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Altered Expression of ${\beta}_3$ Integrin on Sclerotic Aortic Valves in a Hypercholesterolemic Rabbit Model (고콜레스테롤혈증을 유발한 토끼의 대동맥 판막에서 ${\beta}_3$ Integrin 발현의 변화)

  • Park, Chan-Beom;Kim, Young-Du;Choe, Mi-Sun;Jin, Ung;Moon, Seok-Whan;Kim, Yong-Han;Kim, Chi-Kyung;Jo, Keon-Hyon;Kweon, Jong-Bum
    • Journal of Chest Surgery
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    • v.41 no.6
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    • pp.687-694
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    • 2008
  • Background: Although aortic valve sclerosis causes no significant hemodynamic alterations, it is associated with an increased risk of cardiovascular death and myocardial infarction. However, the role of ${\beta}_3$ integrin in aortic valve sclerosis remains unclear. Material and Method: Twenty male New Zealand rabbits were divided into two groups. Group 1 rabbits (n=10) received a normal chow diet, while group 2 (n=10) rabbits received a diet containing 1% cholesterol for 12 weeks. After the rabbits were euthanized, their aortic valves and ascending aortas were excised for analysis. Result: Total serum cholesterol ($2,148.3{\pm}1,012.5\;mg/dL$ versus $53.7{\pm}31.8\;mg/dL$, p<0.05), triglyceride ($240.4{\pm}218.3\;mg/dL$ versus $31.6{\pm}6.4\;mg/dL$, p<0.05), and low density lipoprotein (LDL)-cholesterol($2,065.3{\pm}960.9\;mg/dL$ versus $29.1{\pm}30.9\;mg/dL$, p<0.05) levels were significantly higher in the cholesterol diet group compared with the normal diet group. Myofibroblasts and macrophages were more highly expressed in the aortic valve leaflets of rabbits in the cholesterol diet group than of those in the normal diet group. A real-time polymerase chain reaction revealed decreased ${\beta}_3$ integrin mRNA levels in the hypercholesterolemic aortic valves and aortas. Conclusion: The present study shows that hypercholesterolemia induces aortic valve sclerosis. These findings suggest that alterations in ${\beta}_3$ integrin may playa role in the development of aortic valve sclerosis.

Potential Reproductive Toxicity Study of p53 Expressing Adenoviral Vector in Mice (아데노바이러스 유전자치료벡터의 생식독성 연구)

  • Rhee, Gyu-Seek;Kwack, Seung-Jun;Kim, Soon-Sun;Lee, Rhee-Da;Seok, Ji-Hyun;Chae, Soo-Young;Chung, Soo-Youn;Kim, Seung-Hee;Lee, Seung-Hoon;Park, Kui-Lea
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.151-158
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    • 2007
  • The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

Application of Molecular Biological Technique for Development of Stability Indicator in Uncontrolled Landfill (불량매립지 안정화 지표 개발을 위한 분자생물학적 기술의 적용)

  • Park, Hyun-A;Han, Ji-Sun;Kim, Chang-Gyun;Lee, Jin-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.2
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    • pp.128-136
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    • 2006
  • This study was conducted for developing the stability parameter in uncontrolled landfill by using a biomolecular investigation on the microbial community growing through leachate plume. Landfill J(which is in Cheonan) and landfill T(which is in Wonju) were chosen for this study among a total of 244 closed uncontrolled landfills. It addressed the genetic diversity of the microbial community in the leachate by 165 rDNA gene cloning using PCR and compared quantitative analysis of denitrifiers and methanotrophs with the conventional water quality parameters. From the BLAST search, genes of 47.6% in landfill J, and 32.5% in landfill T, respectively, showed more than 97% of the similarity where Proteobacteria phylum was most significantly observed. It showed that the numbers of denitrification genes, i.e. nirS gene and cnorB gene in the J site are 7 and 4 times higher than those in T site, which is well reflecting from a difference of site closure showing 7 and 13 years after being closed, respectively. In addition, the quantitative analysis on methane formation gene showed that J1 spot immediately bordering with the sources has the greatest number of methane formation bacteria, and it was decreased rapidly according to distribute toward the outer boundary of landfill. The comparative investigation between the number of genes, i.e. nirS gene, cnorB gene and MCR gene, md the conventional monitoring parameters, i.e. TOC, $NH_3-N,\;NO_3-N,\;NO_2-N,\;Cl^-$, alkalinity, addressed that more than 99% of the correlation was observed except for the $NO_3-N$. It was concluded that biomolecular investigation was well consistent with the conventional monitoring parameters to interpret their influences and stability made by leachate plume formed in downgradient around the uncontrolled sites.

Reproducibility Evaluation of Deep inspiration breath-hold(DIBH) technique by respiration data and heart position analysis during radiation therapy for Left Breast cancer patients (좌측 유방암 환자의 방사선치료 중 환자의 호흡과 심장 위치 분석을 통한 Deep inspiration breath-hold(DIBH) 기법의 재현성 평가)

  • Jo, Jae Young;Bae, Sun Myung;Yoon, In Ha;Lee, Ho Yeon;Kang, Tae Young;Baek, Geum Mun;Bae, Jae Beom
    • The Journal of Korean Society for Radiation Therapy
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    • v.26 no.2
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    • pp.297-303
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    • 2014
  • Purpose : The purpose of this study is reproducibility evaluation of deep inspiration breath-hold(DIBH) technique by respiration data and heart position analysis in radiation therapy for Left Breast cancer patients. Materials and Methods : Free breathing(FB) Computed Tomography(CT) images and DIBH CT images of three left breast cancer patients were used to evaluate the heart volume and dose during treatment planing system( Eclipse version 10.0, Varian, USA ). The signal of RPM (Real-time Position Management) Respiratory Gating System (version 1.7.5, Varian, USA) was used to evaluate respiration stability of DIBH during breast radiation therapy. The images for measurement of heart position were acquired by the Electronic portal imaging device(EPID) cine acquisition mode. The distance of heart at the three measuring points(A, B, C) on each image was measured by Offline Review (ARIA 10, Varian, USA). Results : Significant differences were found between the FB and DIBH plans for mean heart dose (6.82 vs. 1.91 Gy), heart $V_{30}$ (68.57 vs. $8.26cm^3$), $V_{20}$ (76.43 vs. $11.34cm^3$). The standard deviation of DIBH signal of each patient was ${\pm}0.07cm$, ${\pm}0.04cm$, ${\pm}0.13cm$, respectively. The Maximum and Minimum heart distance on EPID images were measured as 0.32 cm and 0.00 cm. Conclusion : Consequently, using the DIBH technique with radiation therapy for left breast cancer patients is very useful to establish the treatment plan and to reduce the heart dose. In addition, it is beneficial to using the Cine acquisition mode of EPID for the reproducibility evaluation of DIBH.

Inhibitory Effect of Steviol and Its Derivatives on Cell Migration via Regulation of Tight Junction-related Protein Claudin 8 (스테비올 및 그 유도체의 세포연접 관련 클라우딘 8 발현 조절을 통한 세포이동 저해효과)

  • Choi, Sun Kyung;Cho, Nam Joon;Cho, Uk Min;Shim, Joong Hyun;Kim, Kee K.;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.403-412
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    • 2016
  • The tight junction, one of Intercellular junctions, performs a variety of biological functions by bonding adjacent cells, including the barrier function to control the movement of the electrolyte and water. Recent studies have revealed that unusual expression of tight junction-related genes have been shown to be related in cancer development and progression. Recently, there are many reports that control of tight junction proteins expression is closely related to the skin moisture. In this study, we are focusing on the regulating mechanism of tight junction-associated genes by the steviol and its derivatives. Steviol, used as a sweetner, is known to chemical compound isolated from stevia plant. The MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) assay was carried out in HaCaT cells (human keratinocyte cell line) in order to determine the cytotoxicity. As a result, while steviol showing cytotoxicity from $250{\mu}M$, steviol derivatives are not cytotoxic more than $250{\mu}M$ concentration. We have observed a change in the tight junction protein via quantitative real-time PCR. Claudin 8 among tight junction proteins is only significantly reduced up to 30% in the presence of steviol. In addition, cell migration was inhibited by steviol, not by stevioside and rebaudioside. Finally, we could observe that steviol, not stevioside and rebaudioside, is able to increase the skin barrier permeability through the transepithelial electric resistance (TEER) measurements. These results suggest that the steviol and its derivatives are specifically acts on the tight junction related gene expression, but steviol derivatives are more suitable as a cosmetic material.

The variability of tumor motion and respiration pattern in Stereotactic Body RadioTherapy(SBRT) for Lung cancer patients (RPM SystemTM을 이용한 호흡 관찰의 유용성 평가)

  • Park, hyun jun;Bae, sun myeong;Baek, Geum Mun;Kang, tae young;Seo, Dong Rin
    • The Journal of Korean Society for Radiation Therapy
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    • v.28 no.1
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    • pp.17-25
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    • 2016
  • Purpose : The purpose of this study is to evaluate the variability of tumor motion and respiration pattern in lung cancer patients undergoing Stereotactic Body RadioTherapy(SBRT) by using On-Board imager (OBI) system and Real-time Position Management (RPM) System. Materials and Methods : This study population consisted of 60 lung cancer patient treated with stereotactic body radiotherapy (48 Gy / 4 fractions). Of these, 30 were treated with gating (group 1) and 30 without gating(group2): typically the patients whose tumors showed three-dimensional respiratory motion > 10 mm were selected for gating. 4-dimensional Computed Tomography (4DCT). Cone Beam CT (CBCT) and Fluoroscopy images were used to measure the tumor motion. RPM system was used to evaluate the variability of respiration pattern on SBRT for group1. Results : The mean difference of tumor motion among 4DCT, CBCT and Fluoroscopy images in the cranio-caudal direction was 2.3 mm in group 1, 2. The maximum difference was 12.5 mm in the group 1 and 8.5 mm in group 2. The number of treatment fractions that patient's respiration pattern was within Upper-Lower threshold on SBRT in group 2 was 31 fractions. A patient who exhibited the most unstable pattern exceeded 108 times in a fraction Conclusion : Although many patients in group 1 and 2 kept the reproducibility of tumor motion within 5 mm during their treatment, some patients exhibited variability of tumor motion in the CBCT and Fluoroscopy images. It was possible to improve the accuracy of dose delivery in SBRT without gating for lung cancer patient by using RPM system.

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Conditioned Media of RAW 264.7 Cells Stimulated with Phellinus linteus Extract Regulates the Epithelial-mesenchymal Transition in Prostate Cancer Cells (상황버섯에 의해 활성화된 RAW 264.7 대식세포주 배양액의 인간 전립선암 세포주의 epithelial-mesenchymal transition 조절)

  • Kang, Taewoo;An, Hyun-Hee;Park, Sul-Gi;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Ji-Won;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.29 no.8
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    • pp.904-915
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    • 2019
  • Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

Gene Expression Analysis of Immune Cell Activation Markers in Extracts of Platycodon grandiflorum Containing Medicinal Herbs (도라지 추출액과 한약재 함유 도라지 추출액에 의한 면역세포 활성 표지유전자 발현 분석)

  • Kang, Shin Ae;Chun, Sung Sik;Kang, Shin Kwon;Chung, Young Chul;Cheon, Eun Woo;Cho, Sang Uk;Jung, Kyung Hwa;Ahn, Soon Cheol;Yu, Hak Sun
    • Journal of Life Science
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    • v.24 no.5
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    • pp.567-572
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    • 2014
  • Extracts of Platycodon grandiflorum have been reported to show anti-inflammatory, antioxidant, anti-metastatic, and hepato-protective effects. This study was designed to evaluate T-cell activation and M1/M2 differential macrophage activation by extracts of P. grandiflorum or P. grandiflorum containing various medicinal herbs. Using real-time RT-PCR, we analyzed expression levels of c-fos, and CD40L (T-cell activation markers) in splenocytes and iNOS, Ym1, and ARG1 in RAW 246.7 cells after treatment of CC (hot water extract of P. grandiflorum), MAEK (hot water extract of P. grandiflorum [82%] and six different plants), and HWAL (hot water extract of P. grandiflorum [7%] and eight different plants. The results showed that MAEK significantly elevated the expression of T-cell activation markers of splenocytes, with the c-fos gene activated more than 10-fold and the CD40L gene activated more than 6-fold. Although CD40L was significantly increased by CC and HWAL, the increase was only about 2-fold. In addition, CC and HWAL did not significantly activate the expression of the c-fos gene. On the other hand, CC elevated the M1 activation marker iNOS, and HWAL elevated the M2 activation marker Ym1 and ARG1 gene expression. In conclusion, MAEK could be used as an immune stimulant because of its ability to activate T cells (elicited c-fos and CD40L gene expression), whereas HWAL could serve as an anti-inflammatory agent because of its differential activation of M2 macrophages.

Isolation of Myrosinase and Glutathione S-transferase Genes and Transformation of These Genes to Develop Phenylethylisothiocyanate Enriching Chinese Cabbage (배추에서 항암물질 phenylethylisothiocyanate의 다량 합성을 위한 myrosinase와 glutathione S-transferase 유전자 분리 및 이를 이용한 형질전환체 육성)

  • Park, Ji-Hyun;Lee, Su-Jin;Kim, Bo-Ryung;Woo, Eun-Teak;Lee, Ji-Sun;Han, Eun-Hyang;Lee, Youn-Hyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.29 no.6
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    • pp.623-632
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    • 2011
  • To increase the anti-carcinogens phenylethylisothiocyanate (PEITC), myrosinase (MYR), and glutathione S-transferase (GST), genes related to PEITC pathway were isolated and the gene expressions were regulated by Agrobacterium transformation. Isolated cDNAs, MYR, and GST genes were 1,647 bp and 624 bp, respectively, and the protein expression was confirmed through pET system. Thereafter, we constructed a sense-oriented over-expressing myrosinase (pBMY) and RNAi down-regulated GST (pJJGST) binary vectors for the Chinese cabbage transformation. After the transformation, thirteen over-expressing transgenic Chinese cabbage plants (IMS) with pBMY and five down-regulated ones (IGA) with pJJGST were selected by PCR analysis. Selected $T_0$ transgenic plants were generated to $T_1$ plants by self-pollination. Based on the Southern blot analysis on these $T_1$ transgenic plants, 1-4 copies of T-DNA were transferred to Chinese cabbage genome. Thereafter, RNA expression level of myrosinase gene or GST gene was analyzed through real-time RT PCR of IMS, IGA, and non-transgenic inbred lines. In case of IMS lines, myrosinase gene was increased 1.03-4.25 fold and, in IGA lines, GST gene was decreased by 26.42-42.22 fold compared to non-transgenic ones, respectively. Analysis of PEITC concentrations using GC-MS it showed that some IMS lines and some IGA lines increased concentrations of PEITC up to 4.86 fold and up to 3.89 fold respectively compared to wild type. Finally in this study IMS 1, 3, 5, 12, and 15 and IGA 1, 2, and 4 were selected as developed transgenic lines with increasing quantities of anti-carcinogen PEITC.

A Study of Tasseled Cap Transformation Coefficient for the Geostationary Ocean Color Imager (GOCI) (정지궤도 천리안위성 해양관측센서 GOCI의 Tasseled Cap 변환계수 산출연구)

  • Shin, Ji-Sun;Park, Wook;Won, Joong-Sun
    • Korean Journal of Remote Sensing
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    • v.30 no.2
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    • pp.275-292
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    • 2014
  • The objective of this study is to determine Tasseled Cap Transformation (TCT) coefficients for the Geostationary Ocean Color Imager (GOCI). TCT is traditional method of analyzing the characteristics of the land area from multi spectral sensor data. TCT coefficients for a new sensor must be estimated individually because of different sensor characteristics of each sensor. Although the primary objective of the GOCI is for ocean color study, one half of the scene covers land area with typical land observing channels in Visible-Near InfraRed (VNIR). The GOCI has a unique capability to acquire eight scenes per day. This advantage of high temporal resolution can be utilized for detecting daily variation of land surface. The GOCI TCT offers a great potential for application in near-real time analysis and interpretation of land cover characteristics. TCT generally represents information of "Brightness", "Greenness" and "Wetness". However, in the case of the GOCI is not able to provide "Wetness" due to lack of ShortWave InfraRed (SWIR) band. To maximize the utilization of high temporal resolution, "Wetness" should be provided. In order to obtain "Wetness", the linear regression method was used to align the GOCI Principal Component Analysis (PCA) space with the MODIS TCT space. The GOCI TCT coefficients obtained by this method have different values according to observation time due to the characteristics of geostationary earth orbit. To examine these differences, the correlation between the GOCI TCT and the MODIS TCT were compared. As a result, while the GOCI TCT coefficients of "Brightness" and "Greenness" were selected at 4h, the GOCI TCT coefficient of "Wetness" was selected at 2h. To assess the adequacy of the resulting GOCI TCT coefficients, the GOCI TCT data were compared to the MODIS TCT image and several land parameters. The land cover classification of the GOCI TCT image was expressed more precisely than the MODIS TCT image. The distribution of land cover classification of the GOCI TCT space showed meaningful results. Also, "Brightness", "Greenness", and "Wetness" of the GOCI TCT data showed a relatively high correlation with Albedo ($R^2$ = 0.75), Normalized Difference Vegetation Index (NDVI) ($R^2$ = 0.97), and Normalized Difference Moisture Index (NDMI) ($R^2$ = 0.77), respectively. These results indicate the suitability of the GOCI TCT coefficients.