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Herpes Simplex Virus Thymidine Kinase Gene Therapy Delivered by Retroviral or Adenoviral Vector in Mouse Model of Lewis Lung Carcinoma (Lewis 폐암 마우스 모델에서 Retroviral Vector나 Adenoviral Vector로 이입된 Herpes Simplex Virus Thymidine Kinase 유전자치료)

  • Kwon, Hee-Chung;Jeong, Jae-Min;Kim, Jung-Hyeon;Ham, Yong-Ho;Seo, Ji-Sook;Lee, Ki-Ho;Kim, Chang-Min;Lee, Han-Soo;Lee, Choon-Taek
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.3
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    • pp.298-309
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    • 2000
  • Background : The antitumor effects of herpes simplex virus thymidine kinase (HSV-tk) and ganciclovir (GCV) strategies for cancer gene therapy have a the following advantages : 1) a direct cytotoxicity to HSV-tk modified cancer cells by GCV 2) a cell death by the local transfer of toxic metabolites from the HSV-tk modified cells to nearby unmodified tumor cells (bystander effect), and 3) in vivo bystander effect such as antitumor-immunity. Retroviral and adenoviral sequences can silence transgene expression in cells and mice. In this study, we investigated the above described advantages of HSV-tk/GCV strategy in Lewis lung cell and mouse lung cancer model using retroviral vector and adenoviral vector. Also, we observed whether the expression of a silenced gene can be reactivated by treating cells with butyrate. Methods : Retrovirus-HSV-tk and adenovirus-HSV-tk vectors were used for the transduction of Lewis lung carcinoma (LLC) cells. The change of HSV-tk expression by butyrate was measured by Western blol The antitumor activities containing bystander effect were observed in vivo (by MTT assay) and in vivo tumor models of various combinations of LLC and LLC-tk. Results : 1. Butyrate induced the enhancement of HSV-tk expression from adenovirally transduced cells but not from retrovirally transduced cells. 2. Both retrovirus-HSV-tk and adenovirus-HSV-tk vectors with GCV treatment were effective for killing of tumor cell in vitro and suppression of LLC tumorigenicity. Bystander effect was responsible for killing of mixture of LLC-tk and LLC in vitro and in vivo-tumorigenicity model. Conclusion : Butyrate could augment adenovirus-mediated HSV -tk gene expression. Cancer gene therapy with HSV-tk suicide gene by retroviral and adenoviral vector seems to be an effective approach for lung cancer therapy.

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Effect of Soil Environment on Diversity and Population of Aerobic Soil Bacteria from Baekdudaegan Mountain Forests in Gyeongsangbuk-do, Korea (경상북도 산림지역의 토양 환경이 호기성 토양 세균의 다양성과 밀도에 미치는 영향)

  • Park, Chul Yeong;Lee, Sun Keun;Kim, Ji Hong;Lee, Sang Yong;Lee, Jong Kyu
    • Journal of Korean Society of Forest Science
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    • v.101 no.3
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    • pp.501-508
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    • 2012
  • This study was carried out to compare species diversity of soil bacteria from Baekdudaegan mountain forests (Bonghwa-gun, Mungyeong-si and Sangju-si) in Gyeongsangbuk-do and to analyze the effects of soil environments on diversity and population of soil bacteria. Soil bacteria were isolated from soil samples by streak plate method, and identified by DNA extaction and 16S rDNA sequence analyses. The population of soil bacteria from the soil samples of Bonghwa-gun was the highest with $5.1{\times}10^5cfu/g$, and followed by those from Mungyeong-si and Sangju-si with $1.9{\times}10^5cfu/g$ and $1.1{\times}10^5cfu/g$, respectively. The population of soil bacteria from surface layer soil was the highest, and then gradually decreased according to soil depth. The increase in population of soil bacteria from soil samples of different sites was correlated with the increase of the altitude of soil sampling site, depth of A horizon, liquid phase among three phases of soil, water content and bulk density of soil. Two hundreds and sixty eight bacterial colonies from Bonghwa-gun were classified into 10 species, 8 genera. One hundred and thirty four bacterial colonies from Mungyeong-si were classified into 15 species, 9 genera. Forty four bacterial colonies from Sangju-si were classified into 5 species, 2 genera. The dominant species (occupancy rate) from Bonghwa-gun and Mungyeong-si were Bacillus weihenstephanensis (36% and 40%, respectively), and Sangju-si was Bacillus cereus (39%). The relationships between soil environment and community structure of soil bacteria were analyzed statistically by using ecological indices. The diversity, evenness and dominance indices of soil bacteria were 6.30, 2.04 and 0.59 in Bonghwa-gun, 9.09, 2.94 and 0.51 in Mungyeong-si, and 4.55, 2.34 and 0.71 in Sangju-si, respectively. The diversity and evenness indices were increased by the increase of water content, drainage condition and gravel content of soil, while the dominance index was decreased.

Evaluation of Renal Oxygenation in Normal Korean Volunteers Using 3.0 T Blood Oxygen Level-Dependent MRI (3.0 T 혈중산소치의존 자기공명영상을 이용한 정상한국인에서의 신장 산소공급의 평가)

  • Hwang, Sung Il;Lee, Hak Jong;Chin, Ho Jun;Chae, Dong-Wan;Na, Ki Young
    • Investigative Magnetic Resonance Imaging
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    • v.17 no.1
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    • pp.19-25
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    • 2013
  • Purpose : Renal blood oxygen level-dependent (BOLD) MRI has been used in the evaluation of renal oxygenation. We tried to provide the normal $R2^*$ value of the human kidney with 3.0 T, and evaluated the differences in $R2^*$ values according to gender and location. Materials and Methods: Twenty-four healthy volunteers underwent BOLD MRI at 3.0 T. Multi gradient echo-echo planar imaging sequence with seventeen echoes was used. After generation of the $T2^*$ map, the $R2^*$ was calculated. The statistical differences in $R2^*$ values between the cortex and medulla, males and females, and the right and left kidney were analyzed. The regional differences of $R2^*$ within the both kidneys were evaluated respectively. Results: BOLD MRI was successful in all participants. No gross artifact interfered with $R2^*$ measurement. The mean $R2^*$ at 3.0 T was $17.1{\pm}2.60s^{-1}$ in the cortex and $27.7{\pm}4.83s^{-1}$ in the medulla (p < 0.001). The $R2^*$ value in the medulla was significantly higher in the male than female volunteers (p = 0.025). There were no statistical differences of $R2^*$ according to the side and location in the kidney (p = 0.197). Conclusion: Renal BOLD MRI can be efficiently performed with 3.0 T MRI. Renal medullary hypoxia is present in normal volunteers. Our results may be used as reference values in the evaluation of pathologic conditions using BOLD MRI.

Measurement of Regional Cerebral Blood Volume in Normal Rabbits on Perfusion-weighted MR Image (MR 관류강조영상에서 정상 가토의 국소 뇌혈류량 측정)

  • 박병래;예수영;나상옥;김학진;이석홍;전계록
    • Investigative Magnetic Resonance Imaging
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    • v.4 no.2
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    • pp.100-106
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    • 2000
  • Purpose : To evaluate the usefulness of cerebral blood flow measurement applied to perfusion weighted image with short-scan time single shot gradient echo-planar technique in measuring cerebral blood volume(rCBV) of normal rabbits. Materials and methods : With 2.1-3.6 kg weighted rabbits, image is acquired when they are in supine position in children positioner. Perfusion weighted image is acquired to 44 seconds per 1 second successively. After 4 seconds later, Gd-DTPA 2ml are injected into int. jugular vein with 2 ml per second and normal saline is also injected after that. Same technique is applied 2 times per 30 minites in same rabbit. After Image is obtained in two part of cerebral cortex at vertex, convexity, in one of basal ganglia with choosing about $3-5{\textrm{mm}^2}$ areas. Curve of signal intensity changes in time sequence is drawn. After this images are transmitted by PC and software IDL, regional cerebral blood volume is measured with imaging processing program made by us. Results : With 22 of 24 rabbits, satisfactory 1-2 signal intensity versus time curve is made. Cerebral blood capacity and contrast media stay time (ST) is measured in two cerebral cortex and basal ganglia refering in parietal cerebral cortex. Mean focal cerebral blood flow capacity ratio in cortex was $0.97{\pm}0.35$ and in basal ganglia, $0.99{\pm}0.37$, mean contrast media stay time in cortex was $9.83{\pm}1.63$ sec and in basal gaiglia, $9.42{\pm}1.14$ sec, but there was no statistically significant difference between two areas ($\rho$=0.05). Conclusion : In cerebral cortex and basal ganglia, there is no difference in mean focal blood volume and mean contrast stay time. Therefore, PWI is useful in cerebral blood flow and early diagnosis, prognosis of cerebral ischemic disease. Hereafter, it is helpful in analysing cerebral blood flow changes with comparison difference in rCBV between normal tissue and ischemic tissue, and that with DWI finding in infarcted patient.

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Gene Expression of Metalloproteinases, Tissue Inhibitors of Metalloproteinases and Cytokines in Adriamycin-induced Cardiomyopathy (아드리아마이신으로 유도된 심근증에서 Metalloproteinase, Metalloproteinase 조직억제자, Cytokine 유전자 발현에 대한 연구)

  • Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.48 no.2
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    • pp.197-203
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    • 2005
  • Purpose : Changes in metalloproteinases(MMP) activity have been demonstrated in several disease states, including rheumatoid arthritis and tumor metastasis. More importantly, increased myocardial MMP activity has been reported to occur in both clinical and experimental forms of dilated cardiomyopathy. There was no report about MMP in adriamycin(ADR)-induced cardiomyopathy. The purpose of this study was to investigate gene expression of MMP and tissue inhibitor of metalloproteinases(TIMP) in ADR-induced cardiomyopathy and clarify the relationship between MMP and cytokines. Methods : Male Sprague-Dawley rats were divided into two groups. The first group was control. The second group was given intraperitoneal injections of ADR(5 mg/kg) twice a week over two weeks. Serum concentrations of MMP, TIMP, interleukin(IL)-6 and tumor necrosis factor(TNF)-${\alpha}$ were measured. RNA extraction was performed from frozen rat hearts. Reverse transcription polymerase chain reaction(RT-PCR) was employed. cDNA Microarray analysis was performed by using a set of 5,184 sequence-verified rat cDNA clones. Results : Serum MMP and TIMP levels were not significantly different between the two groups. IL-6 was $36.8{\pm}2.8pg/mL$ and TNF-${\alpha}$ $2.2{\pm}2.7pg/mL$ in the ADR group. They were significantly higher than in the control group. Serum MMP correlated significantly with TNF-${\alpha}$(r=0.41, P<0.05). There was no gene expression of MMP, IL-6 or TNF-${\alpha}$ in the hearts of both groups. Gene expression of TIMP was significantly depressed in the hearts of the ADR group. Conclusion : These results suggested a potential role for TNF-${\alpha}$ in the regulation of extracellular matrix remodeling in ADR induced cardiomyopathy. Rapid screening of multiple decreased gene expression by DNA chip may be a useful diagnostic test to detect early cardiac injury before developing ADR induced cardiomyopathy.

Seasonal Food Intake Status of the Long-lived Elderly People in Kyungpook Sung-Ju (경북 성주지역 장수노인의 계절별 식품섭취 상태)

  • 백지원;구보경;김규종;이성국;이혜성;이연경
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.691-700
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    • 2000
  • The purpose of this study was to estimate food intakes of the long-lived elderly and to obtain the data for establishing dietary guidelines that may be recommended for the general population for the sake of longvity. The subjects of the study were 224 elderly people of age over 85 years living in Kyungpook Sung-Ju area who have no problem in daily living. The food consumption survey was carried out seasonally by the repeated 24-hr recall method for one year. The subject group for this study was composed of 58 males and 166 females, the average age being 87 years old. Food intakes of the winter were more than any other seasons. The mean daily total food intake per capita was 594.4g, 513.5 g (86.4%) from plant foods and 79.3 g(13.3%) from animal foods. The sequence of high intakes of food groups were cereals, vegetables, fruits, fishes, legumes and meats. Boiled white rice, beef soup, soybean paste soup, soybean paste stew, broiled yellow croaker, kimchi, ra myon and broiled noodles were consumed most frequently. In conclusion, the subjects consumed much more plants foods than animal foods and consumed more natural and seasonal foods than processed foods and also showed food habits of eating small meals. Dietary habits of eating small meals containing abundant amount of fresh plant foods might partially contribute to the longevity of the subjects.

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Structural and Compositional Characteristics of Skarn Zinc-Lead Deposits in the Yeonhwa-Ulchin Mining District, Southeastern Taebaegsan Region, Korea Part I: The Yeonhwa I Mine

  • Yun, Suckew
    • Economic and Environmental Geology
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    • v.12 no.2
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    • pp.51-73
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    • 1979
  • The zinc-lead deposits at the Yeonhwa I mine were investigated in terms of ore-forming geologic setting, structural style of ore control, geometry of individual orebodies, zoning, paragenesis and chemical composition of skarn minerals, as well as metal grades and ratios of selected orebodies. The Yeonhwa I mine is characterized by a large swarm of chimney type massive orebodies with thin skarn envelopes, boldly developed through a thick sequence of Pungchon Limestone, the overlying Hwajeol Formation, and the underlying Myobong Slate of Cambrian age. Nearly 20 orebodies of similar shape, but of varying size are arranged in a V-shaped pattern with northwest and northeast trends, clearly indicating an outstanding ore control by a conjugate system of fractures with these trends. Important orebodies are the Wolam 1, 2, 3, and 5 orebodies in the west, and the Namsan 1, 2, 3. and 5 orebodies in the east, among others. The Wolam 1 orebody, which was observed from the -360 level through the -240, -120, and 0 levels to the surface outcrops (totaling a vertical height of about 500m), shows a vertical variation in skarn mineralogy, ranging from pyroxene-garnet zone on the lower levels. through pyroxene (without garnet) zone on the intermediate levels, and finally to rhodochrosite vein on the upper levels and surface. Microprobe analyses of pyroxene and garnet on a total of 14 mineral grains revealed that pyroxenes are manganoan salitic in most samples, with downward increase of Fe and Mn, whereas garnets are highly andraditic, containing fractions of subordinate grossular with downward decrease of Fe. This indicates a reverse relationship of Fe-contents between pyroxene and garnet with depth. Ore minerals are major sphalerite, subordinate galena, and minor chalcopyrite. Sulfide gangue minerals include major pyrrhotite, and minor pyrite and marcasite of later age. Two types of variational trends in metal grades and ratios with depth are present on the plots of assay data from the Wolam orebodies: one is a steady upward increase in Pb, Zn, and Pb:Zn ratios, with a terminal decline at the top of orebody: the other is an irregular or sinusoidal change. The former is characteristic of chimney-type orebodies, whereas the latter is of vein· shaped orebodies. The Pb grades show large variations among orebodies and from level to level, whereas the Zn grades are relatively constand or less variable.

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Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Cattle Age Prediction by Leukocytes Telomere Quantification (혈액세포의 텔로미어 함량을 이용한 소의 연령예측)

  • Choi, Na-Eun;Kim, Hyun-Sub;Choe, Chang-Yong;Jeon, Gwang-Joo;Sohn, Sea-Hwan
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.367-374
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    • 2010
  • Telomeres at the end of chromosomes consist of tandem repeats of (TTAGGG)n DNA sequence and associated proteins. Telomeres have the essential functions in chromosome stability and genome integrity and are hence related to cell senescence and cancer. This study was carried out to quantify the amount of telomeric DNA and establish age prediction equations by using the quantity of telomeric DNA for cattle. Analysis of the telomere quantity of the lymphocytes was performed at different age, across breeds and between different sexes of cattle. We quantified the amount of telomeric DNA by the Q-FISH technique using the telomeric DNA probe in 460 cattle at age of 1~166 months in Korean Cattle and Holstein breeds. In results, we found that the amount of telomeric DNA decreased gradually with age. The amount of telomeric DNA of Korean Cattle was significantly higher than that of Holstein breed (P<0.01). In addition, the amount of telomeric DNA in male was significantly higher than that in female (P<0.01). Using the relationship between age and the amount of telomeric DNA in cattle, age predicting equations were established as a result of regression analysis. Because sex and breeds influenced telomeric DNA quantity, the age prediction equations were estimated separately in Korean Cattle females and Holstein females. The regression equations were $\hat{Y}$=$38.102X^2$-220.103X + 318.309 (P<0.0001, $R^2$=0.8019) in Korean Cattle females and $\hat{Y}$ = $42.799X^2$ - 199.682X + 242.106 (P<0.0001, $R^2$ = 0.8379) in Holstein females, where the X was quantity of telomeric DNA and Y was predicted age in months. These equations predicted the age of cattle with high significance and accuracy and have high R square values. Thus, it could be possible to scientifically predict the age using the above equations for Korean Cattle and Holstein females.

Comparative Analysis of Signal Intensity and Apparent Diffusion Coefficient at Varying b-values in the Brain : Diffusion Weighted-Echo Planar Image ($T_2^*$ and FLAIR) Sequence (뇌의 확산강조 영상에서 b-value의 변화에 따른 신호강도, 현성확산계수에 관한 비교 분석 : 확산강조 에코평면영상($T_2^*$ 및 FLAIR)기법 중심으로)

  • Oh, Jong-Kap;Im, Jung-Yeol
    • Journal of radiological science and technology
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    • v.32 no.3
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    • pp.313-323
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    • 2009
  • Diffusion-weighted imaging (DWI) has been demonstrated to be a practical method for the diagnosis of various brain diseases such as acute infarction, brain tumor, and white matter disease. In this study, we used two techniques to examine the average signal intensity (SI) and apparent diffusion coefficient (ADC) of the brains of patients who ranged in age from 10 to 60 years. Our results indicated that the average SI was the highest in amygdala (as derived from DWI), whereas that in the cerebrospinal fluid was the lowest. The average ADC was the highest in the cerebrospinal fluid, whereas the lowest measurement was derived from the pons. The average SI and ADC were higher in $T_2^*$-DW-EPI than in FLAIR-DW-EPI. The higher the b-value, the smaller the average difference in both imaging techniques; the lower the b-value, the greater the average difference. Also, comparative analysis of the brains of patients who had experienced cerebral infarction showed no distinct lesion in the general MR image over time. However, there was a high SI in apparent weighted images. Analysis of other brain diseases (e.g., bleeding, acute, subacute, chronic infarction) indicated SI variance in accordance with characteristics of the two techniques. The higher the SI, the lower the ADC. Taken together, the value of SI and ADC in accordance with frequently occurring areas and various brain disease varies based on the b-value and imaging technique. Because they provide additional useful information in the diagnosis and treatment of patients with various brain diseases through signal recognition, the proper imaging technique and b-value are important for the detection and interpretation of subacute stroke and other brain diseases.

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