• KSBB Journal
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• v.23 no.3
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• pp.231-238
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• 2008

The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

• Journal of Korean Society of Forest Science
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• v.84 no.4
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• pp.424-431
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• 1995

This study was conducted to evaluate resistance and physiological responses of Zelkova serrata Makino seedlings to ozone in open-top chamber. One-year-old seedlings of Zelkova serrata were planted in pots in April and grown in greenhouse until August. The plants were transferred into two out-door open-top chambers with a dimension of 2.0 m in diameter and 2.0 m in height. First chamber served as a control and was supplied with ambient air. Ozone was added to the second chamber for 5 hours per day(10.00 AM-15.00 PM) for 23 consecutive days at 0.1 ppm. Each chamber housed 70 pots. Every two, three or five days after initiation of exposure, ten pots were randomly removed from the chamber and determined for the contents of chlorophyll a, b, total chlorophyll and ${\beta}$-carotene in the leaves. Photosynthesis and dark respiration were estimated by measuring $CO_2$ absorption in a gas exchange chamber and oxygen absorption by oxygen monitoring system, respectively. Superoxide dismutase(SOD) activity in the leaves was assayed by a xanthine oxidase method. First visible injury of translucent(water-soaked looking) spots appeared on the leaves 14 days after the initial exposure, and ozone accelerated senescence of old leaves. Contents of chlorophyll a and b decreased by 17%, and 31%, respectively, in ozone treatment two days after exposure. The decrease in chlorophyll b was greater than that of chlorophyll a. Content of ${\beta}$-carotene in ozone treatment decreased by 25% two days after initiation of exposure, but the reduction was recovered with time. Photosynthesis decreased by 45%, and the respiration increased by 28% in the ozone treatment. SOD activity started to increase 4 days after beginning of exposure and increased by 285% 7 days after exposure, and decreased to the level below the control treatment with the advancement of the visible injury.

• Journal of The Korean Society of Grassland and Forage Science
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• v.22 no.4
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• pp.265-272
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• 2002

Forage breeding laboratory of National Livestock Research Institute, R.D.A. has made interspecific hybrids of Lolium multiflorum $\times$ L. pratensis and intergeneric hybrids of Lolium $\times$ Festuca since 1984, and has collected ecotypes of Italian ryegrass since 1991. Growth characteristics of these hybrids and ecotypes were researched, and then these clone lines were named. Among these clone lines, the several clones that have polen fertility, high cold-tolerance, and similar heading time were used for making synthetics, Naehan 6, 7, 8, 9, with polycrossing method in 1997. Field experiments were carried out to compare the mophological and agronomical characteristics and forage productivity and quality of the synthetics with those of Italian ryegrass varieties, Barmultra and Hwasan 101. in Suwon and Yonchun from 1999 to 2000. Heading time of the synthetics were 22th to 24th May that belong to late-mature types to be similar to that of Barmultra and Hwasan 101 in Suwon. The synthetics were 101 to 106 c3n in plant length, medium or thick in thickness of stem, dark peen in leaf color, broad and long in flag leaf, strong in lodging resistance, and excellent in regrowth. Winter survivals of the synthetics were no different from that of Barmultra or Hwasan 101 in Suwon, but better than that of Barmultra or Hwasan 101 in Yonchun where was -10 to -12$^{\circ}C$ of minimum average air temperature in January or February. Dry matter(DM) yields of the synthetics were similar to DM 8,238kg per ha of Barmultra in Suwon, but in Yonchun, were more 7 to 13% than DM 7,291kg per ha of Barmultra. Forage qualities, IVDMD, ADF, NDF and TDN of the synthetics were lower than those of Hwasan 101, but higher than those of Barmultra.

• Korean Journal of Organic Agriculture
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• v.24 no.4
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• pp.833-847
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• 2016

Drought stress is a major agricultural limitation to crop productivity worldwide, especially by which leafy vegetables, plant leaves eaten as vegetable, could be more lethal. The study was carried out to know the effect of drought tolerance plant growth promoting bacteria (PGPB) on water stress of kale seedlings. A total of 146 morphologically distinct bacterial colonies were isolated from bulk soil and rhizosphere soil of leafy vegetables and screened for plant growth promoting microbioassay in greenhouse. Out of them the isolate SB19 significantly promoted the growth of kale seedlings in increasement of about 42% of plant height (14.1 cm), 148% of leaf area ($19.0cm^2$) and 138% of shoot fresh weight (1662.5 mg) attained by the bacterially treated plants compared to distilled water treated control (9.9 cm, $7.7cm^2$, 698.8 mg). Shoot water content of SB19 treated kale seedlings (1393.8 mg) was also increased about 152% compared with control (552.5 mg). The SB19 isolated from bulk soil of kale plant in Iksan, Korea, was identified as species of Bacillus based on 16S rRNA gene sequencing analysis. We evaluated the effect of drought tolerance by the Bacillus sp. SB19 on kale seedlings at 7th and 14th days following the onset of the water stress and watering was only at 7th day in the middle of test. In the survey of 7th and 14th day, there were mitigation effect of drought stress in kale seedlings treated with $10^6$ and $10^7cell\;mL^{-1}$ of SB19 compared to distilled water treated control. Especially, there were more effective mitigation of drought damage in kale seedlings treated with $10^7cell\;mL^{-1}$ than $10^6cell\;mL^{-1}$. Further, although drought injury of bacterially treated kale seedlings were not improved at 14th day compared with 7th day, drought injury of $10^7cell\;mL^{-1}$ of SB19 treated kale seedlings were not happen rapidly but developed over a longer period of time than $10^6cell\;mL^{-1}$ of SB19 or control. The diffidence of results might be caused by the concentration of bacterial suspension. This study suggests that beneficial plant-microbe interaction could be a important role of enhancement of water availability and also provide a good method for improving quality of leafy vegetables under water stress conditions.

• Radiation Oncology Journal
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• v.24 no.4
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• pp.255-262
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• 2006

$\underline{Purpose}$: To evaluate the pathological prognostic factors related to local recurrence after radical surgery and adjuvant radiation therapy in advanced rectal cancer. $\underline{Materials\;and\;Methods}$: Fifty-four patients with advanced rectal cancer who were treated with radical surgery followed by adjuvant radiotherapy and chemotherapy between February 1993 and December 2001 were enrolled in this study. Among these patients, 14 patients experienced local recurrence. Tissue specimens of the patients were obtained to determine pathologic parameters such as histological grade, depth of invasion, venous invasion, lymphatic invasion, neural invasion and immunohistopathological analysis for expression of p53, Ki-67, c-erb, ezrin, c-met, phosphorylated S6 kinase, S100A4, and HIF-1 alpha. The correlation of these parameters with the tumor response to radiotherapy was statistically analyzed using the chi-square test, multivariate analysis, and the hierarchical clustering method. $\underline{Results}$: In univariate analysis, the histological tumor grade, venous invasion, invasion depth of the tumor and the over expression of c-met and HIF-1 alpha were accompanied with radioresistance that was found to be statistically significant. In multivariate analysis, venous invasion, invasion depth of tumor and over expression of c-met were also accompanied with radioresistance that was found to be statistically significant. By analysis with hierarchical clustering, the invasion depth of the tumor, and the over expression of c-met and HIF-1 alpha were factors found to be related to local recurrence. Whereas 71.4% of patients with local recurrence had 2 or more these factors, only 27.5% of patients without local recurrence had 2 or more of these factors. $\underline{Conclusion}$: In advanced rectal cancer patients treated by radical surgery and adjuvant chemo-radiation therapy, the poor prognostic factors found to be related to local recurrence were HIF-1 alpha positive, c-met positive, and an invasion depth more than 5.5 mm. A prospective study is necessary to confirm whether these factors would be useful clinical parameters to measure and predict a radio-resistance group of patients.

• Journal of Chest Surgery
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• v.34 no.11
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• pp.823-830
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• 2001

Background: Paraplegia is a serious complication of thoracic or thoracoabdominal aortic operations, which is related to ischemic injury of the spinal cord induced by low perfusion pressure during cross clamping of the aorta. Ischemic preconditioning of heart or brain with reversible sublethal ischemic injury induces resistance to subsequent lethal ischemia. The aim of this study is to investigate whether ischemic tolerance could be induced by the preconditioning of the spinal cord using swine model. Material and Method: The animals were randomly assigned to three groups: sham group(n=3), control group(n=6) and pre-conditioning group(n=8). In the sham group, we performed the left thoracotomy only without any ischemic injury. In the preconditioning group, the swine received reversible spinal cord ischemic injury by aortic clamping for 20 minutes, whereas control group had no previous aortic cross- clamping. Forty-eight hours later, the aorta was clamped for 30 minutes in both groups. Neurological examination was done 24 hours later, then the animals were euthanized for histopathology and malonedialdehyde(MDA) spectrophotometry assay of the spinal cord. Result: Statistically significant difference in neurological outcome was observed between the control and preconditioning groups at 24 hours after ischemic injury. The incidence of paraplegia and severe paresis was 100% in the control group, and 62.5% in the preconditing group(p=0.028). There was no statistically significant difference in histopathology and MDA assay of the ischemic spinal cord between these two groups with borderline statistical difference in MDA assay(p=0.0745). Conclusion: In the present swine study, ischemic preconditioning could induce tolerance against 30 minute ischemic insult of the spinal cord, although the animals did not completely recover(stand-up or walk). We expect that combining this preconditioning with other currently existing protection methods might lead to a synergistic effect, which warrants further investigation.

• Journal of Chest Surgery
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• v.32 no.5
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• pp.413-421
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• 1999

Background: Ischemia reperfusion injury is known to contribute to the major causes of the early graft failure in lung transplantation. Triiodothyronine (T3) has been suggested to ameliorate ischemia reperfusion injury from both in vivo and in vitro experiments of various organs. Prospecting its beneficial effect for pulmonary allograft preservation, we made a new solution by adding T3 into the extracellular type dextran solution. Material and Method: Twelve adult mongrel dogs underwent left lung allotransplantation. Six donor dogs were flushed with the new solution(Group 1, n=6), and the remaining six were flushed with Euro-Collins solution to serve as controls(Group 2, n=6). Allografts were stored in each preservation solution for 20 hours at 4$^{\circ}C$. Left single lung transplantations were performed. The right pulmonary artery and the right main bronchus were clamped at 15 minutes after the reperfusion and maintained throughout the experiment to evaluate the transplanted left lung function. Result: Arterial carbon dioxide tension was better in group 1 than in group 2 throughout the experiment period and the difference was statistically significant at 2 hours after reperfusion(28.0${\pm}$3.0 mmHg and 53.1${\pm}$17.4 mmHg, p<0.05). The differences of arterial oxygen partial pressure, peak airway pressure and pulmonary vascular resistance showed no statistical significance. The malondialdehyde(MDA) level, measured from tissue obtained at 120 minutes after reperfusion showed no statistically significant difference. The tissue wet/dry ratio of group 1(649${\pm}$27 %) was significantly lower than that of group 2(686${\pm}$71 %, p<0.05). The microscopic examination revealed varying degrees of injury represented mainly by findings such as perivascular neutrophil infiltration, capillary hemorrhage and interstitial congestion. These findings were less severe in group 1 than those in group 2. Conclusion: The new solution demonstrated superior allograft preservation after 20 hour ischemia compared to Euro-Collins solution in canine single left lung transplantation model, these results suggest that T3 might be a promising agent for pulmonary allograft preservation.

• Tuberculosis and Respiratory Diseases
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• v.39 no.5
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• pp.425-432
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• 1992

Background: The prevalence of tuberculosis in Korea decreased remarkably for the past 30 years, while for at least the recent 10 years, the frequency of disease attributable to mycobacteria other than tuberculosis (MOTT) began to increase both in actual numbers and in the proportion of the total burden of mycobacterioses. Method: Twenty nine cases, diagnosed as having pulmonary disease due to MOTT on the basis of repeated isolations of the relevant organisms from sputum specimens and chest X-ray findings from Jan. 1982 throught Dec. 1991 at the Chest Clinic of the Korean Institute of Tuberculosis, were reviewed in terms of clinical features and courses. Results: 1) Nineteen cases (66%) were infected with Mycobacterium avium-intracellulare, 5 cases (17%) with M. fortuitum, 4 cases (14%) with M. chelonei, and 1 cases (3%) with M. szulgai. 2) The ratio of male versus female patients was 1.9:1. 3) The peak incidence was in the fifth decade. 4) Eighteen cases (62%) had moderately advanced disease and 11 cases (38%) had far advanced disease on chest P-A film. 5) The common symptoms were cough (59%), sputum (52%), and dyspnea (34%). 6) All cases had a previous history of tuberculosis. 7) Most of the isolates were highly resistant to the major antituberculosis drugs and 48~52% showed a sensitivity to cycloserine, kanamycin or enviomycin. 8) Of 19 cases which were treated and followed-up for over 12 months, 3 cases attained negative conversion on cultures (2 M. fortuitum infections, 1 M. szulgai infection). Sixteen cases failed in attaining negative conversion on cultures. However, their clinical courses were chronic and indolent despite of persistant culture positivity. Conclusion: Treatment of these patients has been difficult because of the frequency of severe underlying conditions and the natural resistance of most of the nontuberculous mycobacteria to the presently available drugs.

• Journal of Chest Surgery
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• v.38 no.3 s.248
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• pp.197-203
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• 2005

Homograft aortic valve replacement (AVR) has many advantages such as excellent hemodynamic performance, faster left ventricular hypertrophy regression, resistance to infection and excellent freedom of thromboembolism. To find out the results of homograft AVR, we reviewed our surgical experiences. Material and Method: Eighteen patients (male female=16 : 2, mean age=39.3$\pm$16.2 years, range: 14$\~$68 years) who underwent homo-graft aortic valve replacement between May 1995 and May 2004 were reviewed. The number of homografts was 20 (17 aortic and 3 pulmonic homografts) including two re-operations. Ten patients had a history of previous aortic valve surgery. Indications for the use of a homograft were native valve endocarditis (n=7), prosthetic valve endocarditis (n=5), or Behcet's disease (n=8). The homograft had been implanted predominantly as a full root except in one patient in the subcoronary position. Result: Mean follow-up was 41.3 $\pm$ 26.2 months. There was one operative mortality. Postoperative complications included postoperative bleeding in 3 patients, and wound infection in 1. There was no late death. Three patients underwent redo-AVR. The etiology of the three reoperated patients was Behcet's disease (p=0.025). Freedom from reoperation was $87.5\pm8.3\%$, $78.8\pm11.2\%$ at 1, 5 years respectively, In patients with infective endocarditis, there was no recurrence of endocarditis. There was no thromboembolic complication. Conclusion: Although longer term follow-up with larger numbers of patients is necessary, the operative and mid-term results for homograft AVR was good when we took into account the operative risks of Behcet's disease or infective endocarditis. Behest's disease was a risk factor for reoperation after the homograft AVR. We think homograft AVR is the procedure of choice, particularly in patients with infective endocarditis.

• Tuberculosis and Respiratory Diseases
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• v.45 no.3
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• pp.574-582
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• 1998

Background: Bronchofiberscopy is a procedure with a chance of airway irritation and it may cause pathophysiologic changes of respiratory system. So we tried to evaluate the influence of bronchofibercopy on $O_2$ saturation, ABGA and PIT by patient's basal status and procedure type. Method: $O_2$ saturation was measured every 1 minute from the left index finger tip with percutaneous oximetry. ABGA was done before and right after the bronchofiberscopy and PIT was done before and within 10 minutes after the bronchofiberscopy. Results: The mean time for bronchofiberscopy procedure was 14.5mim and $SaO_2$ maximally fall to 89.0 below 8% of the baseline after mean time of 8.4min, which was recovered at the end of the procedure. $SaO_2$ change amount was 8.4 % on Non-$O_2$ supply group, which was lower compared to 6.4 % of the $O_2$-supply group without statistically significance. Biopsy Group and BAL group showed more $SaO_2$ fall than washing only group. The level of $PaO_2$ and FEV1 of the patient didn't influence significantly on $SaO_2$ fall during the procedure. ABGA taken before and after the bronchofiberscopy showed mild fall of $PaO_2$ and mild rise of $PaCO_2$. Whereas PFT showed decrease of FEV1(P<0.05) and increase of RV without changes in airway resistance and pulmonary diffusion capacity. Comparing before and after the bronchofiberscopy, the washing group showed no significant changes on PIT, while the biopsy group and the BAL group showed increase of RV & decrease of $FEV_1$ after the bronchofiberscopy. BAL group showed more changing tendency rather than biopsy group although not statistically significant. Conclusion: Bronchofiberscopy is considered as a relatively safe procedure, but it would be better to be done with $O_2$ supply especially in the patient with low $PaO_2$ and in the case of biopsy and BAL.