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Expression of the 38 kDa Protein of Mycobacterium tuberculosis in M. bovis BCG and Use in the Serodiagnosis of Tuberculosis

  • Cho, Sang-Nae;Kim, Hee-Jin;Lee, Hye-Young;Kim, Seung-Chul;Kim, Joo-Deuk
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.555-559
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    • 1999
  • The 38 kDa protein of Mycobacterium tuberculosis, which was known previously as antigen 5, has been extensively used in the serodiagnosis of tuberculosis. In an attempt to develop and evaluate a serodiagnostic test using the antigen, we expressed the 38 kDa protein in BCG and its seroreactivity was compared to that expressed in Escherichia coli. The coding region of the 38 kDa protein was amplified by PCR, and the gene was cloned into a Mycobacterium-E. coli shuttle expression vector pYMC-his and pQE30 expression vector and expressed in BCG and E. coli, respectively. Both recombinant 38 kDa proteins showed strong seroreactivity against pooled serum from tuberculosis patients. There was no significant difference in seroreactivity between the two recombinant antigens in sera from the far advanced tuberculosis patients. However, of 25 tuberculosis patients graded as "minimal" by chest X-ray, 5 (20.0%) were seropositive by r38 kDa expressed in E. coli, while 8 (32.0%) by that expressed in BCG. Likewise, higher seroreactivity by r38 kDa expressed in BCG was found in sera from the moderately advanced tuberculosis. This study thus indicates that the recombinant 38 kDa expressed in BCG is more effective than that expressed in E. coli in detecting antibodies to the native 38 kDa protein of M. tuberculosis in sera from minimally affected tuberculosis patients.

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Analgesic effects of DA-5018, a non-narcotic agent

  • Kim, Hee-Kee;Bae, Eun-Ju;Shin, Myeong-Soo;Son, Moon-Ho;Kim, Soon-Hoe;Park, No-Sang;Kim, Won-Bae;Junnick Yang
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.235-235
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    • 1996
  • 1. 초산 writhing법에서 DA-5018, morphine, capsaicin 및 acetaminophen의 E $D_{50}$ 은 각각 0.1, 0.3, 1.4 및 45.4 mg/kg이었으며, formalin법에서 DA-5018과 morphine의 E $D_{50}$은 0.17과 4.3 mg/kg이었다. Randall-Selitto법에서 DA-5018, morphine, capsaicin 및 acetaminophen의 E $D_{50}$은 0.66, 3.9, 4.2 및 643 mg/kg이었다. Tail flick법에서 DA-5018, morphine 및 capsaicin의 E $D_{50}$ 은 2.0, 2.6 및 14.2 mg/kg이었고 hot plate법에서 DA-5018과 capsaicin의 E $D_{50}$은 1.7 및 26.5mg/kg이었다. 그러나 열자극에 의한 동통모델에서 acetaminophen은 진통효과가 없었다. 2. Streptozotocin유발 당뇨랫드에 DA-5018 0.2, 0.5 및 1.0mg/kg 또는 capsaicin 10 mg/kg 투여시 1, 3, 7 일째에서 각각 유의성있는 동통역치의 증가를 나타내었으나 ketoprofen(10 mg/kg) 및 desipramin(10 mg/kg) 투여시는 동통역치의 유의성있는 증가가 나타내지 않았다. 관절염 동통모델에서 DA-5018, ketoprofen 및 capsaicin을 투여한 후 7일째에 동통역치를 2배 증가시키는 용량은 각각 0.66, 3.76 및 17.38 mg/kg이었다. 3. 이상의 결과로부터 DA-5018은 morphine 및 capsaicin에 비해 동등 이상의 효력을 갖고 있으며, 비스테로이드성 진통제보다 효능이 우수함을 확인하였다.

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A Study on the Contact Allergic Skin Sensitization of 0.3% DA-5018 Cream, a Non-narcotic Analgesic Agent, with Guinea Pig Maximization Test (Guinea Pig Maximization Test를 이용한 0.3% DA-5018 cream의 접촉성 앨러지에 대한 연구)

  • 강경구;김동환;백남기;김원배;양중익
    • Biomolecules & Therapeutics
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    • v.4 no.4
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    • pp.391-397
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    • 1996
  • The aim of this study was to assess the allergenic potential of 0.3% DA-5018 cream, a non-narcotic analgesic agent, using a guinea pig maximization test. Five male and female guinea pigs in the experimental group were sensitized in two steps. First, ,0.3% DA-5018 cream was injected intradermally, and 7 days later, the material was applied topically. After another 2 weeks test material was applied, the skin response was evaluated by visual observation. Five male and female guinea pigs served as cream base group, negative(ultreated) group or positive (2,4-dinitrochlorobenzene, DNCB) group, respectively. 0.3% DA-5018 cream provoked slight erythema in 1 out of 5 cases in male and female guinea pigs sensitized with 0.3% DA-5018 cream or cream base. The animals challenged with cream base also showed slight erythema in 1/5 female guinea pig sensitized with 0.3% 3A-5018 cream or 2/5 male guinea pjgs sensitized with cream base, respectively. Histologically, however, no indication of skin sensitization was observed in all of these cases. The positive control group was sensitized with 0.1% DNCB suspended in olive oil and challenged with 0.01% and 0.1% DNCB ointment, all the animal showed remarkable skin reactions and obvious skin sensitization reactions in a dose dependent manner. From the challenge test it was evident that 0.3% DA-5018 cream did not elicit positive skin reaction interpreted as delayed hypersensitivity reactions, compared with cream base or untreated control group. These findings indicate that allergenic side effects by 0.3% DA-5018 cream is not likely in the clinical use.

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Properties of hydrolyzed α-lactalbumin, β-lactoglobulin and bovine serum albumin by the alcalase and its immune-modulation activity in Raw 264.7 cell

  • Yu, Jae Min;Son, Ji Yoon;Renchinkhand, Gerelyuya;Kim, Kwang-Yeon;Sim, Jae Young;Nam, Myoung Soo
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.459-470
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    • 2020
  • This study investigated the effects of the proteolytic hydrolysates of α-lactalbumin (LA), β-lactoglobulin (LG) and bovine serum albumin (BSA) by alcalase on inflammatory cytokines. The proteolytic hydrolysates were separated into two fraction of peptides, ≤ 10,000 Da and > 10,000 Da, respectively, because various low molecular weight peptides were generated during the hydrolysis reaction time. Among the hydrolysate peptides, BSA (all types), β-LG (> 10,000 Da), and α-LA (> 10,000 Da) showed an inhibitory activity against thymic stromal lymphopoietin (TSLP) mRNA expression in lipopolysaccharide-induced RAW264.7 murine macrophages. α-LA (> 10,000 Da), β-LG (hydrolysates), and BSA (> 10,000 Da) showed an inhibitory activity against tumor necrosis factor (TNF)-α expression. α-LA (all types), β-LG (hydrolysates, > 10,000 Da), and BSA (> 10,000 Da) showed an inhibitory activity against interleukin-6 (IL-6) expression. α-LA (> 10,000 Da), β-LG (> 10,000 Da), and BSA (all types) showed an inhibitory activity against inducible nitric oxide synthase (iNOS) expression. α-LA (> 10,000 Da), β-LG (> 10,000 Da), and BSA (all types) showed an inhibitory activity against cyclooxygenase (COX)-2 expression. The lowest level of TNF-α production was measured with α-LA (> 10,000 Da) and β-LG (> 10,000 Da) for all types, and a similar low level was measured for all types of BSA. The highest level of IL- 6 production was measured with α-LA (≤ 10,000 Da) among α-LA, β-LG, and IL-6. The low level of NO production was similar with α-LA, β-LG, and BSA but not with α-LA (≤ 10,000 Da). These potential peptides from whey protein hydrolysates could be used for food, medicinal, and industrial applications.

Isolation of Mutant Strains from Keratinase Producing Bacillus subtilis SMMJ-2 and Comparision of Their Enzymatic Properties (Keratinase 생산균 Bacillus subtilis SMMJ-2의 변이주 분리와 효소학적 특성 비교)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • KSBB Journal
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    • v.25 no.5
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    • pp.429-436
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    • 2010
  • Keratinase is widely used in certain industrial applications. The present study sought to improve the culture conditions of Bacillus subtilis SMMJ-2 to facilitate mass production of keratinase. Strain SMMJ-2 was irradiated by ultraviolet light and the resulting isolates were tested for keratinase activity. Isolates displaying elevated keratinase activity were selected and used to determine the optimum temperature (24, 30, 37, 45, $55^{\circ}C$) for bacterial keratinase production during a 4 day incubation period. The highest enzyme activity (55 units/mL/min), from a Bacillus subtilis SMMJ-2 mutant (mutant No. 2) was demonstrated following incubation at $30^{\circ}C$. The effects of carbon and nitrogen sources on keratinase production were confirmed by measuring the enzyme activity from the culture broth of the mutant strain cultured in various media containing different carbon source and nitrogen sources during a 4 day period. The optimal medium composition for producing keratinase consisted of 1% glucose, 0.7% $K_2HPO_4$, 0.2% $K_2HPO_4$, and 1.2% soybean meal. Optimal initial pH and temperature for producing keratinase were 7.0 and $30^{\circ}C$, respectively. Keratinases produced by B. subtilis SMMJ-2 and the mutant No. 2 were purified from the culture broth which used soybean meal as a nitrogen source. Membrane ultrafiltration, DEAE-sephacel ion exchange and Sephadex G-100 gel chromatography were used to purify the enzymes. The purified keratinases from both B. subtilis SMMJ-2 and the mutant No. 2 showed single bands and their molecular weights were estimated as 28 kDa and 42 kDa, respectively on SDS-polyacrylamide gel electrophoresis.

Evaluation of the Genotoxicity of Decursin and Decursinol Angelate Produced by Angelica gigas Nakai

  • Kim, Kang-Min;Kim, Tae-Ho;Park, Yun-Jung;Kim, Ik-Hwan;Kang, Jae-Seon
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.83-87
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    • 2009
  • In this study, we assessed the stability and toxicological safety of Angelica gigas Nakai (A. gigas Nakai) extract, which is comprised of decursin and decursinol angelate (D/DA). D/DA was tested for mutagenicity using Ames Salmonella tester strains (TA102, TA1535, and TA1537) with or without metabolic activation (S9 mix). No increase in the number of revertants was observed in response to any of the doses tested (1.25, 12.5, 125, and $1,250{\mu}/mLg$). In addition, a chromosome aberration test was conducted in the Chinese hamster lung (CHL) cell line. To accomplish this, cells were treated with D/DA (3.28, 13.12, 52.46, and $209.84{\mu}g/mL$) or with Mitomycin C ($0.1{\mu}/mLg$) as a positive control in the case of no metabolic activation or benzo(a)pyrene ($20{\mu}g/mL$) in the case of metabolic activation. No significant increase in chromosome aberrations was observed in response to treatment with any of these concentrations, regardless of activation of the metabolic system. According to these results, we concluded that D/DA did not induce bacterial reverse mutation or clastogenicity in vitro in the range of concentrations evaluated in these experiments.

Mutagenicity Study of DA-3030, A New Recombinant Human G-CSF(rhG-CSF) (새로운 재조합 인 과립구 콜로니 자극인자 DA-3030의 변이원성연구)

  • 강경구;최성학;김옥진;안병옥;백남기;김계원;김원배;양중익
    • Biomolecules & Therapeutics
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    • v.2 no.3
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    • pp.286-291
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    • 1994
  • The mutagenicity of DA-3030(rhG-CSF)was studied by reverse mutation test, chromosome aberration test and micronucleus test. The reverse mutatuon test in bacteria was performed using salmonella typhimurium strain TA100, TA98, TA1535 and TA1537 with rhG-CSF in any of the concentrations(150, 75, 37.5, 18.75, 9.375 and 4,6875 $\mu\textrm{g}$/plate), no increase in the number of revertant colonies in each strain was observed, irrespective of treatment with the metabolic activation system(S-9 mix) The chromosome aberration test was carried out using CHL cells, cell line from chinese hamster lung. With 4 doses(75, 37.5, 18.75 and 9.375 $\mu\textrm{g}$/ml) of rhG-/CSF the cells were treated for 24 or 48 hours in the direct method or for 6 hours followed by 18 hour-expression time in the metabolic activation method. Results of the study showed, by the direct method or metabolic activation method, no trend toward increase in the number of aberrant metaphase. The micronucleus test was carried out using ICR mice at the age of 8 weeks. Three doses(862.5, 1725 and 3450 $\mu\textrm{g}$/kg) of DA-3030 were admintstered intraperitoneally with single shot and bone marrow cells were sampled at 24 hours after administration. Neither the number of polychromatic erythrocytes with micronuclei nor the ratio of normochromatic erythrocytes to polychromatic erythrocytes increased singinficantly in each dose, compared with a vehicle control. These results indicate that rhG-CSF has not mutagenic potential under the condiions.

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Surgical Outcomes of Congenital Atrial Septal Defect Using da VinciTM Surgical Robot System

  • Kim, Ji Eon;Jung, Sung-Ho;Kim, Gwan Sic;Kim, Joon Bum;Choo, Suk Jung;Chung, Cheol Hyun;Lee, Jae Won
    • Journal of Chest Surgery
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    • v.46 no.2
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    • pp.93-97
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    • 2013
  • Background: Minimally invasive cardiac surgery has emerged as an alternative to conventional open surgery. This report reviews our experience with atrial septal defect using the da VinciTM surgical robot system. Materials and Methods: This retrospective study included 50 consecutive patients who underwent atrial septal defect repair using the da VinciTM surgical robot system between October 2007 and May 2011. Among these, 13 patients (26%) were approached through a totally endoscopic approach and the others by mini-thoracotomy. Nineteen patients had concomitant procedures including tricuspid annuloplasty (n=10), mitral valvuloplasty (n=9), and maze procedure (n=4). The mean follow-up duration was $16.9{\pm}10.4$ months. Results: No remnant interatrial shunt was detected by intraoperative or postoperative echocardiography. The atrial septal defects were mainly repaired by Gore-Tex patch closure (80%). There was no operative mortality or serious surgical complications. The aortic cross clamping time and cardiopulmonary bypass time were $74.1{\pm}32.2$ and $157.6{\pm}49.7$ minutes, respectively. The postoperative hospital stay was $5.5{\pm}3.3$ days. Conclusion: The atrial septal defect repair with concomitant procedures like mitral valve repair or tricuspid valve repair using the da VinciTM system is a feasible method. In addition, in selected patients, complete port access can be helpful for better cosmetic results and less musculoskeletal injury.

Structural and Physicochemical Studies on DA-5018, a New Capsaicin Derivative (새로운 Capsaicin 유도체 DA-5018의 구조 및 물리화학적 성질 연구)

  • Kim, Heung-Jae;Lee, Jong-Jin;Lee, Eung-Doo;Shim, Hyun-Joo;Lee, Sang-Deuk;Ok, Kwang-Dae;Kim, Won-Bae;Park, No-Sang
    • Journal of Pharmaceutical Investigation
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    • v.27 no.2
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    • pp.119-123
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    • 1997
  • The physicochemical and structural properties of new capsaicin derivative, DA-5018, were examined. The reference standard of this compound was obtained by the recrystallization. A method for the determination of the dissociation constant of the compound is described. pH-solubility and distribution coefficient were determined by chromatographic method. Fundamental properties on thermal behaviors were investigated by TG, DTA and DSC. Structural analysis based on spectroscopic method coincided with the chemical structure of DA-5018. Approximate dissociation constant of the compound determined by UV spectral method was 9.35. Solubilities and partition coefficients in various pH buffer solution appeared pH-dependency. No crystal transition or further transition was found in the thermal analysis. This compound showed good stability, but pH 13 buffer and acetone made some degradative products.

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Similar Operative Outcomes between the da Vinci Xi® and da Vinci Si® Systems in Robotic Gastrectomy for Gastric Cancer

  • Alhossaini, Rana M.;Altamran, Abdulaziz A.;Choi, Seohee;Roh, Chul-Kyu;Seo, Won Jun;Cho, Minah;Son, Taeil;Kim, Hyung-Il;Hyung, Woo Jin
    • Journal of Gastric Cancer
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    • v.19 no.2
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    • pp.165-172
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    • 2019
  • Purpose: The robotic system for surgery was introduced to gastric cancer surgery in the early 2000s to overcome the shortcomings of laparoscopic surgery. The more recently introduced da Vinci $Xi^{(R)}$ system offers benefits allowing four-quadrant access, greater range of motion, and easier docking through an overhead boom rotation with laser targeting. We aimed to identify whether the $Xi^{(R)}$ system provides actual advantages over the $Si^{(R)}$ system in gastrectomy for gastric cancer by comparing the operative outcomes. Materials and Methods: We retrospectively reviewed all patients who underwent robotic gastrectomy as treatment for gastric cancer from March 2016 to March 2017. Patients' demographic data, perioperative information, and operative and pathological outcomes were collected and analyzed. Results: A total of 109 patients were included in the $Xi^{(R)}$ group and 179 in the $Si^{(R)}$ group. Demographic characteristics were similar in both groups. The mean operative time was 229.9 minutes in the $Xi^{(R)}$ group and 223.7 minutes in the $Si^{(R)}$ group. The mean estimated blood loss was 72.7 mL in the $Xi^{(R)}$ group and 62.1 mL in the $Si^{(R)}$ group. No patient in the $Xi^{(R)}$ group was converted to open or laparoscopy, while 3 patients in the $Si^{(R)}$ group were converted, 2 to open surgery and 1 to laparoscopy, this difference was not statistically significant. Bowel function was resumed 3 days after surgery, while soft diet was initiated 4 days after surgery. Conclusions: We found no difference in surgical outcomes after robotic gastrectomy for gastric cancer between the da Vinci $Xi^{(R)}$ and da Vinci $Si^{(R)}$ procedures.