• Asian-Australasian Journal of Animal Sciences
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• v.13 no.11
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• pp.1568-1575
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• 2000

Growing pigs (N=25; 18 kg) were used to study effects of L-carnitine and protein intake on plasma carnitine, energy and carnitine balance, and carnitine biosynthesis. Corn-soybean meal basal diets containing low or high protein (13.6% or 18%) were formulated so that protein accretion would be limited by metabolizable energy (ME). Each basal diet was supplemented with 0 or 500 mg/kg L-carnitine and limit fed to pigs for 10 d in a balance trial. Final carnitine concentration was compared with weight/age matched pigs measured on d 0 to calculate carnitine retention rates. Supplementation of carnitine increased (p<0.01) plasma free carnitine (by 250%), short-chain (by 160%) and long-chain acyl-carnitine concentrations (by 80%) irrespective of blood sampling time (p<0.01). The proportion of long-chain carnitine esters decreased by 40% (p<0.01) by carnitine supplementation; whereas, the proportion of short-chain acyl-carnitine concentration was not changed (p>0.10). All criteria of energy balance were unaffected by L-carnitine (p>0.10). Total body carnitine retention was increased by 450% over unsupplemented controls (p<0.01). Carnitine biosynthesis rates in pigs fed diets without L-carnitine were estimated at 6.71 and $10.63{\mu}mol{\cdot}kg^{-1}{\cdot}d^{-1}$ in low protein and high protein groups, respectively. In supplemented pigs, L-carnitine absorption and degradation in the intestinal tract was estimated at 30-40% and 60-70% of L-carnitine intake, respectively. High protein feeding effect did not affected plasma carnitine concentrations, carnitine biosynthesis or carnitine retention (p>0.10). We conclude that endogenous carnitine biosynthesis may be adequate to maintain sufficient tissue levels during growth, but that supplemental dietary carnitine (at 500 ppm) sufficiently increased plasma acyl-carnitine and total body carnitine.

• Journal of Microbiology and Biotechnology
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• v.7 no.5
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• pp.318-322
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• 1997

In order to isolate a microorganism having preferential degradation of D-carnitine from DL-carnitine, a bacterium assimilating D-carnitine as a sole carbon and energy source was isolated from soil by enrichment culture and partially identified as Enterobacter sp. Also, a mutant having lessened L-carnitine decomposition rates was selected with nitrosoguanidine mutagenesis, which led to decrease the specific activities of carnitine dehydrogenase (7.6-fold) and ${\beta}$-hydroxybutyrate dehydrogenase (9.5-fold) as compared to the wild strain. Meanwhile, optimal culture conditions for optical resolution of DL-carnitine were investigated. Under optimal conditions, 3.53 g/l L-carnitine was obtained from 20 g/l DL-carnitine, which corresponded to 35.3% L-carnitine yield and 97.9% optical purity.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.5
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• pp.799-805
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• 1999

This study was conducted to investigate the effects of L-carnitine with different levels of lysine on performance of pigs weaned at 21 days of age. A total of 120 pigs were allotted into a $3{\times}2$ factorial design with three different levels of lysine (1.40%, 1,60% and 1.80%) and two levels of L-carnitine (0 and 1,000 ppm). Each treatment had 4 replications with 5 pigs per replicate. Pigs of $22{\pm}1$ days (5.9 kg of body weight) were grouped into a completely randomized block design. Treatments were 1) 1.4-Crt; 1.40% of lysine with 1,000 ppm of L-carnitine, 2) 1.4-N; 1.40% of lysine without L-carnitine, 3) 1.6-Crt; 1.60% of lysine with 1,000 ppm of L-carnitine, 4) 1.6-N; 1.60% of lysine without L-carnitine, 5) 1.8-Crt; 1.80% of lysine with 1,000 ppm of L-carnitine and 6) 1.8-N; 1.80% of lysine without L-carnitine. Growth performance was optimized in pigs fed 1.6% lysine regardless of carnitine addition. For the first 7 days of the experimental period, the best ADG and F/G were found in pigs within the 1.6-Crt group. Carnitine significantly improved (p<0.05) ADG of pigs when the lysine level in the diet was 1.6%. Only in the third week carnitine had a significant influence on growth performance of pigs. A lysine-sparing effect of L-carnitine was not detected in this study. The 1.6-Crt group showed the best proximate nutrient digestibility, and the crude fat and gross energy digestibility were higher when the L-carnitine was added in the diet. Lysine level significantly affected the digestibilities of DM (p<0.001), GE (p<0.001), CP (p<0.01) and C.fat (p<0.05). Carnitine also significantly improved digestibility of nutrients. Lysine level as well as carnitine level affected the amino acids digestibility, however, in 1.8% lysine diet carnitine did not influence on amino acids digestibility. Plasma carnitine content was significant higher (p<0.05) in pigs fed L-carnitine. This indicates the increased biological availability of carnitine within the body. L-carnitine supplementation tended to improve feed utilization during the third week (p<0.10) and during the entire period (p=0.10). Lysine level significantly affected feed utilization of pigs during the third week and entire period (p<0.05). As pigs grew, the lysine requirement was reduced.

• Clinical and Experimental Pediatrics
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• v.45 no.9
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• pp.1083-1089
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• 2002

Purpose : The prevalence of obese children has recently increased. Obesity is known to be associated with complications such as hypertension, fatty liver, hyperlipidemia, and insulin resistance. L-carnitine is an essential cofactor for the transport of long chain fatty acids into mitochondria for ${\beta}$-oxidation. The purpose of this study is to measure serum free fatty acid and carnitine levels, and evaluate the role of L-carnitine as a therapeutic drug in obese children with fatty liver. Methods : Nine obese children, ranging from seven to 18 years of age, and 10 normal children were examined. Serum lipid(total cholesterol, triglyceride, HDL-cholesterol, and LDL-cholesterol) and fatty acid levels were analyzed. Serum total, free, and acyl carnitine levels were performed also by a new enzymatic cycling technique. Results : Long chain fatty acids(myristic acid, palmitoleic acid, palmitic acid, linoleic acid, oleic acid, and stearic acid)were significantly increased in obese children compared to the control group. Total, and acyl carnitine levels were significantly increased in obese children compared to the control group. Conclusion : Serum free fatty acid and carnitine levels were significantly increased in obese children with fatty liver compared to the normal control. This may suggest that L-carnitine can be used as antilipidemic agent to decrease fatty acid and lipid levels for obese children. Prospective studies will investigate serum fatty acid and carnitine levels after treatment of L-carnitine in obese children in the future.

• Clinical and Experimental Pediatrics
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• v.45 no.9
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• pp.1075-1082
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• 2002

Purpose : Obesity is known to be associated with hypertension, dyslipidemia, and fatty liver and is thought to be associated with increased levels of free fatty acids. One of the strategies for decreasing free fatty acid levels is stimulation of hepatic lipid oxidation with L-carnitine. Carnitine is an essential cofactor for transport of long-chain fatty acid into mitochondria for oxidation. This study was designed to evaluate the changes of serum fatty acids and carnitine levels after exogenous injection of L-carnitine. Methods : Sprague Dawley rats were divided into two groups. Group A was control. Group B was given intraperitoneal injection with L-carnitine(200 mg/kg) daily for two weeks. Serum lipid (total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol) and fatty acid levels were analyzed on the first day of the first and second weeks after injection of L-carnitine. Total, free, and acyl carnitine levels also were performed by a enzymatic cycling techniques at the same day intervals. Results : There was no significant difference between the two groups in total cholesterol, HDL-cholesterol, LDL-cholesterol levels before and after the administration of L-carnitine. But triglyceride levels were significantly decreased at the first week in group B compared with group A. Among free fatty acids, linoleic acid showed significant decrement(A group : $131.3{\pm}31.3mg/dL$ vs B group : $90.0{\pm}7.0mg/dL$) at the first week. Total, free, and acyl carnitine levels showed significant increments at all days intervals, but only free carnitine showed significant increments according to cumulative doses of carnitine. Conclusion : Plasma linoleic acid, a long-chain fatty acid, showed significant decrement after administration of L-carnitine in the first week. This may suggest that L-carnitine can be used as an antilipidemic agent for obese patients. A prospective study will investigate obese children in the future.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.2
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• pp.237-242
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• 2001

Colostrum deprived, newborn pigs (N=12, $1.64{\pm}0.05kg$) were used to study the renal threshold of carnitine, and effects of emulsified medium-chain triglyceride (MCT, tri-8:0) feeding on kinetics of plasma carnitine and urinary carnitine excretion. An arterial catheter was inserted through an umbilical artery, and a bladder catheter was inserted via the urachus. Piglets were oro-gastrically gavaged with one of six carnitine levels (0, 60, 120, 180, 240, $480{\mu}mol/kg\;W^{0.75}$) with (+MCT) or without medium-chain triglycerides (-MCT) in 0.9% NaCl solution. Blood was sampled into heparinized tubes at 0, 1, 2, 4, 6, 8, 14, and 20 h after gavage, and urine was collected and pooled into 1 h or 2 h composite samples to determine free- and short-chain carnitine concentrations. Plasma from the 12 newborn piglets before gavage contained $10.6{\pm}1.2{\mu}mol/L$ free carnitine and $7.2{\pm}0.6{\mu}mol/L$ acid-soluble acyl carnitine. The renal threshold for carnitine was similar between the MCT and the +MCT group (42.6 13.1 and $46.4{\pm}2.0{\mu}mol/L$, respectively), but the correlation between plasma free carnitine and urinary excretion was altered. Plasma free carnitine linearly increased with increasing carnitine dosage (-MCT group, $R^2=0.95$, p<0.001; +MCT group, $R^2=0.91$, p<0.001), but was decreased by 50% when medium-chain triglycerides were fed. The peak in plasma free carnitine concentration was depressed by medium-chain triglycerides feeding also. Therefore, the plasma and urinary short-chain/free carnitine ratio of the +MCT group was increased by 100% and 40%, respectively (p<0.01). Feeding of medium-chain triglycerides may delay plasma carnitine elevation via altering the kinetics of absorption. Similarly, the plasma and urinary short-chain/free carnitine ratio were affected by interaction between medium-chain triglycerides and time (p<0.01). The present study suggests that an oral carnitine dose over $480{\mu}mol/kg\;W^{0.75}$ may be needed to reach the free carnitine renal threshold within a short period, especially when provided together with medium-chain triglyceride.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.601-605
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• 1998

For the resolution of L-carnitine from DL-carnitine, resting cells of Enterobacter sp. NH-104, which had a higher capacity of D-carnitine decomposition, were harvested at maximal specific activity of D-carnitine decomposition of 47.05 unit/mg cell. The cells were frozen at $-80^{\circ}C$ to assess functions as enzyme sources. Optimal concentration of cells and DL-carnitine were 17 g/$\ell \; and \; 20 g/\ell$, respectively, and reaction buffer was best at 75 mM of Tris. HCl. Optimal temperature and pH were $36^{\circ}C$ and 8.2, respectively. When the reaction at optimal conditions was carried out for 14 h, the optical purity was 98.21 %, and the quantity and yield of remaining L-carnitine were 4.432 g/$\ell$ and 44.32%, respectively.

• Preventive Nutrition and Food Science
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• v.3 no.3
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• pp.251-255
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• 1998

Rat testis known to contain all of the enzymes required for carnitine biosynthesis also contains high concentration of calmodulin, a protein which may or may not contain trimethyllysine, the major substrate in carnitine biosynthesis. The purpose of this study was to investigate the levels of carnitine and the state of calmodulin N-methylation in rat testes, and to discuss the possibility of the involvement of calmodulin incarnitine biosynthesis. Nonesterified carnitine , acid soluble acyl carnitine, and acid insoluble acyl carnitine of ra tests were 273 nmole, 62nmole, and 4 nmole/g tissue, respectively. Total carnitine level was 339 nmole/g testes tissue. Calmodulin purified from rat tests was assayed for methylation potential using N-methyltransferase from the rat testes. Rat testes calmodulin showed no 3H-methyl incorporation indicating that the calmodulin was trimethylated already by endogenous calmodulin N-methyltransferase. Amino acid composition analysis revealed that the rat testes calmodulin containd one mole of trimethyllysine per mole of calmodulin. These data suggest that testes calmodulin could provide the trimethyllysine needed for the synthesis of carnitine in the rat tests.

• Journal of Nutrition and Health
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• v.36 no.7
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• pp.720-728
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• 2003

This study was conducted to evaluate changes in plasma concentration and urinary excretion of carnitine, as well as plasma lipid level and fatty acid composition, caused by short term supplementation of carnitine in humans. Ten healthy male subjects (21.2 $\pm$ 0.5 years old) received oral carnitine supplementation (4 g/day) as tablets for two weeks. Fasting blood and random urine samples were collected from each subject both prior to and at the end of carnitine supplemention program. Following the 2 weeks of carnitine supplementation, plasma total carnitine (TCNE) concentration increased 20% (85.1 $\pm$ 7.4 vs 67.3 $\pm$ 9.1 $\mu$ mol/1, p> 0.05), while urinary excretion of total carnitine increased ten times compared to the value measured prior to the supplementation (3051 $\pm$ 692 vs 278 $\pm$ 90.1 $\mu$ mol/g creatinine, p < 0.01). Non-esterified carnitine (NEC) comprised from 71 to 88% of TCNE in plasma, and from 32 to 40% of TCNE excreted in the urine. Two weeks of carnitine supplementation in healthy adults significantly elevated plasma level of acid soluble acylcarnitine (ASAC) which is esterified mostly with short chain fatty acids (21.6 $\pm$ 1.6 $\mu$ mol/l) compared to the value measured prior to the supplementation (6.4 $\pm$ 0.8 $\mu$ mol/l) (p < 0.05). Carnitine supplementation significantly increased plasma HDL-cholesterol level (p < 0.05), and decreased the atherogenic index (p < 0.05), but failed to cause any significant change in plasma levels of total cholesterol, triglyceride, and free fatty acids. Plasma triglyceride and phospholipid fatty acid compositions were not significaly affected as well by the oral supplementation of carnitine in subjects with normal range of blood lipid levels.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.8
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• pp.1263-1272
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• 1999

The effects of carnitine in diets with or without added fat (5% lard or soybean oil) were evaluated in 72 Large White ${\times}$ Landrace ${\times}$ Duroc pigs weaned at 35 days of age. Pigs were fed a 1.30% lysine corn-soybean basal diet+15% dried whey+4% fish meal with carnitine at 0 or 50 mg/kg and either 0% added fat, 5% soybean oil or 5% lard for 6 weeks in a $2{\times}3$ factorial trial (6 treatments, 3 pens per treatment, 4 pigs per pen). Addition of carnitine increased average daily gain (ADG) and average daily feed intake (ADFI) in the second two weeks of the six-week trial and overall, but had no significant effect on feed per gain (F/G). Lard alone depressed ADG (p<0.05) in the last two weeks of the trial and overall, but the ADG for pigs fed lard+carnitine was similar to the control. Lard reduced feed intake in the first two weeks of the trial (p<0.05). Carnitine reduced the percentage of pigs with poor (ADG<375 g/d) growth (15 vs 40%; p<0.05). The greater uniformity of growth was most evident in low-weaning-weight pigs in the second period (16 vs 62%, p<0.005). Addition of fat did not produce any positive effect on uniformity and had no interaction with carnitine on uniformity. Carnitine addition increased serum total carnitione and short-chain acyl-carnitine levels (p<0.05), but did not modify free carnitine levels. Serum carnitine levels were lower at weaning than at 14, 28, or 39 days after weaning (p<0.05). Carnitine increased serum protein levels on day 14 (p<0.05). Addition of fat in the form of soybean oil or lard did not improve piglet growth performance. Addition of 50 mg/kg of carnitine to the diet of weanling pigs enhanced postweaning performance.