• Title/Summary/Keyword: P/C Based Method

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Development and validation of enzyme-linked immunosorbent assay for anti-mouse pertussis immunoglobulin G using international reference anti-Bordetella pertussis mouse serum NIBSC 97/642

  • Kyu-Ri Kang;Yi-Hyeon Kwon;Gyu-Won Cho;Gi-Sub Choi;Joon-Hwan Ji;Hyun-Mi Kang;Soo-Young Lee;Jin-Han Kang
    • Clinical and Experimental Vaccine Research
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    • v.13 no.3
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    • pp.242-252
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    • 2024
  • Purpose: In this study, an in-house enzyme-linked immunosorbent assay (ELISA) was developed and validated. The titer of ELISA was calculated using the reference line (RFL) method based on the standard curve drawn using the international reference anti-mouse serum NIBSC (National Institute for Biological Standards and Control) 97/642. Materials and Methods: In the development step, signal to noise was depicted to select the buffers that showed the most appropriate ratio. In the validation step, standard range, precision, dilution linearity, and specificity were confirmed, and RFL and parallel line (PLL) methods were compared in precision and dilution linearity. Results: Coating concentration for plate was achieved at 0.1 ㎍/mL for pertussis toxin (PT), 0.15 ㎍/mL for filamentous hemagglutinin antigen (FHA), and 0.25 ㎍/mL for pertactin (PRN). The signal to noise ratio was 22.02 for PT, 14.93 for FHA, and 8.02 for PRN with 0.25% goat serum in phosphate-buffered saline (PBS) as a dilution buffer, and 2% skim milk in PBS as a blocking buffer. Based on the precision results, we assessed the lower limit of quantification by 1, 0.2, and 1.5 EU/mL concentration for PT, FHA, and PRN which met the ICH (International Council for Harmonization) M10 criteria of a 25% accuracy and total error of 40%. In specificity, homologous serum was spiked into heterologous serum and the accuracy met the criteria. There was no difference in the results between RFL and PLL calculations (p-value=0.3207 for PT, 0.7394 for FHA, 0.2109 for PRN). Conclusion: ELISA validated with RFL calculation method in this study is a relatively accurate assay for mouse humoral immunogenicity test.

Physico-chemical Properties and Utilization of Sarcoplasmic Proteins for the Determination of End-point Cooking Temperatures of Ground Pork Hams Containing Salt and Fat (식염 및 지방을 함유한 분쇄돈육의 이화학적 성상 및 최종가열온도 측정을 위한 근장단백질의 이용)

  • Kang, S.M.;Chin, K.B.;Cho, S.H.;Lee, J.M.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.83-90
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    • 2004
  • Processed meals, such as a ground meat and hamburger patty, are required to ensure that no pathogens remain in the final products. However, there was no rapid method available to verify that the recommended end-point cooking temperature(EPT) was reached. Thus, the objective of this study was to rapidly determine EPT of ground pork hams using sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SOS-PAGE), based on the disappearance of sarcoplasmic proteins after cooking. Fresh pork hams were added two levels of salt(0, 2%) and fat(15, 25%) combinations, and stored in refrigerator overnight, and cooked to internal cooking temperatures of $64^{\circ}C$ to $74^{\circ}C$ with $2^{\circ}C$ increments. Cooked pork hams were measured cooking loss(CL, %), protein solubility(PS) and SOS-PAGE. CL(%) was reduced with the addition of 2% salt, as compared to the control, regardless of fat contents. It was also increased with increasing eooking temperature. Protein solubility was affected by the cooking temperature, resulting in reduced PS up to $64^{\circ}C$(P < 0.05), but remained constant higher than $68^{\circ}C$. In SOS-PAGE analysis, protein bands with the molecular weights of 36 and 66 kDa were affected by the addition of salt and fat combinations. regardless of treatments. These protein fractions were decreased gradually with increased cooking temperatures up to $68^{\circ}C$ ${\sim}$ $70^{\circ}C$ and might be good indicators for the determination of EPT in ground pork hams.

MICROLEAKAGE OF COMPOSITE RESIN RESTORATION ACCORDING TO THE NUMBER OF THERMOCYCLING (열순환 횟수에 따른 복합레진의 미세누출)

  • Kim, Chang-Youn;Shin, Dong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.32 no.4
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    • pp.377-384
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    • 2007
  • Present tooth bonding system can be categorized into total etching bonding system (TE) and self-etching boding system (SE) based on their way of smear layer treatment. The purposes of this study were to compare the effectiveness between these two systems and to evaluate the effect of number of themocycling on microleakage of class V composite resin restorations. Total forty class V cavities were prepared on the single-rooted bovine teeth and were randomly divided into four experimental groups: two kinds of bonding system and another two kinds of thermocycling groups. Half of the cavities were filed with Z250 following the use of TE system, Single Bond and another twenty cavities were filled with Metafil and AQ Bond, SE system. All composite restoratives were cured using light curing unit (XL2500, 3M ESPE, St. Paul, MN, USA) for 40 seconds with a light intensity of $600mW/cm^2$. Teeth were stored in distilled water for one day at room temperature and were finished and polished with Sof-Lex system. Half of teeth were thermocycled 500 times and the other half were thermocycled 5,000 times between $5^{\circ}C$ and $55^{\circ}C$ for 30 second at each temperature. Teeth were isolated with two layers of nail varnish except the restoration surface and 1 mm surrounding margins. Electrical conductivity (${\mu}A$) was recorded in distilled water by electrochemical method. Microleakage scores were compared and analyzed using two-way ANOVA at 95% level. From this study, following results were obtained: There was no interaction between variables of bonding system and number of thermocycling (p = 0.485). Microleakage was not affected by the number of thermocycling either (p = 0.814). However, Composite restoration of Metafil and AQ Bond, SE bond system showed less microleakage than composite restoration of Z250 and Single Bond, TE bond system (p = 0.005).

A Relations of Bone Mass Promoting Behaviors for Prevention of Osteoporosis and Multidimensional Health Locus of Control Scale Cluster (골다공증 예방을 위한 골량증진행위와 건강통제위 유형과의 관계)

  • Yeoum, Soon-Gyo
    • Women's Health Nursing
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    • v.3 no.2
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    • pp.208-223
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    • 1997
  • This study was made to suggest the nursing strategies for promoting the behaviors about bone mass health behaviors in order to prevent middle aged women's osteoporosis. This study was a descriptive-correlational design that also concerned to the types which improve bone mass promoting behaviors by inspecting patterns of health locus of control method out of recognizable variables of health improving models influencing on these bone mass promoting behaviors. For these purpose, data were collected by self reported questionnaire in middle school, from 158 women living in Seoul. The measuring tools used in this study about bone mass promoting behaviors and multidimensional health locus of control, were developed by author on the basis of literature review and analyzed by SPSS-PC window, into pearson's correlation, ANOVA, multiple regression, cluster analysis. Data was analyzed as follows. 1. 6 Multidimensional health locus of control scale clusters were existed. : a)cluster I (pure internal), b)cluster II(pure chance), c) cluster III(Believer in control), d), cluster IV(Type VI), e)cluster V(yea sayer), f) cluster VI(nay sayer). There were no findings of the powerful others external cluster and double external cluster. 2. The higher the value of internal health locus of control was, the better the bone mass promoting behaviors were(r=.2891, $p=.00^{**}$). The higher the value of chance external health locus of control was, the worse the bone mass promoting behaviors were(r=-.1367, $p=.00^{**}$). 3. On the basis of these relationships, 6 clusters were significantly different in the bone mass promoting behaviors(F=2.27, $p=.05^*$). The value of bone mass promoting behaviors was ranked the order of type VI>believer in control>pure internal>yea sayer>nay sayer>pure chance external highly. 4. Bone mass promoting behaviors were not significantly different as to age. Suggestion. Based on the results from the study, I would like to make some suggestions as follows. 1) To delay the loss of bone mass in middle aged women, the study on the cluster of the multidimensional health locus of control should be conducted repeatedly. 2) The tool of multidimensional health locus of control should be developed through a qualitative method adjusted on Korean' health culture.

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Characteristics of the gene resources and selected strains of Agrocybe cylindracea (버들송이버섯(Agrocybe cylindracea)유전자원 및 선발계통의 특성)

  • Heo, Byong-Soo;Yoo, Young-Jin;Seo, Sang-Young;Choi, Kyu-Hwan;Choi, Young Min;Kwon, Seog-Ju;Jang, Kab-Yeul
    • Journal of Mushroom
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    • v.17 no.2
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    • pp.52-63
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    • 2019
  • Agrocybe spp. belongs to the Agaricales order, Bolbitiaceae family, and Agrocybe genus. In Korea, so far, it has been cultivated through bottle cultivation; therefore, this study was conducted for the development of a new cultivar using the bag cultivation method for quantitative improvement. Thirty-three gene resources of Agrocybe spp. were collected and their quantity and characteristics of bag cultivation were examined. Next, 5 kinds of crossing parents were selected based on the cultivation period and shape of the fruit body. Seven strains were selected through 3 cross combinations. The 7 selected strains and the comparative cultivar 'Cham' were cultivated in a bag. As a result, the cultivation period was 49 days for 'JBAC15-1' and 50 days for 'JBAC15-6' which are 4 days and 3 days less than the cultivation period of the comparative cultivar 'Cham'(53 days), respectively. Cultivation periods of other strains except for 'JBAC15-1' and 'JBAC15-1' were longer than that of the comparative cultivar'Cham'. The best ratio of primordia formation among the selected strains was found to be that of 'JBAC15-1' with 96.1% followed by 'JBAC15-6' with 94.5%. These rates were 3.1% and 1.5% higher than the ratio of primordia formation of the comparative cultivar 'Cham', which is found to be 93.0%. The quantity was maximum in the 'JBAC15-1' cultivar with 176.8 g per bag followed by 'JBAC15-6' with 168.7 g per bag. The quantities were 10% and 5% more than the comparative cultivar 'Cham' with 160.7 g per bag. Based on these results, 'JBAC15-1' and 'JBAC15-6', which had shorter cultivation periods and more quantities than the comparative cultivar 'Cham' were finally selected. For the selected strains of 'JBAC15-1' and 'JBAC15-6', mycelial growth was observed to be optimal on PDA medium and the optimum temperature was $27.5^{\circ}C$. The optimum pH was pH 5 for 'JBAC15-1' and pH 6 for 'JBAC15-6'. The color of the pileus of the fruit body was dark brown in 'JBAC15-1' and 'Cham' and light brown in 'JBAC15-6'. The pileus was hemispherical in shape in both 'JBAC15-1' and 'Cham'. However, the colors of the stem were different - light brown in 'JBAC15-1', white in 'JBAC15-6', and ivory in the comparative cultivar 'Cham'.

Monitoring of Quality Characteristics of Chungkookjang Products during Storage for Shelf-life Establishment (청국장 제품의 유통기한 설정을 위한 저장중의 품질 특성 monitoring)

  • Kim, Dong-Myung;Kim, Seong-Ho;Lee, Jin-Man;Kim, Ji-Eun;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.132-139
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    • 2005
  • The major obstacle in the popularization of Chungkookjang is the short shelf-life of $2{\sim}3$ months and some problems concerning storage including the growth of molds even in the products even within shelf-life. To solve these problems we conducted a research to improve its storage by using the vacuumed packaging and sanitary method through seed culture, innoculation and sterilization. For the optimization of storage time, temperature and sterilization temperature, we measured viable cell numbers of bacteria and fungi, amount of gas outbreak and contents of amino type nitrogen and monitored these experimental results by response surface methodology of SAS program, so that we could observe the quality changes of Chungkookjang during shelf-life. Especially fungi, which are the biggest troublemaker in Chungkookjang shelf-life, couldn't be detected from the generally and vacuum-packed samples; also, viable cell numbers were highly influenced by sterilization temperature and in vacuum-packed samples. In the case of vacuum-packed samples, amount of gas outbreak was highly influenced by sterilization temperature of its storage conditions and it was higher in generally packed samples as compared to vacuum-packed samples even at any storage conditions. The changes of pH in generally and vacuum-packed samples were highly influenced by the storage temperature. As the temperatures of storage and sterilization were higher and the storage time was longer, so the amount of gas outbreak was accordingly lower. These results showed that amino type nitrogen contents in generally and vacuum-packed samples were systematically influenced by the temperature, storage time and sterilization temperature. Also the result showed that the change of amino type nitrogen contents during storage was less in vacuum-packed samples than in general ones. Based on the above results, we can produce Chungkookjang products with extended shelf-life of as far as 6 months without any quality change using sanitary manufacturing method, vacuumed packaging condition, sterilization in $70^{\circ}C$ for 60 minutes and storage under $10^{\circ}C$ during shelf-life. According to this research, we have the possibility to greatly increase the goods value of Chungkookjang by developing the manufacture processing and packaging.

Effects of chromium chloride addition on coloration and mechanical properties of 3Y-TZP (크롬염화물 첨가에 따른 지르코니아 색상 및 물리적 성질 변화에 관한 연구)

  • Oh, Gye-Jeong;Seo, Yoon-Jeong;Yun, Kwi-Dug;Lim, Hyun-Pil;Park, Sang-Won;Lee, Kyung-Ku;Lim, Tae-Kwan;Lee, Doh-Jae
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.2
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    • pp.120-127
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    • 2011
  • Purpose: The purpose of this study was to examine the effects of chromium chloride addition on coloration, mechanical property and microstructure of 3Y-TZP. Materials and methods: Chromium chloride was weighed as 0.06, 0.12, and 0.25 wt% and each measured amount was dissolved in alcohol. $ZrO_2$ powder was mixed with each of the individual slurry to prepare chromium doped zirconia specimen. The color, physical properties and microstructure were observed after the zirconia specimen were sintered at $1450^{\circ}C$. In order to evaluate the color, spectrophotometer was used to analyze the value of $L^*$, $C^*$, $a^*$ and $b^*$, after placing the specimen on a white plate, and measured according to the International Commission on Illumination (CIE) standard, Illuminant D65 and SCE system. The density was measured in the Archimedes method, while microstructures were evaluated by using the scanning electron microscopy (SEM) and XRD. Fracture toughness was calculated Vickers indentation method and indentation size was measured by using the optical microscope. The data were analyzed with 1-way ANOVA test (${\alpha}$ = 0.05). The Tukey multiple comparison test was used for post hocanalysis. Results: 1. Chromium chloride rendered zirconia a brownish color. While chromium chloride content was increased, the color of zirconia was changed from brownish to brownish-red. 2. Chromium chloride content was increased; density of the specimen was decreased. 3. More chromium chloride in the ratio showed increase size of grains. 4. But the addition of chromium chloride did not affect the crystal phase of zirconia, and all specimens showed tetragonal phase. 5. The chromium chloride in zirconia did not showed statistically significant difference in fracture toughness, but addition of 0.25 wt% showed a statistically significant difference (P<.05). Conclusion: Based on the above results, this study suggests that chromium chlorides can make colored zirconia while adding in a liquid form. The new colored zirconia showed a slight difference in color to that of the natural tooth, nevertheless this material can be used as an all ceramic core material.

Association of PINX1 but not TEP1 Polymorphisms with Progression to Hepatocellular Carcinoma in Thai Patients with Chronic Hepatitis B Virus Infection

  • Sriprapun, Methee;Chuaypen, Natthaya;Khlaiphuengsin, Apichaya;Pinjaroen, Nutcha;Payungporn, Sunchai;Tangkijvanich, Pisit
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2019-2025
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    • 2016
  • Hepatocellular carcinoma (HCC) is major health problem with high mortality rates, especially in patients with hepatitis B virus (HBV) infection. Telomerase function is one of common mechanisms affecting genome stability and cancer development. Recent studies demonstrated that genetic polymorphisms of telomerase associated genes such as telomerase associated protein 1 (TEP1) rs1713449 and PIN2/TERF1-interacting telomerase inhibitor 1 (PINX1) rs1469557 may be associated with risk of HCC and other cancers. In this study, 325 patients with HCC and 539 non-HCC groups [193 healthy controls, 80 patients with HBV-related liver cirrhosis (LC) and 266 patients with HBV-related chronic hepatitis (CH)] were enrolled to explore genetic polymorphisms of both SNPs using the allelic discrimination method based on MGB probe TaqMan real time PCR. We demonstrated that all genotypes of both genes were in Hardy-Wienberg equilibrium (P>0.05). Moreover, there was no significant association between rs1713449 genotypes and HCC risk, HCC progression and overall survival (P>0.05). Interestingly, we observed positive association of rs1469557 with risk of HCC when compared with the LC group under dominant (CC versus CT+TT, OR=1.89, 95% CI= 1.06-3.40, P=0.031) and allelic (C versus T alleles, OR=1.75, 95% CI=1.04-2.94, P=0.033) models, respectively. Moreover, overall survival of HCC patients with CC genotype of rs1469557 was significantly higher than non-CC genotype (Log-rank P=0.015). These findings suggest that PINX1 rs1469557 but not TEP1 rs1469557 might play a role in HCC progression in Thai patients with LC and be used as the prognosis marker to predict overall survival in HCC patients.

The Plasma Concentrations and Systemic Toxicity of Lidocaine after Maximal or Supramaximal Recommended Doses of Epidural Administration (경막외 Lidocaine의 최대사용량과 혈중농도에 대한 고찰)

  • Park, Han-Suk;Chung, Chan-Jong;Chin, Young-Jhoon
    • The Korean Journal of Pain
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    • v.12 no.1
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    • pp.36-42
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    • 1999
  • Background: The current maximal recommended doses of lidocaine are 7 mg/kg with $5\;{\mu}g/ml$ of epinephrine. But in clinical practice, sometimes more doses of lidocaine are required to produce adequate regional anesthesia. Method: Twenty-two healthy women patients were divided into two groups and pretreated with valium 5 mg p.o., morphine 5 mg i.m., and midazolam 2 mg i.v. before operation. Of these, 7 mg/kg of 2% lidocaine with $5\;{\mu}g/ml$ of epinephrine were given to 11 patients epidurally. Initial 3 ml of epinephrine mixed lidocaine was given as a test dose and remaining doses were given 5 ml/30 sec with 3 min intervals. Radial arterial blood were drawn at 5, 10, 15, 20, 30, 45, 60, 90, 120 min to measure plasma lidocaine concentrations. After confirming all of the peak plasma concentrations of 7 mg/kg lidocaine were absolutely under $5\;{\mu}g/ml$, the other 11 patients were given 10 mg/kg of 2% lidocaine with $5\;{\mu}g/ml$ of epinephrine epidurally and blood samplings were taken according to the same method of 7 mg/kg group. The peak plasma concentration ($C_{max}$), time to reach to $C_{max}$ ($T_{max}$), time to reach to $T_4$, maximal sensory block level, systemic toxicity, and vital sign changes were investigated. Result: $C_{max}$ was significantly higher in 10 mg/kg group ($5.1{\pm}1.3\;{\mu}g/ml$) than 7 mg/kg group($3.3{\pm}0.5\;{\mu}g/ml$), but $T_{max}$ ($10.5{\pm}2.7$ min vs $10.9{\pm}3.1$ min) was not different. Time to reach $T_4$ was significantly shorter in 10 mg/kg group ($9.5{\pm}2.7$ min) than 7 mg/kg group ($12.7{\pm}3.2$ min) but maximal sensory block level ($T_{3.7{\pm}0.7}$ vs $T_{2.7{\pm}1.0}$) was not different. In four patients of 10 mg/kg group, peak plasma concentrations exceeded $5\;{\mu}g/ml$, but no systemic toxicities appeared. No significant vital sign changes were observed. Conclusion: The current maximal recommended doses of lidocaine, merely based on body weight are not always appropriate. Further studies are needed to determine more precise guideline of maximal doses that include various pharmacokinetic components.

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Identification and Fermentation Characteristics of Lactic Acid Bacteria Isolated from Dongchimi as Starter for Radish Juice (무 쥬스 제조를 위한 starter로써 동치미에서 분리한 유산균의 동정 및 발효 특성)

  • Kim, Jung-Hee;Kim, Jong-Il
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.307-314
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    • 1999
  • This study was aimed at the investigation of the possibility of the addition of lactic acid bacteria as "starter"for the preparation of radish juice. Forty strains of lactic acid bacteria were isolated from dongchimi that was fermented by a traditional method. The isolates were assorted into 5groups, Leuconostoc mesenteroides subsp. mesenteroides (J-9), Lacobacillus brevis (J-12), Lactobacillus fermentum (J-7), Lactobacillus sake (J-20), and Lactobacillus plantarum (J-39). Leuconostoc mesenteroides was predominated in the sample of dongchimi with frequency of 52.5%. Each of the strain, which exhibited the beat growth in the species, was selected in the 5species, and investigation of the fermentation characteristcis was carried out. The fermentation were performed for 9 days at 25${\circ}C$ after the inoculation of 0.3% ($10^{6}$ cfu/㎖) to each ultra-filtrated radish juice. The pH, total acidity, content of non-volatile organic acids were examined during the fermentation period. Lactobacillus plantarum showed the highest growth rate and the growth rate of Lactobacillus sake was the lowest. The pH (6.3-6.36) and total acidity (0.09-1.0 %) fo the ultrafiltrated radish juice before fermentation were changed to 3.2-4.3 and 0.65-1.2% after 9days, respectively. The changes of the pH and total acidity were related with the growth of the lactic acid bacteria; the better growth of lactic acid bacteria, the more rapid decrease of pH and increase of the total acidity. when the amount of non-volatile organic acids were estimated during fermentation, citric acid, malic acid, malonic acid, and succinic acid were decreased in all cases. However, the content of lactic acid increased with the progression of fermentation. L. mesenteroides (J-9), L. brevis (J-12) and L. fermentum (J-7) were chosen for the candidates of the starter for the lactic fermentation of radish juice based on the biochemical analysis and sensory evaluation.

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