• Natural Product Sciences
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• 제16권4호
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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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• 제30권11호
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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.

• 미생물학회지
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• 제50권2호
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• pp.137-151
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• 2014

댐의 존재가 하천 생태계에 미치는 영향을 연구하기 위해 다양한 연구가 지금까지 수행되어 왔지만, 댐이 하류의 탈질화에 미치는 영향은 잘 알려져 있지 않다. 대한민국 낙동강의 댐 원류에서 탈질화 효소 활성도(잠재 탈질율)와 탈질균 분포(nirS, nirK, nosZ 유전자를 표지유전자로 사용)를 조사하였다. 자갈 혹은 모래로 채워진 하천의, 갈대가 우거진 하변지역과 강바닥의 침전물을 채취하여 조사하였다. 이 실험의 가설은 다음과 같다. (i) 하천 침전물의 N과 C의 사용유효량이 높을수록 대조군에 비해 미생물 군집의 탈질화 작용이 더욱 증진한다, (ii) 하천생태계마다 상이하게 나타나는 잠재 탈질율 간의 차이는 탈질 미생물의 양에 비례한다. 30여 년간 댐에 의해 수문학적으로 큰 차이가 있었고 또한 댐 하류의 저서에 무기질소와 용존유기탄소 농도가 대조군에 비해 매우 높았음에도 불구하고, 탈질균 군집의 양과 잠재 탈질율은 하천 간에 큰 차이가 없었다. 하지만 nirS 유전자와 nosZ 유전자의 양과 잠재 탈질율은 댐 하류에 존재하는 자갈이 많은 하변과 모래가 많은 하천 바닥에서 홍수빈도와 계절별 온도변동에 관련하여 크게 증가함을 알 수 있었다. nirK 유전자는 모든 시료에서 발견되지 않았다. Canonical correspondence analysis (CCA) 분석결과는 탈질균 군집 양과 영양염류 가용도와 잠재 탈질율 사이에는 약한 상관관계가 있음을 보여주었다.

• 생명과학회지
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• 제12권5호
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• pp.505-514
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• 2002

생쥐에서 최근 합성된 4가지 종류의 tropinone유도체들의 항경련 효과를 조사하기 위하여 pentylenetetrazole (PTZ) 및 Maximal Electroshock (MES)로 유발된 경련에 tropinone 유도체들이 경련상태에 미치는 효과를 관찰하였다. Troponine 유도체로는 화학구조가 다른 4가지 종류를 사용하였다(T-1:2,4-dipywolylmethenylnortropinone, T-2:2,L di phenylme thenylnortropinone, T-3 : 2,Ldifurfurylmetheny- Inortropinone, 74 : 2,4-dimetho xyphenylmethenylnortro- pinone). nZ 25 mg/kg을 복강 내로 투여 후 전신성 경련을 유발하였으며 tropinone 유도체를 전처치한 후 PTZ에 의한 경련의 변화를 관찰하였다. 대조군에 비하여 T-1과 T-2는 경련정도에 변화가 없었으나 T-3과 T-4는 유의하게 경련정도를 약화시켰다. PTZ에 의한 경련의 시작 시간은 T-4에서 유의하게 지연되어 T-4가 PTZ에 의한 경련에 항경련 효과가 있음을 나타내었다. MES로 경련을 유발한 경우에 있어서는 T-1이 경련정도를 유의하게 약화시켰으며 경련 후 회복시간도 T-1에서 가장 빨리 회복되는 특성을 보였다. 따라서 T-1이 MES에 의한 경련유발에 항경련 효과가 있음을 나타내었다. Troponine 유도체에 의한 경련 억제 효과와 경련과 동반되어 증가한다고 알려진 neuronal nitric oxide synthase (nNOS) 발현과의 관계를 알아보기 위하여 조직 단백질에서 Western blot을 하였다. 대조군에 비하여 PTZ 및 MES에 의해 경련을 유발한 생쥐에서 모두 해마부 및 전뇌피질부에서 nNOS가 증가하였다. Tropinone 유도체를 투여하지 않고 경련을 유발시킨 대조군에 비하여 tropinone 유도체를 투여한 군에서도 모두 nNOS의 발현이 해마부 및 전뇌피질부에서 증가되었다. MES로 경련을 유발한 생쥐에서 대조군에 비하여 T-1 및 T-4는 피질부에서 nNOS가 감소했으나 나머지군에서는 감소가 없었다. 이 상의 결과를 토대로 tropinone 유도체들은 경련유발의 자극 조건에 따라 항경련 효과가 다르게 나타났으며, PTZ유 발경련에서 2,4-dimethoxyphenylmethenylnortropinone의 항경련 효과가 가장 크고, MES 유발경련에서는 2,4-dipyrrolylmethenylnortropinone의 항경련 효과가 가장 크게 나타났다.

• Molecular & Cellular Toxicology
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• 제4권1호
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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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• 제20권1호
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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.

• 생물정신의학
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• 제19권1호
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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.

• 한국지하수토양환경학회지:지하수토양환경
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• 제19권4호
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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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• 제17권4호
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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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• 제18권1호
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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.