• Tuberculosis and Respiratory Diseases
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• v.39 no.6
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• pp.517-525
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• 1992

Background: Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent, and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis. Method: To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB-1, -2), and 123 base pair DNA, a part of IS6110 fragment, which multiple copies are known to exsist PCR one genome (the primers are Sal I-1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results. Results: 1) Using TB-1, -2 primers, PCR was positive in 73.1% (19/26) of culture positive sputa, in 12.5% (1/8) of culture negative. but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology. 2) Using Sal I-I, -2 primers, PCR was positive in 94.1% (32/34) of culture positive sputa, in 23.1% (6/26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14/16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology. Conclusion: PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.547-558
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• 1997

Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

• Tuberculosis and Respiratory Diseases
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• v.44 no.6
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• pp.1353-1365
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• 1997

Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majority of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to host. In the previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF, and the probability was shown that the acquired resistance to TNF might be associated with synthesis of some protective protein. Understanding the mechanisms of TNF -resistance in TNF-$\alpha$ cDNA transfected cancer cells would be. an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of MnSOD, an antioxidant enzyme, in the acquired resistance to TNF of TNF-$\alpha$ cDN A transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164(murine fibrosarcoma cell line), NCI-H2058(human mesothelioma cell line), A549(human non-small cell lung cancer cell line), ME180(human cervix cancer cell line) cells using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF-$\alpha$ cDNA transfected cells(WEHI164-TNF, NCIH2058-TNF, A549-TNF, ME180-TNF) and the changes of MnSOD mRNA expressions with Northern blot analysis. Results : The MnSOD mRNA expressions of parental cells and genetically modified cells of WEHI164 and ME180 cells(both are naturally TNF sensitive) were not significantly different The MnSOD mRNA expressions of genetically modified cells of NCI-H2058 and A549(both are naturally TNF resistant) were higher than those of the parental cells, while those of parental cells with exogenous TNF were also elevated. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ cDNA transfection may not be associated with the change in the MnSOD expression, but the difference in natural TNF sensitivity of each cell may be associated with the level of the MnSOD expression.

• Tuberculosis and Respiratory Diseases
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• v.45 no.2
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• pp.397-403
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• 1998

Background: Little information is available concerning the value of bronchoscopy in patients with a lymphocytic exudative pleural effusion in which percutaneous pleural biopsy have been regarded as cornerstone in investigating the etiology. Recently, a few reports suggest that bronchoscopy may be more effective diagnostic method in patients with unexplained pleural effusion accompanied by hemoptysis or other roentgenographic abnormalities, such as mass, infiltrate, atelectasis. Method: Mter initial examinations of sputum and pleural fluid through thoracentesis in 112 patients(male 75 cases, female 37 cases, mean age 53.2 years) who were admitted for evaluation of the cause of pleural effusion, we performed bronchoscopy and closed pleural biology in most patients with undiagnosed lymphocytic exudate and compared the diagnostic yield of both invasive methods according to hemoptysis or other roentgenographic abnormalities, and investigated the sole diagnostic contribution of bronchoscopy. Results: Tuberculosis(57 cases, 51%) was the most common cause of pleural effusion. Percutaneous pleural biopsy showed more diagnostic yield than bronchoscopy regardless of presence or absence of other clinical or radiologic abnormalities. In 25 cases with unknown etiology after pleural biopsy, additional diagnostic yield by bronchoscopy was 36 % (4/11) in patients with associated features and only 7 % (1/14) with lone effusion, and, as the sole mean for diagnsosis in all patients with pleural effusion, was only 4.5%(5/12). Conclusion : In a region of high prevalence of tuberculosis as a cause of pleural effusion, percutaneous pleural biospy is more effective method when invasive method is required for confirmative diagnosis of unexplained lymphocytic exudative pleural effusion, and bronchoscopy is unlikely to aid in the diagnosis of lone pleural effusion.

• Journal of The Korean Association For Science Education
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• v.21 no.2
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• pp.357-384
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• 2001

In this paper, we described practical teaching-learning plans based on three different theoretical approaches to Integrated Science Education (ISE): a knowledge centered ISE, a social problem centered ISE, and an individual interest centered ISE. We believe that science teachers can understand integrated science education through this paper and they are able to apply simultaneously our integrated science teaching materials to their real instruction in classroom. For this we developed integrated science teaching-learning plans for the topic of energy which has a integrated feature strongly among integrated science subject contents. These modules were based upon the teaching strategies of 'Energy' following each integrated directions organized in the previous paper (Three Strategies for Integrated Science Teaching of "Energy" Applying Knowledge, Social Problem, and Individual Interest Centered Approaches) and we applied instruction models fitting each features of integrated directions to the teaching strategies of 'Energy'. There is a concrete describing on the above three integrated science teaching-learning plans as follows. 1. For the knowledge centered integration, we selected the topic, 'Journey of Energy' and we tried to integrate the knowledge of physics, chemistry, biology, and earth science applying the instruction model of 'Free Discovery Learning' which is emphasized on concepts and inquiry. 2. For the social problem centered integration, we selected the topic, 'Future of Energy' to resolve the science-related social problems and we applied the instruction model of 'Project Learning' which is emphasized on learner's cognitive process to the topic. 3. For the individual interest centered integration, we selected the topic, 'Transformation of Energy' for the integration of science and individual interest and we applied the instruction model of 'Project Learning' centering learner's interest and concern. Based upon the above direction, we developed the integrated science teaching-learning plans as following steps. First, we organized 'Integrated Teaching-Learning Contents' according to the topics. Second, based upon the above organization, we designed 'Instructional procedures' to integrate within the topics. Third, in accordance with the above 'Instructional Procedures', we created 'Instructional Coaching Plan' that can be applied in the practical world of real classrooms. These plans can be used as models for the further development of integrated science instruction for teacher preparation, textbook development, and classroom learning.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.207-216
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• 2010

Objective: This study was performed to clarify the role of HomeoboxA (HOXA) and its related signaling molecules in the decidualization of primary cultured endometrial cells. Methods: Human endometrial tissues were obtained by curettage of hysterectomy specimens from patients with conditions other than endometrial diseases. Tissues were minced and digested with Trypsin-EDTA for 20 min, $37^{\circ}C$. Cells were cultured with DMEM/F12 medium in $37^{\circ}C$, 5% $CO_2$ incubator for 24 hrs. Cells were treated with HOXA10 siRNA and added transforming growth factor (TGF)-${\beta}1$ (10 ng/mL) for 48 hrs to induces decidualization in vitro. Reverse transcription polymerase chain reaction analysis was accomplished to observe the expression of HOXA10, prolactin, cyclooxygenase (COX)-2, peroxisome proliferatoractivated receptor (PPAR)-$\gamma$, and wingless-type MMTV integration site family (Wnt). Results: HOXA10 expression was increased (1.8 fold vs. non-treated control) in TGF-${\beta}1$ treated cells. Decidualization marker, prolactin, was significantly increased in TGF-${\beta}1$ treated cells compared with HOXA10 siRNA treated cells. Endometrial cell differentiation marker, COX-2 was down-regulated by HOXA10 siRNA even if cells were treated with TGF-${\beta}1$. Wnt4 was down-regulated by treated with HOXA10 siRNA, this expression patters was not changed by TGF-${\beta}1$. Expression of PPAR-$\gamma$ was down regulated by TGF-${\beta}1$ in regardless of HOXA10 siRNA treatment. Conclusion: TGF-${\beta}1$ which is induced by progesterone in endometrial epithelial cells may induces stromal cell decidualization via HOXA10 and Wnt signaling cascade.

• The Korean Journal of Mycology
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• v.41 no.3
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• pp.172-180
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• 2013

Grey mold infection rate in tomato was investigated with the inoculation of dead flowers on Botrytis selective media. The grey mold infection rate of flower after fruiting were higher in the order of after 45 days, after 25 days, and fruiting day with 100%, 87% and 65%, respectively. The number of infected flowers were increased with time increase after the flowering before fruiting. BSM (Botrytis selective medium) was used to check grey mold infection rate depending on the flowering stage and cultivar. Grey mold infection rate depending on the flowering stage was similar in all the beef-tomato cultivar as 1.5~5% at preflowering, 1.5~45% at flowering and 75~90% at fruiting. On the other hand, cherry tomato cultivar "KoKo" had lower infection rates of 0~3.5% at pre-flowering, 10~30% at flowering and 20~50% at fruiting. These resulted from the fact that beaf-tomato cultivar have much bigger flowers and larger amount of pollens compared to those of cherry tomato cultivar. The amounts of falling pollens of Botrytis spp. were checked for beaf-tomato cultivar and cherry tomato cultivar using BSTM. The amounts of falling pollens were increased as growth period was extended, and the amount of spores increased rapidly during the outbreak of grey mold. Twelve field trials in Buyeo and Iksan areas showed that Fluazinam, and Diethofencarb+Carbendazim were effective fungicides to control tomato grey mold, and these results were similar to those of field trials with BSTM. This is the first report of Fluazinam as a effective fungicide for the control of grey mold of tomato even though it has not been registered yet for the control of gray mold in tomato.

• The Korean Journal of Zoology
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• v.18 no.2
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• pp.71-86
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• 1975

The G-banding patterns of normal and of delayed spiralized chromosomes by BUdR were investigated in three established cell lines of dwarf hamsters. The results obtained were as follows: 1. The number of G-bands appeared in Chinese hamster T-233 cell line was 65. The centromeric dark band was found in No.1 chromosome and weakly stained bands were also observed in part of the centromeric regions of Nos. 2, 3, 8 and $X_2$ chromosomes. Two homologous X chromosomes were found in different banding patterns. Terminal dark bands were shown in No. 1 chromosome. No conspicuous bands appeared in No. 10 chromosome. 2. Eighty four bands appeared in Armenian hamster Y-1249 cell line. Centromeric dark bands were observed in Nos. 5 and 10 chromosomes and moderatly stained bands were also found in near the centromeric region of the long arms of Nos. 7 and 9 chromosomes. Two isomorphic X chromosomes were also distinguished by their banding patterns. 3. In Y-1313 Armenian hamster cell line, the bands were 69. No centromeric dark bands were observed in this cell line, but moderatly stained bands appeared in the centromeric area in the long arm of No. 9 chromosome. The banding patterns of these two cell lines of Armenian hamster were quite different and readily distinguished. Only No. 8 chromosome showed similar G-banding patterns. Although Nos. 5, 7 abd 8 chromosomes revealed the same number of bands in these two cell lines, the location and staining intensity were quite different. 4. Chromosomes of Nos. 1, 2, 6, $X_1$ and $X_2$ in T-233 cell line and of 1, 4, 7, 8, 9, $X_1$ and $X_2$ in both cell lines of Armenian hamster were found to be elongated due to the inhibition of mitotic spiralization by BUdR. G-banding patterns of these chromosomes were found to be identical to those of normal chromosomes in these cell lines.

• The Korean Journal of Zoology
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• v.23 no.2
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• pp.89-108
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• 1980

Comparative studies on the parafollicular cells of the some mammalian species from five different orders were carried out; i.e., man from Primates, cattle, pig, and black goat from Artiodactyla, dog from Carnivora, rabbit from Lagomorpha, rat, mouse, and squirrel from Rodentia. For this study, various special techniques for the parafollicular cells, including Grimelius' silver impregnation method (Sawicki and Bajko, 1974), Singh's argentaffin method (Singh, 1964), HCl-toluidine blue stain (Sawicki, 1971), and HCl-lead hematoxylin stain (Solcia et al., 1969), were applied. Authors obtained the following results: 1. Number of parafollicular cells in the same area of thyroid tissues are significantly different from species to species. Number of cells were largest in dog and less cells were found in the following orders; rat, squirrel, mouse, rabbit, cattle, pig, black goat and finally the smallest number in man. 2. Distribution of parafollicular cells within thyroid gland are significantly different from portion to portion in case of cattle, rabbit, squirrel and mouse, but it is not significant in dog, man, pig, black goat and rat (see Table 1-1 and 1-2). 3. In dog, clustered parafollicular cells are located usually in the interfollicular space, and groups of parafollicular cells are located in the para-and/or inter-follicular positions in rabbit. But in the other animals parafollicular cells are found solitarily in the intra-and/or para-follicular positions. 4. The shape of parafollicular cells shows oval to round contour in dog, but it is polymorphic, for example, spindle, conical, oval, round or elongated with cytoplasmic processes, in the other animals. 5. Size of parafollicular cells is larger in cattle, dog and pig, smaller in rat, mouse and squirrel, and medium-size in rabbit, man and black goat. 6. Parafollicular cells of pig, cattle, dog and squirrel are observed to contain densely packed granules, whereas those of mouse, rat and man contain relatively scanty granules. 7. Parafollicular cells of all the mammals show more or less positive reaction to Grimelius' argyrophile silver impregnation method, HCl-toluidine blue stain and HCl-lead hematoxylin stain, whereas they show negative reaction to argentaffin method (see Table 2). 8. Considering the above finding, it is concluded that there are species differences in the distribution, location and shape of parafollicular cells, and infer that preferable staining method should be selected for reliable detection of parafollicular cells, beacuse staining methods applied on the cells in this study show variable reactions according to species.

• The Korean Journal of Zoology
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• v.27 no.2
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• pp.117-125
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• 1984

Mosquitoes were collected with light trap at a pig shed in the vicinity of Daegu city from mid-April to the end of November in 1981 and 1982. A total of 12,942 and 118,061 individuals were collected in 1981 and 1982 respectively. The collection comprised 77% females and 23% males in 1981, and 96% females and 4% males in 1982. The catches were classified into following 7 species: Culex (Culex) pipiens pallens, C. (C.) tritaeniorhynchus summorosus, Anopheles (Anopheles) sinensis, C. (C.) vagans, C. (C.) oritntelis, C. (C.) bitaeniorhynchus, Aedes (Aedimorphus) vexans nipponii. The former three species showed distinct seasonal prevalence. Arranged in the descending order in size of the catches, in 1981, Culex (Culex) pipiens pallens was 44.9% of the total collection (at sex ratio of 0.85), Anopheles (Anopheles) sinensis 42.9% (0.05), Culex (Culex) tritaeniorhynchus summorosus 12.1% (0.00). On the contrary, Culex (Culex) tritaeniorhynchus summorosus 70.4% (0.00), Anopheles (Anopheles) sinensis 25.2% (0.05), Culex (Culex) pipiens pallens 4.4% (2.19) in 1982. The monthly percentages of collected mosquitoes to the total collection were 0.1% (in 1981) and 0.0% (in 1982) in May; 3.5%, 1.3% in June; 50.0%, 33.9% in July; 37.1%, in August; 8.8%, 11.9% in September; 0.5%, 0.8% in October and 0.0% in November. As for seasonal prevalence, mosquitoes appeared in May in both years and began to increase in number from the first week of June in 1981, but from the second week of May, ahead of three weeks in 1982. The highest peak time in 1982 was the second week of August, two weeks later than the fifth week of July in 1981. Culex (Culex) pipiens pallens showed the maximum activity for the fifth week of July in 1981, but for the third week of July, ahead of two weeks in 1982. Culex (Culex) tritaeniorhynchus reached the highest peak for the second week of August in both years. Anopheles (Anopheles) sinensis showed the maximum activity for the fifth week of July in 1981, but for the third week of July, two weeks earlier in 1982. The highest peak times of three main species were compared respectively as folows. Culex (Culex) pipiens pallens had the highest peak time in common with Anopheles (Anopheles) sinensis, Culex (Culex) tritaeniorhynchus summorosus showed the maximum activity for the second week of August in 1982.