• Biotechnology and Bioprocess Engineering:BBE
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• 제6권2호
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• pp.128-132
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• 2001

Bacterial vaginosis can be treated by restoring the normal vaginal flora using lactobacilli. Lactobacillus crispatus KLB46 that was isolated from the human vagina has a string antimicrobial activity and was grown in a batch and in a continuous fermentor. During batch cultivation, the maximum specific growth rate of L. crispatus KLB 46 was 0.63h(sup)-1 and the highest viable cell count (1.9$\times$10(sup)9 CFU/mL) was obtained at pH 5.5. L. crispatus KLB 46 did not grow well at either pH 3.5 or 7.5. During continuous cultivation, the highest viable cell count (1.53$\times$10(sup)9 CFU/mL) was obtained at a dilution rate of 0.32h(sup)-1, and was 7.33$\times$10(sup)11 CFU L(sup)-1 h(sup)-1, that is approximately 5 times higher than that obtained from batch culture.

• 미생물학회지
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• 제48권4호
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• pp.309-313
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• 2012

본 연구는 multiplex PCR 방법을 적용하여 질 내부에 서식하는 유산균을 동시에 그리고 신속하게 검출하는 방법을 개발하는 것을 목적으로 수행되었다. 세균의 감염에 의한 질염 증상이 없는 건강한 20대 여성 166명으로부터 질 분비물을 채취하였다. 건강한 여성의 질 내부에 서식하는 것으로 확인된 Lactobacillus 속에 속하는 6종의 유산균을 종 특이 multiplex PCR primer를 제작하기 위해 선정하였다. Multiplex primer I은 L. iners, L. crispatus, L. gasseri를 선택적으로 검출하기 위해 특성화하였고 multiplex primer II는 L. acidophilus, L. jensenii, L. fermentum를 선택적으로 검출하기 위해 특성화하였다. Multiplex PCR 기술을 이용하여 분석한 결과 L. crispatus (77%)가 가장 높은 빈도로 검출되었고 L. acidophilus (57%)과 L. jensenii (57%)는 상대적으로 높은 빈도로 검출되었다. L. iners (59%)는 L. acidophilus (57%)와 L. jensenii (57%) 보다 높은 빈도로 검출되었지만 항생제 치료 과정 후 또는 세균성 질염의 경우에도 발견되는 균종으로 건강한 여성에게서 주로 서식하는 종이라고 결론 내리기는 어렵다. 결론적으로, 특정 유산균의 특이적인 primer를 이용한 multiplex PCR 기술은 질의 건강상태의 변화 또는 질의 질병으로부터 회복 과정을 예측할 수 있는 도구로서 유용할 것으로 생각된다.

• KSBB Journal
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• 제29권2호
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• pp.81-86
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• 2014

Lactobacilli, the dominant species of microorganisms in the vaginal flora of healthy women, play important roles to prevent bacterial vaginosis and other sexually transmitted diseases. In this study, we carried out studies on stress adaptation prior to various stress treatment. We found that heat or salt adapted KLB46 showed higher cell viability than non adapted upon heat stress at $60^{\circ}C$ for 20 min. When chloramphenicol was added during the adaptation process, heat tolerance was abolished. This result suggested that de novo protein synthesis was essential during adaptation.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.918-922
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• 2001

In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

• KSBB Journal
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• 제31권1호
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• pp.40-45
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• 2016

Vaginitis, also known as vaginal infection and vulvovaginitis, is an inflammation of the vagina and possibly vulva. The three main kinds of vaginitis are bacterial vaginosis, vaginal candidiasis, and trichomoniasis. The purpose of this study was to characterize Lactobacillus plantarum MG989 and L. fermentum MG901 isolated from the vaginas of healthy Korean women in terms of their inhibitory activity against the vaginitis associated pathogens such as Gardnerella vaginalis and Candida albicans. Co-culture experiments showed that the two Lactobacillus strains MG989 and MG901 significantly reduced the viability of G. vaginalis and C. albicans. Also, the two strains were resistant to bile acid up to 1% and their autoaggregation rates were as high as 83.33%. Further studies are underway to demonstrate that the two strains can be applied as pharmaceutical agents for recovering healthy vaginal ecosystem.

• KSBB Journal
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• 제16권6호
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• pp.626-631
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• 2001

본 연구에서 L. crispatus KLB46을 저온 전처리함으로서 제제화 과정에 받게 되는 얼림과 녹임, 건조 스트레스뿐만 아니라, 여러 다른 환경 스트레스에 대한 내성이 증진된다는 것을 확인하였다. 그리고 내성 증진에 신규 단백질 합성이 필요함을 확인하였으며 나아가, 저온 충격 유전자 (csp)를 확인하였다. 따라서 이 균주를 제제화 하기 위한 방법으로 저온 전처리를 이용할 경우 생균력 유지에 큰 효과를 얻을 수 있다고 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2069-2074
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• 2014

Purpose: To appraise the frequency of cervical cytological abnormalities in a population at normal risk via analysing the archive records of cytology for the period of approximately 9,5 years, comparing them with patient demographic charecteristics, and discuss the results for women under age of 35. Materials and Methods: A total of 32,578 cases of Pap smears were retrieved and analysed from our archive included the Pap tests performed between January 2001 and April 2010 at the Early Cancer Screening, Diagnosing and Education Center by the consent of three pathologists via utilizing the Bethesda System Criteria 2001 and the results were compared with some demographical characteristics. Results: Our rate of the cervical cytological abnormality was 1.83%, with ASCUS in 1.18%, LSIL in 0.39, HSIL in 0.16%, AGUS in 0.07%, squamous cell carcinoma in 0.02%, and adenoarcinoma in 0.006%. Cytological abnormalities were detected mostly in those with higher age, lower parity, and premenopausal period whereas the smoking status was without influence. Bacterial vaginosis (5.6%) was the most frequent infectious finding (Candida albicans 2.7%; Actinomyces sp. 1.3%; and Trichomonas vaginalis 0.2%) detected on the smears. The rate of abnormal cervical cytology was 9.5% among the women aged between 30-34. Conclusions: Early detection of the cervical abnormalities by means of the regular cervical cancer screening programmes is useful to attenuate the incidence, mortality, and morbidity of cervical cancer. Our prevalence of the cytological abnormalities was much lower than the one in Western populations in general but very similar to those reported from other Islamic countries that may be explained by the conservative lifestyle and the lower prevalence of HPV in Turkey. A remarkable rate of abnormal cervical cytology of women aged 30-34 was pointed out in the present study.

• Journal of Microbiology and Biotechnology
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• 제12권2호
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• pp.312-317
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• 2002

Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

• KSBB Journal
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• 제18권4호
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• pp.294-300
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• 2003

본 연구에서는 L. fermentum KLB12을 열 전처리 함으로서 제제화 과정 동안 거치게 되는 열 스트레스에 대한 내성이 증진됨을 확인하고, 최적의 열 전처리 조건을 수행하였다. 또한 열 전처리 뿐만 아니라, 저온과 열 전처리 조건에도 열 스트레스에 대한 간섭 효과를 확인하였다. 그리고 내성 증진에 신규 단백질 합성이 필요함을 확인하였으며 나아가, 2-D electrophoresis를 통하여 7개의 신규 단백질을 확인하였다. 따라서 이 균주를 제제화하기 위한 방법으로 열 전처리를 이용할 경우 생균력 유지에 큰 효과를 얻을 수 있다고 사료된다.

• Journal of Preventive Medicine and Public Health
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• 제49권3호
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• pp.165-175
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• 2016

Objectives: The present study was undertaken to study the maternal risk factors for preterm birth (PTB) and low birth weight (LBW) with a special emphasis on assessing the proportions of maternal genitourinary and periodontal infections among Indian women and their association with adverse pregnancy outcomes. Methods: A hospital-based prospective study comprising 790 pregnant women visiting the obstetrics clinic for a routine antenatal check-up was undertaken. Once recruited, all study participants underwent clinical and microbiological investigations for genitourinary infections followed by a dental check-up for the presence of periodontitis. The study participants were followed up until their delivery to record the pregnancy outcomes. Infectious and non-infectious risk factors for PTB and LBW were assessed using univariate and multivariate Cox regression analysis. Independent risk factors for PTB and LBW were reported in terms of adjusted relative risk (ARR) with the 95% confidence interval (CI). Results: Rates of PTB and LBW in the study population were 7.6% and 11.4%, respectively. Previous preterm delivery (ARR, 5.37; 95% CI, 1.5 to 19.1), periodontitis (ARR, 2.39; 95% CI, 1.1 to 4.9), Oligohydramnios (ARR, 5.23; 95% CI, 2.4 to 11.5), presence of Nugent's intermediate vaginal flora (ARR, 2.75; 95% CI, 1.4 to 5.1), gestational diabetes mellitus (ARR, 2.91; 95% CI, 1.0 to 8.3), and maternal height <1.50 m (ARR, 2.21; 95% CI, 1.1 to 4.1) were risk factors for PTB, while periodontitis (ARR, 3.38; 95% CI, 1.6 to 6.9), gestational hypertension (ARR, 3.70; 95% CI, 1.3 to 10.8), maternal height <1.50 m (ARR, 2.66; 95% CI, 1.3 to 5.1) and genital infection during later stages of pregnancy (ARR, 2.79; 95% CI, 1.2 to 6.1) were independent risk factors for LBW. Conclusions: Our study findings underscore the need to consider screening for potential genitourinary and periodontal infections during routine antenatal care in developing countries.