Ornithine alpha-ketoglutarate and creatine effects on growth and plasma metabolites of nursery pigs.
(33/1042)
Four experiments were conducted to determine the effect of dietary ornithine alpha-ketoglutarate (OKG) and creatine monohydrate on growth performance and plasma metabolites of nursery pigs. In each experiment, treatments were replicated with four to five pens of four to six pigs each. Each experiment lasted from 3 to 4 wk and Phase I (1.6% Lys) and Phase II (1.3 to 1.5% Lys) diets were fed for 9 to 16 d each. In Exp. 1, pigs (4.7 kg and 15 d of age) were fed diets containing 0, .10, or .75% OKG. Daily gain during a 13-d Phase I period and ADFI during Phase I and overall (29 d) were increased (P < .10) in pigs fed .75% OKG. Gain:feed ratio was not affected (P > .10) by diet. In Exp. 2, pigs (7.1 kg and 23 d of age) were fed 0 or .50% OKG during Phase I only. During Phase I, II, and overall, ADG and ADFI were not affected (P > .10) by OKG supplementation, but gain:feed was decreased during Phase I (P < .04), Phase II (P < .08), and overall (P < .04). Plasma urea N (PUN), glucose, and NEFA concentrations were not affected (P > .10) by OKG supplementation in this experiment. In Exp. 3, pigs (5.8 kg and 20 d of age) were fed diets containing 0, .10, or .50% creatine. Creatine tended to linearly decrease ADG (P = .11) and plasma albumin (P = .12) and PUN (P < .10) concentrations in Phase II (d 12 to 26). In Exp. 4, 850 mg of OKG or 750 mg of creatine was provided daily by oral capsule to pigs 4 d before weaning to 2 d after weaning. Pigs within a litter received either no capsule or capsules containing OKG or creatine. After weaning, pigs that received no capsule before weaning received no treatment, .50% creatine, or .50% OKG in the nursery diet. Pigs that received OKG before weaning received no treatment or .50% OKG, and pigs that received creatine before weaning received no treatment or .50% creatine in the nursery diet. Pigs weighed 3.9 kg 4 d before weaning and 4.9 kg at weaning at an average age of 20 d. The OKG provided by capsule decreased ADG (P < .02) and ADFI (P < .09) during Phase II. The OKG did not affect (P > .10) plasma NEFA, glucose, or urea N concentrations. Creatine added to the nursery diet increased (P < .02) ADFI and decreased (P < .10) gain:feed during Phase II and overall. Creatine in the nursery diet also increased (P < .01) PUN, but it did not affect plasma glucose or NEFA concentrations. Creatine and OKG have variable effects on growth performance and plasma metabolites of nursery pigs. (+info)
Bioactive products of arginine in sepsis: tissue and plasma composition after LPS and iNOS blockade.
(34/1042)
Blockade or gene deletion of inducible nitric oxide synthase (iNOS) fails to fully abrogate all the sequelae leading to the high morbidity of septicemia. An increase in substrate uptake may be necessary for the increased production of nitric oxide (NO), but arginine is also a precursor for other bioactive products. Herein, we demonstrate an increase in alternate arginine products via arginine and ornithine decarboxylase in rats given lipopolysaccharide (LPS). The expression of iNOS mRNA in renal tissue was evident 60 but not 30 min post-LPS, yet a rapid decrease in blood pressure was obtained within 30 min that was completely inhibited by selective iNOS blockade. Plasma levels of arginine and ornithine decreased by at least 30% within 60 min of LPS administration, an effect not inhibited by the iNOS blocker L-N(6)(1-iminoethyl)lysine (L-NIL). Significant increases in plasma nitrates and citrulline occurred only 3-4 h post-LPS, an effect blocked by L-NIL pretreatment. The intracellular composition of organs harvested 6 h post-LPS reflected tissue-specific profiles of arginine and related metabolites. Tissue arginine concentration, normally an order of magnitude higher than in plasma, did not decrease after LPS. Pretreatment with L-NIL had a significant impact on the disposition of tissue arginine that was organ specific. These data demonstrate changes in arginine metabolism before and after de novo iNOS activity. Selective blockade of iNOS did not prevent uptake and can deregulate the production of other bioactive arginine metabolites. (+info)
Attenuation of myocardial ischemia/reperfusion injury by superinduction of inducible nitric oxide synthase.
(35/1042)
BACKGROUND: Nitric oxide (NO) has been implicated as a mediator in myocardial ischemia/reperfusion (I/R) injury, but its functional properties have been conflicting. We investigated whether NO has a protective role against I/R injury. METHODS AND RESULTS: Using endothelial NO synthase knockout (eNOS KO) mice, inducible NOS KO mice, the NO donor S-nitroso-N-acetylpenicillamine (SNAP), and the NOS inhibitor N-iminoethyl-L-ornithine (L-NIO), we performed studies of isolated perfused hearts subjected to 30 minutes of global ischemia followed by reperfusion. After 60 minutes of reperfusion, nitrite levels in the coronary effluent in the SNAP and eNOS KO groups were significantly elevated compared with other groups. Immunoblot and immunohistochemistry showed that iNOS was markedly induced in the eNOS KO hearts. Under spontaneous beating conditions during reperfusion, increased NO activity was correlated with a prevention of the hyperdynamic contractile response and enhanced myocardial protection, as evidenced by a reduction in myocardial injury and infarct size. During prolonged reperfusion, SNAP-treated hearts were able to preserve contractile functions for 180 minutes, whereas L-NIO-treated hearts showed a sustained deterioration in contractility. CONCLUSIONS: NO protects against I/R injury by preventing the hyperdynamic response of isolated perfused hearts during early reperfusion. In the eNOS KO hearts, a paradoxical increase in NO production was seen, accompanied by a superinduction of iNOS, possibly due to an adaptive mechanism. (+info)
Involvement of glutamine, arginine, and polyamines in the action of ornithine alpha-ketoglutarate on macrophage functions in stressed rats.
(36/1042)
The ability of ornithine alpha-ketoglutarate (OKG) to enhance macrophage cytotoxicity in stress situations has been described, but the mechanisms involved remain unclear. It is known that OKG administration generates glutamine (GLN), arginine (ARG), and polyamines. This study will (1) evaluate the effect of OKG on tumor necrosis factor alpha (TNF-alpha) secretion and nitric oxide (NO*) production in macrophages from glucocorticoid (DEX)-treated rats, and determine whether these effects can be reproduced by GLN or ARG supplementations, and (2) use in vivo metabolic inhibitors methionine sulfoximine (inhibitor of GLN synthetase), S-methylthiourea (inhibitor of inducible nitric oxide synthase), and difluoromethylornithine (inhibitor of ornithine decarboxylase) to assess the roles of GLN, ARG, and polyamines in OKG action. Controls received a mixture of nonessential amino acids (NEAA). GLN, ARG, and OKG all restored TNF-alpha secretion by macrophages of glucocorticoid-treated rats. The same results were obtained with GLN and ARG supplementation. However, the use of inhibitors clearly showed that OKG does not modulate TNF-alpha secretion by GLN, ARG, or polyamine pathways. We also observed that OKG enhanced NO* release by stimulated macrophages (DEX-OKG, 1.77 +/- 0.64 vs. DEX-NEAA, 0.29 +/- 0.29 nmol/ 10(6) cells, P < 0.05). Using inhibitors, it appears that this action of OKG is probably mediated via polyamine synthesis and GLN. However, an oral administration of an equimolar amount of GLN failed to reproduce the OKG-mediated effect, possibly because OKG generates more GLN in the systemic circulation than GLN itself when these substances are given orally. Our results underline the complexity of the mechanism of action of OKG, which can differ according to the functions of even a single cell type. (+info)
Arginase activity in human breast cancer cell lines: N(omega)-hydroxy-L-arginine selectively inhibits cell proliferation and induces apoptosis in MDA-MB-468 cells.
(37/1042)
L-Arginine is the common substrate for two enzymes, arginase and nitric oxide synthase (NOS). Arginase converts L-arginine to L-ornithine, which is the precursor of polyamines, which are essential components of cell proliferation. NOS converts L-arginine to produce NO, which inhibits proliferation of many cell lines. Various human breast cancer cell lines were initially screened for the presence of arginase and NOS. Two cell lines, BT-474 and MDA-MB-468, were found to have relatively high arginase activity and very low NOS activity. Another cell line, ZR-75-30, had the highest NOS activity and comparatively low arginase activity. The basal proliferation rates of MDA-MB-468 and BT-474 were found to be higher than the ZR-75-30 cell line. N-Hydroxy-L-arginine (NOHA), a stable intermediate product formed during conversion of L-arginine to NO, inhibited proliferation of the high arginase-expressing MDA-MB-468 cells and induced apoptosis after 48 h. NOHA arrested these cells in the S phase, increased the expression of p21, and reduced spermine content. These effects of NOHA were not observed in the ZR-75-30 cell line, which expresses high NOS and relatively low arginase. The effects of NOHA were antagonized in the presence of L-ornithine (500 microM), which suggests that in MDA-MB-468 cell line, the arginase pathway is very important for cell proliferation. Inhibition of the arginase pathway led to depletion of intracellular spermine and apoptosis as observed by terminal deoxynucleotidyl transferase (Tdt)-mediated nick end labeling assay and induction of caspase 3. In contrast, the ZR-75-30 cell line maintained its viability and its L-ornithine and spermine levels in the presence of NOHA. We conclude that NOHA has antiproliferative and apoptotic actions on arginase-expressing human breast cancer cells that are independent of NO. (+info)
Dietary arginine requirement of juvenile red drum (Sciaenops ocellatus) based on weight gain and feed efficiency.
(38/1042)
Increasing aquacultural production of red drum (Sciaenops ocellatus) has prompted the determination of many of this species' nutritional requirements. However, limited information is available concerning its amino acid requirements, especially for arginine. Therefore, a feeding trial was conducted with juvenile red drum to determine their quantitative dietary requirement for arginine. Experimental diets contained 35 g crude protein/100 g from red drum muscle and crystalline amino acids. Incremental levels of arginine were added to the diets in place of a mixture of glycine and aspartic acid to maintain all diets isonitrogenous. All diets were fed in triplicate to juvenile red drum for 7 wk. Graded levels of arginine significantly (P < 0.05) affected weight gain, feed efficiency, protein efficiency ratio (PER) and plasma arginine levels of the fish. Based on least-squares regression of feed efficiency and PER data, the minimum requirement (+/- SEM) of red drum for arginine was estimated at 1.44 (+/- 0.15) and 1.48 (+/- 0.12) g/100 g diet (4.11 and 4.23 g/100 g dietary protein), respectively. The arginine requirements estimated from weight gain data were 1.75 (+/- 0.18) g/100 g diet or 5.0 g/100 g dietary protein. These values are similar to those reported for other carnivorous fish species. (+info)
N-Acetylglutamate synthase of Escherichia coli regulation of synthesis and activity by arginine.
(39/1042)
N-Acetylglutamate synthase, the first enzyme of arginine biosynthesis, was stabilized in crude extracts from Escherichia coli. At 4 degrees the enzyme lost less than 5% of activity per day. L-Arginine repressed the formation of N-acetylglutamate synthase. Under conditions of genetic or physiological derepression, a specific activity of approximately 50 nmol per min per mg of protein was measured. No activity (i.e. less than 0.2 nmol per min per mg of protein) could be detected in extracts from cells grown under conditions of repression, whereas an intermediate level was found in cell cultivated on minimal medium. In a 6-fold purified preparation L-arginine inhibited the enzyme. Of 11 precursors and analogues of arginine tested only O-[L-norvalyl-5]-isourea inhibited N-acetylglutamate synthase as strongly as L-agrinine. (+info)
Canavanine-mediated depletion of polyamine pools in Escherichia coli: effect of head morphogenesis and DNA synthesis.
(40/1042)
We have found that L-canavanine inhibited the synthesis of polyamines in T4-infected Escherichia coli. These polyamines are known to be required for T4 DNA synthesis and may be involved in phage morphogenesis. The new data indicate that the inhibition of polyamine synthesis is not primarily responsible for the L-conavanine-mediated inhibition of DNA synthesis nor does it seem to be involved in the induction of lollipops. L-Canavanine does influence the relative amounts of putrescine and spermidine found in the phage particle, but it does not influence the amount of DNA phosphate neutralized by polyamines. (+info)