• Journal of Pharmaceutical Investigation
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• v.37 no.5
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• pp.297-303
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• 2007

The purposes of this study were to clarify the involvement of P-glycoprotein (P-gp) in the efflux of levosulpiride in knockout mice that lack the mdr1a1b gene and to evaluate the relationship between the genetic polymorphisms in MDR1 gene (exon 21) and levosulpiride disposition in healthy Korean subjects. After oral administration ($10\;{\mu}g/g$) of levosulpiride to mdr1a/1b(-/-) and wild-type mice, plasma and brain samples were obtained at 45 min. We also investigated the genotype for MDR1 (exon 21) gene in humans using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. A single oral dose of 25 mg levosulpiride was administered to 58 healthy subjects, who were based on the MDR1 genotype for the G2677T SNP. Blood samples were taken up to 36 hr after dosing. The concentrations of levosulpiride in mouse plasma and brain were statistically significant difference between the two animal groups (P<0.05). In addition, the average brain-to-plasma concentration ratio (Kp) of levosulpiride was 3.4-fold (P<0.01) higher in the mdr1a/1b(-/-) mice compared with the wild-type mice. We also found that the values of $AUC_{0-{\infty}$, partial AUC ($AUC_{0-4h}$) and $C_{max}$ were significantly different between homozygous 2677TT subjects and the subjects with at least one wild-type allele (GG and GT subjects, P=0.012 for $AUC_{0-{\infty}$; P=0.008 for $AUC_{0-4h}$; P=0.038 for $C_{max}$). The results confirm that levosulpiride is a P-gp substrate in vivo, and clearly demonstrate the effect of SNP 2677G>T in exon 21 of the MDR1 gene on levosulpiride disposition.

• Journal of Pharmaceutical Investigation
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• v.39 no.5
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• pp.345-351
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• 2009

Organic cation transporters (OCTs) are important for absorption, elimination of many endogenous small organic cations as well as a wide array of drugs and environmental toxins. This gene is located in a cluster on chromosome 6 and OCTs are in major organs such as intestine, liver, kidney, brain and placenta. Therefore, expression levels and function of OCTs directly affect plasma levels and intracellular concentrations of drugs and thereby determine therapeutic response. The aim of this study was to investigate the frequency of the SNPs on OCT1 (C181T and C1022T) and OCT2 (G808T) to analyze haplotype frequency in healthy Korean population. Human subjects have been genotyped for OCT1 (C181T for 195 subjects and C1022T for 825 subjects), using polymerase chain reaction-based diagnostic tests (RFLP). And for OCT2 (G808T), a total of 861 subjects have been genotyped, using pyrosequencing method. Haplotype was statistically inferred using an algorithm based on the expectation-maximization (EM). OCT1 C181T genotyping showed 100% homozygous wild-type (C/C). OCT1 C1022T genotyping showed wild-type (C/C), heterozygous (C/T) and homozygous mutant-type (T/T) and each accounted for 72.1, 24.5 and 3.4%, respectively. OCT2 G808T genotyping results also showed homozygous wild-type (G/G), heterozygous (G/T) and homozygous mutant-type (T/T) and each took 81.8, 17.9 and 0.3%, respectively. Based on these genotype data, haplotype analysis between OCT1 C181T and OCT1 C1022T has proceeded. The result has revealed that linkage disequilibrium between alleles is not obvious (P=0.0122).

• Korean Journal of Agricultural Science
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• v.39 no.2
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• pp.219-226
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• 2012

The acyl-CoA dehydrogenase, C-2 to C-3 short chain (ACADS) gene is known to be related with fat metabolism, especially coverts the fat to the energy sources in cattle. In human, the mutations in this gene cause SCAD deficiency, which is one of the fatty acid metabolism disorders. The ACADS gene is located on bovine chromosome 17. The objective of this study was to identify SNPs in Hanwoo ACADS gene and identify the relationships with economic traits. In this study, two SNPs, T1570G SNP in exon 2 and G13917A SNP in exon 4, were observed. Moreover, in the coding region, 2 missense mutations, T (Cys) ${\rightarrow}$ G (Trp) mutation at 1570 bp and G (Arg) ${\rightarrow}$ A (Gln) mutation at 13917 bp, were observed. These mutations were subjected to the PCR-RFLP for typing 198 Hanwoo animals. The observed genotype frequency for T1570G was 0.135 (TT), 0.860 (TG) and 0.005 (GG), respectively. Also, 0.900 (GG) and 0.100 (GA) were observed for the G13917A mutation. The association of these SNPs with four economic traits, CW (Carcass Weight), BF (Backfat Thickness), LMA (Longissimus Muscle Area), MS (Marbling Score), were also observed. The results indicated that no significant results were observed in all four traits (P>0.05). This might indicate that further studies are ultimately needed to use the SNPs in ACADS gene in lager populations for effectively used for the marker assisted selection.

• Korean Journal of Agricultural Science
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• v.40 no.3
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• pp.215-220
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• 2013

Fatty acids (FAs) were considered in activating nuclear hormone receptors that play significant roles in the cellular lipid metabolism by the regulation of several genes. Previously, fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD) genes have been known to regulating the FA metabolism. In this study, associations of FASN and SCD genes with fatty acid (FA) composition in broilers were investigated. Tissue samples from 95 Cobb 500 broilers were used for DNA extraction. The g.1222 A>G SNP located in intron 42 of FASN gene and 2 SNPs in SCD gene, one in exon 2 (g.3728A>G) and the other in exon 4 (g.12903G>A), were subjected for genotyping using PCR-RFLP method. One of the SNPs in SCD gene, SNP g.3728A>G had significant association with myristoleic acid (C14:1; P<0.05), palmitic acid (C16:0; P<0.05), palmitoleic acid (C16:1; P<0.05) and saturated FA (SFA; P<0.05). However, the SNP g.1222A>G in FASN gene had only suggestive association with arachidic acid (C20:0; P=0.08). The findings in this study suggest that the SNP in exon 2 of SCD gene can be used as a molecular marker for selecting birds having desirable FA composition in broilers.

• The Journal of Korean Medicine
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• v.26 no.1 s.61
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• pp.85-92
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• 2005

Objective: A number of candidate genes have been in implicated in the pathogenesis of obesity in humans. Tumor necrosis factor-alpha $(TNF-{\alpha})$ is expressed primarily in adipocytes, and elevated levels of this cytokine have been linked to obesity and insulin resistance. Recently, the A allele of a polymorphism at position 308 in the promoter region of $TNF-{\alpha}$ (G-308A) has been shown to increase transcription of the gene in adipocytes. Therefore, we designed this study to test whether obese and non-obese subjects differ in $TNF-{\alpha}$ genotype distribution, and how the genotypes affect anthropometric parameters, including degrees of body mass index (BMI). Methods : The study included 153 obese but otherwise healthy women ($BMI{\geq}kg/m^2$, range 25-54.7, age range 15-40 years) and 82 non-obese healthy women ($BMI, age range 15-40 years). Total fat mass and percent body fat were determined by dual-energy X-ray absorptiometry. Genomic DNA was extracted and used for Ncol restriction fragment length polymorphism (RFLP) based genotyping of $TNF-{\alpha}$. Results: No differences were observed for allelic and genotype frequencies between the obese ($BMI{\geq}25$) and non-obese women. Also, no association of TNF-(l polymorphism was observed with body mass index (BMI) for genotype in obese women. In addition, age, pertent body fat, BMI, and cholesterol levels did not differ by $TNF-{\alpha}$ genotype. However, waist-to­hip ratio (WHR) was significantly lower in subjects with $TNF-{\alpha}$ GA or AA genotype (0.94 0.07 vs. 0.920.03, P<0.005). Conclusion: These results suggest that $TNF-{\alpha}$ promoter polymorphism at position-308 is not a significant factor for BMI, but affects the WHR in obese healthy women from Koreans.

• The Journal of Korean Medicine
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• v.20 no.4
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• pp.16-22
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• 2000

This study intends to report the significance of several experimental results obtained from analysing the genes extracted from the plants and herbal medicine such as P. temalta (Thunb.) Breit, A. amurense var serratum Nakai, A. erubescens (Wall.) Schott, Pinelliae Tuber and Arisaematis Tuber, mainly by the method of RAPD(randomly amplified polymorphic DNA) and the method of RFLP(restriction fragment length polymorphism) on ITS(internal transcribed spacer) region. Genomic DNA could be extracted from both original plants and dried materials. DNA fragments of P. temata kind and A. amurense kind showed the same aspect separately within the same species under the method of RAPD using random primer, while various aspects(polymorphism) were discovered among different species. In RAPD analysis by uniprimer, common bands were extracted from all types of P. temata in the case of uniprimer #4, which were distinguished from the kind of A. amurense. Other polymorphic bands appeared in between different A. amurense species as well. In the case of uniprimer #11, particular band came out in the kind of P. temata. On the other hand, in the case of uniprimer #5, #6, and #8, various bands(polymorhism) were revealed in both kinds of P. temata and A. amurense. Although further study is needed to ascertain whether these results are due to the differences of species, kinds, or growing place, the results could be used as a scientific method of identifying the substitutes for A. amurense genus. The author believes that as if P. temata class of plants used in this experiment are different among themselves in terms of the shape, size and property, those are clearly a class of P. temata or belong to the same genus.

• The Journal of Korean Medicine
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• v.28 no.1 s.69
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• pp.51-62
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• 2007

Objectives . The types of Sasang constitutional medicine (SCM) have definite effect on response to herbal drugs. The majority of human P45O dependent xenobiotic metabolism is carried out by polymorphic enzymes which can cause abolished, altered or enhanced metabolism. Therefore, we evaluated the relation of major CYP2C19, 2D6 polymorphism with Sasang types. Methods : 214 healthy subjects were recruited with informed consent; 172 among them had Sasang diagnosis by QSCC2. CYP2D6, 2C19 polymorphism were determined by PCR-RFLP method. Results : None of the Sasang types showed significant difference in CYP2D6, 2C19 polymorphism. However, the Tae-um type showed relatively low frequency of CYP2D6 $^{*}$10/$^{*}$10 polymorphisms with low activity (p=0.110). In the So-yang type, specific $^{*}$3/$^{*}$3 genotype which is a poor metabolizer of CYP2C19$^{*}$3 was detected (p=0.078).Conclusion . These results suggest that the Tae-um type which is said to have high liver function in SCM has the tendency of high drug-metabolizing enzyme activity. With further study, the CYP polymorphism could serve as a scientific tool for SCM diagnosis.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1826-1835
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• 2008

An autotrophic denitrification reactor (ADR-l) and a heterotrophic denitrification reactor (HDR-2) were operated to remove nitrate and nitrite in an anoxic environment in raw sewage. The $NO_3$-N removal rate of ADR-l was shown to range from 52.8% to 78.7%, which was higher than the $NO_3$-N removal rate of HDR-2. Specific denitrification rates (SDNR) of ADR-l and HDR-2 were 3.0 to 4.0 and 1.1 to $1.2\;mgNO_3$-N/gVSS/h, respectively. From results of restriction fragment length polymorphism (RFLP) of the 16S rRNA gene, Aquaspirillum metamorphum, Alcaligenes defragrans, and Azoarcus sp. were $\beta$-Proteobacteria that are affiliated with denitritying bacteria in the ADR-l. Specifically, Thiobacillus denitrificans was detected as an autotrophic denitrification bacteria. In HDR-2, the $\beta$-Proteobacteria such as Denitritying-Fe-oxidizing bacteria, Alcaligenes defragrans, Acidovorax sp., Azoarcus denitrificans, and Aquaspirillum metamorphum were the main bacteria related to denitrifying bacteria. The $\beta$-and $\alpha$-Proteobacteria were the important bacterial groups in ADR-l, whereas the $\beta$-Proteobacteria were the main bacterial group in HDR-2 based on results of fluorescent in situ hybridization (FISH). The number of Thiobacillus denitrificans increased in ADR-l during the operation period but not in HRD-2. Overall, the data presented here demonstrate that many heterotrophic denitritying bacteria coexisted with autotrophic denitrifying bacteria such as Thiobacillus denitrificans for nitrate removal in ADR-l. On the other hand, only heterotrophic denitritying bacteria were identified as dominant bacterial groups in HDR-2. Our research may provide a foundation for the complete nitrate removal in raw sewage of low-COD concentration under anoxic condition without any external organic carbon or the requirement of post-treatment.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.173-177
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• 2000

연구목적: 한국여성의 다낭성 난포증후군 환자에서 황체형성 호르몬 exon 2 유전자의 변이를 탐색하여 이들 변이와 질환과의 관련성 여부를 밝히고자 하였다. 연구재료 및 방법: 21명의 다낭성 난포증후군 환자를 대상으로 황체형성 호르몬 exon 2(Trp8Arg;TGG to CGG and Ile15Thr; ATC to ACC)의 변이를 탐색하였다. 혈액에서 Genomic DNA를 추출하여 PCR로 증폭한 후 RFLP 방법으로 변이형을 구분하였다. 결과: 황체형성 호르몬 exon 2의 변이형이 다낭성 난포증후군 환자에서 28.6%로 이미 조사된 바 있는 대조군의 16.7% 보다 약간 높게 나타났으나 통계적으로 유의한 차이는 없었다(p>0.05). 결론: 황체형성 호르몬 exon 2의 변이가 한국인의 다낭성 난포증후군 발병과 관련이 있는지를 밝히기 위해 더 많은 개체에 대한 연구가 요구된다.