• Asian-Australasian Journal of Animal Sciences
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• v.29 no.10
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• pp.1490-1499
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• 2016

In this study, 69 lactobacilli isolated from Tibetan Qula, a raw yak milk cheese, were screened for their potential use as probiotics. The isolates were tested in terms of: Their ability to survive at pH 2.0, pH 3.0, and in the presence of 0.3% bile salts; tolerance of simulated gastric and intestinal juices; antimicrobial activity; sensitivity against 11 specific antibiotics; and their cell surface hydrophobicity. The results show that out of the 69 strains, 29 strains (42%) had survival rates above 90% after 2 h of incubation at pH values of 2.0 or 3.0. Of these 29 strains, 21 strains showed a tolerance for 0.3% bile salt. Incubation of these 21 isolates in simulated gastrointestinal fluid for 3 h revealed survival rates above 90%; the survival rate for 20 of these isolates remained above 90% after 4 h of incubation in simulated intestinal fluid. The viable counts of bacteria after incubation in simulated gastric fluid for 3 h and simulated intestinal fluid for 4 h were both significantly different compared with the counts at 0 h (p<0.001). Further screening performed on the above 20 isolates indicated that all 20 lactobacilli strains exhibited inhibitory activity against Micrococcus luteus ATCC 4698, Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 19115, and Salmonella enterica ATCC 43971. Moreover, all of the strains were resistant to vancomycin and streptomycin. Of the 20 strains, three were resistant to all 11 elected antibiotics (ciprofloxacin, erythromycin, tetracycline, penicillin G, ampicillin, streptomycin, polymyxin B, vancomycin, chloramphenicol, rifampicin, and gentamicin) in this study, and five were sensitive to more than half of the antibiotics. Additionally, the cell surface hydrophobicity of seven of the 20 lactobacilli strains was above 70%, including strains Lactobacillus casei 1,133 (92%), Lactobacillus plantarum 1086-1 (82%), Lactobacillus casei 1089 (81%), Lactobacillus casei 1138 (79%), Lactobacillus buchneri 1059 (78%), Lactobacillus plantarum 1141 (75%), and Lactobacillus plantarum 1197 (71%). Together, these results suggest that these seven strains are good probiotic candidates, and that tolerance against bile acid, simulated gastric and intestinal juices, antimicrobial activity, antibiotic resistance, and cell surface hydrophobicity could be adopted for preliminary screening of potentially probiotic lactobacilli.

• Journal of Life Science
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• v.10 no.3
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• pp.262-272
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• 2000

As a part of investigation for basic epidemiology of diarrheogenic disease, we attempted isolation of Escherichia coli from patients with diarrhea. Seven hundred and twenty-one strains of E. coli were isolated from 1,239 patients with diarrhea. Seasonal distribution of patient with diarrhea was shown the most high at August (18.2%). Age group distribution of patient was shown the most high at children (54.6%, 2 to 10 years old). The serotypes of 721 E. coli isolates were in order of serotype O44 (16.8%), O153 (8.6%), O1 (7.5%), O166(5.7%), O8 and O86a (4.7%), and O125 (4.6%). The supernates cultured 36 strains among 721 E. coli isolates were indicated cytotoxicity against monolayered Vero cells. All of the isolates were susceptible to amikacin. The isolates were resistant in order of novobiocin (99.0%), moxalactam (97.1%), carbenicillin (96.1%), tetracycline (90.4%), ampicillin (85.9%), gentamicin (84.0%), streptomycin (78.4%), cephalothin (46.6%) and polymyxin B (4.2%). In the antibiotic resistant patterns, 125 kinds of multiple resistance patterns of E. coli isolates were detected. The highest resistant pattern was ampicillin-carbenicillin-chloramphenicol-cephalothin-erythro-mycin- gentamicin-moxalactam-novobiocin-penicillin G-streptomycin-tobramycin-tetracycline-tri methoprim type (24.3%).

• Korean Journal of Food Science and Technology
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• v.37 no.3
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• pp.456-464
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• 2005

Antimicrobial drug-resistance is natural response to antimicrobial stress based on selection, which weakens chemotherapy effect. Introduction of large numbers of chemotherapeutic agents to clinical practice has generated strains of microorganisms that survive and multiply in vivo with high-drug concentrations. Methicillin-resistant Staphylococcus aureus (MRSA), bacteria found in normal daily life, can be easily ingested through milk vegetables, and meats, etc. MRSA emerged in many port of the world, increasing complex clinical problems. Therefore, new agents are needed to treat MRSA. Glycyrrhiza uralensis was extracted using 80% MeOH to investigate its antimicrobial activity against MRSA stains KCCM 11812, 40510, and 40512 through bacterial measurement, disc diffusion, and O.D. methods, MIC values, MRSA gene expression investigation, and scanning electron microscope observation. Results revealed MecA, Mecl, MecRI, and FemA were the most highly manifested MRSA genes. Methanolic extract of G. uralensis significantly inhibited MRSA and thus could be used in development of antibacteria.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.5
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• pp.436-446
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• 2006

The purpose of this study was to isolate and identify the bacteria in chronic maxillary sinusitis (CMS) lesions from 3 patients and to determine the antimicrobial susceptibility of them against 10 antibiotics. One of them was odontogenic origin and the others were non-odontogenic origin. Pus samples were collected by needle aspiration from the lesions and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, CO2, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene (16S rDNA) nucleotide sequencing method. To test the sensitivity of the bacteria isolated from the maxillary sinusitis lesions against seven antibiotics, penicillin G, amoxicillin, tetracycline, ciprofloxacin, cefuroxime, erythromycin, clindamycin, and vancomycin, minimum inhibitory concentration (MIC) was performed using broth dilution assay. Our data showed that enterobacteria such as Enterobacter aerogenes (30%), Klebsiella pneumoniae (25%), and Serratia marcescens (15%) were predominately isolated from the lesion of non-odontogenic CMS of senile patient (70 year old). Streptococcus spp. (40.3%), Actinomyces spp. (27.4%), P. nigrescens, M. micros, and P. anaerobius strains were isolated in the lesion of odontogenic CMS. In the lesion of non-odontogenic CMS, Streptococcus spp. (68.4%), Rothia spp. (13.2%), and Actinomyces sp. (10.5%) were isolated. The susceptibility pattern of 10 antibiotics was determined according to the host of the bacteria strains ratter than the kinds of bacterial species. Even though the number of CMS was limited as three, these results indicate that antibiotic susceptibility test must be accompanied with treatment of CMS. The combined treatment of two or more antibiotics is better than single antibiotic treatment in the presence of multidrug-resistant bacteria in the CMS lesions.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.34 no.5
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• pp.562-570
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• 2008

Most purulent maxillofacial infections are of odontogenic origin. Treatment of infection includes the surgical intervention, such as incision and drainage, and adjunctive treatment. The use of high-dose antibiotics is also indicated. The choice of an antibiotics should be based on the knowledge of the usual causative microbes and the results of antibacterial sensitivity test. We have undertaken clinical studies on 119 patients in Dept. of Oral and Maxillofacial Surgery, Inha University Hospital from January 2000 to December 2007. Many anaerobic microbes are killed quickly when exposed to oxygen. Thus the needle aspiration techniques and the transfer under inert gas were used when culturing. The aim of this study was to obtain informations for the bacteriologic features and the effective antimicrobial therapy against maxillofaical odontogenic infections. The obtained results were as follows: 1. The most frequent causes of infections were odontogenic (88.3%), and in odontogenic cause, pulpal infections were the most common causes(53.8%). 2. The buccal and submandibular spaces (respectively 23.5%) were the most frequent involved fascial spaces, followed by masticator spaces (14.3%). 3. The most common underlying medical problems were diabetes (17.6%), however the relation with prognosis was not discovered. 4. The complications were the expiry, mediastinitis, necrotizing fasciitis, orbital abscess, and osteomyelitis. 5. The most common admission periods were 1-2 weeks, and the most patients were discharged within 3 weeks. However, patients who admitted over 5 weeks were about 10%. 6. A total of 99 bacterial strains (1.1 strains per abscess) was isolated from 93 patients (78.2%). The most common bacterium isolated was Streptococcus viridans (46.2%), followed by $\beta$-hemolytic group streptococcus (10.1%). 7. Penicillins (penicillin G 58.3%, oxacillin 80.0%, ampicillin 80.0%) have slightly lower sensitivity. Thus we recommend the antibiotics, such as glycopeptides (teicoplanin 100%, vancomycin 100%) and quinolones (ciprofloxacin 90.0%) which have high susceptibility in cases in which peni cillin therapy failed or severe infections.

• Pediatric Infection and Vaccine
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• v.21 no.1
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• pp.65-70
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• 2014

Although congenital syphilis can be prevented with prenatal screening, the disease remains problematic. Currently, there are no cases that describe hematuria and pneumonia related to congenital syphilis. We report a case of congenital syphilis that involved nephrotic syndrome and pneumonia alba in a 22-day-old male infant whose mother did not receive adequate prenatal care. The congenital syphilis diagnosis was confirmed with a serologic test and the patient recovered with penicillin treatment. Clinical findings may be subtle in neonates and delayed recognition occurs frequently, thus complete prenatal screening is critical for congenital syphilis prevention. Immediate serologic testing should be performed to obtain a differential diagnosis if an infant is delivered by a mother that has not received appropriate prenatal examinations.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.67-67
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• 2003

Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

• Journal of fish pathology
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• v.9 no.2
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• pp.195-201
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• 1996

Ninety-six isolates of Edwardsiella tarda recovered from outbreaks of Edwardsiellosis in cultured eels(Anguilla japonica) in Kunsan, were examined for drug susceptibility, distribution and transferabilities of R plasmid. All of the E. tarda isolates examined were sensitive to gentamicin(GM), streptomycin(SM), norfloxacin(NF), and amikacin(AK). But most isolates were resistant to sulfadimethoxine(SD, 86 strains), ampicillin(AM, 84 strains), penicillin G(PM, 80 strains), nalidixic acid (NA, 67 strains), oxytetracycline(OT, 44 strains), and oxolinic acid(OA, 37 strains). Twenty different combinations of drug resistance patterns were observed : the frequently encountered pattern was SD-AM-PM-NA-OA(16 strains), SD-AM-PM-NA(14 strains), SD-AM-PM-NA-OT-OA(12 strains), SD-AM-PM-OT(10 strains), and SD-AM-PM-NA-OT(8 strains). Transferable R plasmids were found out to be carried in 78 out of 94 resistant strains, indicating that these isolates carry conjugally transferable R plasmids associated with single or multiple drugs. The frequently observed transferarble R plasmids were AM(8 strains), AM-PM-NA(8 strains), Am-SD(6 strains), PM(6 strains), and SD(6 strains) These results suggest that high dose of various antibacterials might have already been introduced to eel culture system leading to the acquirement of multi-drug resistance to wide range of antibacterials.

• Food Science of Animal Resources
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• v.28 no.5
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• pp.587-595
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• 2008

In order to develop a new starter for fermented milk, 1037 bacterial strains were isolated from raw milk. The strain that showed excellent acid producing and angiotensin converting enzyme (ACE) inhibitory activity (88.6%) was selected and identified as a Lactobacillus zeae based on the result of API carbohydrate fermentation pattern and 16S rDNA sequence. Lactobacillus zeae RMK354 was investigated further to study its physiological characteristics. It showed strong ACE inhibitory activity compared with commercial LAB starters tested. The optimum growth temperature of L. zeae RMK354 was $40^{\circ}C$ and it took 10 hr to reach pH 4.3 under this condition. L. zeae RMK354 showed more sensitive to penicillin-G, bacitracin, novobiocin, in a comparison of 14 different antibiotics, and showed most resistance to polymyxin B and vancomycin. It showed higher esterase and leucine arylamidase activities compared with 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed inhibitory activity against Salmonella Typhimurium with the rate of 60%. Based on these and previous results, L. zeae RMK354 could be an excellent starter culture for fermented milk with high level of ACE inhibitory activity.

• Journal of Periodontal and Implant Science
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• v.32 no.4
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• pp.865-873
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• 2002

The primary cause of tooth loss after 30 years of age is periodontal disease. Destruction of alveolar bone by periodontal disease is done by bone resorbing activity of osteoclasts. Understanding differentiation and activation mechanism of osteoclasts is essential for controling periodontal disease. The purpose of this study is to identify the possible effects of Vitamin D and cytokines affecting osteoclasts and its precursor cells. Four to six week-old mice were killed and humerus, radius, tibia and femur were removed aseptically and washed two times with Hank's solution containing penicillin-streptomycin and then soft tissue were removed. Bone marrow cells were collected by 22 gauge needle. Cells were cultured in Hank's solution containing 1 mg/ml type II collagenase, 0.05% trypsin, 41mM EDTA. Supernatant solution was removed 5 times after 15 minutes of digestion with above mentioned enzyme solution, and remained bone particles were maintained in alpha-MEM for 15 minutes and $4^{\circ}C$ temperature. Bone particles were agitated for 1 minute and supernatant solution containing osteoclast precursor cells were filtrated with cell stainer. These separated osteoclast precursor cells were dispensed with 100-mm culture dish by $1{\times}10^7$ cells unit and cultured in ${\alpha}$- MEM containing 20 ng/ml recombinant human M-CSF, 30 ng/ml recombinant human soluble osteoclast differentiation factor and 10% fetal calf serum for 2 and 7 days. Total RNA of osteoclast precursor cells were extracted using RNeasy kit. One ${\mu}g$ of total RNA was reverse transcribed in $42^{\circ}C$ for 30 minutes using SuperScriptII reverse transcriptase. Expression of transcribed receptors of each hormone and cytokine were traced with 1 ${\mu}l$ of cDNA solution by PCR amplification. Vitamin D receptor WAS found in cells cultured for 7 days. TNF-${\alpha}$ receptor was found in cells cultured for 2 days and amount of receptors were increased by 7 days. IL-1 type I receptor was not found in cells cultured 2 and 7 days. But, IL-1 receptor type II was found in cells cultured for 2 days. TGF-${\alpha},{\beta}$type I receptor was found in cells cultured 2 and 7 days, and amount of receptors were increased by 7 days of culture. These results implies Vitamin D and cytokines can affect osteoclasts directly, and affecting period in differentiation cycle of osteoclasts is different by Vitamin D and cytokines.