Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.
Park, Jun-Ik;Kim, Hak-Bong;Kim, Mi-Ju;Lee, Jae-Won;Bae, Jae-Ho;Park, Soo-Jung;Kim, Dong-Wan;Kang, Chi-Dug;Kim, Sun-Hee
Journal of Life Science
/
v.19
no.9
/
pp.1209-1217
/
2009
Many cancer cells are sensitive to the TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis. However, some cancer cells show either partial or complete resistance to TRAIL. Human colon carcinoma KM12 cells have been shown to be insensitive to TRAIL-induced apoptosis. To overcome TRAIL resistance in KM12 cells, we targeted key anti-apoptotic molecules involved in the modulation of TRAIL resistance in the cells, and evaluated the effects of quercetin as a TRAIL sensitizer in the cells. We found that quercetin acted in synergy with TRAIL to enhance TRAIL-induced apoptosis in KM12 cells by the down-regulation of c-FLIP and DNA-PKcs/Akt and up-regulation of death receptors (DR4/DR5), which led to the enhancement of TRAIL-mediated activation of caspases and subsequent cleavage of PARP, as well as up-regulation of Bax. These findings suggest that the DNA-PKcs/Akt signaling pathway, as well as c-FLIP, play essential roles in regulating cells in the escape from TRAIL-induced apoptosis. Based on these results, this study provides a potential application of quercetin in combination with TRAIL in the treatment of human colon cancer.
Bensulfuron concentrations of $10^{-7}$, $10^{-6}$, $10^{-5}$ and $10^{-4}M$ were applied to agar medium on susceptible (cv. KH 17854 and cv. IR 1846) and tolerant (cv. Chinsurah Boro II and IR 14252) rice cultivars were grown for microscopic inspection. Susceptible cultivars showed the decrease in shoot and root growth at the concentration of $10^{-5}M$ while ones showed no difference. Such a tendency was also observed from microscopic inspection in the elongation zone of shoot meristematic tissue. Seedlings grown in soil for 10 days were transfered to distilled water containing only bensulfuron solutions. There were significant differences between cultivars in terms of supression of shoot meristematic activity and swelling of cell volume. Observations of those cells made it clear that especially susceptible cultivars showed the irregular cell layering, vacuolation, cell swelling and partial damage in membrane of shoot tissue. The major response of root tips of susceptible cultivars showed the disorganization of cortex, rupture and contraction of membrane, inhibition of cell division, swelling and emergence of lateral root while tolerant ones showed no such responses.
A serial study on differential response in absorption and translocation of $^{14}C$-oxyfluorfen was conducted by use of a group of selected rice cultivars as the tolerant or the susceptible to oxyfluorfen. Trial 1. Differential response in absorption and translocation of selected cultivar group. The susceptible cultivar group has reached as higher rate as 209%, 193%, 344%, 204% and 152% of the tolerant cultivar group in root absorption rate per unit dry weight, lower-shoot absorption rate, higher shoot absorption rate, whole amount of absorption, and the rate of translocation from bottom to shoot, respectively. Trial 2. Differential Response in Absorbtion and translocation of selected cultivar as affected by exposed portion and time. ${\bullet}$ Regardless of cultivar, the rate of root absorption has effectively realized but the translocation was significantly limited. ${\bullet}$ Lower portion of shoot has also achieved a higher amount of absorption but the translocation was very limited. ${\bullet}$ By relaying the exposure time. the amount of root absorption was ted increasing, and persistent absorption was rather effective in the susceptible (cv. Mushakdanti) cultivar than the tolerant (cv. Chokoto). ${\bullet}$ Translocation of oxyfluorfen from shoot to root was tended to easily attaining in both cultivars.
Eighty-eight strain3 o~methicillin resistant Stopllylococcus awecis were Isolated from pus (64.7%); spuhm (26.2%), blood, fluid, andurine of 83 patients at Dong-A Hospital in P~~san to invesligate theil-coagulase typ- Ing, and multi-drug resistaut ppattems. The presence of niec A gene confe~~ing melhicillin resistance was tested by polymerase chain reaction (PCR) with uwo mec A gene specific primers using purified clromosonlal DNA as templates. DNA fragments of expected size wel-e detected frorn 86 strains, but not from two strains. !i coagulase typmg, the 86 isolates were assigned to 5 coagulase lypes, I, 11, lll. 1V, VI, VII, VIlI, but there was no isolate helong lo type V. The most abundant coagulase type was type TI(50 %), lollowed by type IV Rest ofthe coagulase types were ininor; ranging fmm 4.5 to 12.5 '% Most of the type I1 ~netlucillin resistant Stapl\ulcorneryiococcus nwem (MRSA) strams were isolated from the generd sulzely ward, but major strains of type IV were Isolated from the otorhinolq~ngology of the hospital's outpatient clinic center. All of the 88 st~nins were sensitive to vancomycin and teicoplanin, but 71 (81%) strains showed multi-drug resitant to penicillin, cephalotl~n, eiythroinycin, gentan~ycin, imipenem, clindamycin, ciprofloxacin and ooxacillin. Yo relationship was found between the antibiotic resistance pattems aud the coagulase typing patterns.
Jung, Hye-In;Jung, So Young;Park, Indal;Bae, Il Kwon
Journal of Life Science
/
v.26
no.2
/
pp.220-225
/
2016
Coagulase-negative staphylococci (CNS) have recently become the bacteria most frequently found in clinical infections. The aim of this study was to investigate the prevalence, antimicrobial susceptibilities, and molecular characteristics of CNS isolates from dental clinic environments in Busan, Korea. One hundred and fifty-four samples were collected from 10 dental clinics and dental hospitals in Busan from December 2014 to January 2015. Species were identified by matrix-assisted laser desorption/ionization–time-of-flight. Antimicrobial susceptibility was determined by disk diffusion methods. A polymerase chain reaction was performed to detect mecA, mupA gene, and SCCmec types. Of the 154 samples, 10(6.5%) isolates were identified as CNS (5 Staphylococcus epidermidis, 2 Staphylococcus capitis, 2 Staphylococcus, and 1 Staphylococcus haemolyticus). Among the 10 isolates, 6 were resistant to penicillin, 5 were resistant to gentamicin, 3 were resistant to tetracycline, and 2 were resistant to cefoxitin and erythromycin. However, clindamycin, ciprofloxacin, teicoplanin, and trimethoprim-sulfamethoxazole resistant isolates were not present. Genes encoding mecA were detected in 4 (2 S. warneri and 2 S. haemolyticus) isolates, and mupA in 1 (S. epidermidis) isolate. One methicillin-resistant CNS (S. warneri) isolate was determined as being of the SCCmec type I. It is concluded that CNS resistant to various antimicrobial agents was widely distributed in dental clinic environments in Korea.
Inwon HWANG;Sang-Ha KIM;Taewon JUNG;Young-Kwon KIM;Sunghyun KIM
Korean Journal of Clinical Laboratory Science
/
v.56
no.2
/
pp.115-124
/
2024
The emergence and spread of Staphylococcus aureus, which is resistant to quinolone antibacterial agents, has made it difficult to treat infectious diseases. Accordingly, this study examined the molecular epidemiological characteristics of quinolone-resistant S. aureus (QRSA) to obtain helpful data for treatment. Mutations in mecA and SCCmec typing, gyrA, and gyrB genes were investigated for QRSA strains isolated from the blood culture specimens at a general hospital in Daejeon Metropolitan City. The ciprofloxacin-resistant strains in SCCmec typing were II (44 strains, 73%), IVa (five strains, 8%), III, and V (one strain, 2%); the non-typeable strains (11 strains, 18%), and levofloxacin (LVX) and moxifloxacin (MXF) strains were II (44 strains, 73%), IVa (five strains, 8%), III, and V (one strain, 2%); the non-typeable strains were 10 (17%). In both gyrA and gyrB regions, there were 58 mutations, or 96.7%. In LVX, there were 56 mutations or 93.3%, and in MXF, there were 57 mutations or 95%. Twelve mutations, six mutations each in gyrA and gyrB, were identified for the QRSA strain. The resistance rate for the quinolone antibiotics of QRSA studied was approximately 98%, and 12 mutations, six each in gyrA and gyrB, were identified in the QRSA strain. Therefore, the rational use of antibiotics needs to be improved.
Kim, Tae-Hyung;Kim, Kyoung-A;Lim, Chae-Man;Lee, Sang-Do;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
Tuberculosis and Respiratory Diseases
/
v.45
no.3
/
pp.509-518
/
1998
Background: Short-course chemotherapy for 6 months is well established for pulmonary tuberculosis. However, little is known about the efficacy of the short-course chemotherapy for tuberculous pleural effusion. Tuberculous pleural effusion itself may be self-limiting without any treatment, but about two thirds of the patients with tuberculous pleural effusion may subsequently develop pulmonary tuberculosis within 5 years. After completing treatment for tuberculous pleural effusion. prolonged follow-up is necessary for evaluating the efficacy of the treatment There is still no report on the efficacy of 6-month regimens for tuberculous pleural effusion in Korea, where the incidence of tuberculous disease and drug resistance is high. We studied the efficacy of 6 month short-course chemotherapy comparing with 9 month chemotherapy. Method : Retrospective study was done through medical record review in 238 patients with tuberculous pleural effusion who admitted to Asan Medical Center during May 1989-May 1993. The diagnosis of tuberculous pleural effusion was made by bacteriologic or histopathologic study. Results: Among 238 patients, 38 patients were dropped out during follow-up period. In 2 patients, second line drugs were prescribed according to known drug resistance results. And, in 23 patients, treatment longer than 9 months was done due to accompanying extrapulmonary tuberculosis or durg resistance. In 8 patients, treatment regimen was changed due to hepatotoxicity. Remaining 167 cases (70.2%) completed the treatment as scheduled ; 6 month chemotherapy in 88 cases and 9 month chemotherapy in 79 cases. In 60 patients (35.9%) with pleural effusion only in chest X-ray finding, sputum smear or culture for M.tuberculosis was positive in 6 cases (10.0%), and in 63 patients (37.7%) with radiologically inactive pulmonary tuberculosis, sputum smear or culture was positive in 18 cases (28.6%). In 44 patients (26.3%) with radiologically active pulmonary tuberculosis, the sputum smear or culture was positive in 24 cases (54.5%). In 6-month chemotherapy group (n=88), during mean 23 months (range; 1~61months) follow-up period, pulmonary tuberculosis developed in 1 case (1.4%). In 9-month chemotherapy group(n=79), during mean 23 months (range; 3~70months) follow-up period, pulmonary tuberculosis developed in 2 cases (2.5%). All the cases who developed pulmonary tuberculosis also showed active pulmonary tuberculosis on initial chest X-ray before treatment Conclusion: In patients with tuberculous pleural effusion, the incidence of pulmonary tuberculosis after 6 month chemotherapy showed no difference from that after 9 month chemotherapy. Thus, 6 month short-course chemotherapy seems to be an effective treatment for tuberculous pleural effusion.
Noh, Sung Hoon;Yu, Ka Young;Kim, Jung Soo;Hwang, Pyoung Han;Jo, Dae Sun
Clinical and Experimental Pediatrics
/
v.52
no.7
/
pp.791-797
/
2009
Purpose : We aimed to investigate the clinical manifestations of and antibiotic resistance in culture-proven childhood salmonellosis. Methods : Clinical manifestations and laboratory data of and antibiotic use in subjects with culture-proven childhood salmonellosis, who were treated at the Chonbuk National University Hospital between September 1998 and August 2008, were analyzed retrospectively. Patients with underlying diseases or concomitant illnesses were excluded. Results : We assessed a total of 72 patients. There were 68 stool culture-positive cases, 7 blood culture-positive cases, and 3 both stool culture- and blood culture-positive cases. Salmonella group D was the most frequent pathogen in stool (63.9%) and blood (71.4%) cultures. Salmonella typhi was isolated in 1 case. Of the 72 patients, 45 (62.5%) were male children, of which 29 (40.3%) were aged <3 years. The patients most commonly presented with diarrhea (90.2%) and fever (83.3%). Leukocytosis (leukocyte count, >$15,000/{\mu}L$) and leukopenia (leukocyte count, <$4,000/{\mu}L$) were detected in 8.3% and 5.6% of the patients, respectively. Elevated serum C-reactive protein concentration (>5 mg/dL) and erythrocyte sedimentation rate (>20 mm/h) were observed in 88.9% and 58.3% of the patients, respectively. Fifty-two (85.2%) of 61 patients who had undergone antibiotic treatment received a third-generation cephalosporin as definitive antibiotic therapy. Multidrug resistance rate was 40.0 % in the first 5 years of the study and 71.4% in the last 5 years. No fatalities occurred in this series. Conclusion : Children with culture-proven salmonellosis showed relatively benign clinical outcomes. Appropriate antibiotic treatment of <2 weeks is probably adequate for those without a suppurative focus of infection. The incidence of antibiotic resistant isolates was recently seen to increase.
A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.
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