• Asian-Australasian Journal of Animal Sciences
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• v.29 no.10
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• pp.1483-1489
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• 2016

Most of amino acid (AA) digestibility values for feed ingredients are obtained using pigs cannulated in the distal ileum. The ileal-cannulated pig model uses pigs older than six weeks due to difficulties related to implanting the T-cannula in distal ileum of younger pigs and complications during the post-surgical recovery. However, to properly formulate the diet of weaned pigs, the nutritive value of feed ingredients should be determined with younger pigs. Thus, 25 weaned pigs were used to determine the apparent total tract digestibility (ATTD) of nutrients, energy, and apparent ileal digestibility (AID) and standardized ileal digestibility (SID) ileal AA digestibility of broken rice (BR), with or without multicarbohydrase (MC) and phytase (Phy) supplementation. Piglets were weaned at 23 d of age and individually housed in digestibility cages until 45 d of age. The trial consisted of 7 d of adaptation to the experimental diets and 3 d of excreta (feces and urine) collection. Ileal digesta was collected at slaughter (about 6 weeks of age). A completely randomized experimental design was used to determine the effects of MC and Phy. Reference diets (RD, 5% casein) was replaced by 30% of BR with or without MC, Phy, or MC+Phy. The RD was used to quantify endogenous AA losses. BR with Phy supplied had increased the ATTD of dry matter (p<0.05) and SID of histidine (p = 0.05), arginine, leucine, lysine, valine, alanine, and proline (p<0.05). BR with MC had been increased digestible energy and protein and SID for histidine (p<0.05). There was no interaction between Phy and MC on the BR nutrient digestibilities. Standardized amino acid digestibilities of BR, without enzymes, were lower than those values reported in the literature. The MC and Phy improved the digestibility of some nutrients and energy of BR in post-weaned piglet diets.

• Biomolecules & Therapeutics
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• v.13 no.3
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• pp.190-197
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• 2005

Advanced glycation end products (AGE) have been implicated in the pathogenesis of diabetic complications including nephropathy. However, the role of AGE in the activation of mesangial cells cultured under high glucose has not been elucidated. The effects of aminoguanidine, which prevents formation of AGE and protein cross-linking, on the synthesis of $TGF-{\beta}1$ and fibronectin by rat mesangial cells cultured under high glucose for 2 weeks were examined and compared with the effects of $N^G$-nitro-L-arginine methyl ester (NAME), a selective nitric oxide synthase inhibitor, because aminoguanidine also inhibits the inducible nitric oxide synthase. Culture of mesangial cells in 30 mM (high) glucose for 2 weeks induced 1.5-fold (ELISA) and 1.9-fold (Western blot analysis) increase in AGE in the culture media compared to 5.6 mM (control) glucose. Northern blot analysis revealed 1.5-fold increase in $TGF-{\beta}1$ and 1.7-fold increase in fibronectin mRNA expression in cells cultured under high glucose compared to control glucose. Increases in mRNA expression were followed by increased protein synthesis. Mink lung epithelial cell growth inhibition assay revealed 1.4-fold increase in $TGF-{\beta}1$ protein in high glucose media compared to control. Fibronectin protein also increased 2.1-fold that of control glucose by Western blot analysis. Administration of aminoguanidine suppressed AGE formation in a dose dependent manner and at the same time suppressed $TGF-{\beta}1$ and fibronectin synthesis by mesangial cells cultured in both control and high glucose. In contrast, NAME did not affect high glucose-induced changes. These findings support a role for AGE in high glucose-induced upregulation of $TGF-{\beta}1$ and fibronectin synthesis by mesangial cells.

• Journal of Preventive Medicine and Public Health
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• v.32 no.4
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• pp.459-466
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• 1999

Objectives:eurotoxicity is mediated by nitric oxide(NO) in the rat primary neuronal cultures and assess the effect of $Mn^{2+}$ on the N-methyl-D aspartate(NMDA) receptors. Methods: We have used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)assay to examine the effect of cytotoxicity of $MnCl_2$ in neuronal cells , NO production was determined by measuring nirites, a stable oxidation product of NO. The neurons in the rat that contains neuronal nitric oxide synthase(nNOS) were examined by immunofluorescence and confocal microscopy. The effects of $Mn^{2+}$ on the NMDA receptors was assesed by the whole cell voltage clamp technique. Results: We showed that the NO release and NOS expression was increased with 500uM $MnCl_2$ treatment and an NOS inhibitors, $N^G-nitro-L-arginine$, prevented neurotoxicity elicited by manganese. In the electrophysiological study, $Mn^{2+}$ does not block or activate the NMDA receptors and not pass through the NMDA receptors in a neurons of basal ganglia. Conclusions: It is concluded that manganese neurotoxicity in basal ganglia was partially mediated by nitric oxide in the cell culture model.

• Restorative Dentistry and Endodontics
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• v.24 no.1
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• pp.212-221
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• 1999

The periapical response to injury is a complex interaction of inflammatory, immune, neural, vascular and synthetic activity. Nitric oxide(NO), synthesized by nitric oxide synthetase(NOS) from L-arginine, is becoming recognized as an important bio-regulatory molecule in a variety of tissue, but little is known about its possible role in periapical tissue. The purpose of this study was to investigate the expression of nitric oxide synthetase(NOS) in tooth follicle, periapical abscess, granuloma and cyst. The expression of NOS in periapical lesions was evaluated by immunohistochemical staining for $NOS_2$, and $NOS_3$. The immunoreactivity was evaluated by staining intensity, and inflammatory cell infiltration. Correlationship between the periapical lesion in immunoreractivity were statistically analyzed by SPSS. The degree of $NOS_2$ and $NOS_3$ expression in periapical abscess was higher than in any other periapical lesions, and stastically significant. The expression degree of $NOS_2$ and $NOS_3$ was not correlated with periapical abscess and granuloma, but expression of $NOS_2$ showed very significant in periapical cyst. The increased expression of $NOS_2$ and $NOS_3$ was correlated with inflammatory cell infiltration degree of the periapical cyst. These results suggested that NO should play an important role in progress and/or mediation of periapical lesions.

• The Journal of Korean Medicine
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• v.21 no.3
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• pp.20-30
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• 2000

Objective : This experiment was performed in order to study the effect of an aqueous extract of Salviae radix root(SRRAE) on NO production and NOS gene induction from macrophages Methods : To investigate dose-dependent effects of SRRAE for NO release on the $rIFN-{\gamma}-treated$ macrophages, the cells were incubated for 6 hrs in a medium containing $rIFN-{\gamma}$ (5 U/ml), stimulated with SRRAE and incubated in a CO2 incubator. The cells were treated with 5 U/ml $rIFN-{\gamma}$ plus 100 g/ml of SRRAE, Then, the cells were incubated with various concentrations of NGMMA at $37^{\circ}C$ for 48 hrs, Results : SRRAE had no effect on NO production by itself, whereas recombinant $interferon-{\gamma}(rIFN-{\gamma})$ alone showed modest activity, When SRRAE was used in combination with $rIFN-{\gamma}$, there was a marked cooperative induction of NO production in a dose-dependent manner. The optimal effect of SRRAE on NO production was shown at 6hrs after treatment with $rIFN-{\gamma}$. The SRRAE-induced production of NO was inhibited by NG-monomethyl- L-arginine(NGMMA) and arginase. $rIFN-{\gamma}$ in combination with SRRAE showed a marked increase of the expression of the inducible NOS(iNOS) gene. In addition, the effect of SRRAE was mainly dependent on the SRRAE-induced tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ secretion. Conclusions : SRRAE induces NO production from macrophages as a result of SRRAE-induced $TNF-{\alpha}$ secretion. SRRAE may provide a second signal for synergistic induction of NO production in macrophages already induced to express iNOS gene by $rIFN-{\gamma}$.

• The Journal of Korean Medicine
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• v.27 no.4
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• pp.74-83
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• 2006

Ssanghwatang (SHT) has been known to prove effective in the treatment for erectile dysfunction (ED), and its modified formula is widely used in clinical practice. However, its fundamental mechanism of action is not clearly known. It is well known that endothelial cells can achieve the relaxation of vascular smooth muscles by the release of nitric oxide (NO). NO is synthesized by the enzyme NO synthase (NOS) from L-arginine and oxygen. It is widely accepted that NO plays an important role in the relaxation of corpus cavernous smooth muscle and vasculature. In addition, in terms of the penile erection, the NO/cGMP pathway is more potent than the PCE1/cAMP pathway. The main purpose of the present study was to investigate the mechanism of the erectile effects of SHT by focusing on its direct effects on corpus cavernous smooth muscle cells. We investigated the NOS activity, nitrite concentration and cGMP levels in rat corpus cavernous smooth muscle cell lines activated by SHT extracts. Furthermore, we evaluated the effect of SHT extracts on penile smooth muscle relaxation following oral administration of SHT extract powder to rats by the dosage of 1 g/kg over fifteen days. As a result, we found that SHT stimulated NO release. NOS activity and cGMP levels were increased by SHT respectively. Furthermore, SHT relaxed the corpus cavernous smooth muscle. These results are consistent with the concept that penile erection by SHT is carried out through the NO/cGMP pathway. In conclusion, the present study shows that SHT increases the NOS activity, synthesizes NO and augments the cGMP, which mediates penile erection. Further determination of the SHT mechanism related with the NO/cGMP pathway strongly indicates that SHT can be used as a remedy for erectile impotence.

• BMB Reports
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• v.34 no.3
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• pp.194-200
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• 2001

Secreted arginase from Evernia prunastri thallus has been purified 616-fold from the incubation medium. Purified arginase was resolved as only one peak in a capillary electrophoresis with a pI value of 5.35. The protein contained high amounts of acidic amino acids, such as Asx and Glx, and a relatively high quantity of Ser and Gly. The molecular mass of native, purified arginase was estimated as about 26 kDa by SE-HPLC. Substrate saturated kinetic showed a typical Michaelis-Menten relationship with a K_m value of 3.3 mM L-arginine. Atranorin behaved as a mixed activator of the enzyme (apparent $K_m$ = 0.96 mM); whereas evernic and usnic acid were revealed as non competitive inhibitors (apparent $K_m$ values were 3.16 mM and 3.05 mM, respectively). Kinetics of atranorin binding indicated that saturation was reached from 0.18 ${\mu}mol$ of the total atranorin and the occurrence of multiple sites for the ligand. This agrees with a possible aggregation of several enzyme subunits during the interaction process. A value of binding sites of about 12 was obtained. The binding of evernic acid was saturated from 23 nmol of total phenol. The number of binding sites was about 5. The loss of the binding ability of evernic acid could be interpreted as a single negative cooperatively. Usnic acid behaves in a similar way to evernic acid, although the binding saturation occurs at $0.14\;{\mu}moles$ of the ligand. This binding appears to be unspecific, and has 28 usnic acid binding sites to the protein.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.5
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• pp.305-314
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• 2005

Diabetes mellitus is associated with vascular complications, including an impairment of vascular function and alterations in the reactivity of blood vessels to vasoactive substances in various vasculature. In the present study, the authors have observed endothelin-B ($ET_B$) receptor agonist-induced relaxation in precontracted mesenteric arterial segments from streptozotocin (STZ)-induced diabetic rats, which was not shown from control rats or in other arterial segments from diabetic rats. Accordingly, the goal of this study was to investigate in what way STZ-induced diabetes altered reactivity of the mesenteric arterial bed and to examine the causal relaxation, if any, between this $ET_B$ receptor-mediated relaxation and endothelial paracrine function, especially nitric oxide (NO) production. The relaxation induced by $ET_B$ agonists was not observed in mesenteric arteries without endothelium. The relaxation to $ET_B$ agonists was completely abolished by pretreatment with BQ788, but not by BQ610. $N_{\omega}-nitro-L-arginine$ methyl ester and soluble guanylate cyclase inhibitors, methylene blue or LY83583 significantly attenuated the relaxant responses to $ET_B$ agonists, respectively. When the expression of eNOS and iNOS was evaluated on agarose gel stained with ethidium bromide, the expression of eNOS mRNA in diabetic rats was significantly decreased, but the expression of iNOS was increased compared with control rats. Furthermore, the iNOS-like immunostaining was densely detected in the endothelium and slightly in the arterial smooth muscle of diabetic rats, but not in control rats. These observations suggest that $ET_B$ receptor may not play a role in maintaining mesenteric vascular tone in normal situation. However, the alterations in $ET_B$ receptor sensitivity were found in diabetic rats and lead to the $ET_B$ agonist-induced vasorelaxation, which is closely related to NO production. In the state of increased vascular resistance of diabetic mesenteric vascular bed, enhanced NO production by activation of iNOS could lead to compensatory vasorelaxation to modulate adequate perfusion pressure to splanchnic area.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.10
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• pp.1422-1428
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• 2000

This investigation was conducted to evaluate the shrimp flesh (SF) and shrimp by-product (SB) of the most abundant shrimp species (Metapenaeus affinis, Penaeus semisulcatus and Penaeus monodon) caught in Central Vietnam, with the emphasis on yield, gross and amino acid (AA) composition and effect of heat treatment. The results showed that the mean edible SF and SB (head and shells with tail) yields of the three shrimp species averaged 56.7 and 43.3%, respectively, of the total wet body weight, with the M. affinis generating the highest by-product yield (45.7%) and P. semisulcatus (40.6%) the lowest. Significant differences in dry matter (DM), crude protein (CP) and ash content were found between SF and SB. The DM content of SF (21.5%) was lower than of SB (24.9%) and the ash content (on a DM basis) of the SB in all shrimp species was more than three times that of the SF (p<0.05), whereas the CP content was almost twice as high in the SF as compared with the SB (p<0.05). The SB of the three species contained (on a DM basis) between 44.0 and 49.8% CP (p<0.05) and between 13.5 and 18.1% chitin (p<0.05). The Ca content of SB differed also between species (p<0.05). On average, the sum of AA in SB corresponded to 89.3% of the CP and essential AA accounted for about 50% of the total AA. The most abundant AA were arginine, aspartic and glutamic acids, which accounted for 33% of the total AA. Minor, but significant differences in some AA concentrations of SB between species were observed (p<0.05). With the exception of the DM and ether extract content, all other chemical constituents of entire shrimp, SF and SB were not significantly affected by heat treatment (p>0.05).

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.7
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• pp.1100-1107
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• 2007

The effects of offering broilers phosphorus-adequate diets containing 10.0 and 11.8 g/kg lysine, without and with 500 FTU/kg exogenous phytase, on growth performance and nutrient utilisation were determined. Each of the four experimental diets was offered to 6 replicates of 10 birds from 7 to 28 days of age. Effects of treatment on performance, apparent metabolisable energy, apparent ileal digestibility of amino acids and bone mineralisation were examined. Both additional lysine and phytase supplementation improved (p<0.05) weight gain and feed efficiency, with interactions (p<0.05), as phytase responses were more pronounced in lysine-deficient diets. Phytase improved (p<0.05) apparent metabolisable energy, which was independent of the dietary lysine status. Bone mineralisation, as determined by percentage toe ash, was not affected by treatment, which confirms the phosphorus-adequate status of the diets. Phytase increased (p<0.05) the apparent ileal digestibility of the sixteen amino acids assessed. Unexpectedly, however, the dietary addition of 1.8 g/kg lysine, as lysine monohydrochloride, increased (p<0.05) the ileal digestibility of lysine per se and also that of isoleucine, methionine, phenylalanine, valine, aspartic acid, glutamic acid and tyrosine. In addition, there were significant interactions (p<0.05) between additional lysine and phytase supplementation for arginine, lysine, phenylalanine, aspartic acid, glutamic acid, glycine and serine digestibilities, with the effects of phytase being more pronounced in lysine-deficient diets. The possible mechanisms underlying the increases in amino acid digestibility in response to additional lysine and the interactions between lysine and microbial phytase in this regard are discussed. Also, consideration is given to the way in which phytate and phytase may influence ileal digestibility of amino acids.