• 미생물학회지
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• 제30권6호
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• pp.564-569
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• 1992

녹황색 세균인 Chlorobium limicola f. thiosulfatophilum NCIB 8327을 glutamate 가 질소원으로 포함된 변형 Pfennig 배지에서 배양한 다음 균체를 얻고, glutamine synthetase 효소를 초원심분리, DEAE-Sepharose CL-6B 이온교환 크로마토그래피, Sephacryl S-300 젤여과 크로마토그래피, 분취용 액체크로마토그래피의 과정을 통해 2% 수율에 46.3배의 정제배수로효소를 분리정제하였다. UV-VIS 흡수스펙트럼과 polycrylamide 젤 정기영동을 통해분리된 효소의 순수도는 확인하였다. Sephacryl S-3000 젤을 이용하여 얻은 효소의 분자량은 280 kDa 정도였으며, SDS-polyacrylamide 젤 전기영동 결과, 30 kDa 정도의 분자랴ㄹ을 갖는 단위체의 심합체로 추정된다. 이효소의 최적 온도와 pH 는 각각 $30^{\circ}C$ 와 pH 7.0 이었으나, 두기질 L-glutamine 과 hydroxylamine-HCI 에 대한 km 값은 각각 27.9 mM 과 0.92 mM 이었다. 이 효소의 활성은 alamine, glycine, tryptophan 등의 아미노산에 의해 상당한 저해를 받는 것으로 나타났으며, asparagin, lysine, leucine, valine 등에 의해서는 거의 영향을 받지 않았다.

• Journal of Acupuncture Research
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• 제31권4호
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• pp.57-70
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• 2014

Objectives : The purpose of this study is to examine if Clematidis Radix(CR) pharmacopuncture may be effective to the neuropathic pain(mechanical allodynia) in a rat model of neuropathic pain. Methods : To produce the model of neuropathic, L5 spinal nerve was ligated by 6-0 silk thread. After neuropathic surgery, the author examined if the animals exhibited the plantar withdrawal response of allodynia. The plantar withdrawal response was assessed by dynamic plantar aesthesiometer three days after the neuropathic surgery, CR pharmacopuncture was injected at $BL_{23}$ 1time/week for 6 weeks. After that, the author examined the plantar withdrawal response of rats' leg by dynamic plantar aesthesiometer. And also the author examined mGluR5, Bax, Bcl-2, Bax/Bcl-2 ratio in spinal cord, and c-Fos. Also the author observed the change of aspartate aminotransferase(AST), alanine aminotransferase(ALT) count in the blood serum of neuropathic rats. Results : 1. The withdrawal response of allodynia decreased in the PT3, PT4 group as compared with control group. 2. The mGluR5 increased in the PT1, PT2, PT3, and PT4 group. 3. The Bax and Bax/Bcl-2 ratio decreased in the PT4 group. 4. The c-Fos increased in the PT1 group, and decreased in the PT4 group. 5. The changes of AST in blood serum decreased in the every group excluded control group and the changes of ALT in blood serum isn't shown the signigicant change. Conclusions : These results are suggested that CR pharmacopuncture at BL23 decreased mechanical allodynia, can act on anti-apoptotic, neuroprotective effects and liver fuction in the model of neuropathic pain.

• Bulletin of the Korean Chemical Society
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• 제31권9호
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• pp.2497-2502
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• 2010

Arg13 is a conserved active-site residue in all known Pi class glutathione S-transferases (GSTs) and in most Alpha class GSTs. To evaluate its contribution to substrate binding and catalysis of this residue, three mutants (R13A, R13K, and R13L) were expressed in Escherichia coli and purified by GSH affinity chromatography. The substitutions of Arg13 significantly affected GSH-conjugation activity, while scarcely affecting glutathione peroxidase or steroid isomerase activities. Mutation of Arg13 into Ala largely reduced the GSH-conjugation activity by approximately 85 - 95%, whereas substitutions by Lys and Leu barely affected activity. These results suggest that, in the GSH-conjugation activity of hGST P1-1, the contribution of Arg13 toward catalytic activity is highly dependent on substrate specificities and the size of the side chain at position 13. From the kinetic parameters, introduction of larger side chains at position 13 results in stronger affinity (Leu > Lys, Arg > Ala) towards GSH. The substitutions of Arg13 with alanine and leucine significantly affected $k_{cat}$, whereas substitution with Lys was similar to that of the wild type, indicating the significance of a positively charged residue at position 13. From the plots of log ($k_{cat}/{K_m}^{CDNB}$) against pH, the $pK_a$ values of the thiol group of GSH bound in R13A, R13K, and R13L were estimated to be 1.8, 1.4, and 1.8 pK units higher than the $pK_a$ value of the wild-type enzyme, demonstrating the contribution of the Arg13 guanidinium group to the electrostatic field in the active site. From these results, we suggest that contribution of Arg13 in substrate binding is highly dependent on the nature of the electrophilic substrates, while in the catalytic mechanism, it stabilizes the GSH thiolate through hydrogen bonding.

• 한국수정란이식학회지
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• 제10권2호
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• pp.131-138
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• 1995

Essential and non-essential amino acids supplemented to culture medium stimulate mammalian embryo development in vitro. Amino acids such as glycine, taurine and alanine are concentrated in the lumen of oviduct and uterus and it can he thought that these amino acids may have physiological role on fertilization and embryo development. Our aim of this experiment was to investigate the effects of essential and non-essential amino acids, taurine or glycine supplemented to fertilization medium on the cleavage and subsequent in vitro development of bovine oocytes matured and fertilized in vitro. Immature oocytes were obtained from slaughtered Holstein cows and heifers and matured in TCM199 containing 10% fetal calf serum, 2.5 $\mu$g /mL of FSH and LH and 1 $\mu$g / mL of estradiol with granulosa cells in vitro. After maturation, oocytes were coincubated with sperm in fertilization medium supplemented with Minimum Essential Medium (MEM) essential and non-essential amino acids, taurine (3.75 mM) or glycine (10 mM) for 30 hours in vitro. Inseminated oocytes were cultured in synthetic oviduct fluid medium (SOEM) containing MEM essential, non-essential amino acids and 1 mM glutarnine up to 8 days after fertilization.Supplementation of fertilization medium with MEM essential and non-essential amino acids lowered significantly (p<0.05 and p<0.001) the cleavage rate after 30 hours of IVF (53.3%) and at Day 3 (62.7%: Day 0: the day of I VF) compared to control (64.3% and 77.3%, respectively). Subsequent developmental rates to morulae (Mo) and expanding blastocysts (ExBL) also significantly decreased (p<0.001 and p<0.05 for Mo and ExBL) when oocytes were coincubated with sperm in the medium containing MEM amino acids. Taurine added to fertilization medium have not increased the cleavage rate over the control, whereas glycine showed significantly lower (p<0.01) cleavage rate at Day 3 than that of taurine, but there was no significant difference in the developmental rates to Mo and ExBL of bovine embryos irrespective of the supplementation of taurine or glycine to fertilization medium. In conclusion, supplementation of fertilization medium with essential and non-essential amino acids, taurine or glycine has no beneficial effect on in vitro cleavage and development of bovine oocytes matured and fertilization in vitro.

• Clinical and Experimental Pediatrics
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• 제59권5호
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• pp.231-238
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• 2016

Purpose: Lipopolysaccharide-binding protein (LBP) is a 65-kDa acute phase protein, derived from the liver, which is present in high concentrations in plasma. Data regarding the association between circulating plasma LBP levels and obesity-related biomarkers in the pediatric population are scarce. We aimed to determine whether there was a difference in plasma LBP levels between overweight/obese and normal-weight adolescents and to assess the correlation of circulating LBP levels with anthropometric measures and obesity-related biomarkers, including insulin resistance, liver enzyme levels, and lipid profiles. Methods: The study included 87 adolescents aged 12-13 years; 44 were overweight/obese and 43 were of normal-weight. We assessed anthropometric and laboratory measures, including body mass index (BMI), blood pressure, insulin resistance, liver enzyme levels, and lipid profiles. Plasma LBP levels were measured using an enzyme-linked immunosorbent assay. Results: The mean age of the participants was $12.9{\pm}0.3$ years. Circulating plasma LBP levels were significantly increased in overweight/obese participants compared with those in normal-weight participants ($7.8{\pm}1.9{\mu}g/mL$ vs. $6.0{\pm}1.6{\mu}g/mL$, P<0.001). LBP levels were significantly and positively associated with BMI, systolic blood pressure, aspartate aminotransferase, alanine aminotransferase, total cholesterol, low density lipoprotein-cholesterol, fasting glucose and insulin, and insulin resistance as indicated by the homeostatic model assessment of insulin resistance (HOMA-IR) (all P<0.05). In multivariate linear regression analysis, BMI and HOMA-IR were independently and positively associated with plasma LBP levels. Conclusion: LBP is an inflammatory biomarker associated with BMI and obesity-related insulin resistance in adolescents. The positive correlation between these parameters suggests a potentially relevant pathophysiological mechanism linking LBP to obesity-related insulin resistance in adolescents.

• Radiation Oncology Journal
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• 제11권1호
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• pp.109-117
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• 1993

To analyze biochemical changes of liver function following combined radiotherapy and hyperthermia, we reviewed retrospectively 37 patients with hepatocellular carcinoma treated with radiotherapy and hyperthermia between July 1988 and December 1990 at Department of Radiation Oncology, Yonsei University College of Medicine. Mean age was 52.7 years and male to female ratio was 11:1. The patients were classified as follows; to A and B group by Child's classification, to M and L group by irradiated volume, and subclassified into BM, BL, AM and AL group according to the combination of Child's classification and irradiated volume. Radiation dose to the primary tumor was 3060 cGy with daily 180 cGy, 5 fraction per week using 10 MV or 4 MV linear accelerator. Hyperthermia (Thermotron RF-8) was performed more than 4 times in all patients. Biochemical parameters including albumin (Alb), total bilirubin (T. Bil), aspartate aminotransferase (AST or SGOT), alanine aminotransferase (ALT or SGPT), and alkaline phosphatase (ALP) were regularly followed from 1 week before the treatment to 3 months after the treatment. The results are summerized as follows; 1) In all the patient, mean ALP level peaked at 1 month, decreased at 2 months, slightly increased at 3 months after the treatment. Mean SGOT and SGPT levels peaked at 1 month after the treatment. Mean T. Bil level increased continuously and highest at 3 months after the treatment. Mean Alb level did not show significant changes.; 2) Mean ALP level retured to normal level at 3 month after the treatment in A but increased in B group and the differences were statistically significant (p<0.01). Mean SGOT and SGPT levels peaked 1 month in A and 2 months after the treatment in B group. All the biochemical parameters did not show significant difference between M and L group. Mean ALP level increased at 3 months after the treatment in BM and BL groups and decreased in AM and AL groups. Mean SGOT level increased at 3 months after the treatment in BL groups.; 3) Hepatic failure occurred within 3 months after the treatment in 4 patients, all of whom were in BL group. It is suggested that pre-treatment liver function and irradiated volume influence biochemical changes of liver in patients with hepatocellular carcinoma following combined radiotherapy and hyperthermia, and this treatment modality appears generally to be safe but might cause hepatic failure particularly in patient with poor liver function and large treatment volume.

• 한국식품과학회지
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• 제44권3호
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• pp.367-372
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• 2012

5종의 약용식물을 첨가한 발효홍삼(FRGM) 추출물의 항산화활성 및 ${\alpha}$-glucosidase 저해 활성을 in vitro에서 검토하였고, 또한 STZ로 당뇨를 유발시킨 흰쥐에서 FRGM 추출물의 효능을 살펴보았다. 그 결과, FRGM 추출물은 총 폴리페놀 및 총 플라보노이드 함량이 각각 $22.41{\pm}3.51{\mu}g/mg$, $16.80{\pm}4.22{\mu}g/mg$으로 존재하였고, DPPH 소거 활성을 가지고($RC_{50}=95.57{\pm}7.40{\mu}g/mL$), 또한 높은 ${\alpha}$-glucosidase 저해 활성을 나타내었다. STZ 투여에 의해 증가된 혈청 내 ALT와 AST의 활성이 FRGM 추출물의 투여로 감소하였으며, 간 조직중의 글루타치온 함량은 STZ 투여군에서는 감소되었다가 FRGM 투여시 유의적인 증가를 보였고, 지질과산화물 함량은 STZ 투여시 증가되었다가 FRGM 추출물의 투여시 유의적으로 감소되었다. 또한 간 조직중의 SOD, CAT, GP 그리고 GR의 활성도 STZ 투여로 유의적으로 감소되었다가, FRGM 추출물의 투여로 이들 항산화효소 활성이 유의적으로 증가하였다.

• 한국양식학회지
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• 제19권3호
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• pp.149-156
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• 2006

본 연구에서는 담수 및 해수사육 감성돔을 대상으로 $T_3$가 처리된 사료(10 mg/kg사료)를 90일간 공급하면서 어체의 스트레스, 삼투압조절 및 성장을 조사하였다. 혈장 $T_3$ 농도는 60일째에 FN, FT이 SN, ST에 비해 낮았으며, 또한 FN은 FT보다 유의하게 낮았다. 혈장 cortisol은 60일째를 제외하고는 실험구 사이에서 차이가 없었다 60일째에 FT $(21.0{\pm}3.3ng/ml)$는 $FN(5.1{\pm}1.4ng/ml)$보다 4배정도 높았으며, 또한 유의한 차이는 인정되지 않았지만 ST가 SN에 비해 다소 높은 것으로 나타났다. FN, FT의 혈장 glucose는 실험기간동안 SN, ST에 비해 낮은 경향을 보였으나, 담수 및 해수사육 감성돔에서 호르몬 처리에 의한 차이는 나타나지 않았다. 해수사육 감성돔의 AST 및 ALT는 실험기간동안 차이가 없었으나, 실험개시시 FN, FT의 AST및 ALT는 SN, ST보다 높았다. 또한 FN, FT의 AST의 해수수준으로 회복은 각각 실험종료시와 30일째에 해수수준으로 나타나 호르몬을 처리한 FT가 더 빨랐다. 그러나, 담수 및 해수사육 감성돔의 삼투압조절 향상 및 성장촉진을 위한 외인성 $T_3$ 공급 효과는 나타나지 않았다.

• 대한수의학회지
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• 제46권1호
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• pp.7-12
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• 2006

We have studied pharmacokinetics of a new anti-human immunodeficiency virus (HIV) agent VP-0501 and its amino acid prodrug VP-0501AL which is designed to improve oral bioavailability. After oral administration at 100 mg/kg dose in rats (n = 4), VP-0501 was not detectable in plasma (<50 ng/ml), while after the administration of VP-0501AL, VP-0501 was quantitatively detected, at least for 8 hrs, with Cmax of ca. $2.5{\mu}g/ml$ and AUC of $8hr^{\ast}{\mu}g/ml$. When VP-0501 was intravenously administered at 50mg/kg, this compound appeared at a marginal level in plasma with AUC of $2hr^{\ast}{\mu}g/ml$, $t_{1/2}$ of 2 hr, $C_0$ of $0.7{\mu}g/ml$, and MRT of 3 hr. On the other hand, with intravenous VP-0501AL at the same dose, both the prodrug VP-0501AL and its metabolite VP-0501 appeared comparatively at higher level in the plasma: pharmacokinetic parameters of VP-0501AL including $Vd_{\beta}$, AUC, $t_{1/2,{\beta}}$, $C_0$, $CL_{tot}$, and MRT were ca. 2 L/kg, $70hr^{\ast}{\mu}g/ml$, 2 hr, $180{\mu}g/ml$, 0.7 L/hr/kg, and 1 hr, respectively. These results demonstrate that attachment of amino acid alanine to VP-0501 is an effective approach for improvement of its oral bioavailability. Therefore, VP-0501AL is expected to become a new highly bioavailable and potent anti-AIDS drug candidate/lead compound.

• 한국미생물·생명공학회지
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• 제38권4호
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• pp.467-474
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• 2010

일반 청국장과 한약재를 첨가한 CKJ-DD, CKJ-RG, CKJDD+RG, CKJ-RED의 기호도 및 기능성을 측정하였다. 색도분석 결과, 5가지 청국장 모두 브라운 계열이었으며, a값 (redness), b값(yellowness)은 5가지 청국장에서 모두 유의적인 차이를 나타냈다. 유리아미노산 분석 결과, 구수한 맛을 나타내는 Aspartic acid, Glutamic acid의 함량이 CKJ보다 한약재를 첨가한 청국장(CKJ-DD, CKJ-RG, CKJ-DD+RG, CKJ-RED)이 더 많은 함량을 나타났으며, 단맛을 나타내는 alanine, glycine, lysine 또한 CKJ에 비해 한약재를 첨가한 청국장(CKJ-DD, CKJ-RG, CKJ-DD+RG, CKJ-RED)이 3~4.5배 더 많은 함량을 나타냈다. 반면, 한약재의 고유한 향에 의해서 쓴맛을 나타내는 leucine은 CKJ에 비해 CKJ-DD, CKJ-RED이 2배의 함량을 더 나타냈다. 한편 혈당강하 조절기전과 관련된 항당뇨 활성 결과에서는 농도 의존적으로 혈당상승억제효과가 증가하는 경향을 나타내지만, CKJ과 한약재를 첨가한 청국장(CKJ-DD, CKJ-RG, CKJ-DD+RG, CKJRED CKJRED) 간의 큰 유의적인 차이는 나타나지 않았다. 또한 5종의 청국장의 기능성을 알아보기 위해, 총 피놀릭성분의 함량 분석 결과, CKJ에 비해 한약재를 첨가한 청국장(CKJDD, CKJ-RG, CKJ-DD+RG, CKJ-RED)의 총 피놀릭 함량이 유의적으로 증가하는 경향을 나타냈다. 항산화 활성(Peroxyl radical 소거능)을 측정한 결과, $10\;{\mu}g/mL$ 농도에서 대조군인 CKJ과 비교하여 CKJ-DD은 비슷한 활성을 보였으나 CKJ-RG는 1.5배, CKJ-DD+RG은 3배 높은 항산화 활성을 나타냈다. 이상의 결과로 다양한 한방청국장 제품의 표준화 및 제품화를 위한 다양한 지표들을 확인 할 수 있었다.