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The Effects of Bee Venom and Melittin Solution on PGE2, COX-2, and NF-kB Dependent Luciferase Activity in RAW 264.7 Cells (봉약침액(蜂藥鍼液)과 Melittin 약침액(藥鍼液)이 RAW 264.7 세포의 PGE2, COX-2 및 NF-kB에 미치는 영향(影響))

  • Jeong, Il-kook;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.21 no.6
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    • pp.19-36
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    • 2004
  • Objective : The purpose of this study was to investigate the effect of Bee Venom and Melittin Solution on the lipopolysaccharide(LPS) and sodium nitroprusside(SNP)-induced expression of prostaglandin $E_2(PGE_2)$, cyclooxygenase-2(COX-2), nuclear factor kappa B($NF-{\kappa}B$) and nuclear factor kappa B($NF-{\kappa}B$) dependent luciferase activity in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of PGE2 was determined by determination of $PEG_2$, COX-2 was by western blotting with corresponding antibodies, $NF-{\kappa}B$ was by gel mobility shift assay method and $NF-{\kappa}B$ dependent luciferase activity was investigated by luciferase assay in RAW 264.7 cells. Results : 1. LPS and SNP-induced expression of $PEG_2$ was significant after 24hour. 2. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $PEG_2$ and, the $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $PEG_2$ compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom could not significantly inhibit SNP-induced expression of $PEG_2$ compared with control. 3. The $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of COX-2 compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom inclined to decrease LPS and SNP-induced expression of COX-2 compared with control. 4. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ compared with control, respectively. 5. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $NF-{\kappa}B$ dependent luciferase activity and the 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. 6. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS + IFN-${\gamma}$, TNF-${\alpha}$ and LPS + TNF-${\alpha}$-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. Conclusions : These results suggest the inhibitory action of bee venom and melittin solution on the inflammatory mediators such as $PEG_2$, COX-2 and $NF-{\kappa}B$.

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The Bone Regenerative Effects of Chitosan on the Calvarial Critical Size Defectin Sprague Dawley Rats (백서 두개골 결손부에서 키토산의 골조직 재생 유도 효과)

  • Jung, Ui-Won;Suh, Jong-Jin;Choi, Seong-Ho;Choi, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.851-870
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    • 2000
  • The major goals of periodontal therapy is the functional regeneration of periodontal supporting structures already destructed by periodontal disease as well as the reduction of signs and symptoms of progressive periodontal disease. There have been many efforts to develop materials and therapeutic methods to promote periodontal wound healing. There have been increasing interest on the chitosan made by chitin. Chitin is second only to cellulose as the most abundant natural biopolymer. It is a structural component of the exoskeleton of invertebrates(e.g., shrimp, crabs, lobsters), of the cell wall of fungi, and of the cuticle of insects. Chitosan is a derivative of chitin made by deacetylation of side chains. Many experiments using chitosan in various animal models have proven its beneficial effects. The aim of this study is to evaluate the osteogenesis of chitosan on the calvarial critical size defect in Sprague Dawley rats. An 8 mm surgical defect was produced with a trephine bur in the area of the midsagittal suture. The rats were divided into two groups: Untreated control group versus experimental group with 50mg of soluble chitosan gel. The animals were sacrificed at 2, 4 and 8 weeks after surgical procedure. The specimens were examined by histologic, histomorphometric and radiodensitometric analyses. The results are as follows: 1. The length of newly formed bone in the defects was $102.91{\pm}25.46{\mu}m$, $219.46{\pm}97.81{\mu}m$ at the 2 weeks, $130.95{\pm}39.24{\mu}m$, $212.39{\pm}89.22{\mu}m$ at the 4 weeks, $181.53{\pm}76.35{\mu}m$ and $257.12{\pm}51.22{\mu}m$ at the 8 weeks in the control group and experimental group respectively. At all periods, the means of experimental group was greater than those of control group. But, there was no statistically significant difference between the two groups. 2. The area of newly formed bone in the defects was $2962.06{\pm}1284.48{\mu}m^2$, $5194.88{\pm}1247.88{\mu}m^2$ at the 2 weeks, $5103.25{\pm}1375.88{\mu}m^2$, $7751.43{\pm}2228.20{\mu}m^2$ at the 4 weeks and $8046.20{\pm}818.99{\mu}m^2$, $15578.57{\pm}5606.55{\mu}m^2$ at the 8 weeks in the control group and experimental group respectively. At all periods, the means of experimental group was greater than those of control group. The experimental group showed statistically significant difference to the control group at the 2 and 8 weeks. 3. The density of newly formed bone in the defects was $14.26{\pm}6.33%$, $27.91{\pm}6.65%$ at the 2 weeks, $20.06{\pm}9.07%$, $27.86{\pm}8.20%$ at the 4 weeks and $22.99{\pm}3.76%$, $32.17{\pm}6.38%$ at the 8 weeks in the control group and experimental group respectively. At all periods, the means of experimental group was greater than those of control group. The experimental group showed statistically significant difference to the control group at the 2 and 8 weeks. These results suggest that the use of chitosan on the calvarial defects in rats has significant effect on the regeneration of bone tissue in itself

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Enzymatic Preparation and Antioxidant Activities of Protein Hydrolysates from Tenebrio molitor Larvae (Mealworm) (갈색거저리 유충 단백가수분해물의 제조 및 항산화 활성)

  • Yu, Mi-Hee;Lee, Hyo-Seon;Cho, Hye-Rin;Lee, Syng-Ook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.4
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    • pp.435-441
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    • 2017
  • The present study was carried out to evaluate the applicability of Tenebrio molitor larvae (mealworm) as a health functional food material in order to contribute to the development of the domestic insect industry and health functional food industry. Protein hydrolysates were prepared from mealworm powder by enzymatic hydrolysis using five different proteases (alcalase, bromelain, flavourzyme, neutrase, and papain), and the hydrolysates were then tested for their antioxidant activities. Based on available amino group contents and sodium dodecyl sulphate-polyacrylamide gel electrophoresis analyses, mealworms treated with alcalase ($4,781.39{\mu}g/mL$), flavourzyme ($5,429.35{\mu}g/mL$), or neutrase ($3,155.55{\mu}g/mL$) for 24 h showed high degree of hydrolysis (HD) value, whereas HD values of bromelain ($1,800{\mu}g/mL$) and papain-treated ($1,782.61{\mu}g/mL$) mealworms were much lower. Protein hydrolysates showing high HD values were further separated into > 3 kDa and ${\leq}3kDa$ fractions by a centrifugal filter system and then lyophilized, and the production yields of the low molecular weight protein hydrolysates (${\leq}3kDa$) by alcalase, flavourzyme, and neutrase were 42.05%, 26.27%, and 30.01%, respectively. According to the RC_{50} values of the protein hydrolysates (${\leq}3kDa$) obtained from three different antioxidant analyses, all three hydrolysates showed similar antioxidant activities. Thus, alcalase hydrolysates showing the highest production yield of low molecular weight protein hydrolysates were further tested for their inhibitory effects on peroxidation of linoleic acid by measuring thiobarbituric acid values, and the results show that peroxidation of untreated linoleic acid increased dramatically during 6 days of incubation. However, pretreatment with the hydrolysates ($100{\sim}800{\mu}g/mL$) significantly inhibited linoleic acid peroxidation in a dose-dependent manner over 6 days.

EFFECT OF LASER IRRADIATION AND FLUORIDE APPLICATION ON REMINERALIZATION OF ERODED PRIMARY DENIAL ENAMEL (침식된 유치 법랑질에 대한 레이저 조사 및 불소도포의 재광화 효과)

  • Yang, Young-Sook;Kim, Dae-Eup;Ra, Ji-Young;Lee, Kwang-Hee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.262-268
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    • 2006
  • The purpose of this in vitro study was to evaluate the remineralizing effects of Nd : YAG laser irradiation and fluorides application on primary tooth enamel eroded by acidic drink. The materials were 30 sound primary teeth with intact smooth enamel surfaces. They were demineralized with Coca-cola at $37^{\circ}C$ for 12 hours and then irradiated by Nd: YAG laser with 6W power, $50mJ/cm^2$ energy density. and 20Hz pulse repetition. After laser irradiation, teeth were treated by three kinds of fluorides : (1) 0.05% NaF solution, (2) 1.23% APF gel and (3) 0.1% fluoride varnish. Diagnodent scores and microhardness (VHN) were measured before and after the each treatment. The results were as follows: 1. Diagnodent scores decreased to 23.51% from the initial after demineralization, and then increased to 37.37% after laser irradiation, and to 51.34% after fluoride treatment. There were significant differences between the total scores of initial, demineralization, laser irradiation and fluoride treatment (P<0.05). There was no significant difference between scores after fluoride treatment according to fluoride types. 2. Microhardness(VHN) decreased to 33.58% from the initial after demineralization and then increased to 43.99% after laser irradiation, and to 51.38% after fluoride treatment. There were significant differences between the total scores of initial, demineralization, laser irradiation and fluoride treatment (P<0.05). There was no significant difference between scores after fluoride treatment according to fluoride types.

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Detection of Pyrazinamide-Resistant Mycobacterium tuberculosis is by PCR-SSCP of pncA Gene (pncA 유전자 PCR-SSCP법을 이용한 결핵균 Pyrazinamide 내성의 진단)

  • Shim, Tae-Sun;Kim, Young-Whan;Chin, Jae-Yong;Lim, Chae-Man;Lee, Sang-Do;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.6
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    • pp.1178-1187
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    • 1998
  • Background : Recently the incidence of tuberculosis is increasing in many countries and control of the disease is further threatened by the emergence of multi-drug resistant tuberculosis. So rapid detection of drug resistance is very important. Pyrazinamide (PZA) is a first-line chemotherapeutic agent for tuberculosis. Now in Korea, we perform PZase activity test instead of actual pyrazinamide susceptibility test for the detection of PZA resistant M. tuberculosis. Recently the pncA gene, encoding the PZase of M. tuberculosis, was completely sequenced. And it was reported that the mutation of pncA gene would be associated with PZA resistance of M. tuberculosis. Therefore we performed this study to evaluate the possibility for the rapid detection of PZA resistant M. tuberculosis using PCR-SSCP of pncA gene. Method : 44 cultured clinical isolates of M. tuberculosis, BCG Tokyo strain. BCG French strain, and one M. bovis isolate were studied. We used H37Rv as the reference strain, The PZase activity test was done at the reference laboratory of Korean Tuberculosis Institute. DNA was extracted by bead-beater method and 561 bp fragment including pncA gene was amplified by PCR. The PCR product were digested by BstB I enzyme. SSCP was done using MDE gel. Of the 44 strains of M. tuberculosis, 22 strains were PZase-positive and other 22 strains were PZase negative. Results : Of the 22 PZase positive strains, 18 strains(82%) showed the same mobility compared with that of H37Rv and 4(18%) showed different mobility. Of the 22 PZase-negative strains, 19(86%) strains showed the same mobility pattern compared with that of H37Rv and 3(14%) showed different mobility. Naturally PZA-resistant BeG-French strain, BCG-Tokyo strain, and one M. bovis isolate showed the same band pattern each other, but their mobility were different from that of H37Rv. The results of PZase activity test and PCR-SSCP of pncA of M. tuberculosis were statistically significantly correlated each other (p<0.01). Conclusion : The PCR-SSCP after BstB I restriction of pncA gene of M. tuberculosis may be a useful method for the rapid detection of PZA-resistant M. tuberculosis.

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Rapid detection of Rifampicin- resistant M, tuberculosis by PCR-SSCP of rpoB gene (결핵균의 rpoB유전자 PCR-SSCP법에 의한 Rifampicin 내성의 신속 진단)

  • Shim, Tae Sun;Yoo, Chul-Gyu;Han, Sung Koo;Shim, Young-Soo;Kim, Young Whan
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.6
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    • pp.842-851
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    • 1996
  • Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

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The Appraisal for Effectiveness of School-based Oral Health Program in Daegu, Korea (대구광역시 일부초등학교 구강보건실의 계속구강건강관리 평가)

  • Park, Ji-Hye;Lee, Young-Eun;Kim, Ji-Young;Kim, Hye-Young;Choi, Youn-Hee;Song, Keun-Bae
    • Journal of dental hygiene science
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    • v.11 no.6
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    • pp.521-526
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    • 2011
  • The purpose of this study was to evaluate the long term effect of school-based oral health program on decrease of dental caries among elementary school children. The subjects of this study were total 727 students of one elementary school in Deagu, Korea. They had received school-based oral health programs for more than one year since 2004. This school-based oral health program included regular oral examination, fluoride mouth-rinsing, pit-and-fissure sealing, APF gel application, tooth brushing instruction and take the xylitol tablet after lunch during school days. The subjects' oral health status were examined by two calibrated dentists every year from 2004 to 2009. The DMFS index of $1^{st}$ grade to $6^{th}$ grade experimental and control group was 0.07, 0.39, 0.55, 0.67, 0.81, 1.79 and 0.31, 0.86, 1.07, 1.04, 2.71, 3.08 respectively. It's significantly different in $5^{th}$ grade(p<0.001) and $6^{th}$ grade(p=0.018) when it compare with control group. This study partially showed effect of school-based oral health programs on decrements of dental caries. This program should be expanded widely among elementary school in Korea. Xylitol tablet also recommended to included in school-based oral health programs.

Time Resolution Improvement of MRI Temperature Monitoring Using Keyhole Method (Keyhole 방법을 이용한 MR 온도감시영상의 시간해상도 향상기법)

  • Han, Yong-Hee;Kim, Tae-Hyung;Chun, Song-I;Kim, Dong-Hyeuk;Lee, Kwang-Sig;Eun, Choong-Ki;Jun, Jae-Ryang;Mun, Chi-Woong
    • Investigative Magnetic Resonance Imaging
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    • v.13 no.1
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    • pp.31-39
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    • 2009
  • Purpose : This study proposes the keyhole method in order to improve the time resolution of the proton resonance frequency(PRF) MR temperature monitoring technique. The values of Root Mean Square (RMS) error of measured temperature value and Signal-to-Noise Ratio(SNR) obtained from the keyhole and full phase encoded temperature images were compared. Materials and Methods : The PRF method combined with GRE sequence was used to get MR temperature images using a clinical 1.5T MR scanner. It was conducted on the tissue-mimic 2% agarose gel phantom and swine's hock tissue. A MR compatible coaxial slot antenna driven by microwave power generator at 2.45GHz was used to heat the object in the magnetic bore for 5 minutes followed by a sequential acquisition of MR raw data during 10 minutes of cooling period. The acquired raw data were transferred to PC after then the keyhole images were reconstructed by taking the central part of K-space data with 128, 64, 32 and 16 phase encoding lines while the remaining peripheral parts were taken from the 1st reference raw data. The RMS errors were compared with the 256 full encoded self-reference temperature image while the SNR values were compared with the zero filling images. Results : As phase encoding number at the center part on the keyhole temperature images decreased to 128, 64, 32 and 16, the RMS errors of the measured temperature increased to 0.538, 0.712, 0.768 and 0.845$^{\circ}C$, meanwhile SNR values were maintained as the phase encoding number of keyhole part is reduced. Conclusion : This study shows that the keyhole technique is successfully applied to temperature monitoring procedure to increases the temporal resolution by standardizing the matrix size, thus maintained the SNR values. In future, it is expected to implement the MR real time thermal imaging using keyhole method which is able to reduce the scan time with minimal thermal variations.

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Inorganic-organic nano-hybrid; Preparation of Nano-sized TiO$_2$ Paste Trapped OMC Nano-emulsion and it's Application for Cosmetics (OMC Nano-emulsion을 포집하고 있는 Nano-TiO$_2$-Paste의 합성과 화장품의 응용)

  • Byung Gyu, Park;Jong Heon, Kim;Jin Hee, Im;Kyoung Chul, Lee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.181-187
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    • 2004
  • Preparations of mesoporous materials using various templates and their applicability have been intensively investigated for many years. We studied on synthesizing mesoporous Ti02 with pores in which sensitive compounds having weak physico-chemical properties such as thermal or UV irradiation and low solubility in solvent are trapped. Prior to trapping OMC in the pores of mesoporous titania, OMC was nano-emulsified in O/W system using Lecithin. Thereafter the OMC was trapped in the pores of mesoporous titania using sol-gel method. Main focus of this work is to prepare OMC-trapped mesoporous titania and to trace the stability and solubility of nano-emulsified OMC in the pores of mesoporous titania, and compared with that of mesoporous silica. OMC-trapped mesoporous Inorganic-Organic hybrid titania showed higher factors in sun protecting and a skin penetration phenomenon was reduced.

Identification and Characterization of Wild Yeasts Isolated from Korean Domestic Grape Varieties (국산 포도로부터 분리한 야생효모의 동정 및 특성)

  • Choi, Sang-Hoon;Hong, Young-Ah;Choi, Yoon-Jung;Park, Heui-Dong
    • Food Science and Preservation
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    • v.18 no.4
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    • pp.604-611
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    • 2011
  • Several wild yeasts were isolated from Korean grape varieties before and during spontaneous fermentation. Among them, four strains were isolated based on the alcohol content and flavor production in wine after fermentation of apple juice. In this study, the four yeast strains were identified and characterized. PCR-restriction fragment length polymorphism analysis of ITS I-5.8S-ITS II region with restriction endonuclease Hae III and Hinf I resulted in that all the strains showed a typical pattern of Saccharomyces cerevisiae. Pulse field gel electrophoresis showed three different chromosome patterns with a same band between strains SS89 and SS812. When ITS I-5.8S-ITS II sequences of the four strains were compared with one another, they were similar to those of Saccharomyces cerevisiae CBS 4054 type strain. Identity of the sequences was higher than 97% with those of the type strain. Phylogenetic analysis showed based on the sequences showed they were genetically closed to the type strain. The four identified strains were tested in a medium containing 200 ppm potassium metabisulfite, and the MM10 and WW108 inhibition rates resulted at up to 24 h. The four strains were tested at an incubation temperature of $30^{\circ}C$. The 30% sugar concentration in the medium (w/v) showed the highest growth in 36 h, especially in the case of SS89, which was close to growth 40. The four strains were tested in an 8% ethanol medium (v/v). Alcohol tolerance was initially kept in the incubation process. The strains began to adapt, however, to the exceeded resistance. The four strains showed the lowest inhibition rate at 24 h.