• Journal of Korean Society on Water Environment
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• v.31 no.6
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• pp.599-609
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• 2015

In order to investigate why OSA (oxic-settling-anaerobic) process produces less sludge than CAS (conventional activated sludge) process, sequential cultivation through 1^st aerobic-anaerobic-2^nd aerobic conditions, were carried out. Then, the intracellular concentrations of adenosine triphosphate (ATP), nicotinamide adenine dinucleotide (NAD and NADH), and nicotinamide adenine dinucleotide phosphate (NADP and NADPH) were monitored for these three stages. Results showed that the concentrations of these energy substances rapidly decreased through time in both aerobic and anaerobic conditions but the anaerobic culture contained the lower energy level than aerobic culture. The 2^nd aerobic culture that experienced anaerobic condition showed lower concentration of these energy substances than those of the 1^st aerobic culture. Meanwhile, the anaerobic culture corresponding to the sludge holding stage of OSA was subjected to different soluble chemical oxygen demand (SCOD) levels, detention time, and temperature to evaluate the effects of these variations on the energy level difference between the 1^st and 2^nd aerobic stages. The lower the SCOD concentration, the longer detention time; and the higher temperature in the anaerobic stage tended to further reduce the intracellular level of the 2^nd aerobic culture. On the average, the intracellular energy level of the anaerobic and 2^nd aerobic stage were 57.73% and 39.12% of the 1st aerobic culture, respectively. These indicated that the insertion of an anaerobic stage between two aerobic stages could lower the intracellular energy levels, hence the lower the sludge in OSA than CAS process. Moreover, manipulation of the operating conditions of the intervening anaerobic stage can change intracellular energy levels thereby controlling sludge production.

• Tuberculosis and Respiratory Diseases
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• v.50 no.1
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• pp.94-105
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• 2001

Background : Examining the biological susceptibility of Mycobacterium tuberculosis to pyrazinamide (PZA) in vitro is very difficult as PZA is inactive under normal culture conditions. The biological susceptibility test, an enzyme assay for Pzase activity, and a genetic test for pncA gene mutations, were performed in order to predict PZA resistance. Methods : 28 cultured clinical isolates of Mycobacterium tuberculosis were tested. The biological susceptibility was performed by the absolute concentration method using Lowenstein-Jensen media. The PZase activity was tested by means of Wayne's method. A 710-bp region includes the entire open reading frame of pncA was amplified and sequenced. Results : All six strains with positive PZase activity exhibited no pncA mutations with one strain showing a false resistance in the biological susceptibility test. Among the 22 strains with no PZase activity, 21 exhibited showed pncA mutations. In the biological susceptibility test, 20 strains were resistant, and one was susceptible, and the other flied to test. The mutation types varied with ten missense, one silent and one nonsense mutation 1 slipped-strand mispairing, and 6 frameshift mutations. Three strains had an adenine to guanine mutation at position -11 upstream of the start codon. Conclusion : The mutation at the pncA promotor region is frequent at -11 upstream position. Automatic sequencing of pncA is a useful tool for rapid and accurate detection of PZA resistant M. tuberculosis, and for demonstrating the epidemiological relatedness of the PZA resistant M. tuberculosis strains.

• Tuberculosis and Respiratory Diseases
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• v.60 no.4
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• pp.451-463
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• 2006

Background : Reactive oxygen species (ROS) take center stage as executers in ventilator-induced lung injury (VILI). The protein with DNA-damage scanning activity, poly (ADP-ribose) polymerase-1 (PARP1), signals DNA rupture and participates in base-excision repair. Paradoxically,overactivation of PARP1 in response to massive genotoxic injury such as ROS can induce cell death through ${\beta}$ -nicotinamide adenine dinucleotide ($NAD^+$) depletion, resulting in inflammation. The purpose of this study is to investigate the role of PARP1 and the effect of its inhibitor in VILI. Methods : Forty-eight male C57BL/6 mice were divided into sham, lung protective ventilation(LPV), VILI, and PARP1 inhibitor (PJ34)+VILI (PJ34+VILI) groups. Mechanical ventilator setting for the LPV group was $PIP\;15cmH_2O$ + $PEEP\;3cmH_2O$ + RR 90/min + 2 hours. The VILI and PJ34+VILI groups were ventilated on a setting of $PIP\;40cmH_2O$ + $PEEP\;0cmH_2O$ + RR 90/min + 2 hours. As a PARP1 inhibitor for the PJ34+VILI group, 20 mg/Kg of PJ34 was treated intraperitoneally 2 hours before mechanical ventilation. Wet-to-dry weight ratio and acute lung injury (ALI) score were measured to determine the degree of VILI. PARP1 activity was evaluated by using an immunohistochemical method utilizing biotinylated NAD. Myeloperoxidase (MPO) activity and the concentration of inflammatory cytokines such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, and IL-6 were measured in bronchoalveolar lavage fluid (BALF). Results : In the PJ34+VILI group, PJ34 pretreatment significantly reduced the degree of lung injury, compared with the VILI group (p<0.05). The number of cells expressing PARP1 activity was significantly increased in the VILI group, but significantly decreased in the PJ34+VILI group (p=0.001). In BALF, MPO activity, $TNF-{\alpha}$, $IL-1{\beta}$, and IL-6 were also significantly lower in the PJ34+VILI group (all, p<0.05). Conclusion : PARP1 overactivation plays a major role in the mechanism of VILI. PARP1 inhibitor prevents VILI, and decreases MPO activity and inflammatory cytokines.

• Clinical and Experimental Pediatrics
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• v.52 no.4
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• pp.476-480
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• 2009

Purpose : The purpose of this study was to investigate the polymorphisms of the TNF-alpha promotor gene, its susceptibility to Kawasaki disease (KD) and to assess whether the TNF-alpha promotor gene polymorphism was related the risk of coronary artery lesions (CALs). Methods : From January 2003 to January 2007, 51 children (30 boys and 21 girls) with KD and 48 children forming an age-matched control group were studied. DNA from the peripheral blood of all the children was sampled, and the DNA polymorphisms of the 5' flanking regions of the TNF-alpha promoter gene at position -308 [guanine (G) to adenine (A)] were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Then, the relationship between KD and the TNF-alpha promotor gene polymorphisms was evaluated. Results : The A allele frequency of the -308 site of the TNF-alpha promotor gene was 17.6% (9/51) for children with KD and 6.8% (3/48) for the control group children, but this result was not statistically significant. Twenty-four patients experienced CALs within 60 days after the onset of symptoms. KD children with TNF-alpha -308 A allele had lower frequencies of CALs (12.5% versus 22.2%, P>0.05). Conclusion : The DNA polymorphism of the -308 site TNF-alpha gene was not associated with susceptibility to KD and a risk of CALs. Multicenter, large-scale randomized controlled trials are needed for further study.

• Korean Journal of Microbiology
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• v.49 no.2
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• pp.105-111
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• 2013

The Erm family of adenine-$N^6$ methyltransferases (MTases) is responsible for the development of resistance to macrolide-lincosamide-streptogramin B antibiotics through the methylation of 23S ribosomal RNA. Recently, it has been proposed that well conserved amino acids in ErnC' located in concave cleft between N-terminal 'catalytic' domain and C-terminal 'RNA-binding' domain interacts with substrate RNA. We carried out the site-directed mutagenesis and studied the function of the ErmSF R237 mutant in vitro and in vivo. R237 amino acid residue is located in the concave cleft between two domains. Furthermore this residue is very highly conserved in almost all the Erm family. Purified mutant protein exhibited only 51% enzyme activity compared to wild-type. Escherichia coli with R237A mutant protein compared to the wild-type protein expressing E. coli did not show any difference in its MIC (minimal inhibitory concentration) suggesting that even with lowered enzyme activity, mutant protein was able to efficiently methylate 23S rRNA to confer the resistance on E. coli expressing this protein. But this observation strongly suggests that R237 of ErmSF probably interacts with substrate RNA affecting enzyme activity significantly.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.5
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• pp.686-695
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• 2018

Objective: The objective of this study was to investigate the effects of dietary choline supplementation on hepatic lipid metabolism and gene expression in finishing pigs with intrauterine growth retardation (IUGR). Methods: Using a $2{\times}2$ factorial design, eight normal birth weight (NBW) and eight IUGR weaned pigs were fed either a basal diet (NBW pigs fed a basal diet, NC; IUGR pigs fed a basal diet, IC) or a diet supplemented with two times more choline than the basal diet (NBW pigs fed a high-choline diet, NH; IUGR pigs fed a high-choline diet, IH) until 200 d of age. Results: The results showed that the IUGR pigs had reduced body weight compared with the NBW pigs (p<0.05 from birth to d 120; p = 0.07 from d 120 to 200). Increased (p<0.05) free fatty acid (FFA) and triglyceride levels were observed in the IUGR pigs compared with the NBW pigs. Choline supplementation decreased (p<0.05) the levels of FFAs and triglycerides in the serum of the pigs. The activities of malate dehydrogenase and glucose 6-phosphate dehydrogenase were both increased (p<0.05) in the livers of the IUGR pigs. Choline supplementation decreased (p<0.05) malate dehydrogenase activity in the liver of the pigs. Gene expression of fatty acid synthase (FAS) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation decreased (p<0.05) FAS and acetyl-CoA carboxylase ${\alpha}$ expression in the livers of the IUGR pigs. The expression of carnitine palmitoyl transferase 1A (CPT1A) was lower (p<0.05) in the IC group than in the other groups, and choline supplementation increased (p<0.05) the expression of CPT1A in the liver of the IUGR pigs and decreased (p<0.01) the expression of hormone-sensitive lipase in both types of pigs. The gene expression of phosphatidylethanolamine N-methyltransferase (PEMT) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation significantly reduced (p<0.05) PEMT expression in the liver of the IUGR pigs. Conclusion: In conclusion, the lipid metabolism was abnormal in IUGR pigs, but the IUGR pigs consuming twice the normal level of choline had improved circulating lipid parameters, which could be related to the decreased activity of nicotinamide adenine dinucleotide phosphate-generating enzymes or the altered expressions of lipid metabolism-related genes.

• Journal of the Korean Chemical Society
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• v.42 no.5
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• pp.506-511
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• 1998

When the spermine, which is one of the polyamines containing cation in vivo, binds to DNA, it can increase the stability of DNA. At the same time, it can cause B-form to Z-form transformations of DNA. However, because we cannot determine the binding geometry of the spermine to DNA by using spectroscopic methods, nobody can show the accurate binding mechanism of a DNA-spermine complex. Thus, we used DAPI as a spectroscopic probe of spermine, which binding geometry was well known. At the result of base selective binding geometry of spermine to synthetic DNA, the concentration of spermine gets higher, it grows the hydrophobic environment of DAPI which bound the minor groove of adenine-thymine base pair. Simultaneously, spermine seems to bridge the backbones around the minor groove of $poly[d(A-T)_2]$. So that, the intensity of fluorescence spectrum of that shows sudden increasement. In guanine-cytocine base pair, $poly[d(G-C)_2]$, we can suppose that spermine bind to the major groove of that, shoving out the DAPI which is partially intercalated between the base pocket across the major groove of it. In both cases, spermine doesn't show the base selectivity against to DNA.

• Korean Journal of Acupuncture
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• v.24 no.4
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• pp.221-231
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• 2007

목 적 : 본 연구의 목적은 시상하부에서 침처치에 대한 nitric oxide synthase (NOS)발현을 nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d)를 이용한 조직화학 염색법으로 관찰하였다. 실험방법 : 동물은 Balb/c (wild type) 와 Stat 4 knockout (KO) 생쥐를 사용하였다. 염증유도는 1% carrageenan 용액 (20ul/마리)을 발 뒤꿈치 표피에 주사하였고, 침 처치는 족삼리 (ST36)에 시침하였다. 침 처치 후 5시간까지 부종율을 부종측정기로 측정하였으며, 마지막으로 부종을 측정한 후 동물을 희생하여 뇌를 적출하여 고정하였다. 침에 대한 효과를 확인하기 위하여 NADPH-d 반응의 조직염색을 실시하였다. 염증유도와 그룹간의 유의성 검증은 one-way ANOVA를 사용하였다. 결 과 : 대조군인 Balb/c와 실험군인 stat4 KO 생쥐를 carrageenan으로 염증을 유도시에 대조군은 90%이상 유도된 반면, Stat4 KO 그룹은 50% 정도의 염증만이 유도되었다. 염증을 유도한 생쥐의 족삼리에 침 처치시 대조군은 1시간에서 약 40%정도 감소하였고 (P<0.05), Stat4 KO 실험군은 유의한 염증 감소율을 보이지 않았다. 시상하부의 lateral hypothalamic area (LHA)와 paraventricular nucleus (PVN)부위의 침에 대한 효과를 NADPH-d 에 양성으로 반응하는 세포수로 비교하여 다음과 같은 결과를 얻었다. (1) 대조군에서 염증 유도시 시상하부의 PVN는 NADPH-d 양성세포수가 감소하였고, LHA에서는 증가하였다. (2) 염증을 유도한 대조군에 침을 처치시 PVN은 세포수가 증가하였고, LHA에서는 감소하는 경향을 보였다. (3) 염증을 유도한 Stat4 KO 군에서는 시상하부의 PVN과 LHA부위 모두에서 NADPH-d 양성세포수가 감소하였고, 염증유도그룹에 침을 처치시 PVN과 LHA부위 모두에서 세포수가 증가함을 관찰 할 수 있었다. (4) 대조군과 실험군 모두에 salicylic acid로 비교하였더니 염증유도 효과 및 NADPH-d 세포 수에서 침 처치와 비슷한 결과를 나타내었다. 결 론 : 침은 염증을 유도한 생쥐에서 염증 감소에 유의한 효과가 있다. 염증을 유도한 Balb/c 와 Stat4 KO 생쥐에 침을 처치 시 시상하부의 NADPH-d 발현이 LHA부위와 PVN에서 서로 다르게 나타나는 것으로 나타난다. 이러한 현상은 침 효과가 시상하부의 위치에 대한 작용이 다르기 때문이라고 생각된다.

• Journal of Life Science
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• v.22 no.6
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• pp.751-759
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• 2012

Microbulbifer sp. was used to acquire the degrading products from Ecklonia cava (DPEC) and the products were investigated to determine the physiological activities. Firstly, 2,2-diphenyl-1-picrylhydrazyl (DPPH) activity and superoxide dismutase (SOD) assay were about 84.1% and 89.6% at 2.5 mg/ml, respectively. In addition, nitrite scavenging ability was shown to be 56.3% at 0.5 mg/ml on pH 1.2. ${\alpha}$-Glucosidase inhibitory activity was increased in a dose-dependent manner and was about 58.7% at 2.5 mg/ml. To determine the influence of DPEC on alcohol metabolism, the generating activity of reduced-nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) were measured. Facilitating rates of ADH and ALDH activities by DPEC were 123.3% and 215.2% at 2.5 mg/ml, respectively. For analyses of anti-wrinkling and whitening effects, its elastase and tyrosinase inhibitory activities were measured and were about 73.1% and 42.2% at 2.5 mg/ml, respectively. These results indicated that DPEC has valuable biological attributes owing to its antioxidant, nitrite scavenging, and alcohol metabolizing activities and ${\alpha}$-glucosidase, elastase, and tyrosinase inhibitory activities.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.396-400
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• 2015

To investigate the optimal conditions for shoot organogenesis in Astragalus sinicus L., hypocotyl explants were cultured in Murashige & Skoog's (MS) medium supplemented with 0.1, 1.0, 2.0, or 4.0 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) for 6 weeks. 2,4-D concentration significantly effected morphogenesis: some produced calli with adventitious shoots and roots, some produced calli with adventitious roots, some produced only calli, and some produced deep-brownish calli with roots. The formation of calli with shoots and/or roots was observed at lower levels of 2,4-D, whereas calli without shoots or with deep-brownish roots were formed after treatment with higher levels of 2,4-D. Also, a shoot organogenesis ability of callus clones was observed after treatment with medium with 0.1 or 1.0 mg/L 2,4-D grown in MS medium with combinations of benzyl adenine (BA) and 2,4-D for 4 weeks. Medium with a combination of BA and 2,4-D was effective for shoot formation, whereas root organogenesis from calli decreased. The greatest amount of shoot formation was obtained when calli were cultured in MS medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BA. Upon shoot transfer into 1/2 MS basal medium, plantlets developed, and the plantlets grew well in soil in a greenhouse.