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Establishment of Embryonic Stem Cell Line from Isolated Blastomeres from Mouse Preimplantation Embryos (생쥐 초기 배아에서 분리한 할구를 이용한 배아줄기세포주 확립)

  • Lim, Chun Kyu;Sung, Ji Hye;Choi, Hye Won;Cho, Jae Won;Shin, Mi Ra;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.1
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    • pp.25-33
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    • 2006
  • Objective: The aim of this study was to investigate whether embryonic stem (ES) cells can be established from isolated blastomeres of mouse embryos. Methods: Blastomeres were separated from mouse (C57Bl/6J) 2- or 4-cell embryos. Isolated blastomeres or whole 4-cell embryos were co-cultured with mitosis-arrested STO feeder cells in DMEM supplemented with recombinant murine leukemia inhibitory factor and ES-qualified fetal bovine serum. After the tentative ES cell lines were maintained from isolated blastomeres or whole embryos, some of them were frozen and the others were sub-cultured continually. Characteristics of tentative ES cell lines as were evaluated for specific genes expressions with immunocytochemistry and RT-PCR. Results: One ES cell line (3.0%) was established from isolated blastomere of 2-cell embryo and one cell line (4.0%) from isolated two blastomeres of 4-cell embryo. And five cell lines (16.7%) were established from whole 4-cell embryos. Both cell lines from isolated blastomere and whole embryo expressed mouse ES cell specific markers such as SSEA-1, Oct-4 and alkaline phosphatase. Marker genes of three germ layers were expressed from embryoid bodies of both cell lines. Conclusion: This study suggests that mouse ES cells could be established from isolated blastomeres, although the efficiency is lower than whole embryos. This animal model could be applied to establishment of autologous human ES cells from biopsied blastomeres of preimplantation embryos in human IVF-ET program.

Growth and Predictive Model of Wild-type Salmonella spp. on Temperature and Time during Cut and Package Processing in Cold Pork Meats (냉장돈육 가공공정 온도와 시간에서의 Wild-type Salmonella spp.의 성장특성 및 예측모델)

  • Song, Ju Yeon;Kim, Yong Soo;Hong, Chong Hae;Bahk, Gyung Jin
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.7-12
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    • 2013
  • This study presents the influence on growth properties determined using a novel predictive growth model of wild-type Salmonella spp. KSC 101 by variations in the temperature and time during cut packaging in cold, uncooked pork meat. The experiment performed for model development included an arrangement of different temperatures ($0^{\circ}C$, $5^{\circ}C$, $10^{\circ}C$, $15^{\circ}C$, and $20^{\circ}C$) and time durations (0, 1, 2, and 3 hours) that reflect actual pork-cut and packaging processes. No growth was observed at $0^{\circ}C$ and $5^{\circ}C$, whereas some growth was observed at $10^{\circ}C$, $15^{\circ}C$, and $20^{\circ}C$, with a mean increase of only 0.34 log CFU/g. The growth observed at $20^{\circ}C$ was more robust than that observed at $15^{\circ}C$, but the difference was not statistically significant (p > 0.05). However, compared with PMP (Pathogen Modeling Program), the wild-type Salmonella spp. KSC 101 showed a more rapid growth. We used the Gompertz 4 parameter equation as the primary model, and the exponential decay formula as the secondary model. The estimated $R^2$ values were 0.99 or higher. The developed model was evaluated by comparison of the experimental and predictive values, and the values were in agreement with the ${\pm}0.5$ log CFU/g, although the RMSE (Root mean square error) value was 0.103, which indicates a slight overestimation. Therefore, we suggest that the developed predictive growth model would be useful as a tool for evaluating sanitation criteria in pork cut-packaging processes.

Development of Detection Method for Oilfish (Ruvettus pretiosus and Lepidocybirium flavobrunneum) as a Food Materials not Usable in Foods (식품원료로 사용금지 대상인 기름치 (기름갈치꼬치 및 흑갈치꼬치) 판별법 개발)

  • Park, Yong-Chjun;Kim, Mi-Ra;Jung, Yong-Hyun;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.50-55
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    • 2013
  • Since 1 June 2012, it is prohibited to sell oilfish as a food material but there are still many illegal cases of selling oilfish as if it is tuna or grilled Patagonian toothfish. So it is absolutely crucial to construct the system to distinguish the real food material from oilfish. There are two sorts of oil fish called Ruvettus pretiosus and Lepidocybirium flavobrunneum involved in Percifomes order and Gempylidae class. 16S DNA gene region in mitochondria was selected to design the specific primers. For design species-specific primer, the theoretical experiment were performed for the sequences of R. pretiosus, L. flavobrunneum, Thunnus thynnus, Thunnus albacores, Makaira mitsukurii and Xiphias gladius, registered at the Gene bank from the National Centre for Biotechnology Information, using BioEdit 7.0.9.0. program. Through the analysis of the result from experiments, it was possible to design the 4 kinds of primers to distinguish R. pretiosus and L. flavobrunneum. As a comparison group, 3 kinds of tuna and 4 kinds of billfishes were selected and experimental verification was performed. As a result, for R. pretiosus and L. flavobrunneum, R.P-16S-006-F/R.P-16S-008-R and L.F-16S-004-F/L.F-16S-006-R primers were selected eventually and PCR condition was established. In addition, 178bp and 238bp of PCR products were confirmed from the established condition and non-specific band was not amplified among similar species. Therefore, the species-specific primers developed in this study would be very useful and used in various ways such as internet shopping mall and illegal distributions with fast and scientific results.

A Study of the Effect of a Mixture of Hyaluronic Acid and Sodium Carboxymethyl Cellulose ($Guardix-sol^{(R)}$) on the Prevention of Pericardial Adhesion (Hyaluronic Acid와 Sodium Carboxymethyl Cellulose 혼합용액($Guardix-sol^{(R)}$)의 섬유막유착 방지 효과에 관한 연구)

  • Lee, Song-Am;Kim, Jin-Sik;Kim, Jun-Seok;Hwang, Jae-Joon;Lee, Woo-Surng;Kim, Yo-Han;Cho, Yang-Kyu;Chee, Hyun-Keun
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.596-601
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    • 2010
  • Background: This study was designed to evaluate the efficacy of a mixture of hyaluronic acid and sodium carboxymethyl cellulose ($Guardix-sol^{(R)}$) on experimental pericardial adhesion. Material and Method: Thirty rats were divided into 2 groups of 15 rats each and pericardial mesothelial injury was induced during surgery by abrasion. In the control group, blood and normal saline were administered into pericardium; in the test group, blood and HA-CMC solution were administered. Pericardial adhesions were evaluated at 2 weeks (n=5), 4 weeks (n=5), and 6 weeks (n=5) after surgery. The severity of adhesions was graded by macroscopic examination, and the adhesion tissue thickness was analyzed microscopically with Masson trichrome stain and an image processing program. Result: The test group had significantly lower macroscopic adhesion scores ($2.9{\pm}0.6$ : $3.9{\pm}0.4$, p<0.000) compared with the control group. For microscopic adhesion tissue thickness, the test group had lower scores compared with the control group, but this difference was not statistically significant ($91.73{\pm}49.91$ : $117.67{\pm}46.4$, p=0.106). Conclusion: We conclude that an HA-CMC solution ($Guardix-sol^{(R)}$) reduces the formation of pericardial adhesions in this animal model.

Association of SNP Markers on Chromosomes 3 and 9 with Body Weight in Jeju Horses (제주마에서 3번 및 9번 염색체상의 단일염기변이와 생체중과의 관련성 연구)

  • Kim, Nam Young;Yang, Young Hoon;Park, Nam Geon;Yang, Byoung Chul;Son, Jun Kyu;Shin, Sang Min;Woo, Jae Hoon;Shin, Moon Cheol;Yoo, Ji Hyun;Hong, Hyun Ju;Park, Hee Bok
    • Journal of Life Science
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    • v.28 no.7
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    • pp.795-801
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    • 2018
  • This study was conducted to investigate the association of single nucleotide polymorphism (SNP) markers on equine chromosomes (ECA) 3 and 9 with body weight in Jeju horses. We used DNA samples and body weight data of 320 horses provided by the Livestock Promotion Agency, Jeju Special Self-Governing Province, and the Korean Racing Association, respectively. We genotyped all the experimental animals using nine SNP markers located on ECA 3 (BIEC2-808466, BIEC2-808543, BIEC2-808967, and BIEC2-809370) and ECA 9 (BIEC2-1105370, BIEC2-1105372, BIEC2-1105377, BIEC21105505, and BIEC2-1105840). These markers were selected due to their effects on body conformation traits in horses. The joint effect of the genotypes of the two SNP markers (BIEC2-808467 and BIEC2-1105377) regarding body weight were also evaluated. The estimated breeding value (EBV) of body weight was obtained as the dependent variable for association analyses using a linear mixed model. Significant associations were detected between SNP markers (BIEC2-808543, BIEC2-808967, BIEC2-809370, BIEC2-1105370, BIEC2-1105372, and BIEC2-1105377) and the body weight EBV. In addition, the joint genotype effect of the BIEC2-808467 and BIEC2-1105377 on the body weight EBV was significant. These results indicate that the SNP markers, which showed their significant effects on body conformation, can be used as genetic markers to improve the efficiency of the selective breeding program for the body weight traits in Jeju horses.

Protective effects of Gastrodia rhizoma and steamed & fermented Gastrodiae rhizoma with anti-oxidant efficacy and suppression of NFκB signaling pathway on LPS-induced liver injury (LPS로 유발한 간손상 마우스에서 항산화 및 항염증 효능을 통한 천마와 증숙 발효 천마의 간보호 효과)

  • Kim, Hyoung-Jin;Kwon, O Jun;Lee, Ah Reum;Roh, Seong-Soo;Seo, Young-Bae
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.179-188
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    • 2016
  • This study is aimed to evaluate the protective effect of Gastrodiae rhizoma and steamed, dried & fermented Gastrodiae rhizoma on Lipopolysaccharide (LPS)-induced hepatic injury in the mice model. Sample was selected to GR0F0 (not processed gastrodia rhizome) and GR6F4 (fermented with Saccharomyces cerevisiae before steamed and dried 6 times) based on 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid, and High-performance liquid chromatography analysis. Mice were randomly divided into 4 groups - Normal group, vehicle group (LPS treated), GR0F0 group (fed GR0F0 before LPS treated) and GR6F4 group (fed GR6F4 before LPS treated) with 6 mice in each group. GR0F0 group and GR6F4 group were fed each extract 200 mg/kg/day during 8 days. LPS 20 mg/kg injected to the experimental groups as abdominal injection. We measured aspartate aminotransferase, alanine amino-transferase in serum. GR0F0 and GR6F4 showed a significant decrease compared to the vehicle group. As a result of measuring the ROS, GR6F4 group showed a significant reduction in both the serum and liver tissues compared to the vehicle group. GR0F0 group showed a significant reduction only in the liver tissues. Activator protein-1, cyclooxygenase-2, and Inducible nitric oxide synthase were significantly decreased GR0F0 group and GR6F4 group. But tumor necrosis factor alpha only showed a significant reduction in GR6F4 group. GR0F0 and GR6F4 groups against liver damage in mice with LPS. That showed significant effects on anti-oxidant and anti-inflammatory action. The effects of GR6F4 group showed superior results compared to GR0F0 group. Therefore, Steamed, dried & fermented Gastrodia rhizoma was might have a protective effect on liver injury.

Design of the Condenser and Automation of a Solar Powered Water Pump (태양열 물펌프의 운전 자동화 설계)

  • Kim Y. B.;Son J. G.;Lee S. K.;Kim S. T.;Lee Y. K.
    • Journal of Animal Environmental Science
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    • v.10 no.3
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    • pp.141-154
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    • 2004
  • The solar powered water pump is very ideal equipment because solar power is more intensive when the water is more needed in summer and it is very helpful in the rural area, in which the electrical power is not available. The average so]ar radiation energy is 3.488 kWh/($m^2{\cdot}day$) in Korea. In this study, the automatic control logic and system of the water pump driven by the radiation energy were studied, designed, assembled, tested and analyzed for realizing the solar powered water pump. The experimental system was operated automatically and the cycle was continued. The average quantity of the water pumped per cycle was about 5,320 cc. The cycle time was about 4.9 minutes. The thermal efficiency of the system was about $0.030\%$. The pressure level of the n-pentane vapour in flash tank was 150$\%$450 hPa(gauge) which was set by the computer program for the control of the vapour supply. The pressure in the condenser and air tank during cycles was maintained as about 600 hPa and 1,200 hPa respectively. The water could be pumped by the amount of 128kg/($m^2{\cdot}day$) with the efficiency of $0.1\%$ and the pumping head of 10 m for the average solar energy in Korea.

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Improvement of Pregnancy Rate in Preimplantation Genetic Diagnosis with FISH Procedure by the Laboratory Optimization and Experiences (형광직접보합법을 이용한 착상전 유전진단 기법의 최적화와 경험 축적에 의한 임신율의 향상)

  • Lim, Chun-Kyu;Min, Dong-Mi;Lee, Hyoung-Song;Byun, Hye-Kyung;Park, So-Yeon;Ryu, Hyun-Mee;Kim, Jin-Young;Koong, Mi-Kyoung;Kang, Inn-Soo;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.29-39
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    • 2004
  • Objectives: This study was performed to evaluate the laboratory system for successful PGD using fluorescence in situ hybridization (FISH) and the clinical outcome of PGD cycles in five years experiences. Methods: A total of 181 PGD-FISH cycles of 106 couples were performed, and diagnosed chromosome normality in the preimplantation embryos. The laboratory and clinical data were classified by the following optimization steps, and statistically analyzed. Phase I: Blastomere biopsy with two kinds of pipettes, removal of cytoplasmic proteins without treatment of pepsin and culture of biopsied embryos with single medium; Phase II: Blatomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with single medium; Phase III: Blastomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with sequential media. Results: A total of 3, 209 oocytes were collected, and 83.8% (2, 212/2, 640) of fertilization rate was obtained by ICSI procedure. The successful blastomere biopsies were accomplished in 98.6% (2, 043/2, 071) of embryos, and the successful diagnosis rate of FISH was 94.7% (1, 935/ 2, 043) of blastomeres from overall data. Embryo transfers with normal embryos were conducted in 93.9% (170/181) of started cycles. There was no difference in the successful rate of biopsy and diagnosis among Phase I, II and III. However, the pregnancy rate per embryo transfer of Phase III (38.8%, 26/67) was significantly (p<0.05) higher than those of Phase I (13.9%, 5/36) and Phase II (14.9%, 10/67). Conclusions: The laboratory optimization and experience for the PGD with FISH procedure can increase the pregnancy rate to 38.8% in the human IVF-ET program. Our facility of PGD with FISH provides the great possibility to get a normal pregnancy for the concerned couples by chromosomal aberrations.

Effect of Breast-feeding Education and Follow-up care on the Breast-feeding Rate and the Breast-feeding Method - Focused on Home Visit and Phone Counselling - (모유수유교육과 추후간호방법이 산모의 모유수유실천율과 모유수유방법에 미치는 효과 - 가정방문과 전화상담을 중심으로 -)

  • Park, Sook-Hee;Koh, Hyo-Jung
    • Women's Health Nursing
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    • v.7 no.1
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    • pp.30-43
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    • 2001
  • This was a pre-experimental study to confirm the breast-feeding ability and effect of follow up care on the breast-feeding rate and the breast-feeding method by the mother-infant breast-feeding assessment tool for the mothers who got the breast-feeding education. The subjects were 46 mothers who experienced normal delivery of infants at a college hospital in K-city, Kyungsangbuk-do, from July 1 to October 21, 2000. The instrument for data obtainment were The Mother-Infant Breast-Feeding Assessment Tool of Johnson et al. (1999), and The Breast- Feeding Method Measurement Tool of Jeong, Geum-hee(1997). This instrument was reliable, showing Cronbach $\alpha$.751. This study classified them into 3 groups: at high risk for breast-feeding failure, at risk for breast-feeding problems, and at low risk for breast-feeding failure by the mother-infant breast-feeding assessment tool on the day of discharge from the hospital after delivering individual breast-feeding education to the subjects. This study investigated the breast-feeding rate and the breast-feeding method through mail questionnaire at the four week after childbirth, and through the phone counselling and the home visit for follow up care at the first week and the second week after childbirth. The sixth week after childbirth, this study investigated the breast-feeding rate by phone. The data analyzed the hypothesizes by $x^2$-test, paired t-test, ANOVA, Wilcoxon signed rank test, Wilcoxon rank sum test and trend analysis using SPSS/PC+ WIN 10.0 program. The results were as follows : 1) Hypothesis 1-1, "there won't be any difference the breast-feeding rate of a group at risk for breast-feeding failure by the time elapsed" was supported through constant the breast-feeding rate, because changes in the breast-feeding rate by the time elapsed after childbirth wasn't statistically significant(t= -1.501, p=.270). Hypothesis 1-2, "there won't be any difference the breast-feeding rate of group at low risk for breast-feeding failure by the time elapsed" was supported through constant the breast-feeding rate, because changes in the breast-feeding rate by the time elapsed after childbirth wasn't statistically significant(t=-1.732, p=.225). 2) Hypothesis 2-1, "there won't be any difference between the breast-feeding method of group at risk for breast-feeding failure for four weeks after childbirth and just after childbirth” was rejected, because the mean point of post test appeared to be higher than that of pre test(t=-7.267, p=.000). Hypothesis 2-2, "there won't be any difference between the breast-feeding method of the group at low risk for breast-feeding failure for four weeks after childbirth and just after childbirth" was rejected, because the mean point of post test appeared to be higher than that of pre test(t=-2.501, p=.012). 3)The 3rd hypothesis, "there won't be any difference between breast-feeding method of groups at risk for breast-feeding problems and at low risk for breast-feeding failure at the 4th week after childbirth and just after childbirth" didn't show any difference between the breast-feeding method of groups at risk for breast-feeding problems and at low risk for breast-feeding failure in the advance test(t=-1.521, p=.130) but there was difference between them in post test (t=-2.012, p=.044). As a result, the 3rd hypothesis was supported by pre test, but it was rejected by post test. In conclusion, this study confirmed breast- feeding education and follow up care just after childbirth were effective for the breast-feeding rate and method. Accordingly, it is proposed that successful nursing intervention of breast-feeding to be necessary by continuously providing follow up care through the mother-infant breast-feeding assessment tool as well as to execute individual breast-feeding education to mothers just after childbirth.

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Sputum Smear Conversion During mDOT (Modified Directly Observed Treatment) (변형된 복약확인 치료(mDOT) 수행 후의 균음전율에 대한 연구 부제: 보건소 균양성 폐결핵 환자를 대상으로)

  • Hwang, Taik Gun;Kim, Soon Deok;Yoo, Se Hwa;Shin, Yoo Chul
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.5
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    • pp.485-494
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    • 2004
  • Background : To assess the effects of mDOT implementation on sputum smear conversion for AFB (Acid fast bacilli) positive pulmonary tuberculosis patients, modified Directly Observed Treatment (mDOT) was started on October $8^{th}$ 2001 at a health center in Seoul. mDOT was defined through weekly interviewing and supervising of a patient by a supervisor (doctor, nurse, or lay health worker). The sputum smear conversion of a mDOT group was compared with that of a self-medication (self) group. Methods : This study included 52 AFB positive pulmonary tuberculosis patients registered at a health center in Seoul between October $8^{th}$ 2001 and April $23^{rd}$ 2002. 24 and 28 patients were enrolled in the mDOT and self medication groups, respectively. Paired (1:1) individual matching, by gender, extent of disease, relapse and age-matching variables, was performed between the two groups, resulting in 20 paired matches. This prospective study was planned as an unblinded, non-randomized quasiexperimental pilot project. Outcomes were identified from results of sputum smear examinations for AFB in both groups at 2 weeks, and 1 and 2 months. The paired matching data were analyzed using the SAS program version 8.1 by McNemar test. Results : At the end of 2 weeks of treatment, the sputum smear conversion of the mDOT group was somewhat higher than that of the self medication group (78.57 vs. 50%, p-value=0.289), and after 1 month of treatment no statistically significant difference was shown between the two groups (83.33 vs. 50, p-value=0.125). At the end of 2 months of treatment (initial intensive phase), the sputum smear conversions of the mDOT and self groups were 95 and 75%, respectively (p-value=0.219). Conclusions : The implementation of mDOT did not result in clinically significant increases in the sputum smear conversion at 2 weeks, and 1 and 2 months compared with that of the self medication group. However, the increases experienced might contribute to diminishing the infectious period of AFB positive patients, and this approach may act as a guide for a specific group of patients. In this study, mDOT was performed for one hundred percent of the intensive treatment phase. It can also be an effective treatment for pulmonary tuberculosis patients, and may be useful for some high risk tuberculosis patients.