• Tuberculosis and Respiratory Diseases
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• v.46 no.5
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• pp.618-627
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• 1999

Background : Rifampicin (RFP) is a key component of the antituberculous short-course chemotherapy. Usually the RFP resistant M.tuberculosis is also resistant to isoniazid (INH), so the RFP resistance is the marker of multi-drug resistant (MDR) tuberculosis. But unusual cases of mono-RFP-resistant tuberculosis have been recently reported with increasing frequency, especially associated with HIV infection in western countries. Therefore, we conducted a retrospective study to investigate the frequency, causes, and the clinical characteristics of mono-RFP-resistant tuberculosis in Korea. Methods : Of the bacteriologically confirmed and susceptibility-proven 699 pulmonary tuberculosis patients (921 isolates) who visited Asan Medical Center from January 1990 to August 1997, eighteen patients with INH-susceptible and RFP-resistant tuberculosis were evaluated. Previous history of tuberculosis, antituberculous drug compliances, associated systemic illness, drug susceptibility patterns, and clinical outcomes were analysed. And rpoB gene sequencing was done in 6 clinical isolates of M. tuberculosis. Results : The mean age of 18 patients was $43{\pm}14$ years, and the sex ratio is 12:6 (M : F). Sixteen (89%) patients had previous history of tuberculosis. None had diagnosed gastrointestinal disorders, and 2 HIV tests that were performed came out negative. Susceptibility tests were done repeatedly in eleven patients, and six (55%) were mono-RFP resistant repeatedly while five (45%) evolved to MDR tuberculosis. Eight (44%) patients were cured, six (33%) failed, three (17%) were lost to follow-up, and the other one is now on treatment. rpoB gene sequencing showed 5 mutations, codon 531 TCG to TIG mutation in 4 isolates and 526 CAC to TAC in 1 isolate. Conclusion : The clinical characteristics of mono-RFP resistant tuberculosis were similar to that of MDR tuberculosis in Korea where the HIV infection rate is lower than western countries. But some patients with mono-RFP-resistant tuberculous could be cured by primary drug regimens including RFP, suggesting that mono-RFP-resistant tuberculous is a different entity from MDR tuberculosis.

• Journal of Life Science
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• v.24 no.3
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• pp.242-251
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• 2014

The objective of this study was to isolate a photosensitizer from Pueraria thunbergiana leaves that induces apoptosis in SK-HEP-1 cells. Column chromatography and thin layer chromatography were used to isolate active compounds from extracts of P. thunbergiana leaves. The structures of the isolated compounds were determined by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. A substance, named M4-3, was purified from the leaves of P. thunbergiana using various chromatography methods, and the absorbance of the substance was measured. The absorbance was highest at 410 nm, suggesting that the M4-3 substance was a different compound from chlorophyll a and b, which absorb at 410, 502, 533, and 607 nm. Further analyses revealed that the M4-3 compound was a $13^2$-hydoxy pheophorbide, a methyl ester with a molecular weight of 662. M4-3 was identified as a derivative compound of pheophorbide, with a structure that magnesium comes away from the porphyrin ring. The results of the analysis of the cytotoxicity of the M4-3 substance against the SK-HEP-1 cells revealed that it inhibited rates of cell growth by 40% and 80% at a concentration of 0.04 ${\mu}M$ and 0.08 ${\mu}M$, respectively. The M4-3 compound was found to be a photosensitizer for cytotoxicity because it was appeared only in light condition as examining activity in different irradiation conditions (light condition and nonlight condition) under the same concentration. Analysis of morphological changes in the cells following cell death induced by exposure to the M4-3 substance reveled representative phenomena of apoptosis (nuclear condensation, vesicle formation, and fragmentation of DNA). The induction of apoptosis was attributed to the compound's photodynamic activity.

• 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.

• Clinical and Experimental Pediatrics
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• v.48 no.11
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• pp.1193-1200
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• 2005

Purpose : Invasive bacterial infection is a major cause of morbidity and mortality in children. Previously, we reported etiology of invasive infections in healthy children in 1985-1995. This study was performed to update etiology of invasive bacterial infections in the previously healthy children. Methods : We reviewed medical records of 98 episodes of invasive bacterial infections in immunocompetent children at the Seoul National University Children's Hospital in 1996-2004. Results : The frequent pathogens identified over all age groups were Streptococcus pneumoniae (33%) and Staphylococcus aureus(33%). The proportion of Salmonella species and Haemophilus influenzae has been declined to 4% each from 23% and 14%, respectively, compared to previous study. S. agalactiae was the most common isolate in the infants ${\leq}3$ months. Among the infants and children aged 3 months to 2 years and children of 2-5 years, S. pneumoniae(57%, 52%, respectively, in each group) was the most common isolates followed by S. aureus(17% and 24%, respectively). S. aureus was the most common isolates(73%) in children >5 years. Primary bacteremia was the most common clinical diagnosis(27%). S. pneumoniae was responsible for 42% of primary bacteremia, 50% of meningitis, and 69% of bacteremic pneumonia and empyema. S. aureus accounted for 80% of bone and joint infections. The case fatality rate was 8.1% for all invasive infections. Conclusion : We reviewed frequency of bacterial agents of invasive infections in children. The data may be useful for pediatricians to select adequate empirical antibiotics in the management of invasive bacterial infections.

• Clinical and Experimental Pediatrics
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• v.46 no.3
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• pp.265-270
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• 2003

Purpose : To characterize the infants under 3 months of age with urinary tract infections(UTIs), and especially patients with bacteremia or meningitis Methods : Hospital records of all the infants under 3 months of age discharged from our hospital for 69 consecutive months with the diagnosis of initial episode of UTI were reviewed. UTI was defined when patients had fever with pyuria, and had urine culture results of ${\geq}10^5$ colony forming units/mL from a bag specimen. Patients with previously known urologic abnormality or immunodeficiency were excluded. Nosocomial infections were also excluded from the study. Results : The male:female ratio was 35 : 6. Of the urine cultures, 40(97.6%) yielded single pathogen, one yielded two pathogens. Escherichia coli was the predominant isolate from the urine. Five patients(12%) also had bacteremia. Pathogens isolated from the blood cultures were E. coli(4) and Enterococcus faecalis(1). No patient had culture-positive meningitis or cerebrospinal fluid pleocytosis. Clinical or laboratory findings between patients with and without bacteremia were not different significantly. The rate of vesicoureteral reflux(VUR) was 44%. The sensitivity of ultrasound for detection of VUR was 38%; specificity was 50%. Conclusion : Clinical and laboratory data were not helpful for identifying patients with bacteremia at the time of presentation. Consequently, blood cultures need to be obtained from all febrile infants under 3 months of age with UTIs. A large-scale study including the indication of lumbar puncture for infants with a febrile UTI and study of evaluation and treatment of infants under 3 months of age with UTIs are required.

• Korean Journal of Veterinary Service
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• v.24 no.3
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• pp.223-241
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• 2001

From September 1996 to September 1999, 419 Korean-native calves with diseases under 6-month old collected from Kyonggi, Chungcheong, Chonlla and Kyongsang were examined by clinical, microbiological, parasitic, hematologic and histopathological mean. Among them, 124 cases were tested about the neutralization antibodies against infectious bovine rhinotracheitis virus(IBRV), Parainfluenza-3 virus(PI-3V), bovine uiral diarrhea virus(BVDV), bovine ephemeral fever virus(BEFV). In calf diseases in the survey, enteric diseases(72.8%) were most frequently involved and the following orders were taken by respiratory(17.4%) and reproductive (5.0%) disorders. In the causative pathogens associated with calf diseases and motality, 48.4% was induced by bacteria origin and also 35.6% by viral agents. Calf mortality was up to 76.3% in the cae of bacterial diseases and 55.7% in viral diseases. In bacterial diseases, frequent disorders were composed of colibacillosis(52.7%), salmonellosis(13.8%), pasteurellosis(12.8%) and campylobacteriosis(3.9%) and their mortalities showed 73.8% in colibacillosis, 73.0% in pasteurellosis, 67.9% in salmonellosis and 50.0% in campylobacteriosis (50.0%). Among the outbreaks of viral diseases, there were BVD(22.8%), bovine rotavirus infection(20.8%), bovine coronavirus infection(16.8%), bovine respiratory syncytial virus infection(15.4%), IBR(15.4%). Akabane disease(4.7%) and Chuzan diseases(3.4%). Interesting results through this studies were obtained the first isolate to Chuzan virus and Ainovirus in Korea which could be promised the development for diagnostic method and vaccines soon. Calf mortality to Akabane and Chuzan diseases was 100%. Main parasitic diseases were occupied by coccidiosis and babesiosis and their mortality of babesiosis was 20.0%. Other diseases were abomasal impaction(6.7%) and toxicosis(4.5%). The mortality of abomasal impaction was 89.3%. In some causes with malformations(1.9%) were confirmed as anasarca, derodidymus, polymelia, humerus hypoplasia, and tracheal collapse. Calf diseases had mostly been occurred in one month old grout (52.5%) and its prevalence was 25.1% in two to three month old group and 22.4% in four to six month old group. In calf mortality by age, there were 37.9% in one month old group, 18.1% in two and three month old group, and 13.8% in four to six month old group, respectively. The older the age of calf, the less the prevalence of calf enteric diseases. Respiratory diseases in calves to be tested frequently occurred in one to two month old group (41.4%). In one month old calves, the prevalence of enteric disease was 80.0%(p<0.05) and that of reproductive and respiratory disease was 9.5% and 8.2%, respectively. In two month old and four to six month old, enteric disease was 65.7% and 63.8% and respiratory disease was 28.6% and 26.6%. Seasonal prevalence and mortality of Korean-native calf diseases were not a significant difference. Prevalence of calf diseases in summer(31.5%) frequently occurred to compare that in winter(20.3%). Abortion and malformation in calves frequently occurred in spring. Hematological values in 84 calves with clinical signs showed mild to marked leukocytosis. Also, there was slight increase in hematocrit, platelet, mean corpuscular volume and mean plasma volume, but all of those were included the higher level to normal ranges. Calves with respiratory signs showed slightly erythrocytosis. One hundred seventy three calves without clinical signs were not significant different to ill cases in hematological values, but number of platelets was in higher normal range. In 125 calves, 84.8% was taken the antibody to IBRV, but 72% with the antibody had recorded the titer level lower than log$_2$5. The neutralizing antibody levels of higher than $log_{2}5$ to PI-3V and BVD virus were 60.8% and 67.2% cases, respectively. There were the cases of 57.6% had the neutralizing antibody level lower than log$_2$5 to BEFV.

• Journal of Veterinary Clinics
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• v.21 no.1
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• pp.7-14
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• 2004

Bacteria that are resistant to several different groups of antibiotics have increased during the past few years. The importance of surveillance of antimicrobial resistance is now widely recognized. Unfortunately, this development has not been documented continuously in veterinary medicine in Korea. Therefore, the clinical prevalence and trend of antimicrobial susceptibility of aerobic isolates were investigated in this study. Total 121 isolates of aerobic bacteria were isolated from clinical specimens of dogs and cats at Veterinary Medical Teaching Hospital of Seoul National University from May 2001 to October 2002. Among them, the most common isolated species was Staphylococcus spp. (48 isolates), followed by E.coli (26 isolates), Enterococcus spp. (21 isolates), Klebsiella pneumoniae (9 isolates), Streptococcus spp. (6 isolates), Enterobacter cloacae (3 isolates), Pseudomonas aeruginosa (3 isolates), Corynebacterium xerosis (2 isolates), Chryseomonas spp. (2 isolates), and Providencia stuartii (1 isolate). The susceptibility of isolates to antibiotics was determined by the disk diffusion method. Gram-positive bacterial isolates were showed high susceptibilities to amikacin, amoxacillin/clavulanate, ceftazidime, and oxacillin, while Gram-negative bacterial isolates were showed high susceptibilities to amikacin and ceftazidime. Staphylococcus spp. were showed high susceptibilities to amikacin, amoxicillin/clavulanate, ceftazidime, cephalothin, and oxacillin. Streptococcus spp. and E.coli were showed high susceptibilities to amikacin and ceftazidime. Of the 48 staphylococci, seven Methicillin Resistant staphylococci were observed (14.6%), distributed among S. auricularis (1), S. hemolyticus (2), S. sciuri (1), S. saprophyticus (1), S. warneri (2) isolates. One strain of E.coli and one strain of Corynebacterium xerosis were resistant to all antibiotics tested. And, resistance trends between the you 2000 (from July 1999 to September 2000) and 2002 (from May 2001 to October 2002) were compared. Resistance to antibiotics was increased in both Gram-positive and Gram-negative bacterial isolates (p＜ 0.05). The resistance rates of Staphylococcus spp., E.coli and Klebsiella pneumoniae to all antibiotics tested were also increased (p＜0.05). This study investigated increasing resistance between the year 2000 and 2002 in Veterinary Medical Teaching Hospital of Seoul National University. Surveillance resistance is helpful to alert to veterinarian and select of appropriate therapy. Antimicrobial susceptibility surveillance of isolates should urgently be continued in veterinary medicine.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.1
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• pp.1-6
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• 2000

Dentinogenesis imperfecta is an inherited disorder of dentin formation, usually exhibiting an autosomal dominant mode of transmission. Type I dentinogenesis imperfecta occurs in patients afflicted with osteogenesis imperfecta. Type II dentinogenesis imperfecta is not associated with osteogenesis imperfecta. Type III dentinogenesis imperfecta (Brandywine type) occurs in a racial isolate area in the state of Maryland. In all three types, teeth of both dentitions are affected with variable clinical appearances. The teeth are opalescent with the color ranging from bluish-gray to brown to yellowish. The dentin is abnormally soft, providing inadequate functional support to the overlying enamel. Although the enamel is normal, it fractures or chips away easily, exposing the occlusal and incisal dentin. The exposed soft dentin often undergoes rapid and severe functional attrition. The teeth exhibit bulb-shaped crowns with constricted cementoenamel junctions and thin roots. The teeth will exhibit varying stages of obliteration of the coronal and root pulpal chambers. The cementum, periodontal ligament and supporting alveolar bone appear normal. The enamel is normal. The mantle dentin remains nearly normal, whereas the remaining dentin is severely dysplastic. The dentinal tubules are disoriented, irregular, widely spaced, and usually larger than normal.

• Korean Journal of Veterinary Service
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• v.20 no.1
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• pp.103-125
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• 1997

An epidemiologic study on pleuropneumonia in the slaughter pigs(Chonan and Asan area, Chungnam province, Korea) during the period of January 1994 through December 1995 was conducted. Isolation of A pleuropneumoniae was attempted in 425 pigs with pneumonic lesions. Biochemical properties, antimicrobial susceptibility, serotypes and pathogenicity of isolated A pleuropneumoniae were investigated. In addition, outer membrane protein(OMP) of the Isolates were extracted to determine its properties and immunogenicity in both mice and piglets The results obtained through this study were summarized as followed ; 1. Of 3, 395 slaughter pigs, pleuropneumonia was observed in 425 pigs(10.6%). A pleuropneumoniae was isolated from 22 pigs(5.2%) out of 425 pigs with pneumonic lesions. The biochemical properties of all isolates were same as those of reference A pleuropneumoniae strain. Among 22 isolates, 9, 1 and 12 isolates were serovar 2, 3 and 5, respectively. 2. The results of antimicrobial susceptibility test revealed that the isolates showed high susceptibility to ciprofloxacin and cephalothin, moderate susceptibility to amikacin, gentamicin, kanamycin and streptomycin, and low susceptibility to erythromycin, tylosin and sulfadimethoxin. 3. The isolates were varied in pathogenicity to mice. Median lethal dose of LE9402(serovar 2) and LE9511(serovar 5) were $9.2{\times}10^7$ CFU and $2.8{\times}10^7$%CFU, respectively. Specific pneumonic lesions were observed from the infected mice with clinical signs. Bacteria recovery rate was high in the lung, but low In heart blood and tracheas. 4. Serovar 2 was found to be more pathogenic than serovar 5 in guinea pig. Mortality on guinea pigs inoculated with serovar 2($5.4{\times}10^8-5.4{\times}10^6$CFU) and serovar 5($2.8{\times}10^8-2.8{\times}10^6$ CFU) was 20~40% and 40~80%, respectively. A severe hemorrhagic lesions and focal pneumonic lesions were observed from dead guinea pigs. Bacteria recovery rate was relatively higher in the lung than that of other organs. 5. In the SDS-PAGE analysis, OMP-enriched fractions of both isolates and reference strains contain common peptide bands equivalent to molecular weight of 17, 27, 42, 52 and 95Kd. In addition to common peptide bands, the bands which are specific to each isolate were also observed. The profiles of Sephadex G25 fractions showed 3 major peaks. The common peptide bands which were observed by SDS-PAGE of the crude OMPs were found in the peaks 1 and 2. 6. The OMPs extracted from serovar 2(LE9402) and serovar 5(LE9511) provided high level of protection in mice(70~80%) and pigs(100%). All animals inoculated with OMPs were seroconverted, showing micro-agglutination titer of 640 to 1280.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1179-1185
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• 2008

The immune-stimulating activities of Bordetella bronchiseptica antigens containing dermonecrotoxin (BBD) loaded in chitosan microspheres (CMs) have already been reported in vitro and in vivo with a mouse alveolar macrophage cell line (RAW264.7) and mice. Therefore, this study attempted to demonstrate the successful induction of mucosal immune responses after the intranasal administration of BBD loaded in CMs (BBD-CMs) in colostrum-deprived pigs. The BBD was introduced to the CMs using an ionic gelation process involving tripolyphosphate (TPP). Colostrum-deprived pigs were then directly immunized through intranasal administration of the BBD-CMs. A challenge with a field isolate of B. bronchiseptica was performed ten days following the final immunization. The BBD-specific IgG and IgA titers, evident in the nasal wash and serum from the vaccinated pigs, increased with time (p<0.05). Following the challenge, the clinical signs of infection were about 6-fold lower in the vaccinated pigs compared with the nonvaccinated pigs. The grades for gross morphological changes in the turbinate bones from the vaccinated pigs were also significantly lower than the grades recorded for the nonvaccinated pigs (p<0.001). Therefore, the mucosal and systemic immune responses induced in the current study would seem to indicate that the intranasal administration of BBD-CMs may be an effective vaccine against atrophic rhinitis in pigs.