• Natural Product Sciences
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• v.16 no.4
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• pp.259-264
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• 2010

The essential oil fraction was obtained from the underground parts o of Ligusticum chuanxiong (Umbelliferae) by steam distillation, and its main components, Z-ligustilide and butylidene phthalide, were isolated by column chromatography. Its essential oil fraction and the isolated main components were examined for effects on their anti-inflammatory properties in RAW 264.7 macrophage cells to develop a new natural anti-inflammatory drug. The results showed that the L. chuanxiong essential oil fraction and its main components, Z-ligustilide and butylidene phthalide, inhibited the production of nitric oxide significantly in lipopolysaccharide (LPS)-treated RAW 264.7 cells. LPS-induced interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-$\alpha$) production was also decreased in a dose-dependent manner. In addition, western blot analysis revealed that the L. chuanxiong essential oil fraction and also its main components, Z-ligustilide, and butylidene phthalide reduced the expression levels of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS).

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1688-1696
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• 2020

Soil physical and chemical characteristics, soil potential denitrification rates (PDR), community composition and nirK-, nirS- and nosZ-encoding denitrifiers were studied by using MiSeq sequencing, quantitative polymerase chain reaction (qPCR), and terminal restriction fragment polymorphism (T-RFLP) technologies base on short-term (5-year) tillage field experiment. The experiment included four tillage treatments: conventional tillage with crop residue incorporation (CT), rotary tillage with crop residue incorporation (RT), no-tillage with crop residue retention (NT), and rotary tillage with crop residue removed as control (RTO). The results indicated that soil organic carbon, total nitrogen and NH₄⁺-N contents were increased with CT, RT and NT treatments. Compared with RTO treatment, the copies number of nirK, nirS and nosZ in paddy soil with CT, RT and NT treatments were significantly increased. The principal coordinate analysis indicated that tillage management and crop residue returning management were the most and the second important factors for the change of denitrifying bacteria community, respectively. Meanwhile, this study indicated that activity and community composition of denitrifiers with CT, RT and NT treatments were increased, compared with RTO treatment. This result showed that nirK, nirS and nosZ-type denitrifiers communities in crop residue applied soil had higher species diversity compared with crop residue removed soil, and denitrifying bacteria community composition were dominated by Gammaproteobacteria, Deltaproteobacteria, and Betaproteobacteria. Therefore, it is a beneficial practice to increase soil PDR level, abundance and community composition of nitrogen-functional soil microorganism by combined application of tillage with crop residue management.

• Korean Journal of Microbiology
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• v.50 no.2
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• pp.137-151
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• 2014

Various studies have been conducted to investigate effects of dams on river ecosystems, but less information is available regarding damming impacts on downstream denitrification. We measured denitrification enzyme activity (potential denitrification rate) and denitrifier abundances (using nirS, nirK, and nosZ as markers) in dammed headstreams of the Nakdong River in South Korea. Sediments in Phragmites-dominated riparian areas and in-stream areas across streams (dammed vs. reference) with different streambed materials (gravel and sand) were sampled occasionally. We hypothesized that (i) the higher available N and C contents in sediments downstream of dams foster larger denitrifier communities than in the reference system and (ii) differences in potential denitrification rates across the systems correspond with denitrifier abundances. Despite 30 years of different hydrological management with dams and greater inorganic N and DOC contents in sediments downstream of dams, compared to the references, abundances of denitrifier communities and potential denitrification rates within the whole sediment were not significantly different across the systems. However, nirS and nosZ denitrifier abundances and potential denitrification rates were considerably increased in specific sediments downstream of dams (gravelly riparian and sandy in-stream) with regard to flooding events and seasonal temperature variation. nirK was not amplified in all sediments. Canonical correspondence analyses (CCA) revealed that the relationship between abundances of denitrifier communities and nutrient availabilities and potential denitrification rates was a weak one.

• Journal of Life Science
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• v.12 no.5
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• pp.505-514
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• 2002

The aim of this study was to evaluate the anticonvulsant activity of four agents derived from tropinone (T-1: 2,4-dipyrrolylmethenylnortropinone, T-2: 2,4-diphenylmethenylnortropinone, T-3 : 2,4-difurfurylmethenylnortropinone, T-4 : 2,4-dimethoxyphenylmethenylnortropinone) in NIH Swiss mouse. Pentylenetetrazole (nZ) was injected via intraperi-toniurn in mouse and Maximal Electroshock (MES) stimulation was through both conjunctivas by electrodes. Tropinone derivatives were treated at 15 minutes before PTZ or MES procedure. PIZ of 25 mg/kg induced generalized seizure in mouse, effects of tropinone derivatives on PTZ-induced seizure were monitored. Compared with control group, T-4 decreased seizure grade most effectively. Also T-4 increased onset time of PTZ-induced seizure. This result showed that T-4 is most effective on PTZ-induced seizure. In MES-induced seizure, T-1 decreased seizure grade and recovery time. nNOS expression in hippocampus and cortex were increased in nZ- and MES-induced seizure animals compared with control. Pretreatment of tropinone derivatives in PTZ-induced seizure did not affected nNOS expression in brain tissues, but T-1 and T-4 decreased nNOS expression in cortex of MES-induced seizure animals. These findings suggest that tropinone derivatives have specific anticonvulsant activities according to PTZ- and MES-induced seizure. 2,4- dimethoxyphenylmethenylnoroopinone is most effective in PTZ-induced seizure and 2,4-di methoxyphenylmethenylnortropinone is most effective in MES-induced seizure.

• Molecular & Cellular Toxicology
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• v.4 no.1
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• pp.11-15
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• 2008

The expression of neuronal nitric oxide synthase (nNOS) is regulated by various spliced first exons (exon 1a-1i), sharing differentially common exon 2 in diverse human tissues. The highly complex structure and regulation of human nNOS gene gave limitations of information for the precise mechanism of nNOS regulation. In the present study, we report that the repeats of polymorphic dinucleotides $(GT)^nA(TG)^n$ repeats located in just upstream to the exon 1f in human nNOS gene play suppressive role in transcription, as shown in the characteristics of Z-DNA motif in other genes. In neuronal and trophoblast cells transfected transiently with luciferase construct without dinucleotide repeats at the 5'-flanking region of exon 1f in nNOS gene, the luciferase activity was increased markedly. However, the presence of the dinucleotide repeats dramatically suppressed the luciferase activity to the basal level, and which was dependent on the length of $(GT)^n$ and $(TG)^n$ repeats. More importantly, we found the polymorphisms in the length of dinucleotide repeats in human. Furthermore, we show for the first time here that there is a significant association of the lengths of polymorphic dinucleotide $(GT)^n$ and $(TG)^n$ repeats with the risk of schizophrenia.

• Environmental Engineering Research
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• v.20 no.1
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• pp.51-57
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• 2015

As expected, the expression of denitrifying genes in a Typha wetland (relatively stagnant compared to other ponds), showing higher nitrogen removal efficiency in summer, was affected by temperature. The abundance and gene transcripts of nitrate reductase (narG), nitrite reductase (nirS), nitric oxide reductase (norB), and nitrous oxide reductase (nosZ) genes in seasonal sediment samples taken from the Acorus and Typha ponds of free surface flow constructed wetlands were investigated using quantitative polymerase chain reaction (Q-PCR) and quantitative reverse transcription PCR (Q-RT-PCR). Denitrifying gene copy numbers ($10^5-10^8$ genes $g^{-1}$ sediment) were found to be higher than transcript numbers-($10^3-10^7$ transcripts $g^{-1}$ sediment) of the Acorus and Typha ponds, in both seasons. Transcript numbers of the four functional genes were significantly higher for Typha sediments, in the warm than in the cold season, potentially indicating greater bacterial activity, during the relatively warm season than the cold season. In contrast, copy numbers and expression of denitrifying genes of Acorus did not provide a strong correlation between the different seasons.

• Korean Journal of Biological Psychiatry
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• v.19 no.1
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• pp.45-52
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• 2012

Objectives : This study was designed to investigate the usability of IVA + Plus (Continuous Performance Test) and Neuro-Guide [Quantitative electroencephalography (EEG) normative database] as an auxiliary diagnostic tools for attention-deficit hyperactivity disorder (ADHD). Methods : The scores of IVA + Plus and resting EEG were obtained from 34 elementary school-aged children. Also, the Korean ADHD Rating Scale (K-ARS) and the Diagnostic Interview Schedule for Children version IV (DISC-IV) was done for the parent of them. From the result of the DISC-IV, we divided them into three groups, ADHD Not Otherwise Specified (NOS), and Normal Control (NC). Using NeuroGuide, the z-scores of relative power for delta (1-4 Hz), theta (4-8 Hz), alpha (8-12 Hz), and beta (12-25 Hz) were calculated. Then the correlation and variance analysis were done to investigate the differences between three groups. Results : The scores of IVA + Plus were negatively correlated with the K-ARS. IVA + Plus have successfully discriminated the ADHD from NC and NOS. The z-scores of relative power of delta and theta were positively correlated with the K-ARS. The z-scores of relative power of alpha and beta were negatively correlated with the K-ARS. Conclusions : The IVA + Plus and NeuroGuide QEEG test are expected to be used as the valuable tools for diagnosing ADHD accurately.

• Journal of Soil and Groundwater Environment
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• v.19 no.4
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• pp.62-69
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• 2014

In the results of monitoring nitrate concentration in more than 8,000 groundwater wells around agro-livestock, the average and maximum nitrate concentration was 9.4 mg/L and 101.2 mg/L, respectively. Since about 31% of the monitoring wells was exceed the quality standard for drinking water, nitrate control such as remediation or source regulation is required to conserve safe-groundwater in South Korea. Typical nitrate-treatment technologies include ion exchange, reverse osmosis, and biological denitrification. Among the treatment methods, biological denitrification by indigenous microorganism has environmental and economic advantages for the complete elimination of nitrate because of lower operating costs compared to other methods. Major mechanism of the process is microbial reduction of nitrate to nitrite and nitrogen gas. Three functional genes (nosZ, nirK, nirS) that encode for the enzyme involved in the pathway. In this work, we tried to develop simple process to determine possibility of natural denitrification reaction by monitoring the functional gene. For the work, the functional genes in nitrate-contaminated groundwater were monitored by using PCR with specific target primers. In the result, functional genes (nosZ and nirK) encoding denitrification enzymes were detected in the groundwater samples. This method can help to determine the possibility of natural-nitrate degradation in target groundwater wells without multiplex experimental process. In addition, for field-remediation application we selected nitrate-contaminated site where 200~600 mg/L of nitrate is continuously detected. To determine the possibility of nitrate-degradation by stimulated-natural attenuation, groundwater was sampled in two different wells of the site and nitrate concentration of the samples was 300 mg/L and 616 mg/L, respectively. Fumarate for different C/N ratio was added into microcosm bottles containing the groundwater to examine denitrification rate depending on carbon concentration. In the result, once 1.5 times more than amount of fumarate stoichiometry required was added, the 616 mg/L of nitrate and 300 mg/L of nitrate were completely degraded in 8 days and 30 days. The nitrite, byproduct of denitrification process, was also completely degraded during the experimental period.

• Preventive Nutrition and Food Science
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• v.17 no.4
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• pp.274-279
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• 2012

For the quantitative analysis of genetically modified (GM) maize in processed foods, primer sets and probes based on the 35S promoter (p35S), nopaline synthase terminator (tNOS), p35S-hsp70 intron, and zSSIIb gene encoding starch synthase II for intrinsic control were designed. Polymerase chain reaction (PCR) products (80~101 bp) were specifically amplified and the primer sets targeting the smaller regions (80 or 81 bp) were more sensitive than those targeting the larger regions (94 or 101 bp). Particularly, the primer set 35F1-R1 for p35S targeting 81 bp of sequence was even more sensitive than that targeting 101 bp of sequence by a 3-log scale. The target DNA fragments were also specifically amplified from all GM labeled food samples except for one item we tested when 35F1-R1 primer set was applied. A reference plasmid pGMmaize (3 kb) including the smaller PCR products for p35S, tNOS, p35S-hsp70 intron, and the zSSIIb gene was constructed for real-time PCR (RT-PCR). The linearity of standard curves was confirmed by using diluents ranging from $2{\times}10^1{\sim}10^5$ copies of pGMmaize and the $R^2$ values ranged from 0.999~1.000. In the RT-PCR, the detection limit using the novel primer/probe sets was 5 pg of genomic DNA from MON810 line indicating that the primer sets targeting the smaller regions (80 or 81 bp) could be used for highly sensitive detection of foreign DNA fragments from GM maize in processed foods.

• The Korean Journal of Physiology and Pharmacology
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• v.18 no.1
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• pp.25-32
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• 2014

Nitric oxide (NO) is recognized as a mediator and regulator of inflammatory responses. NO is produced by nitric oxide synthase (NOS), and NOS is abundantly expressed in the human dental pulp cells (HDPCs). NO produced by NOS can be cytotoxic at higher concentrations to HDPCs. However, the mechanism by which this cytotoxic pathway is activated in cells exposed to NO is not known. The purpose of this study was to elucidate the NO-induced cytotoxic mechanism in HDPCs. Sodium nitroprusside (SNP), a NO donor, reduced the viability of HDPCs in a dose- and time-dependent manner. We investigated the in vitro effects of nitric oxide on apoptosis of cultured HDPCs. Cells showed typical apoptotic morphology after exposure to SNP. Besides, the number of Annexin V positive cells was increased among the SNP-treated HDPCs. SNP enhanced the production of reactive oxygen species (ROS), and N-acetylcysteine (NAC) ameliorated the decrement of cell viability induced by SNP. However, a soluble guanylate cyclase inhibitor (ODQ) did not inhibited the decrement of cell viability induced by SNP. SNP increased cytochrome c release from the mitochondria to the cytosol and the ratio of Bax/Bcl-2 expression levels. Moreover, SNP-treated HDPCs elevated activities of caspase-3 and caspase-9. While pretreatment with inhibitors of caspase (z-VAD-fmk, z-DEVD-fmk) reversed the NO-induced apoptosis of HDPCs. From these results, it can be suggested that NO induces apoptosis of HDPCs through the mitochondria-dependent pathway mediated by ROS and Bcl-2 family, but not by the cyclic GMP pathway.