• Archives of Pharmacal Research
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• v.29 no.2
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• pp.145-151
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• 2006

We investigated the effects of Ginsenoside $R_e$ on human sperm motility in fertile and asthenozoospermic infertile individuals in vitro and the mechanism by which the Ginsenosides play their roles. The semen samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Spermatozoa were separated by Percoll and incubated with 0, 1, 10 or $100\;{\mu}M$ of Ginsenoside $R_e$. Total sperm motility and progressive motility were measured by computer-aided sperm analyzer (CASA). Nitric oxide synthase (NOS) activity was determined by the $^{3}H$-arginine to $^{3}H$-citrulline conversion assay, and the NOS protein was examined by the Western blot analysis. The production of sperm nitric oxide (NO) was detected using the Griess reaction. The results showed that Ginsenoside $R_e$ significantly enhanced both fertile and infertile sperm motility, NOS activity and NO production in a concentration-dependent manner. Sodium nitroprusside (SNP, 100 nM), a NO donor, mimicked the effects of Ginsenoside $R_e$. And pretreatment with a NOS inhibitor $N^{w}$-Nitro-L-arginine methyl ester (L-NAME, $100\;{\mu}M$) or a NO scavenger N-Acetyl-L-cysteine (LNAC, 1 mM) completely blocked the effects of Ginsenoside $R_e$. Data suggested that Ginsenoside $R_e$ is beneficial to sperm motility, and that induction of NOS to increase NO production may be involved in this benefit.

• Journal of Photoscience
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• v.9 no.2
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• pp.205-208
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• 2002

Nitric oxide (NO) is a newly described transmitter involved with cell to cell communication that is generated in biologic tissues by specific types of nitric oxide synthase (NOS), which metabolize L-arginine and molecular oxygen to citrulline and nitric oxide. In the skin. NO has been reported to play an important role in such diseases as psoriasis, atopic dermatitis, and contact dermatitis, as well as act as an important modulator in UVB-induced erythema. Ultraviolet B irradiation to the skin evokes an increase in NO production in the epidermis through two pathways; induction of inducible NOS, mediated by inflammatory cytokines, and elevation of constitutive neuronal NOS activity. In a cell culture system, it has been demonstrated that NO functions as a melanogen after being produced in keratinocytes in response to UVB-irradiation. NO-stimulated melanogenesis in melanocytes is mediated by the cGMP/PKG pathway. In this study, up-regulation of tyrosinase gene expression by NO-stimulation and the involvement of NO in UVB-induced pigmentation were examined. In NO-induced melanogenesis, protein synthesis and tyrosinase activity increased along with an up-regulation of tyrosinase gene expression. In an animal model, UVB-induced pigmentation in skin was suppressed by sequential daily treatments with a specific inhibitor of NOS. Thus, NO plays an important role in UVB-induced pigmentation, where its function as a melanogen is considered to be one of the mechanisms. Together with its role in the development of erythema, NO contributes to the total protective response of skin against UVB-irradiation.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.185-185
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• 1996

Nitric oxide synthase(NO Synthase:E.C.1.14.13.39)는 생체내에서 L-arginine을 기질로 하여 citrulline과 nitric oxide(NO)를 생성하는 효소로서, 최근 연구에 의하면 2/3 부분 간 절제술 후 prereplicative phase동안에 발현되는 것으로 알려져 있다. 한편, 생체내에서 NO Synthase에 상경적 길항제인 methylarginine에 관해서도 수년간 많은 연구가 진행되어 왔다. 이들의 생성 기전은 protein methylation에 의해 생성된 methylated protein이 생체 내에서 분해되어 생성된다고 알려져 있으나, 정확한 기전에 대해서는 아직 논란의 여지가 많다. 따라서, 본 연구에서는 In vivo 실험을 통해 부분 간 절제술 후 시간대별로 간 조직에서의 NO Synthase 활성도와 혈청에서 NO의 최종 대사물인 Nitrite/Nitrate를 측정하였으며, 또한 NO Synthase 조절에 관여하는 세포내 기질인 arginine과 억제 인자인 methylarginine함량을 간 조직 및 혈청에서 측정하고, 세포 신호 전달체계에 관여하는 cyclic GMP 함량을 측정함으로써, 부분 간 절제술 후 간 재생동안에 NO Synthase 활성도와 methylarginine 및 arginine과의 상관 관계를 규명하고, 간 재생동안, 생성된 nitric oxide의 역할을 연구하려한다.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1681-1691
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• 2017

This study investigated the effect of proline addition on the salt tolerance of Tetragenococcus halophilus. Salt stress led to the accumulation of intracellular proline in T. halophilus. When 0.5 g/l proline was added to hyperhaline medium, the biomass increased 34.6% (12% NaCl) and 27.7% (18% NaCl) compared with the control (without proline addition), respectively. A metabolomic approach was employed to reveal the cellular metabolic responses and protective mechanisms of proline upon salt stress. The results showed that both the cellular membrane fatty acid composition and metabolite profiling responded by increasing unsaturated and cyclopropane fatty acid proportions, as well as accumulating some specific intracellular metabolites (environmental stress protector). Higher contents of intermediates involved in glycolysis, the tricarboxylic acid cycle, and the pentose phosphate pathway were observed in the cells supplemented with proline. In addition, addition of proline resulted in increased concentrations of many organic osmolytes, including glutamate, alanine, citrulline, N-acetyl-tryptophan, and mannitol, which may be beneficial for osmotic homeostasis. Taken together, results in this study suggested that proline plays a protective role in improving the salt tolerance of T. halophilus by regulating the related metabolic pathways.

• Applied Biological Chemistry
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• v.47 no.4
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• pp.403-408
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• 2004

We studied active substances like crude cell wall components, crude protein, composing amino acids and free amino acids including orinithine cycle-related amino acids such as asparagine, ornithine and citrullin in fully ripe fruits of Korean native squash, Cucurbita moschata Poir. Crude protein content of 'Jeju 2' was the highest with $2,830\;{\mu}g/g$, while 'Sangju' was the lowest with $1,319\;{\mu}g/g$. Regarding the contents of crude cell wall components, fruit 'Kanghaw' was the highest with 2,961 mg% while 'Namhea' was the lowest with 1,582 mg%. Pectin contents of crude cell wall components were the highest in 'Kanghaw' (2,198 mg%) followed by 'Jeju 2' (2,178 mg%) and 'Jeju l' (1,461 mg%). The main contents of amino acids in squash were glutamic acid, aspartic acid, lysine, leucine and valine, which comprised to be more than 50% of total amino acid contents. Especially, in 'Jeju 2' aspartic acid and threonine were not detected. In fully ripe fruits, a total of 34 kinds of free amino acids were detected including 8 kinds of essential amino acids (histidine, isoleucine, leucine, lysine, phenylalanine, methionine, threonine and valine). More than 50% of the total free amino acids were aspartic acid and asparagine, and also all varieties were detected in ornithine, citrullin, and arginine, which are related to Ornithine cycle. There was a big difference in the contents of arginine in all varieties whereas the contents of ornithine and citrullin were very similar. 'Teaan' 29.34% was 7 times higher than 'Namhea' 4.30% in regards to arginine contents.

• Food Science and Preservation
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• v.10 no.3
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• pp.302-307
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• 2003

In the present study, we investigated the quality characteristic of old pumpkin extract treated with enzymes. As a results, all the groups treated with pectinase were better in quality characteristic than control group and the group treated with 0.15％(w/w) pectinase was specially great. All the groups treated with simultaneous pectinase and cellulase were higher in the extraction rate than the groups treated with pectinase or cellulase. The experimental groups were divided into non-treated control(I) and three treatment groups(II- IV) for optimum condition of enzyme treatment. The II and IV groups were treated with 0.15％(w/w) pectinase and 0.15％(w/w) cellulase, respectively, and the ill group was treated with both 0.15％(w/w) petinase and 0.05％(w/w) cellulase. Yield for old pumpkin extract of the III group (86.94％) was higher than that of other groups, but there were no significant difference among the groups in soluble solide content and pH of the extract. Reducing sugar and total sugar contents in the ill group were 2.81％ and 4.60％, respectively. Total carotene content in the II group (5.36 mg％) was higher than other groups. Old pumpkin extracts in all the groups showed nitrite-scavenging ability to pH 1.2, 3.0 and 4.0. Total free amino acid content in the III group (176.7 mg％) was higher than other groups. Citrulline contents in the II and III groups were detected 1.66 and 1.41 mg％, respectively but the contents in other groups were not detected.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1997.04a
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• pp.90-90
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• 1997

Natural products, which have been used for the treatment of hypertension, diuresis and nephritis in traditional oriental medicine, were selected for the screening of nitric oxide (NO) production in endothelial cells and kidney tissues in vitro as well as in vivo by measuring the conversion of [$\^$14/C]-L-arginine to [$\^$14/C]-L-citrulline, a coproduct of the enzyme reaction with NO. Confluent monolayer of endothelial cells were used for the screening of 16 natural products. Among the natural products, Zizyphus jujuba and Codonopsis pilosula stimulated endothelial NO synthase activity. Thus, both confluent monolayer of endothelial cells and kidney homogenates (glomeruli, cortical tubules, meudllae) were treated with Zizyphus jujuba and Codonopsis pilosula (final concentration 10 $\mu\textrm{g}$/$m\ell$) and NO releases were compared with those by receptor - dependent agonists, bradykinin and ADP and receptor - independent calcium ionophore A23187 in vitro. In rat experiment, NO releases in glomeruli, cortical tubules and medullae and plasma renin activity were assessed after intraperitoneal injection of Zizyphus jujuba and Codonopsis pilosula (10 mg/kg/day for 4 days). As a result, both Zizyphus jujuba and Codonopsis pilosula significantly increased NO releases in cultured endothelial cells, kidney tissues in vitro as well as in vivo. Stimulation of NO releases by Zizyphus jujuba and Codonopsis pilosula was similar to those by receptor - dependent agonists, bradykinin and ADP and receptor - independent calcium ionophore A23187 in cultured endothelial cells. However, plasma renin activity was not influenced by these two natural products. In conclusion, stimulatory effects of Zizyphus jujuba and Codonopsis pilosula on NO release in kidney may contribute their hypotensive effects and antinephritic action possibly by increasing renal blood flow.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.7
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• pp.927-935
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• 2012

This study was performed to develop functional vinegar by using cucumbers through two stages of fermentation. The alcohol content was maximized (7.8%) after 6-days of alcohol fermentation at $25^{\circ}C$ by adjusting the initial sugar concentration to $15^{\circ}Brix$, and vinegar with an acidity of 5.8% was obtained after 12-days of acetic acid fermentation at $30^{\circ}C$. The major sugars in the produced vinegar were glucose and fructose, which were present in concentrations of 3,067.26 and 395.73 mg%, respectively. The major organic acids were acetic acid and succinic acid, which were present in concentrations of 4,410.5 and 841.11 mg%, respectively. The total free amino acid content of the cucumber vinegar was 181.45 ${\mu}g/mL$ and citrulline, valine, aspartic acid, asparagine, and ornithine were the major amino acids. The inorganic components included various alkaline elements, such as K, Ca, and Mg. In addition, experimental methods to assess the DPPH and $ABTS^+$ radical-scavenging ability, reducing power, and ${\beta}$-carotene bleaching activity showed that the cucumber vinegar had strong antioxidant properties. The total polyphenol content, which are the major components responsible for the antioxidant activities of the cucumber vinegar, was 40.14 mg/100 mL. The cucumber vinegar showed significantly higher hepatic aldehyde dehydrogenase activity when compared to the alcoholic control (negative) and the marketing drink (positive), resulting in decreased plasma acetaldehyde concentrations in rats. These results demonstrate that cucumber vinegar possesses antioxidant properties and holds great promise for use in preventing hangovers.

• Journal of Yeungnam Medical Science
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• v.13 no.2
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• pp.159-172
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• 1996

대기오염물질이면서 동시에 생체내 화학반응의 산물이기도 한 nitric oxide(NO)는 그 생체내 분포가 광범위하고 생리적 역할이 다양하여, 최근의 생명과학 분야에서 가장 크게 주목받는 몇가지 연구대상 중 하나이다. 세포에서의 NO 산생은 nitric oxide synthase (NOS)에 의해 촉매되는데, 이들은 brain form (bNOS, neuronal; nNOS, NOS I), inducible form (iNOS), 및 endothelial form(eNOS)로 구분되는데, 이중 bNOS(nNOS)와 eNOS는 inducible form에 대비되는 constitutive form(cNOS)에 해당하므로 각각 ncNOS 와 ecNOS로도 불리운다. NOS는 아미노산인 L-arginine을 산소와 결합시켜 L-citrulline으로 변환시키면서 NO를 유리하고, 이 NO는 세포내의 guanylate cyclase를 활성화하여 cyclic GMP를 생산하거나, superoxide(O2-) 및 수소이온과 차례로 결합하여 반응성이 매우 높은 수산화기(-OH)를 발생시켜 세포독작용을 유발하기도 한다. 정상상태에서 뇌혈관내피세포의 ecNOS로 부터 유리된 NO는 혈관을 확장시켜 신경세포에 대한 산소공급을 원활히 유지해 주지만, 순환장애를 일으켰을 때는 뇌조직내의 iNOS로부터 대량의 NO가 유출되어 신경세포의 손상을 가져온다. 호흡기에서는 NO가 기도평활근을 이완시키고 폐혈류를 개선하므로, 미숙아나 성인의 호흡장애시에 소량의 NO를 흡입시키면 oxygenation을 호전시킬 수 있다. 그러나 대기오염이나 흡연 등으로 대량의 NO를 흡입할 경우 치명적인 폐부종이나 methemoglobin혈종을 일으킬 수 있다. 순환계에서는 cNOS가 혈관을 확장시켜 조직의 혈류를 유지하는데 일익을 담당한다. 세균내 독소(lipopolysaccharide; LPS)나 각종 명역조절물질들이 혈관내피세포와 혈관평활근세포로 부터 과다한 NO를 유리시키면 혈압이 급격히 떨어져 순환허탈상태에 빠지게 된다. 심장에서는 관상혈관 내피세포의 eNOS가 심근의 혈류를 유지해 주지만 허혈이나 세균내독소 또는 면역조절물질 등에 의하여 심근세포나 침윤된 대식세포의 iNOS로 부터 과량의 NO가 유리되면 심근세포의 손상이 초래된다. 신장에서는 내피세포의 cNOS에 의하여 사구체여과가 조절되고 있는데, 세균내독소나 면역 조절물질 등에 의하여 사구체관막세포(mesangial cell)등의 iNOS로 부터 과량의 NO가 유리되면 신조직과 사구체의 손상을 초래한다. 위와 같이 대부분의 장기에서 ecNOS는 조직의 혈류를 유지하는 역할을 하며, iNOS는 애초 세균 등 침입자에 대한 세포독작용이 그 존재 목적이라고 풀이할 수 있겠으나 일종의 부작용으로 자체조직의 손상을 초래하게 되는 것으로 본다. 따라서 NO와 관련된 각종 병변의 치료를 위해서는 NOS의 비선택성 억제제인 arginine 유도체 보다는 iNOS에 대한 선택적 억제제인 S-methylisothiourea(SMT), aminoethylisothiourea(AETU), aminoguanidine (AMG), agmatine, L-canavanine, transforming growth factor b1(TGF-b1) 등의 사용을 검토해 보는 것이 타당할 것으로 사료된다.

• Korean Journal of Head & Neck Oncology
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• v.17 no.2
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• pp.155-161
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• 2001

Background and Objectives: Nitric oxide (NO) is generated in mammalian tissue by the conversion of L-arginine to L-citrulline. This reaction is catalyzed by nitric oxide synthase (NOS). NO is an important bioactive agent and a signalling molecule that mediates a variety of biologic actions such as vasodilation, neurotransmission, host defense, and iron metabolism but increased NO production may also contribute to the pathogenesis of a various of disorders, including cancer. Before now, the role of NO in thyroid gland is still investigated and it was supposed that NO mediate the angiogenesis in tumor growth. Others journal and works identified the expression of iNOS that involve by neutrophil and eNOS that involve in part in the vascular remodeling and to understand the role of NO in human thyroid gland. But authors revealed only eNOS in thyroid neoplasm. iNOS was identifed by inflammation in fault. Materials and Methods: Western blot analysis was performed, using a polyclonal antibody against eNOS (Rabbit polyclonal IgG). Using the same antibody, the distribution of eNOS was examined in 15 formalin-fixed paraffin embedded samples by immunohistochemistry. By NADPH consumption rate, NOS activity was estimated at nodular hyperplasia. Results: Western blot analysis exhibited that eNOS was significantly elevated in thyroid papillary carcinoma, compared to that in nodular hyperplasia and normal tissue. Immunohistochemistry showed that the immunoreacitivity was present more significantly in thyroid follicular epithelial cell layer than vascular endothelial cell. NOS activity increased in nodular hyperplasia. Conclusions: Thyroid papillary cancer without neutrophil invasion expressed only eNOS. The endothelial localization of eNOS may play an important role in pathogenensis of human thyroid nodular hyperplasia and the follicular localization of thyroid papillary carcinomas.