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A Randomized, Double-Blind Study to Assess the Efficacy and Safety of Oral LB20304 (Gemifloxacin) at Doses of 160mg or 320mg (Equivalent to 200mg or 400mg of the Mesylate Salt) Once Daily for 7 Days for the Treatment of Acute Exacerbations of Chronic Bronchitis (AECB) in Korean Adult Population (한국인에서 만성기관지염의 급성악화를 치료하기 위한 LB20304(Gemifloxacin) 160mg 또는 320mg 1일 1회 7일간 투여의 유효성과 안전성에 대한 연구)

  • Kim, Young Whan;Shim, Young-Soo;Kim, Won Dong;Shim, Tae Sun;Kang, Hong Mo;Choi, ByoungWhui;Kim, Jae-Yeol;Kwon, O Jung;Kim, Hojoong;Kim, Ju Ock;Jung, Ki-Suck;Hyeon, In Gyou;Mo, Eun Kyung;Lee, Seung-Joon;Nam, Gui Hyun;Lee, Kye Young;Park, Jae Seuk
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.69-87
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    • 2003
  • Background : LB20304(gemifloxacin) is a new fluoroquinolone antibacterial agent with excellent activity against both Gram-negative and Gram-positive organisms. In vitro studies using clinical isolates have shown gemifloxacin to be highly active against penicillin-resistant strains of S. pneumoniae and in contrast to other reference quinolones, gemifloxacin retained good activity against clinical isolates of S. pneumoniae that were resistant to other members of the quinolone class. Therefore, gemifloxacin is thought to be effective in treating acute bacterial exacerbation of chronic bronchitis(AECB). The objective of this study was to evaluate the efficacy and safety of oral gemifloxacin at doses of 160mg or 320mg once daily for 7 days for the treatment of AECB in Korean adult population. Methods : This was a randomized, multicenter, double-blind, parallel group Phase II study to assess the clinical and antibacterial efficacy and safety of oral gemifloxacin for the treatment of AECB. Treatment Group A (67 patients) took oral gemifloxacin 160mg once daily for seven days and treatment Group B (70 patients) took oral gemifloxacin 320mg once daily for seven days. Results : The demographic profiles of the two treatment groups were similar. The clinical response at follow-up was 84.2% in the gemifloxacin 160-mg group, and 88.7% in the gemifloxacin-320 mg group, showing no statistically significant difference between two treatment groups(p=0.49). The clinical response at the end of therapy was 96.5% in the 160-mg group, and 96.4% in the 320-mg group. The bacteriological response at the end of therapy and follow-up were 81.8% and 78.9%, respectively, in the 160-mg group, and 86.4% and 84.2%, respectively, in the 320-mg group, showing no statistically significant difference between two treatment groups(p=0.68 and 0.68, respectively). S. pneumoniae(12 isolates) and H. influenzae(10 isolates) were the most prevalent pathogens. The MICs were lower for gemifloxacin than other quinolones against these key pathogens, and for S. pneumoniae, the MICs for gemifloxacin were considerably lower(${\leq}0.03$ ug/mL) than those for other quinolones, beta-lactams and macrolides. In the period on-therapy plus 30 days post-therapy, a total of 18 patients(26.9%) in the gemifloxacin 160mg group and 22 patients(31.4%) in the 320mg group reported at least one adverse event(AE). The most frequently reported AE was abdominal pain(3/67 patients, 4.5%) in the gemifloxacin 160mg group and increased level of hepatic enzyme(5/70 patients, 7.1%) in the 320mg group. The overall AE profiles for the two treatment groups were similar. Two out of 67 patients(3.0%) in the gemifloxacin 160mg group and 1/70 patients(1.4%) in the 320mg group reported at least one serious AE, however, none of which was considered by the investigator to be of suspected or probable relationship to study medication. Conclusion : The results of this study showed that gemifloxacin at doses of 160mg or 320mg once daily for 7 days in the treatment of acute exacerbations of chronic bronchitis(AECB) in adult Koreans was a very effective and safe treatment both clinically and bacteriologically.

The Evaluation of Difference according to Image Scan Duration in PET Scan using Short Half-Lived Radionuclide (단 반감기 핵종을 이용한 PET 검사 시 영상 획득 시간에 따른 정량성 평가)

  • Hong, Gun-Chul;Cha, Eun-Sun;Kwak, In-Suk;Lee, Hyuk;Park, Hoon;Choi, Choon-Ki;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.1
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    • pp.102-107
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    • 2012
  • Purpose : Because of the rapid physical decay of the short half-lived radionuclide, counting of event for image is very limited. In this reason, long scan duration is applied for more accurate quantitative analysis in the relatively low sensitive examination. The aim of this study was to evaluate the difference according to scan duration and investigate the resonable scan duration using the radionuclide of 11C and 18F in PET scan. Materials and Methods : 1994-NEMA Phantom was filled with 11C of $30.08{\pm}4.22MBq$ and 18F of $40.08{\pm}8.29MBq$ diluted with distilled water. Dynamic images were acquired 20frames/1minute and static image was acquired for 20minutes with 11C. And dynamic images were acquired 20frames/2.5minutes and static image was acquired for 50minutes with 18F. All of data were applied with same reconstruction method and time decay correction. Region of interest (ROI) was set on the image, maximum radioactivity concentration (maxRC, kBq/mL) was compared. We compared maxRC with acquired dynamic image which was summed one bye one to increase the total scan duration. Results : maxRC over time of 11C was $3.85{\pm}0.45{\sim}5.15{\pm}0.50kBq/mL$ in dynamic image, and static image was $2.15{\pm}0.26kBq/mL$. In case of 18F, the maxRC was $9.09{\pm}0.42{\sim}9.48{\pm}0.31kBq/mL$ in dynamic image and $7.24{\pm}0.14kBq/mL$ in static. In summed image of 11C, as total scan duration was increased to 5, 10, 15, 20minutes, the maxRC were $2.47{\pm}0.4$, $2.22{\pm}0.37$, $2.08{\pm}0.42$, $1.95{\pm}0.55kBq/mL$ respectively. In case of 18F, the total scan duration was increased to 12.5, 25, 37.5, and 50minutes, the maxRC were $7.89{\pm}0.27$, $7.61{\pm}0.23$, $7.36{\pm}0.21$, $7.31{\pm}0.23kBq/mL$. Conclusion : As elapsed time was increased after completion of injection, the maxRC was increased by 33% and 4% in dynamic study of 11C and 18F respectively. Also the total scan duration was increased, the maxRC was reduced by 50% and 20% in summed image of 11C and 18F respectively. The percentage difference of each result is more larger in study using relatively shorter half-lived radionuclide. It appears that the accuracy of decay correction declined not only increment of scan duration but also increment of elapsed time from a starting point of acquisition. In study using 18F, there was no big difference so it's not necessary to consider error of quantitative evaluation according to elapsed time. It's recommended to apply additional decay correction method considering decay correction the error concerning elapsed time or to set the scan duration of static image less than 5minutes corresponding 25% of half life in study using shorter half-lived radionuclide as 11C.

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The Evaluation of Reconstruction Method Using Attenuation Correction Position Shifting in 3D PET/CT (PET/CT 3D 영상에서 감쇠보정 위치 변화 방법을 이용한 영상 재구성법의 평가)

  • Hong, Gun-Chul;Park, Sun-Myung;Jung, Eun-Kyung;Choi, Choon-Ki;Seok, Jae-Dong
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.172-176
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    • 2010
  • Purpose: The patients' moves occurred at PET/CT scan will cause the decline of correctness in results by resulting in inconsistency of Attenuation Correction (AC) and effecting on quantitative evaluation. This study has evaluated the utility of reconstruction method using AC position changing method when having inconsistency of AC depending on the position change of emission scan after transmission scan in obtaining PET/CT 3D image. Materials and Methods: We created 1 mL syringe injection space up to ${\pm}2$, 6, 10 cm toward x and y axis based on central point of polystyrene ($20{\times}20110$ cm) into GE Discovery STE16 equipment. After projection of syringe with $^{18}F$-FDG 5 kBq/mL, made an emission by changing the position and obtained the image by using AC depending on the position change. Reconstruction method is an iteration reconstruction method and is applied two times of iteration and 20 of subset, and for every emission data, decay correction depending on time pass is applied. Also, after setting ROI to the position of syringe, compared %Difference (%D) at each position to radioactivity concentrations (kBq/mL) and central point. Results: Radioactivity concentrations of central point of emission scan is 2.30 kBq/mL and is indicated as 1.95, 1.82 and 1.75 kBq/mL, relatively for +x axis, as 2.07, 1.75 and 1.65 kBq/mL for -x axis, as 2.07, 1.87 and 1.90 kBq/mL for +y axis and as 2.17, 1.85 and 1.67 kBq/mL for -y axis. Also, %D is yield as 15, 20, 23% for +x axis, as 9, 23, 28% for -x axis, as 12, 21, 20% for +y axis and as 8, 22, 29% for -y axis. When using AC position changing method, it is indicated as 2.00, 1.95 and 1.80 kBq/mL, relatively for +x axis, as 2.25, 2.15 and 1.90 kBq/mL for -x axis, as 2.07, 1.90 and 1.90 kBq/mL for +y axis, and as 2.10, 2.02, and 1.72 kBq/mL for -y axis. Also, %D is yield as 13, 15, 21% for +x axis, as 2, 6, 17% for -x axis, as 9, 17, 17% for +y axis, and as 8, 12, 25% for -y axis. Conclusion: When in inconsistency of AC, radioactivity concentrations for using AC position changing method increased average of 0.14, 0.03 kBq/mL at x, y axis and %D was improved 6.1, 4.2%. Also, it is indicated that the more far from the central point and the further position from the central point under the features that spatial resolution is lowered, the higher in lowering of radioactivity concentrations. However, since in actual clinic, attenuation degree increases more, it is considered that when in inconsistency, such tolerance will be increased. Therefore, at the lesion of the part where AC is not inconsistent, the tolerance of radioactivity concentrations will be reduced by applying AC position changing method.

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A Study on Urinary Cadmium Concentration and Renal Indices of Inhabitant in an Abandoned Mine Area (폐광지성(廢鑛地城) 주민(住民)의 요중(尿中) 카드뮴 농도(濃度)와 현기능평가(賢機能評價))

  • Park, Jung-Duck;Park, Chan-Byung;Choi, Byung-Sun;Kang, Eun-Yong;Hong, Yeon-Pyo;Chang, Im-Won;Chun, Byung-Yeol;Yeh, Min-Hae
    • Journal of Preventive Medicine and Public Health
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    • v.31 no.3 s.62
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    • pp.424-439
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    • 1998
  • Urinary cadmium is used as a sensitive indicator for internal Cd dose, and increased excretion of $N-acetyl-\beta-D-glucosaminidase(NAG)$, $\beta_2-microglobulin(MG)$ and total protein are useful indices for renal dysfunction by chronic exposure to Cd. The target group was 184 inhabitant(82 men and 102 women) in an abandoned mine area known as exposure to low level Cd. The control group was took 160 individuals(64 men and 96 women) in Cd not-exposed area. Urinary Cd concentration was significantly higher in the target group than the control. The geometric mean of urinary Cd for male was $2.56{\mu}g/\ell,\;2.80{\mu}g/g$ creatinine and $2.50{\mu}g/S.G.$ in the target group and $1.19{\mu}g/\ell,\;1.36{\mu}g/g$ creatinine and $1.17{\mu}g/S.G.$ in the control. For female $2.69{\mu}g/\ell,\;3.94{\mu}g/g$ creatinine and $2.63{\mu}g/S.G.$ in the target group and $1.27{\mu}g/\ell,\;1.97{\mu}g/g$ creatinine and $1.25{\mu}g/S.G.$ in the control, respectively. In addition, urinary Cd of the target group had affected by the period of residence and dietary habit for the rice and the vegetables from the target area. These findings suggest the chronic exposure to Cd of the target population. Mean excretion of urinary NAG, $\beta_2-MG$ and total protein were not significant between two groups. In the target group, urinary NAG activity and total protein were significantly correlated with urinary Cd, but $\beta_2-MG$ was not related. Urinary excretion of NAG, $\beta_2-MG$ and total protein were significantly increased in $10\leqq$ than in <2 of urinary Cd level. In $2\sim10$ group of urinary Cd level, the excretion of NAG significantly increased while not showed for $\beta_2-MG$. In present study, urinary excretion of NAG was relatively sensitive than $\beta_2-MG$ in chronic exposure population to low level Cd.

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Radioiodine Therapy of Liver Cancer Cell Following Tissue Specific Sodium Iodide Symporter Gene Transfer and Assessment of Therapeutic Efficacy with Optical Imaging (조직 특이 발현 Sodium Iodide Symporter 유전자 이입에 의한 방사성옥소 간암세포 치료와 광학영상을 이용한 치료효과 평가)

  • Jang, Byoung-Kuk;Lee, You-La;Lee, Yong-Jin;Ahn, Sohn-Joo;Ryu, Min-Jung;Yoon, Sun-Mi;Lee, Sang-Woo;Yoo, Jeong-Soo;Cho, Je-Yeol;Lee, Jae-Tae;Ahn, Byeong-Cheol
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.5
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    • pp.383-393
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    • 2008
  • Purpose: Cancer specific killing can be achieved by therapeutic gene activated by cancer specific promotor. Expression of sodium iodide symporter (NIS) gene causes transportation and concentration of iodide into the cell, therefore radioiodine treatment after NIS gene transfer to cancer cell could be a form of radionuclide gene therapy. luciferase (Luc) gene transfected cancer cell can be monitored by in vivo optical imaging after D-luciferin injection. Aims of the study are to make vector with both therapeutic NIS gene driven by AFP promoter and reporter Luc gene driven by CMV promoter, to perform hepatocellular carcinoma specific radiodiodine gene therapy by the vector, and assessment of the therapy effect by optical imaging using luciferase expression. Materials and Methods: A Vector with AFP promoter driven NIS gene and CMV promoter driven Luc gene (AFP-NIS-CMV-Luc) was constructed. Liver cancer cell (HepG2, Huh-7) and non liver cancer cell (HCT-15) were transfected with the vector using liposome. Expression of the NIS gene at mRNA level was elucidated by RT-PCR. Radioiodide uptake, perchlorate blockade, and washout tests were performed and bioluminescence also measured by luminometer in these cells. In vitro clonogenic assay with 1-131 was performed. In vivo nuclear imaging was obtained with gamma camera after 1-131 intraperitoneal injection. Results: A Vector with AFP-NIS-CMV-Luc was constructed and successfully transfected into HepG2, Huh-7 and HCT-15 cells. HepG2 and Huh-7 cells with AFP-NIS-CMV-Luc gene showed higher iodide uptake than non transfected cells and the higher iodide uptake was totally blocked by addition of perchlorate. HCT-15 cell did not showed any change of iodide uptake by the gene transfection. Transfected cells had higher light output than control cells. In vitro clonogenic assay, transfected HepG2 and Huh-7 cells showed lower colony count than non transfected HepG2 and Huh-7 cells, but transfected HCT-15 cell did not showed any difference than non transfected HCT-15 cell. Number of Huh-7 cells with AFP-NIS-CMV-Luc gene transfection was positively correlated with radioidine accumulation and luciferase activity. In vivo nuclear imaging with 1-131 was successful in AFP-NIS-CMV-Luc gene transfected Huh-7 cell xenograft on nude mouse. Conclusion: A Vector with AFP promoter driven NIS and CMV promoter driven Luc gene was constructed. Transfection of the vector showed liver cancer cell specific enhancement of 1-131 cytotoxicity by AFP promoter, and the effect of the radioiodine therapy can be successfully assessed by non-invasive luminescence measurement.

Potential Study for the Sedimentary Exhalative Pb-Zn Mineralization in Dyusembay Area, Kazakhstan (카자흐스탄 듀셈바이지역의 퇴적분기형 연-아연 광화작용에 대한 잠재력 연구)

  • No, Sang-gun;Lee, Seung-han;Park, Ki-woong;Jeong, Hyeon-guk;Yun, Ji-seong;Kim, Sun-ok;Park, Maeng-eon
    • Economic and Environmental Geology
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    • v.51 no.3
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    • pp.213-222
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    • 2018
  • Metasediment-hosted Pb-Zn mineralized zone has been found in Dyusembay of Kazakhstan. Its petrological properties, metal index, alteration index and redox-sensitivity are compared with those of SEDEX type deposit. Mineralization is developed along foliation of host rock (graphitic phyllite) and controlled by folds and faults; major ore minerals including pyrite, pyrrhotite, sphalerite, and galena are disseminated or interlayered with fine-grained quartz. The margin of the mineralized zone is metamorphosed accompanying sericite and chlorite. Hydrothermal brecciation and Pb-Zn mineralization formed in quartz-calcite stockworks are confirmed at the around of Maytyubin granitoid intrusions. The mineralization is classified into three types according to those of occurrence, paragenesis, chemical composition and isotopic characteristics. Type 1 whose fine-grained pyrite, pyrrhotite and sphalerite are formed in parallel yet discontinuous to well-developed foliations of the host rock; its geochemistry is similar to those of the earlier stage in SEDEX-type mineralization. In case of type 2, the ore minerals of which are concentrated being parallel to a foliation by regional metamorphism, and most of them associated with quartz and muscovite (${\pm}$ biotite) paragenetically. Type 3 is formed in the hydrothermal breccia zone whose ore minerals are controlled by foliation and breccia and developed in quartz ${\pm}$ calcite veins having a form such as stratification, stockwork or veinlets. Host rocks in the mineralized zone indicate homogeneous metamorphic grade and there is no specific alteration zonation. Also, all types (type 1, type 2, and type 3) represent similar REEs patterns, it can be interpreted that these are originated from a same source. Sulphides occurred in mineralized zone indicate a limited range of sulphur isotope values (type 2, ${\delta}^{34}S=-13.3{\sim}-11.7$‰; type 3, ${\delta}^{34}S=-13.9{\sim}-8.2$‰), and a result of geothermometry presents different temperature ranges: type 2($251{\pm}38^{\circ}C{\sim}277{\pm}40^{\circ}C$); type 3($360{\pm}2^{\circ}C$ to $537{\pm}29^{\circ}C$). It is estimated to be due to the effect of metamorphism and Maytyubin granitoid intrusions, respectively. In addition, ternary chart of thorium, scandium, and zircon for discrimination of tectonic setting and redox sensitivity using V/Mo values indicate that hydrothermal sediments put on reduction environment after precipitation, before being affected by metamorphism and intrusion activity. Geochemical data are plotted on a distal trend of SEDEX-type with discrimination plot using SEDEX index. As a result, petrological-geochemical properties demonstrate that Dyusembay Pb-Zn mineralized zone is comparable to distal-type of SEDEX deposit.

A Comparison of Dietary Behaviors According to Gender and Obesity Status of Middle School Students in Jeonju (전주지역 중학생의 성별 및 비만판정에 따른 식행동 비교 연구)

  • Sung, Sun-Hwa;Yu, Ok-Kyeong;Son, Hee-Sook;Cha, Youn-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.995-1009
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    • 2007
  • The purpose of this study was to investigate the dietary habits, behaviors, and food consumption frequency according to gender and obesity level among middle school students in the Jeonju area. Subjects for the questionnaire were 450 middle school students (male 255, female 195) and were classified as either obese students (n=150 or non-obese students (n=299) by the obesity assessment method. The results were analyzed with SAS program (Version 9.1), and were as follows. 1. Dietary behaviors were significantly different in the rate of 'Skipping breakfast (p<0.05)', 'Duration of meal time (min) (p<0.05)' and 'Unbalanced diet (p<0.01)' between males and females. Dietary habits and behaviors also differed significantly for the rate of ‘Taste preferences (p<0.05)’, and 'Unbalanced diet (p<0.01)' between obese students and non-obese students. 2. Food consumption frequency per week was as follows. First, males were significantly higher than females in 'Instant noodle (p<0.05)', 'Milk (p<0.01)', and 'Soda pop (p<0.01)'; on the other hand females were significantly higher than males in 'Chocolate, Candy (p<0.01)'. Second, non-obese students were significantly higher than obese students in 'Instant noodle (p<0.05)', 'Hamburger, Pizza (p<0.05)', and 'Chocolate, Candy (p<001)'. Especially, non-obese male students were higher in 'Instant noodle (p<0.05)' and 'Hamburger, Pizza (p<0.05)'; non-obese female students were higher in 'Chocolate, Candy (p<0.01)'. In conclusion, an action program is needed to encourage healthful dietary behaviors, increased physical activity, and forming good lifelong habits.

Survey of Current Status of the Patients with Home Ventilator in Seoul and Kyunggi Province (가정용 인공호흡기를 사용하는 서울 및 경기 지역 환자의 실태)

  • Ahn, Jong-Joon;Lee, Ki-Man;Shim, Tae-Sun;Lim, Chae-Man;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Koh, Youn-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.5
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    • pp.624-632
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    • 2000
  • Background : Home ventilation can decrease hospital-acquired infection, increase physical activity, improve nutritional status, enhance quality of life, and reduce medical costs. The number of patient using home ventilators has been increasing, particularly in Europe and United States. Although the number of patients with home ventilation has been increasing in Korea, the current status of these patients is not well known. This study was undertaken to obtain basic information upon these patients in addition to evaluating any problems related to patients' home care in our country. Methods : A register of 92 patients with home ventilators in Seoul and Kyunggi Province were obtained from commercial ventilator supply companies. The patients were contacted by phone and 29 of them accepted our visit. Information concerning education about home care before discharge, equipment cost, and problems related to home care were documented. The mode and preset variables of the home ventilator were checked; tidal volume (TV), peak airway pressure, and oxygen saturation were measured. Results : There were 26 males (90%) and their mean age was 48.0 (${\pm}20.1$) years. The underlying diseases were : 21 neuromuscular disorders, 2 spinal cord injuries, 6 chronic lung diseases. Among the caregivers, spouses (n=14) predominated. Education for home care before discharge was performed primarily by intensive care unit nurses and the education for ventilator management by commercial companies. Twenty-five of the 29 patients had tracheostomies. Volume targeted type (VTT ; n=20, 69%) was more frequently used than the pressure targeted type (PTT). Twenty-three of the 29 patients purchased a ventilator privately, which cost 7,450,000 (${\pm}$3,290,000) won for a PTT, and 14,280.000 (${\pm}$3,130,000) won for a VTT. Total cost for the equipment was 11,430,000 (${\pm}$634,000) won. The average cost required for home care per month was 1,120,000 (${\pm}$1,360, 000) won. Conclusion : The commonest underlying disease of the patients was neuromuscular disease. The VTT ventilator was primarily used with tracheostomy. Patients and their families considered the financial difficulties associated with purchasing and maintaining equipment for home care an urgent problem. Some patients were aided by a visiting nurse, however most patients were neglected and left without professional medical supervision.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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Effect of Solvent Fractions from Doenjang on Antimutagenicity, Growth of Tumor Cells and Production of Interleukin-2 (된장 분획물의 항돌연변이 및 암세포 증식 억제효과와 interleukin-2 생성에 미치는 영향)

  • Kim, Kwang-Hyuk;Park, Kun-Young;Lee, Sook-Hee;Lim, Sun-Young
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.791-797
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    • 2007
  • We studied the inhibitory effect of solvent fractions from doenjang on mutagenicity using Salmonella typhimurium TA100 in Ames test. We also investigated the effect of solvent fractions from doenjang on the growth of tumor cells and the production of interleukin-2 (IL-2). The treatment of dichlorormethane and ethylacetate fractions (2.5 mg/assay) from doenjang to Ames test system inhibited aflatoxin B$_1$ (AFB$_1$) induced mutagenicity by 96% and 97%, respectively, and showed a higher antimutagenic effect than other solvent fractions. In case of N-methyl-N'-nitro-N-nitrosoguamidine (MNNG) induced mutagenicity, the ethylacetate fraction showed the highest inhibitory effect (by 75%) among the other sol-vent fractions, although the inhibitory effect was not stronger compared to AFB$_1$ induced mutagenicity. The treatment of dichloromethane and ethylacetate fractions markedly inhibited the growth of Yac-1 (by 80% and 94%, respectively) and sacroma-180 cancer cells (by 60% and 96%, respectively) after 4 days of incubation at 37${\circ}$C. To elucidate the immunological mechanism of antitumor activity of doenjang, spleen cells of Balb/c mouse were exposed to the dichloromethane and ethyl-acetate fractions for 24 hours at 37${\circ}$C . The culture supernatants following the treatment of djchloromethane and ethylacetate factions to spleen cells increased the production of IL-2. These results indicated that the anticarcinogenic effect of doenjang was mediated by the production of IL-2.