sperm fertility. Treatment with pivalic acid or sodium pivalate did not render male hamsters infertile although it appeared to reduce the fertilizing ability of their spermatozoa. These results suggest that the high concentration of L-carnitine present in the lumen of the cauda epididymidis is not required for maturation of sperm chromatin or development of sperm motility.

Effect of short and long-term treatments by a low level of dietary L-carnitine on parameters related to fatty acid oxidation in Wistar rat

This study was designed to examine whether short- and long-term treatments by a low level of dietary l-carnitine are capable of altering enzyme activities related to fatty acid oxidation in normal Wistar rats. Under controlled feeding, ten days of treatment changed neither body weights nor liver and gastrocnemius weights, but succeeded in reducing the weight of peri-epididymal adipose tissues. Triacylglycerol contents were lowered in liver and ketone body concentrations were found slightly more elevated in blood. In the liver, mitochondrial carnitine palmitoyltransferase I (CPT I) exhibited a slightly higher specific activity and a lower sensitivity to malonyl-CoA inhibition, while peroxisomal fatty acid oxidizing system (PFAOS) was found to be less active. Carnitine supplied for one month reduced the mass of the periepididymal fat tissue, but not those of the other studied organs, and produced a slight but non-significant gain in body weight after ten days of treatment. In the liver, CPTI characteristics were comparable in control and treated groups, while PFAOS activity was less in rats receiving carnitine. Data show that l-carnitine at a low level in the diet exerted two paradoxical effects before and after ten days of treatment. Results are discussed in regard to fatty acid oxidation in mitochondria and peroxisomes, and to the possible altered acyl-CoA/acylcarnitine ratio with increased concentrations of l-carnitine in the liver.

Effects of added L-carnitine, acetyl-CoA and CoA on peroxisomal β-oxidation of [U- 14C]hexadecanoate by isolated peroxisomal fractions

(1) During peroxisomal β-oxidation of [U- 14C]hexadecanoate, at concentrations higher than 100 ,μM, long-chain 3-oxoacyl-CoA-esters and 3-oxobutyryl-CoA accumulate. Only 3-oxobutyryl-CoA accumulates at a low concentration of [U- 14C]hexadecanoate. Accumulation of long chain 3-oxoacyl-CoA esters is most extensive when the supply of CoA can be considered limiting for β-oxidation. (2) Added acetyl-CoA was found to inhibit peroxisomal β-oxidation. This inhibition was not significantly relieved by added L-carnitine and carnitine acetyltransferase (EC 2.3.17). (3) Added L-carnitine, at concentrations below 0.2 mM, was found to stimulate peroxisomal β-oxidation of [U- 14C]hexadecanoate by up to 20%, causing the conversion of acetyl-CoA into acetylcarnitine. Higher concentrations of L-carnitine were progressively inhibitory to f3-oxidation. This effect was specific for L-carnitine as both D-carnitine and aminocarnitine neither caused stimulation at low concentrations, nor inhibition at higher concentrations. Added L-carnitine caused accumulation of acylcarnitines of chain-lengths ranging from 4 to 16 carbon-atoms. The inhibition observed with higher concentrations of added L-carnitine is likely due to conversion of [U- 14C]hexadecanoate into [U- 14C]hexadecanoylcarnitine. (4) Low concentrations of added hexadecanoylcarnitine was shown to inhibit peroxisomal β-oxidation by about 15%, while added acetylcarnitine did not inhibit at concentrations up to 100 μM. (5) These data are interpreted to indicate significant control being exerted on flux at the stage of thiolysis either directly by means of CoA availability, or indirectly by means of the rate of acetyl-CoA generation.

Inhibition of flagellar motility of fowl spermatozoa by L‐carnitine: Its relationship with respiration and phosphorylation of axonemal proteins

The action of carnitine in regulating fowl sperm motility was investigated. As the concentration of L-carnitine was increased (0-20 mM), the motility of intact and demembranated fowl spermatozoa was reduced at 30 degrees C. Even the presence of 1 mM CaCl2 before the addition of 10 mM carnitine could not prevent the inhibition of motility at 30 degrees C and 40 degrees C. However, motility was restored by reducing the concentrations of carnitine. Carnitine also inhibited the oxygen consumption and ATP concentrations of intact spermatozoa, and caused a reduction in intracellular free Ca2+ concentrations. Phosphorylation of a 50 kDa protein and dephosphorylation of 24 kDa and 30 kDa proteins of demembranated spermatozoa were observed after the addition of carnitine. In contrast, the flagellar ATPase activity of crude dynein extract was not affected by the addition of carnitine. These results suggest that inhibitory effect of carnitine for motility may be directly on the axonemal phosphoproteins, but not directly on the dynein ATPase activity. The physiological role of carnitine for fowl spermatozoa in the ductus deferens is discussed.

Dietary Carnitine Did Not Influence Performance and Carcass Composition of Broiler Chickens and Young Turkeys Fed Low- or High-Fat Diets

Two experiments were conducted to determine the influence of supplemental dietary carnitine on performance and carcass composition of young turkeys and broiler chickens. Experiments 1 and 2 were done with poults to 21 d of age and broilers to 45 d of age, respectively. Three dietary concentrations of L-carnitine (0, 50, or 100 mg/kg) were fed in a factorial arrangement with two concentrations of animal-vegetable fat (A-V fat), 2.25 or 8% in Experiment 1 and 1 or 5% in Experiment 2. L-Carnitine did not affect (P > .05) BW gain or feed efficiency in either experiment, irrespective of concentration of dietary fat. Similarly, proximate composition of 21-d-old poults and 45-d-old broilers was not changed by L-carnitine. Increasing levels of fat supplementation improved feed efficiency of poults and improved 45-d BW and feed efficiency of broilers. Carcass fat of poults and broilers was increased (P ≤ .05) by supplemental fat at the expense of carcass water and protein.

Uptake and release of free L-carnitine by boar epididymal spermatozoa in vitro and subsequent acetylation rate.

In the male reproductive tract, very high concentrations (mmol l-1) of free L-carnitine and acetyl-L-carnitine are found in the epididymides, seminal plasma and spermatozoa. It has been reported that the uptake of free L-carnitine by spermatozoa might be related to the epididymal maturation of the sperm membrane, since a greater uptake was found by caput than by cauda spermatozoa in vitro. However, the free L-carnitine concentrations estimated inside the gametes were never greater than those of the surrounding medium. In this study, we investigated the mechanism of transport of free L-carnitine and its ester acetyl-L-carnitine, through the plasma membrane of mature and immature epididymal boar spermatozoa. In vitro, we found a passive diffusion of both compounds to the spermatozoa, whatever the maturation stage. The spermatozoa might progress in the epididymal lumen and accumulate high amounts of free L-carnitine. The active uptake of free L-carnitine occurs only across epididymal mucosa. These results are in agreement with those reported on cells of other organs that exchange pharmacological free L-carnitine concentrations (mmol l-1) by a passive mechanism through the plasma membrane. The acetylation of high amounts of free L-carnitine inside the spermatozoa was found only in caudal spermatozoa. This result suggests that oxidative metabolism (producing acetyl CoA) might be more active in mature cells. The acetyl-L-carnitine added to the incubation medium of boar spermatozoa was hydrolysed. Enzymatic activity of the sperm membrane is low and this may partially explain the low concentrations of acetyl-L-carnitine found in the caudal epididymal plasma.

Hypoxia-induced coronary flow changes in the perfused rat heart: Effects of high L-carnitine concentrations

1. 1. Hemodynamic effects of physiological (0.04 and 0.07 mM) and high (8, 15 and 25 mM) L-carnitine (LC) concentrations were tested on the normally oxygenated and hypoxic perfused rat heart. 2. 2. No effect was detected on aerobic hearts, whereas a dose-dependent rise in coronary flow (CF) during both the hyperemic and constrictive phases was observed in hypoxic hearts with 8, 15 and 25 mM LC. This action was apparently unrelated to a resting tension (RT) improvement, which was observed only with 25 mM LC. 3. 3. When 11 mM glucose was replaced by 11 mM mannitol in the perfusion buffer, LC effects on the hyperemic phase were abolished; however, 25 mM LC resulted in CF-values still significantly higher than those detected without the drug, though RT was similar in these glucose-free groups. 4. 4. It may be concluded that LC is ineffective on the perfused rat heart in aerobic conditions, whereas high LC concentrations can enhance CF only during hypoxia, these effects being independent of heart function improvements and partly unrelated to glucose presence.

Modulation of cytokine production by carnitine

The ability of carnitine congeners to modulate cytokine production by human peripheral blood mononuclear cells (PBMC) was investigated. Modulation of cytokine production by PBMC of young (30 years of age or younger) and old (70 years of age or older) normal donors was first compared. The PBMC were collected over Ficoll–Hypaque and incubated in the presence of various concentrations of acetyl L-carnitine for 24 h. Subsequently the supernatants were collected and examined for cytokine production. The presence of cytokines in tissue culture supernatants was examined by ELISA. The cytokines measured included IL-1α, IL-1β, IL-2, IL-4, IL-6, TNFα, GM–CSF, and IFNγ. The results showed that at 50 μg/ml of acetyl L-carnitine the most significant response was obtained for TNFα. In this regard four of five young donors responded, but only one of five old donors responded. More recently these studies were expanded to examine the ability of L-carnitine to modulate cytokine production at higher doses, 200 and 400 μg/ml, in young donors. The results of these studies showed that in addition to TNFα, significant production of IL-1β and IL-6 was observed. These preliminary studies provide evidence that carnitine may modulate immune functions through the production of selected cytokines.

Kinetic analysis of l-carnitine uptake by the choroid plexus

Choroid plexuses from the lateral (LVCP) and fourth ventricles (FVCP) of rats or rabbits were incubated in artificial cerebrospinal fluid (CSF) containing 1 microM [14C]L-carnitine with various concentrations of L-carnitine ranging from 0.01 mM to 1.0 mM. The time course of 1 microM [14C]L-carnitine uptake by the choroid plexus indicated that it increased linearly for the first 15 min. Steady-state levels were reached by 30 min with tissue concentrations more than 20 (FVCP)- to 30 (LVCP)-fold greater than the concentration in the medium. The uptake of [14C]L-carnitine was increased with increasing concentrations of the substrate in the medium and this uptake followed Michaelis-Menten kinetics. The uptake by the rat choroid plexus took place against a concentration gradient via a saturable process and kinetic analysis revealed Km of 32 microM (LVCP) and 34 microM (FVCP) and Vmax of 21 (LVCP) and 17 nmol/ml/min (FVCP), respectively. Ouabain inhibited the uptake by 51% (FVCP) and 48% (LVCP) and hypothermia (0 degrees C) produced inhibition by 97% (FVCP) and 96% (LVCP), respectively. However, the uptake of L-carnitine was not sensitive to probenecid or tyrosine, which indicates the presence of an independent carrier for L-carnitine in the choroid plexus. Similar results were obtained with the rabbit choroid plexus.

Opposite variations of two epididymal components and blood plasma testosterone in two breeds of rams

1. Testicular volume (T Vol), blood plasma testosterone (T) concentration, seminal plasma α-glucosidase (α-G) specific activity, l-carnitine ( l-C) concentration as well as semen characteristics were compared in eight Finnish Landrace (F) and eight Suffolk (S) rams throughout 21 months. 2. Only T Vol and T exhibited a typical seasonal variation in both breeds, whereas l-C, α-G and live sperm output presented a seasonal profile only in S rams. 3. l-C and α-G variations were opposite to those of T in S rams, while they fluctuated in F rams throughout the entire experiment, as did live sperm output. 4. Only the number of ejaculates and T were significantly higher in F rams (3.50 ± 0.08 in 5 min and 7.62 ± 0.40 ng/ml) than in S rams (2.30 ± 0.05 in 5 min and 5.5$¯0.30 ng/ml); these two characteristics might therefore be considered as two indexes of sexual activity in rams. 5. By contrast, among all characteristics measured, only α-G was significantly higher in S rams than in F rams (1.33 ± 0.04 vs 0.77 ± 0.03 mU/mg proteins); this result, as well as seasonal α-G profile present in only S rams, allowed us to conclude that α-G might be considered as an additional index of seasonal reproduction in rams.

The medium-chain carnitine acyltransferase activity associated with rat liver microsomes is malonyl-CoA sensitive

The data presented herein show that both rough and smooth endoplasmic reticulum contain a medium-chain/long-chain carnitine acyltransferase, designated as COT, that is strongly inhibited by malonyl-CoA. The average percentage inhibition by 17 μ m malonyl-CoA for 25 preparations is 87.4 ± 11.7, with nine preparations showing 100% inhibition; the concentrations of decanoyl-CoA and l-carnitine were 17 μ m and 1.7 m m, respectively. The concentration of malonyl-CoA required for 50% inhibition is 5.3 μ m. The microsomal medium-chain/long-chain carnitine acyltransferase is also strongly inhibited by etomoxiryl-CoA, with 0.6 μ m etomoxiryl-CoA producing 50% inhibition. Although palmitoyl-CoA is a substrate at low concentrations, the enzyme is strongly inhibited by high concentrations of palmitoyl-CoA; 50% inhibition is produced by 11 μ m palmitoyl-CoA. The microsomal medium-chain/long-chain carnitine acyltransferase is stable to freezing at −70 °C, but it is labile in Triton X-100 and octylglucoside. The inhibition by palmitoyl-CoA and the approximate 200-fold higher I 50 for etomoxiryl-CoA clearly distinguish this enzyme from the outer form of mitochondrial carnitine palmitoyltransferase. The microsomal medium-chain/long-chain carnitine acyltransferase is not inhibited by antibody prepared against mitochondrial carnitine palmitoyltransferase, and it is only slightly inhibited by antibody prepared against peroxisomal carnitine octanoyltransferase. When purified peroxisomal enzyme is mixed with equal amounts of microsomal activity and the mixture is incubated with the antibody prepared against the peroxisomal enzyme, the amount of carnitine octanoyltransferase precipitated is equal to all of the peroxisomal carnitine octanoyltransferase plus a small amount of the microsomal activity. This demonstrates that the microsomal enzyme is antigenically different than either of the other liver carnitine acyltransferases that show medium-chain/long-chain transferase activity. These results indicate that medium-chain and long-chain acyl-CoA conversion to acylcarnitines by microsomes in the cytosolic compartment is also modulated by malonyl-CoA.

EFFECT OF GENETIC DIABETES AND ALCOHOL ON TISSUE CARNITINE AND INOSITOL CONCENTRATIONS IN MICE

Concentrations of acid-soluble L-carnitine and inositol were determined in heart, kidney, muscle, pancreas, liver, brain and blood of genetically diabetic obese db/db and their nondiabetic control C57BL/6J (CBL) mice. Results were compared to a group of diabetic and CBL mice fed ethanol (ETOH) 4 g/kg daily for 58-64 days. In CBL and db/db mice, heart muscle was found to have the greatest and brain the least content of carnitine. Diabetes caused a significant decrease in hepatic concentration of carnitine but did not affect carnitine concentration of heart, kidney, skeletal muscle, brain and pancreas. ETOH intake had no effect on carnitine content of any of the tissues studied. Free inositol content was highest in brain and lowest in skeletal muscle of CBL and db/db mice; diabetes or ETOH intake did not affect tissue inositol content. Except for liver, neither diabetes nor ETOH intake affects tissue carnitine or inositol concentration.

Influence of ejaculation frequency on semen characteristics in chimpanzees (Pan troglodytes).

Semen samples were obtained by masturbation from 6 chimpanzees and the spontaneously liquefied fraction and the remaining coagulum were studied separately. When semen was collected once or twice a week, large intra-individual variations were observed for all measures. The liquefied fraction represented 26.5 +/- 3.2% (weighted mean +/- s.d.) of the total ejaculate but contained 51.3 +/- 3.8% of all emitted spermatozoa. Fructose concentration was higher in the coagulum than in the liquefied fraction (29.3 +/- 3.0 mumol/ml vs 12.0 +/- 2.7 mumol/ml, P less than 0.001) whereas acid phosphatase was less concentrated in the coagulum than in the liquefied fraction (3.5 +/- 0.3 x 10(3) IU/ml vs 13.0 +/- 0.9 x 10(3) IU/ml, P less than 0.001). L-Carnitine and citrate concentrations did not differ between the two fractions of the ejaculate. When semen collection was repeated every hour for 5 h, the ejaculate volume increased from 2.6 +/- 0.7 to 4.7 +/- 0.6 ml (P less than 0.001), whereas total sperm count decreased from 1278 +/- 872 x 10(6) to 587 +/- 329 x 10(6) (P less than 0.05) between the 1st and the 6th ejaculate. In the spontaneously liquefied fraction, the sperm count decreased from 984 to 369 x 10(6). The 6 successive ejaculates gave a total of 20.2 +/- 7.6 ml and 4278 +/- 2884 x 10(6) spermatozoa. The increase of the ejaculate volume was essentially due to an increase of the volume of the coagulum which closely correlated with total amount of fructose (from seminal vesicles) (r = 0.913, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Regulation of fatty acid oxidation in rat brain mitochondria: Inhibition of high rates of palmitate oxidation by ADP

Regulation of oxidation of [1- 14C]palmitate in rat brain mitochondria has been investigated in purified mitochondria of nonsynaptic origin prepared by use of a Ficoll/sucrose density gradient. The mitochondrial preparation contained considerable Mg 2+-ATPase activity, but was virtually free of contamination with nonmitochondrial fractions. Palmitate oxidation was inhibited by increasing the concentration of ATP in the assay system to near-physiological levels (2 m m), and the inhibition at 2 or 4 m m ATP was analyzed by comparing it with palmitate oxidation at near-maximal rates with low levels of ATP (0.5 or 1 m m). Inhibition was increased by the addition of ADP or by increasing the concentration of Mg 2+ in the assay system, whereas inhibition was decreased by decreasing the concentration of mitochondrial protein or L-carnitine in the assay system. Increasing CoA concentration also had a deinhibitory effect. With 0.5 or 1 m m ATP, however, neither inhibition by added ADP nor protein concentration-dependent inhibition was observed, and the rate of oxidation was saturated with increasing concentrations of Mg 2+, l-carnitine, or CoA. These results indicated that ADP was involved in the inhibition of high rates of palmitate oxidation in the presence of sufficient ATP and l-carnitine. The inhibitory effect of increasing the concentration of mitochondrial protein could be explained by the enhanced amounts of ADP present in the preparation; similarly, increased concentrations of Mg 2+ would provide higher levels of ADP by stimulating the Mg 2+-ATPase reaction. We discuss the possibility that the transport of ADP across the inner membrane of brain mitochondria is coupled to the inhibition of palmitate oxidation.

Cellular and biochemical characteristics of semen obtained from pubertal chimpanzees by masturbation.

Semen characteristics were studied in 6 wild-born chimpanzees with dental ages ranging approximately from 6 to 12 years. The animals formed 2 groups, early pubertal (EP, N = 3, 6-9 years) and late pubertal (LP, N = 3, 11-12 years). Mean body weight, testicular volume and serum androgen concentration were significantly lower in Group EP (32.2 +/- 1.6 kg, 34.0 +/- 7.7 cm3, 2.1 +/- 0.1 ng/ml) than in Group LP (55.7 +/- 5.7 kg, P less than 0.01; 100.5 +/- 11.9 cm3, P less than 0.01; 3.6 +/- 0.7 ng/ml, P less than 0.05). Ejaculates were obtained by masturbation in all subjects. The mean ejaculate volume was lower in Group EP (0.56 +/- 0.20 ml) than in Group LP (3.77 +/- 0.73 ml, P less than 0.01). In Group EP, 2 animals were azoospermic while the third produced semen with means of 57.1 x 10(6) spermatozoa per ml, 20% motility and 40% vitality. These values were low when compared with the mean values of Group LP (376 x 10(6) spermatozoa per ml, 67% motility and 78% vitality). Mean total sperm count was correlated with testicular volume (r = 0.84) and serum androgen concentration (r = 0.96). The mean concentrations of L-carnitine, fructose, citrate and acid phosphatase for the two groups were not significantly different; but, related to the differences in ejaculate volumes, their total amounts in total ejaculate were lower in Group EP than in Group LP. These results suggest that, in chimpanzees, mechanisms of seminal plasma production and ejaculation are functional early in the reproductive life and that the emission of spermatozoa occurs later.

Gamma-Butyrobetaine hydroxylase and the protective role of glutathione peroxidase.

The selenoenzyme glutathione peroxidase in the presence of GSH effectively replaced catalase in the in vitro assay for gamma-butyrobetaine hydroxylase. Quantitatively, glutathione peroxidase was an order of magnitude more efficient than catalase, with maximal activity at less than 0.1 microM glutathione peroxidase in a standard reaction. Glutathione peroxidase prevented the loss of gamma-butyrobetaine hydroxylase during preliminary incubation with ferrous ions but without other substrates as well as in the course of the reaction. Regardless of whether glutathione peroxidase or catalase was present in the assay, the ascorbate concentrations needed to achieve half-maximal rates were similar (about 1 mM). Phosphate stimulated the rate of L-carnitine synthesis. Ferrous ion saturation indicated a pronounced effect of phosphate on the maximal velocity of the enzyme-catalyzed reaction, but its mechanism of action remains to be elucidated. Based on the subcellular distribution of gamma-butyrobetaine hydroxylase, catalase, and glutathione peroxidase, the role of glutathione peroxidase assumes importance. However, initial studies indicated that the assayable activity of liver gamma-butyrobetaine hydroxylase and L-carnitine concentrations in liver, blood plasma, and muscle were not significantly altered in selenium-deficient rats.