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Effect of NaCl on the Stability of Oncolytic Vaccinia Virus (항암 백시니아 바이러스의 안전성에 대한 염화나트륨의 효과)

  • Kim, Seong-Geun;Ran, Gui Shao;Kwon, Hyuk-Chan;Hwang, Tae-Ho
    • Journal of Life Science
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    • v.26 no.1
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    • pp.23-33
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    • 2016
  • Pexa-Vec (JX-594) is a specific cancer-targeted oncolytic and immunotherapeutic vaccinia virus. The purpose of this study was to develop methods to maximize the stability of Pexa-Vec. In short-term instability testing, viral activity was rapidly decreased both at 4℃ and at room temperature (RT), but it was completely restored after sonication followed by vortex. Long-term stability testing of Pexa-Vec in the following liquid formulations was performed: (A) 30 mM Tris/pH 7.6, (B) 30 mM Tris/pH 8.6, (C) 30 mM Tris/pH 7.6, 150 mM NaCl, 15% sucrose, (D) 30 mM Tris/pH 7.6, 15% sucrose, and (E) 30 mM Tris/pH 8.6, 15% sucrose. Viral activity decreased less than 2 log10 at 4℃, and RT was observed in 3 days in B, while viral activity was not decreased even after 4–8 weeks at 4℃ and at 1 week in RT in A, suggesting that neutral pH may be essential to maintain virus stability. The addition of 15% sucrose into A (D) significantly increased viral stability at −20℃, 4℃, or RT, and it was also observed at pH 8.6 (E). The addition of 150 mM NaCl into D (C) significantly increased viral stability in addition to the sucrose effect at 4℃ or RT. Accordingly, the viral activity in formulation C was maintained for 1.5 years at 4℃, and for 1-2 weeks in RT. In conclusion, we propose that formulation C can provide the most adequate condition for the proper storage of vaccinia oncolytic virus.

The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

Antioxidants in Serum and Induced Sputum of COPD Patients (만성폐쇄성폐질환 환자의 혈청 및 유도객담에서의 항산화제)

  • Park, Hyeon-Kwan;Yu, Young-Kwon;Kim, Kyu-Sik;Lim, Sung-Chul;Kim, Young-Chul;Park, Kyung-Ok
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.2
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    • pp.158-170
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    • 2001
  • Background : Although an oxidants and antioxidants imbalance has been considered in the pathogenesis of chronic obstructive pulmonary disease (COPD), there is a paucity of reports focussing on the smoking-induced changes of oxidants and antioxidants in COPD. Method : The concentration of antioxidants (ascorbic acid, uric acid, retinol, and $\alpha$- & $\gamma$-tocopherol) was measured in the serum and induced sputum of 30 healthy controls and 34 stable COPD patients using high performance liquid chromatography (HPLC). The inhibition of lipid peroxidation as an index of antioxidant capacity was measured in the serum by a TBA assay. Results : The serum concentration of ascorbic acid, $\alpha$-tocopherol, and retinol were significantly lower in the patients with COPD than in healthy controls ($484.8{\pm}473.3$ vs $1497.8{\pm}819.2\;pmol/L$, $48.38{\pm}17.34$ vs $73.96{\pm}26.29\;pmol/L$, p<0.001, and $9.51{\pm}8.33$ vs $15.01{\pm}5.88\;pmol/L$, p<0.05, respectively, mean$\pm$SD). However, there were little differences in the ascorbic acid and uric acid concentrations in the induced sputum between the COPD patients and the controls. The induced sputum to serum ratio of ascorbic acid was significantly higher in COPD patients compared with healthy control (0.375 vs 0.085, p<0.05). In the normal controls, the serum ascorbic acid concentration was lower in smokers than in nonsmokers ($1073{\pm}536$ vs $1757{\pm}845\;pmol/L$, p<0.05), but the level was still higher than that of the COPD patients (p<0.05). The serum retinol levels were correlated with $FEV_1$ in COPD patients (r=0.58, p<0.05). The products of lipid peroxidation were increased in normal smokers and COPD compared with norma1 nonsmokers ($115.56{\pm}19.93$ vs $120.02{\pm}24.56$ vs $91.87{\pm}20.71\;{\mu}mol/{\mu}mol$ Pi of liposome, p<0.05). Conclusion : Cigarette smoking may induce the dep1etion of serum antioxidants and this depletion of antioxidants is suggested to play a role in the pathogenesis of COPD.

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Effect on Survival and Developmental Competence of Vitrified Mouse Embryos Using Various Cryoprotectants and Cooling Speeds (생쥐 배아의 유리화 동결에 동결액의 조성과 냉각속도의 영향)

  • Park, Jae-Kyun;Go, Young-Eun;Eum, Jin-Hee;Won, Hyung-Jae;Lee, Woo-Sik;Yoon, Tae-Ki;Lee, Dong-Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.307-319
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    • 2010
  • Objective: Vitrification requires a high concentration of cyroprotectant (CPA) and an elevated cooling speed to avoid ice crystal formation. We have evaluated the effect of different combinations of cooling rate and CPA on embryonic integrity (developmental competence) in order to increase the efficiency of vitrification without impairing embryo viabilit. We hypothesized that the combination of CPA or the increase of cooling rates can reduce the concentration of toxic CPA for vitrification. As consequently, we performed experiments to evaluate the effect of various composition of CPA or slush nitrogen ($SN_2$) on the mouse embryonic development following vitrification using low CPA concentration. Methods: Vitrification of mouse embryos was performed with EM grid using liquid nitrogen ($LN_2$) or $SN_2$ and different composition of CPAs, ethylene glycol (EG) and dimethylsulfoxide (DMSO). After vitrification-warming process, their survival and blastocyst formation rates were examined. For analyzing long-term effect, these blastocysts were transferred into the uterus of foster mothers. Results: Survival and blastocyst formation rates of vitrified embryos were higher in EG+DMSO group than those in EG only. Furthermor, the group using $SN_2$ with a lower CPA concentration showed a higher survival of embryos and developmental rates than group using $LN_2$. Conclusion: The combination of EG and DMSO as CPAs may enhance the survival of mouse embryos and further embryonic development after vitrification. $SN_2$ can generate high survival and developmental rate of vitrified/warmed mouse embryos when a lower concentration of CPA was applied. Therefore, these systems may contribute in the improvement of cryopreservation for fertility preservation.

A Survey of the Basic Protection Manners of Dental Personnels Against Hepatitis B in and around Seoul (수도권 치과의료 종사자들의 B형 간염에 대한 기초방호실태 및 수행에 관한 조사)

  • Yoon, Mi-Sook;Park, Mi-Young
    • Journal of dental hygiene science
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    • v.1 no.1
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    • pp.46-52
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    • 2001
  • The purpose of this study was to examine dental personnels' concern for health, their personal protection manner, and whether or not they were inoculated against hepatitis B and made an inquiry of patients about that. It's ultimately meant to serve as a basis for the preparation of an anti-infection guide for the protection of dental personnels against all sorts of infectious diseases. After a survey was conducted from March 29 through May 9, 2001, the following findings were obtained: (1) The dental personnel group that had worked for 5 to less than 7 years took health examination the most, with 71.4%, before joining the hospital, and 61.5% of those who had served for 9 years or more, the greatest percentage, took medical checkup after joining the hospital(P<0.05). (2) The largest percentage of being inoculated against the anti-B type hepatitis after employment was 71.4% of the dental personnels over 35, and 57.7% of those with 9-year or more working experiences. Therefore, their age and service term made a significant difference(P<0.05). (3) There was no significant gap in personal protection practices between the dental hygienists and nurse aids(P>0.05). But the use of safety goggles for treatment was not properly done, with just 1.48 on the basis of 3 points, whereas the use of mask was scored 2.40 and the use of gloves was scored 1.96. (4) After a narcotic was used once, the leftovers were wasted by 89.7% of the dental hygienists and 70.0% of the nurse aids. More dental hygienists wasted them(P<0.05). (5) For hand washing, antibiotic liquid soap was used by 19.3% of the dental hygienists and 10.0% of the nurse aids, which were both very low percentage. And just 37.2% of the dental hygienists utilized a disposable paper towel to dry their hands, and only 36.0% of the nurse aids used the same(P<0.05).

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Effect of Scrapping Aerial Mycelia and Light on the Production of Macroconidia and Chlamydospores of Cylindrocarpon destructans Causing Root Rot of Panax ginseng (기중균사 제거와 광처리가 인삼 뿌리썩음병균 Cylindrocarpon destructans의 대형분생포자 및 후막포자 생성에 미치는 영향)

  • Cho Dae-Hui;Yu Yun-Hyun;Ohh Seung-Hwan
    • Journal of Ginseng Research
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    • v.23 no.3 s.55
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    • pp.123-129
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    • 1999
  • Under the light condition of 25,000 Lux (12 hrs dark and light cycle) with scrapping treatment of aerial mycelia of Cylindrocarpon destructans on potato dextrose agar (PDA), V-8 juice agar, and ginseng extract agar, production of the macroconidia was increased to $3.7\~8.1$ fold over them produced in the dark. They were also produced $7.7\~18.0$ times more in the liquid cultures under the light condition than under the dark as well. PDA and V-8 juice agar among the tested were the best for the macroconidium production. On PDA, 1,585 $macroconidia/mm^2$ were produced under the light of 25,000 Lux with scrapping treatment of aerial mycelia of C. destructans, which is 3.2 and 1.4 times more than those produced under 3,000 and 10,000 Lux, respectively. Meanwhile, $20\~99$ macroconidia/$mm^2$ were produced by the non-scrapping under the light condition between 3,000 Lux and 25,000 Lux. The macroconidia were, however, lysed at $6\~7$ days after being incubated under the above range of the light. They were consisted of $1\~3$ cells in a macroconidium while $69.4\~100\%$ of them were the two-celled and the number did not seem to be affected by either the scrapping or the light. Production of chlamydospore converted from mycelia of C. destructans seemed to be promoted by the light and the scrapping as well. The 1,285 chlamydospres/$mm^2$ were produced with the light (25,000 Lux), which is 2.8 and 1.2 times more than those with 3,000 and 10,000 Lux, respectively. Scrapping the aerial mycelia of the cultures increased the chlamydospore formation to 1.9, 2.5 and 1.4 times more than the non-scrapping under the light intensity of 3,000 Lux, 10,000 Lux, and 25,000 Lux, respectively. On PDA, 1 to 8 chlamydospore(s) per catena were formed by all treatments tested and $34.2\~58.9\%$ of them was a single chlamydospore, However, the numbers was affected by neither the light ($3,000\~25,000$ Lux) nor the scrapping the aerial mycelia.

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Comparative Analysis of Laparoscopy-assisted Gastrectomy versus Open Gastrectomy (복강경 보조 위절제술과 개복 위절제술의 비교 분석)

  • Lim, Jung Taek;Kim, Byung Sik;Jeong, Oh;Kim, Ji Hoon;Yook, Jeong Hwan;Oh, Sung Tae;Park, Kun Choon
    • Journal of Gastric Cancer
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    • v.7 no.1
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    • pp.1-8
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    • 2007
  • Purpose: There has been increased the number of early gastric cancer and laparoscopy-assisted gastrectomy (LAG), due to early detection through mass screening program. We started the LAG in April 2004 and performed 119 cases of gastric cancer in 2005, so we report a surgical outcome compared with that of open gastrectomy (OG). Materials and Methods: 119 patients underwent LAG in 2005, and for open group, 126 patiens of early gastric cancer were selected sequentially from January 2005 to March 2005. We compared clinicopathologic characteristics, postoperative courses and complications between two groups. Results: There was no significant difference between age, a length of hospital stay, distal resection margin and a number of retrived lymph nodes. The operation time was longer in LAG group (239.2 vs 123.3 mins, P<0.001) and a diet progression was faster in LAG group (first flatus: 3.05 vs 3.70 days, SOW: 2.86 vs 3.22 days, liquid diet: 3.87 vs 4.19 days, soft diet: 4.84 vs 5.26 days, P<0.001). But there was no difference statistically in postoperative discharge date (7.73 vs 8.25 days, P=0.229). The additional requirement of analgesic injection was less frequent in LAG group (2.97 vs 4.92 times, P<0.001). The harvested lymph nodes were similar in both groups (23.9 vs 23.1, P=0.563). A complication rate was lower in LAG group (4.9% vs 9.5%), but there was no statistical significance (P=0.179). There was no mortality in both groups and no conversion to open gastrectomy in the LAG group. Conclusion: LAG can be performed safely and accepted in view of curative procedure in treatment of early gastric cancer. But we need the follow up of long-term period to evaluate the survival rate and recurrence, and a prospective randomized controlled study should be done to establish that LAG will be a standard operation for early gastric cancer.

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Computer Assisted EPID Analysis of Breast Intrafractional and Interfractional Positioning Error (유방암 방사선치료에 있어 치료도중 및 분할치료 간 위치오차에 대한 전자포탈영상의 컴퓨터를 이용한 자동 분석)

  • Sohn Jason W.;Mansur David B.;Monroe James I.;Drzymala Robert E.;Jin Ho-Sang;Suh Tae-Suk;Dempsey James F.;Klein Eric E.
    • Progress in Medical Physics
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    • v.17 no.1
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    • pp.24-31
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    • 2006
  • Automated analysis software was developed to measure the magnitude of the intrafractional and interfractional errors during breast radiation treatments. Error analysis results are important for determining suitable planning target volumes (PTV) prior to Implementing breast-conserving 3-D conformal radiation treatment (CRT). The electrical portal imaging device (EPID) used for this study was a Portal Vision LC250 liquid-filled ionization detector (fast frame-averaging mode, 1.4 frames per second, 256X256 pixels). Twelve patients were imaged for a minimum of 7 treatment days. During each treatment day, an average of 8 to 9 images per field were acquired (dose rate of 400 MU/minute). We developed automated image analysis software to quantitatively analyze 2,931 images (encompassing 720 measurements). Standard deviations ($\sigma$) of intrafractional (breathing motion) and intefractional (setup uncertainty) errors were calculated. The PTV margin to include the clinical target volume (CTV) with 95% confidence level was calculated as $2\;(1.96\;{\sigma})$. To compensate for intra-fractional error (mainly due to breathing motion) the required PTV margin ranged from 2 mm to 4 mm. However, PTV margins compensating for intefractional error ranged from 7 mm to 31 mm. The total average error observed for 12 patients was 17 mm. The intefractional setup error ranged from 2 to 15 times larger than intrafractional errors associated with breathing motion. Prior to 3-D conformal radiation treatment or IMRT breast treatment, the magnitude of setup errors must be measured and properly incorporated into the PTV. To reduce large PTVs for breast IMRT or 3-D CRT, an image-guided system would be extremely valuable, if not required. EPID systems should incorporate automated analysis software as described in this report to process and take advantage of the large numbers of EPID images available for error analysis which will help Individual clinics arrive at an appropriate PTV for their practice. Such systems can also provide valuable patient monitoring information with minimal effort.

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Skin Whitening Effect of Ethyl Acetate Fraction of Adenophora triphylla var. japonica Sprout (잔대(Adenophora triphylla var. japonica)순 아세트산에틸 분획물의 피부 미백 효과)

  • Yoo, Seul Ki;Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Park, Sang Hyun;Kwon, Bong Seok;Lee, Chang Jun;Kang, Jeong Eun;Park, Su Bin;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.352-363
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    • 2017
  • To investigate skin-whitening effect of Adenophora triphylla var. japonica sprout extract, antioxidant activity, inhibitory effect on tyrosinase and melanin synthesis in B16/F10 melanoma cell were examined. Total phenolic content (246.25 mg GAE/g) and total flavonoid content (303.94 mg RE/g) of ethyl acetate fraction from Adenophora triphylla sprout (EFAT) showed the highest contents than other fractions (n-hexane, chloroform and distilled water). Antioxidant activities of EFAT has been evaluated using ABTS, DPPH radical scavenging activities, FRAP and inhibitory effect of lipid peroxidation. EFAT showed excellent radical scavenging activity and inhibitory effect on MDA production. Inhibitory effect of tyrosinase as a major enzyme of melanin synthesis was also measured. In these results, EFAT showed higher inhibitory effect against L-DOPA (51.27%) than L-tyrosine. $IC_{50}$ value on ${\alpha}-glucosidase$ was $41.93{\mu}g/ml$. In B16/F10 melanoma cells, EFAT inhibited melanin synthesis at $200{\mu}g/ml$ concentration (about 42% decrease). Finally, main physiological compounds of EFAT were identified as a rutin and a chlorogenic acid using high performance liquid chromatography.

Pharmacokinetics of ofloxacin in Patients with Multidrug-Resistant Tuberculosis (다제내성결핵 환자에서 ofloxacin의 약동학적 분석)

  • Park, Seung-Kyu;Yoon, Young-Ran;Lee, Woo-Chul;Jun, Hyung-Min;Shon, Ji-Hong;Kim, Kyoung-Ah;Park, Ji-Young;Shin, Jae-Gook
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.2
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    • pp.128-136
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    • 2002
  • Background: There are few studies that have reported on the pharmacokinetic(PK) disposition of fluoroquinolones in patients with multi-drug resistant tuberculosis(MDR-Tb), even though fluoroquinolones are frequently co-prescribed to those patients. In this study, the PK disposition of ofloxacin, a fluoroquinolone, was evaluated in patients with MD R -Tb. Methods: Twenty patients with MDR-Tb were given 2nd line Tb drugs including ofloxacin (300mg twice a day), prothionamide, cycloserine, para-aminosalicylic acid, kanamycin, and streptomycin. The patients were grouped according to their body mass index(BMI) as an index of emaciation (group A : 18.5$\leq$BMI <23, group B : BMI < 18.5). Blood samples were serially drawn and urine samples were collected upto 24 hours after the last dose of those drugs at steady state (over 1 month). The ofloxacin concentrations were determined using HPLC (High Performance Liquid Chromatography). Results: The AUC of ofloxacin in group B was greater than that in group A ($31.4{\pm}8.9{\mu}g/ml{\cdot}h$ vs. $24.1{\pm}6.2{\mu}g/ml{\cdot}h$)(Check the symbols), (p<0.05). The total clearance(Cl/F) of ofloxacin was $0.16{\pm}0.03$ L/h/kg in group A, and $0.14{\pm}0.03$ L/h/kg in group B. The half-lives of ofloxacin in two groups were similar (group A : $5.3{\pm}0.8$ hours, group B : $5.7{\pm}0.9$ hours). In addition, the other PK parameters in two groups were also similar. Conclusions: The pharmacokinetics of ofloxacin in patients with MDR-Tb appears to be comparable with those of normal subjects, and the extent of emaciation appears to have an influence on the pharmacokinetics of ofloxaicn in chronic debilitated MDR-Tb patients.