• 대한환경공학회지
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• 제33권9호
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• pp.662-669
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• 2011

본 연구에서는 메탄의 생물학적 메탄올 전환에 관한 연구를 수행하였다. 바이오가스 중의 메탄은 메탄산화균의 methane monooxygenase (MMO)의 생물학적 촉매반응에 의해 산화되었으며, 인산염, NaCl, $NH_4Cl$, EDTA와 같은 methanol dehydrogenase (MDH)의 활성 저해제를 이용하여 MDH의 활성도를 저해함으로써 메탄올의 전환이 이루어졌다. 메탄산화균은 $35^{\circ}C$, pH 7, 인공 바이오가스($CH_4$ 50%, $CO_2$ 50%) / Air의 부피비가 0.4인 조건에서 메탄 산화 정도가 0.56 mmol로 최대로 나타났다. 인산염 40 mM, NaCl 50 mM, $NH_4Cl$ 40 mM, EDTA $150{\mu}m$ 이하일 때 저해제의 종류에 상관없이 메탄 산화율은 80% 이상을 달성하였다. 한편, 인산염 40 mM, NaCl 100 mM, $NH_4Cl$ 40 mM, EDTA $50{\mu}m$ 주입 시 각각 1.30, 0.67, 0.74, 1.30 mmol의 메탄이 산화되는 동시에 각각 0.71, 0.60, 0.66, 0.66 mmol의 메탄올이 최대로 생성되었다. 이때의 메탄올 전환율은 각각 54.7, 89.9, 89.6 및 47.8%였으며 최대 메탄올 생성 속도는 $7.4{\mu}mol/mg{\cdot}h$였다. 이로부터 대상 저해제로 MDH 활성도를 일반적으로 35% 저해 시에 메탄올 생산량이 최대인 89.9%까지 나타남을 알 수 있었다.

• Tuberculosis and Respiratory Diseases
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• 제49권2호
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• pp.189-197
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• 2000

연구배경 : 폐암은 진단 당시 이미 국소 침윤이나 원격 전이가 된 경우가 많고 이에 대한 적절한 치료법이 없기 때문에 예후가 불량하다. TIMP(tissue inhibitor of metalloproteinase)는 암세포의 침윤 및 전이에 중요한 역할을 하는 metalloproteinase를 억제하는 물질로서 생체 내에 존재하는 전이 억제 물질이다. 본 연구는 아데노바이러스를 이용한 TIMP 유전자 치료법을 개발하여 폐암의 치료에 응용하고자 하였다. 방법 : 폐암세포는 침윤 및 전이 능력이 큰 Calu-6를 사용하였다. TIMP-2 유전자를 pACCMVpLpA에 subcloning 한 후 pJM17과 함께 293 cell에 cotransfection 한 후 homologous recombination을 이용하여 Ad-TIMP-2를 제작하였다. Ad-TIMP-2를 Calu-6 cell에 이입하여 TIMP-2 protein 이 생산되는지를 TIMP-2 ELISA를 이용하여 확인하였고 TIMP-2의 생물학적 활성은 zymography로 확인하였다. Soft agar clonogenic assay로 종양형성능을 평가하였다. Ad-TIMP-2로 처리한 calu-6를 6주간 soft agar에서 키운 후 육안으로 보이는 colony 수를 측정하였다. Matrigel을 이용하여 invasion assay를 시행하여 calu-6의 침윤 능력의 변화를 평가하였다. 결과 : TIMP-2 ELISA 결과, 모세포 calu-6와 Ad-$\beta$-gal 이입 calu-6 그리고 Ad-TIMP-2 이입 calu-6는 각각 0.44, 0.43, 20.7 ${\mu}g/10^6$ cells/72hrs의 TIMP-2를 생산하였다. Zymography 결과 Ad-TIMP-2에 의해 생산된 TIMP-2는 matrix metalloproteinase-2의 gelatin 분해 효과를 억제하여 생물학적 활성을 확인할 수 있었다. Soft agar clonogenic assay 결과, 모세포인 calu-6는 453$\pm$53개, Ad-$\beta$gal과 Ad-TIMP-2 이입된 calu-6는 각각 332$\pm$35, 280$\pm$45개의 colony가 형성되어 유의한 감소를 보이지 못했다. Invasion assay 로 모세포 calu-6 에 대한 침윤율을 평가한 결과, Ad-$\beta$gal과 Ad-TIMP-2가 이입된 calu-6(10moi)는 각각 71$\pm$8.9%, 12$\pm$8.4%의 침윤율을 보였으며 $\beta$-gal 군에 비해 TIMP-2군이 유의하게 침윤율이 낮았다. 결론 : Ad-TIMP-2는 폐암 세포의 종양형성능을 억제하지 못하였으나 침윤은 억제하여 TIMP-2가 폐암 유전자 요법에 이용될 가능성을 제시해 주었다.

• The Korean Journal of Pain
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• 제13권2호
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• pp.164-174
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• 2000

Background: After an injury to tissue such as the skin, hyperalgesia develops. Hyperalgesia is characterized by an increase in the magnitude of pain evoked by noxious stimuli. It has been postulated that in the mechanism of hyperalgesia (especially secondary hyperalgesia) and allodynia, a sensitization of central nervous system such as spinal dorsal horn may contribute to development of hyperalgesia. However, the precise mechanism is still unclear. In the present study, we investigated the roles of N-methyl-D-aspartate (NMDA) receptor and nitric oxide (NO) system in the mechanism of hyperalgesia, and their relations with c-fos expression Methods: Inflammation was induced by injection of complete Freund adjuvant (CFA) into unilateral hindpaw of Sprague-Dawley rat. Behavioral studies measuring paw withdrawal responses by von Frey filaments and paw withdrawal latencies by radiant heat stimuli and stainings of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase and c-fos immunoreactivity were performed. The effects of MK-801, an NMDA receptor blocker and $N^\omega$-nitro-L-arginine (L-NNA), a nitric oxide synthase (NOS) inhibitor were evaluated. Results: 1) Injection of CFA induced mechanical allodynia, mechanical hyperalgesia and thermal hyperalgesia. And it increased the number of NADPH-diaphorase positive neurons and c-fos expression neurons. 2) MK-801 inhibited mechanical hyperalgesia and thermal hyperalgesia induced by CFA and reduced the number of NADPH-diaphorase positive neurons and c-fos expression neurons. 3) L-NNA inhibited the thermal hyperalgesia and reduced the number of NADPH-diaphorase positive neurons, but did not affect the number of c-fos expression neurons. Conclusions: These results suggest that in the mechanism of mechanical hyperalgesia, NMDA receptor but not NO-system is involved and in the case of thermal hyperalgesia both NMDA receptor and NO system are involved. NO system did not affect the expression of c-fos, but c-fos expression and NOS activity were dependent on the activity of NMDA receptor.

• 한국환경농학회지
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• 제11권1호
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• pp.1-8
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• 1992

농약분해에 관여하는 monooxygenase와 esterase의 활성을 저해하는 것으로 알려진 PBO와 TPP를 첨가하여 제조한 신(新)다이아지논입제에 대하여 벼멸구에 대한 살충률을 조사하였고, 아울러 신(新)다이아지논 입제와 두효소를 저해하는 작용기작을 가지고 있는 tricyclazole, carbofuran 그리고 EPN을 혼합하여 제조한 혼합다이아지논입제에 대하여 담수토양중의 잔류경향을 조사하였다. 1. 살균토양과 비살균토양에서 신(新)다이아지논 입제(0.1% 첨가)의 반감기는 4.53일과 2.33일 이었고, 시판품에 비하여 각각 0.74일과 0.45일 주성분의 분해가 지연되었다. 2. 신(新)다이아지논입제(0.1% 첨가)의 벼멸구에 대한 살충률은 추천량에서는 12%, 추천 1/2량에서는 $30{\sim}60%$가 증가되었다. 3. 신(新)다이아지논 입제 (1%)중 PBO 첨가는 토양중 반감기가 0.44일 그리고 PBO-TPP첨가로 0.65일 지연되었다. 4. 혼합다이아지논 입제는 시판품에 비하여 tricyclazole, carbofuran 그리고 EPN첨가로 반감기가 2.61일, 1.04일 그리고 0.43일 지연되었으며, EPN+carbofuran을 첨가하여 제조한 혼합다이아지논 입제는 2.7일 연장되었다.

• The Korean Journal of Physiology and Pharmacology
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• 제3권4호
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• pp.447-454
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• 1999

The production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) are known to be modulated by a variety of factors. Recent study showed that endotoxin-induced NO synthesis and iNOS expression were greatly enhanced by elevation of extracellular glucose concentration in murine macrophages. Although this was suggested to be due to the activation of protein kinase C (PKC) via sorbitol pathway, there was lack of evidence for this speculation. This study was performed to delineate the underlying intracellular mechanisms of glucose-enhancing effect on endotoxin-induced NO production in Raw264.7 macrophages. The levels of NO release induced by lipopolysaccharide (LPS) significantly increased by the treatment of glucose in a concentration dependent manner and also, this effect was observed in LPS-preprimed cells. Concurrent incubation of cells with PKC inhibitors, H-7 or chelerythrine, and LPS resulted in the diminution of NO production regardless of glucose concentration but this was not in the case of LPS-prepriming, that is, chelerythrine showed a minimal effect on the glucose- enhancing effect. PMA, a PKC activator, did not show any significant effect on glucose-associated NO production. Modulation of sorbitol pathway with zopolrestat, an aldose reductase inhibitor, did not affect LPS-induced NO production and iNOS expression under high glucose condition. And also, sodium pyruvate, which is expected to normalize cytosolic $NADH/NAD^+$ ratio, did not show any significant effect at concentrations of up to 10 mM. Glucosamine marginally increased the endotoxin-induced nitrite release in both control and high glucose treated group. 6-diazo-5-oxonorleucine (L-DON) and azaserine, glutamine: fructose- 6-phosphate amidotransferase (GFAT) inhibitors, significantly diminished the augmentation effect of high glucose on endotoxin-induced NO production. On the other hand, negative modulation of GFAT inhibitors was not reversed by the treatment of glucosamine, suggesting the minimal involvement, if any, of glucosamine pathway in glucose-enhancing effect. In summary, these results strongly suggest that the hexosamine biosynthesis pathway and the activation of PKC via sorbitol pathway do not contribute to the augmenting effect of high glucose on endotoxin induced NO production in macrophage-like Raw264.7 cells.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.207-207
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• 2017

The proso millet is vulnerable to lodging due to high plant height and shallow root. A lodging results in a hard mechanical harvesting and yield loss. One of solutions on this problem is inhibition of internode elongation. The objective of this study was to set up use time and dose of prohexadium-calcium, is growth inhibitor. The experimental variety was Ibaekchal. The experiment design was a split-plot design with three replications. The treatments were as follow: Main-plots were 25 and 35 day after sowing(DAS) as use time and sub-plots were 0%, 50%, 100%(diluted solution of 1000 times, $1000{\ell}\;ha^{-1}$), 150% as dose. The amount of nitrogen, phosphate and potassium fertilization were 90, 70, $80kg\;ha^{-1}$, respectively. The size of high ridge and plant spacing were $90{\times}30cm$ and $60{\times}15cm$, respectively. Proso millet was sown on June 9, 2016 by hands and was adjusted at 2 plant per hill. The growth survey of vegetative growth stage was conducted at 1 day before treatment and with one week interval after treatment. Data were collected: (1) grain yield: weight of grain in $kg\;ha^{-1}$, (2) 1000 grain weight: average weight of 1000 grain, (3) plant height: distance from soil to top of panicle or leaf in cm, (4) ear length: distance from top of stem to top of ear in cm, (5) stem diameter: diameter of second internode (6) degree of lodging: percentage of lodging area, etc. Analyses of variance were performed using R version 3.3.1(https://www. r- project. org). The Duncan's multiple range test(DMR) was used to separate treatment means at P < 0.05. There was a significant difference in plant height and number of stem among the use time and dose of prohexadium-calcium during vegetative growth stage. At 25 DAS, the difference with no treatment increased until 25 day after treatment and decreased since then. The difference in number of stem increased until 18 day and decreased since 25 day. At 35 DAS, the difference with no treatment in plant height and number of stem increased until 22 day after treatment and decreased since then. We assumed that the effect of prohexadium-calcium was inhibition of internode elongation and promotion of tillering, continued untel 25day after treatment. At 25 DAS, the degree of lodging deceased to 100%, 30%, 10% and 0% as dose increased. At 35 DAS, the degree of lodging decreased to 100%, 20%, 0% and 0% as dose increased. At 25 DAS, the yield was 2910, 2710, 3190, $2310kg\;ha^{-1}$ among dose. At 35 DAS, the yield was 2750, 2630, 2220, $2050kg\;ha^{-1}$. We recommend that the optimum use time and dose of prohexadium-calcium for proso millet is 1000 times diluted solution of $1000{\ell}$ per ha at 25 day after sowing.

• Applied Microscopy
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• 제25권3호
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• pp.51-62
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• 1995

It has been well known that ischemia and reperfusion injury to skeletal muscle following an acute arterial occlusion causes significant morbidity and mortality. The skeletal muscle, which contains high energy phosphate compounds, has ischemic tolerance. During the ischemia, the ATP is catalyzed to hypoxanthine anaerobically and hypoxanthine dehydrogenase is converted to xanthine oxidase. During reperfusion, the hypoxanthine is catalyzed to xanthine by xanthine oxidase under $O_2$, presence and that results in production of cytotoxic oxygen free radicals. These cytotoxic free radicals, $O_2^-,\;H_{2}O_2,\;OH^-$, are toxic and make lesions in skeletal muscle during reperfusion. The authors perform the present study to investigate the effects of allopurinol, the inhibitor of xanthine oxidase, on reperfused ischemic skeletal muscles by observing the ultrastructural changes of the muscle fibers. A total of 48 healthy Sprague-Dawley rats weighing from 200 g to 250 g were used as experimental animals. Under urethane(3.0mg/kg., IP) anesthesia, lower abdominal incision was done and the left common iliac artery were ligated by using vascular clamp for 1, 2 and 6 hours. The left rectus femoris muscles were obtained at 6 hours after the removal of vascular clamp. In the allopurinol pretreated group, 50mg/kg of allopurinol was administered once a day for 2 days and before 2 hours of ischemia. The specimens were sliced into $1mm^3$ and prepared by routine methods for electron microscopic observations. All preparations were stained with uranyl acetate and lead citrate, and then observed with Hitachi -600 transmission electron microscope. The results were as follows: 1. In 1 hour ischemia/6 hours reperfused rectus femoris muscles of rats, decreased glycogen particles and electron density of mitochondrial matrix and dilated terminal cisternae are seen. In 2 hours ischemia/6 hours repersed rectus femoris muscles of rats, mitochondria with electron lucent matrix, irregularly dilated triad and spheromembranous bodies are observed. In 6 hours ischemia/6 hours reperfused rectus femoris muscles of rats, irregularly arranged myofibrils, and many spheromembranous bodies, fat droplets and lysosome are seen. 2. In 1 hour ischemia/6 hours reperfused rectus femoris muscles of rats pretreated with allopurinol, decreased glycogen particle and dilated cisternae of sarcoplasmic reticulum and triad are observed. In 2 hours ischemia/6 hours reperfused rectus femoris muscles of rats pretreated with allopurinol decreased electron density of mitochondrial matrix and spheromembranous bodies are seen. In 6 hours ischemia/6 hours reperfused rectus femoris muscles of rats pretreated with allopurinol, mitochondria with electron lucent matrix, spheromembranous bodies and dilated cisternae of sarcoplasmic reticulum and terminal cistern are observed. The results suggest that the allopurinol attenuates the damages of the skeletal muscles of rats during ischemia and reperfusion.

• 동의생리병리학회지
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• 제20권2호
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• pp.383-387
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• 2006

To investigate the effect of GyeongshinhaeGihwan 1(GGT1) frequently used as an anti-obesity herbal medicine in oriental medicine on the expression of obesity-related genes, we measured the changes in mRNA levels of these genes by GGT1 in human growth hormone transgenic (hGHTg) obese female rats, and these effects by GGT1 were compared with those of reductil (RD), an anti-obesity drug approved by FDA. Rats received once daily oral administrations of autoclaved water, RD, or GGT1 for 8 weeks. At the end of study, rats were sacrificed and tissues were harvested. Total RNA from adipose tissue, liver and kidney was prepared and the mRNA levels for LPL (lipoprotein lipase), $PPAR{\gamma}$ (peroxisome proliferator activated receptor-gamma), $PPAR{\delta}$ (peroxisome proliferator activated receptor-delta), leptin, $TNF{\alpha}$ (tumor necrosis factor-alpha), and internal standard G3PDH (glyceraldehyde-3-phosphate dehydrogenase) were analyzed by RT-PCR. Compared with control group, $PPAR{\gamma}$ mRNA levels of liver and kidney were decreased in both RD and GGT1 groups, and the effects were more prominent in GGT1 group than in RD group, suggesting that GGT1 is effective in the inhibition of lipid storage by decreasing the $PPAR{\gamma}$ expression. $PPAR{\delta}$ mRNA levels of adipose tissue were increased by RD and GGT1 compared with DW, and the magnitude of increase were higher in GGT1 group than in RD group, indicating that GGT1 stimulates fatty acid oxidation and energy metabolism by activating $PPAR{\delta}$ expression. GGT1 group had higher concentrations of serum leptin, a well-known inhibitor of appetite, than control and RD groups. However, The mRNA levels of leptin, LPL, and $TNF{\alpha}$ were not changed by GGT1. These results indicate that GGT1 can prevent obesity in hGHTg obese female rats by down-regulating and up-regulating the mRNA expression of $PPAR{\gamma}$ and $PPAR{\delta}$, respectively, and that this anti-obesity effects were more pronounced in GGT1 group compared with RD group. In addition, GGT1 seems to inhibit obesity by increasing the circulating leptin levels.

• 한국식품영양과학회지
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• 제29권4호
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• pp.737-742
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• 2000

대두발효식품의 기능성 탐색 연구의 일환으로 Bacillus natto 를 접종한 납두의 발효과정 중 고혈압을 유발하는 angiotension converting enzyme의 저해 peptide를 분리하고 저해효과를 검토함으로서 대두발효식품의 우수성을 입증하기 위한 과학 적 접근을 시도하고자 하였다. 납두는 Bacillus natto균을 이용하여 제조하였고, 2$0^{\circ}C$, 3$0^{\circ}C$, 4$0^{\circ}C$, 5$0^{\circ}C$, 6$0^{\circ}C$에서 0~72시간 동안 배양하면서 protein량, protease activity, ACE 저해율 을 측정하고 저해활성을 가지는 peptide를 정제 후 아미노산 조상을 분석하였다. Bacillus natto에 의한 납두의 발효시간이 경과함에 따라 protein 함량이 증가하여 4$0^{\circ}C$, 60시간에서 최대를 나타낸 후 감소하였다. 발효시간에 따른 protease activity는 4$0^{\circ}C$, 60 시간 배양이 최적의 조건이었으며, 최적 발효조건에 따라 납두를 제조한 후 20 mM sodium phosphate buffer(pH 7.0)를 가해 추출한 추출물을 Amicon membrane YM-3 filtration 과 Sephadex G-10, G-25를 이용한 gel filtration으로 부분정제하였다. 또한 정제한 peptide는 첨가함량이 높아질수록 저해활성은 높게 나타났으며, 1 mg 정도의 peptide 함량으로 74.74%의 저해율 을 나타내었다. 정제한 peptide의 아미노산 조성은 alanine(30.84%), phenylalanine(30.03%), histidine(20.24%) 순서로 그 함량이 높게 나타났다.

• 미생물학회지
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• 제9권3호
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• pp.103-120
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• 1971

We have been studied on brewing sweet starch. We obtained the results as follows ; 1) 5 strains, T-T-2, T-T-4, T-K-2, T-T-18, T-T-1, were the most available in view of fermentative power by capacity of $CO_2$. 2) 5 strains, T-T-4, T-T-2, T-T-1, T-T-3, T-K-2, produced capacity of alcohol more than 5.78%. 3) 6 strains, T-T-2, T-K-2, T-T-4, T-S-2, T-I-3, T-I-1, are available not only taste and flavour, but productive power of alcohol in sweet potato starch. 4) The form of 6 strains are long oval and round and most of them are similar to the other yeast in size. 5) In giant colony the color was cream color and cream buff, and T-K-2 was formed by $15{\times}12mm$ on diameter and by 3.5mm on high. 6) Optimum temperature of most of all strains is 25~ $300^{\circ}C$but T-K-4 is 28-30.deg.C. 7) Optimum pH is 3.4-4.6. 8) T-S-2 was died off at 65.deg.C, the other strains died $60^{\circ}C$. 9( Making Bun-kok with non-heated wheat bran .alpha.-amylase was more increased by 4.5-13.5 mg of glucose in reaction solution and .betha.-amylase more 1.6-3.4ml of N/10-$KMnO_4$ Solution than Bun-kok with heated wheat bran. 10) It seems that mycellium grows better than original in substance containing 0.4 ~ 1.2% of HCl. 11) Making Bun-kok to add 0.8% HCl, .alpha.-amylase was increased 9.93-20.7mg of glucose and .betha.-amylase ws increased 2.6~4.3ml of N/10-$KMnO_4$ solution to reaction solution. 12) 1.2%-HCl, or higher concentration, acts as inhibitor, in the meanwhile the concentration between 0.4~0.8% of HCl acts as activator. 13) We must make Bun-kok for 42 hours, at 28~$30^{\circ}C$) After we made Bun-kok using S-O-II and R-J-I one by one, Bun-kok which mix each other in equal quantity is increased more than original on enzyme acrivity. 15) Oxidation is the best way of refining sweet potato starch in N/10-phosphate buffer solution (pH 7.5). 16) When we prepared sweet potato starch, first pH was 3.0.