Gamble G.;Beaumont B.;Smith H.;Zorn J.;Sanders G.;Merrilees M.;MacMahon S.;Sharpe N.
대한예방의학회:학술대회논문집
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1994.02b
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pp.169-179
/
1994
B-mode ultrasound is being used to assess carotid atherosclerosis in epidemiological studies and clinical trials. Recently the interpretation of measurements made from ultrasound images has been questioned. This study examines the anatomical correlates of B-mode ultrasound of carotid arteries in vitro and in situ in cadavers. Twenty-seven segments of human carotid artery were collected at autopsy. pressure perfusion fixed in buffered 2.5% gluteraldehyde and 4% paraformaldehyde and imaged using an ATL UM-8 (10 MHz single crystal mechanical probe). Each artery was then frozen, sectioned and stained with van Gieson or elastin van Gieson. The thickness of the intima. media and adventitia were measured 'to an accuracy of 0.01 mm from histological sections using a calibrated eye graticule on a light microscope. Shrinkage artifact induced by histological preparation was determined to be 7.8%. Digitised ultra sound images of the artery wall were analysed off-line. The distance from the leading edge of the first interface ($LE_{1}$) to the leading edge of the second interface ($LE_2$) was measured using a dedicated programme. $LE_{1}$-$LE_{2}$ measurements were correlated against histological measurements corrected for shrinkage. Mean values for the far wall were: ultra sound $LE_{1}$-$LE_{2}$ (0.97 mm, S.D. 0.26), total wall thickness (1.05 mm, S.D. 0.37), adventitia (0.35 mm, S.D. 0.16), media (0.61 mm, S.D. 0.18). intima (0.09 mm, S.D. 0.13). Ultrasound measurements corresponded best with total wall thickness, rather than elastin or the intima-media complex. Excision of part of the intima plus media or removal of the adventitia resulted in a corresponding decrease in the $LE_{1}$-$LE_{2}$ distance of the B-mode image. Furthermore. increased wall thickness due to intimal atherosclerotic thickening correlated well with $LE_{1}$-$LE_{2}$ distance of the B-mode images. B-mode images obtained from the carotid arteries in situ in four cadavers also corresponded best with total wall thickness measured from histological sections and not with the thickness of the intima plus media. In conclusion, the $LE_{1}$-$LE_{2}$ distance measured on B-mode images of the carotid artery best represents total wall thickness of intima plus media plus adventitia and not intima plus media alone.
Objective : It has long been known about the osteogenic effect of CPC-HAS on bone tissues. However, it has not been determined the effect of CPC-HAS on cancer cells. The purpose of this study is to screen the CPC-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells. Oligonucleotide microarray and proteomics approaches were employed to screen the differential expression genes. Methods : CPC-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CPC-HAS (0.1, 0.5, 1.5, 10, 20mg/ml) for 24 h. Cell toxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with 1.5mg/ml of CPC-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide Genechip(Human genome Ul33 Plus 2.0., Affimatrix Co.). For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : It has no cytotoxic effects on both HepG2 cell in all concentrations(0.l, 0.5, 1.5, 10, 20mg/ml). In oligonucleotide microarray assay, the number of more than twofold differentially regulated known genes was 23 with 5 up-regulated and 18 down-regulated genes in HepG2 cells. In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. Two down-regulated proteins were aldehyde dehydrogenase 1 and enolase 1, and up-regulated protein was fatty acid binding protein 1 by 1.5mg/ml of CPC-HAS. Discussion : This study showed the screening of CPC-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray and proteomic analysis. The screened genes will be used for the better understanding of the therapeutic effects of CPC-HAS on cancer fields.
Lee Sang Hun;Lee Seung Eon;Lee Si Hyeong;Shin Jo Young
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.2
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pp.528-533
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2004
The immune system acts to protect the host from infectious agents that exist in the environment and from other noxious insults. The immune system has two functional divisions: the innate and the acquired. Both components involve various factors such as cytokines. A number of methodologies exist to assess aspects of immune function. There are large inter-individual variations in many immune functions even among the healthy. Genetics, age, gender, smoking habits, habitual levels of exercise, alcohol consumption, diet, stage in the female menstrual cycle, stress, history of infections and vaccinations, and early life experiences are likely to be important contributors to the observed variation. While it is clear that individuals with immune responses significantly below 'normal' are more susceptible to infectious agents and exhibit increased infectious morbidity and mortality, it is not clear how the variation in immune function among healthy individuals relates to variation in susceptibility to infection. Oriental medicine is an important factor contributing to immune competence. The author investigated the immune enhancement effects of Bojungikkitanggami-bang (BITB). The forced swimming test (FST) has been used as a screening model for new immune enhancement agents. In the present study, the author investigated the effects of BITB on FST and blood biochemical parameters related to fatigue, glucose (Glc); blood urea nitrogen (BUN); lactate dehydrogenase (LDH); creatinine; and total protein (TP). The author found that BITB (1 g/kg) significantly reduced the immobility time in the FST compared to the control. In addition, the contents of Glc, LDH, BUN, TP in the blood serum were increased in BITB (1g/kg)-fed group. Also, the author investigated the effects of BITB on the production of cytokines in human T-cell line, MOLT-4 cells. BITB (1 mg/ml) significantly increased the interferon (IFN)-vproduction compared with media control (about 2.2-fold for IFN-γ) at 24 h. However, BITB has not affect the production of IL-2 and IL-4. In addition, BITB increased the protein expression level of IFN-γ in MOLT-4 cells. Thus, BITB may have therapeutic value in generating or enhancing immune function in a clinical setting.
Geraniol (1), olivetol (2), cannabinoids (3 and 4) and 5-fluorou.a.il (5). were tested for their growth inhibitory effects against SK-MEL-3 cell lines using two different 3-(4,5-dimethylthiazol- 2yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay and sulforhodamine B protein (SRB) assay. These compounds showed inhibitory activity in vitro in the micromolar range against SK-MEL-3 cell lines. In general, the antitumor activity of these compounds (1, 2, 3, 4 and 5) was in a dose-dependent over the micromolar concentration range $1\;to\;100{\mu}M$. The comparison of $IC_{50}$ values of these compounds in tumor cell lines shows that their susceptibility to these compounds decrease in the following order : OLVTL > CBG > CBD > 5-FU > CRNL in MTT assay, CBG > OLVTL > CBD > GRNL > 5-FU in SRB assay. Cannabinoids (3 and 4) and 5-fluorouracil (5) were tested for their cytotoxic effects on NIH 373 fibroblasts using two different MTT assay and SRB assay. These compounds exhibited potent cytotoxic activities in vitro in the micromolar range against NIH 373 fibroblasts. In general, the cytotoxic activities of these compounds (3, 4 and 5) were in a dose-dependent over the micromolar concentration range $1\;to\;100{\mu}M$. The comparison of $CD_{50}$ values of these compounds on NIH 373 fibroblasts shows that their susceptibility to these compounds decrease on the following order ; CBD > 5-FU > CBG in MTT assay and SRB assay. Cannabigerol (3) was shown the least cytotoxic activity on NIH 373 fibroblasts. Cannabigerol (3) exhibited the most growth-inhibitory activity against SK-MEL-3 cell lines.
Seo, Ji-Yeon;Lim, Soon-Sung;Park, Ji-A;Lim, Ji-Sun;Kim, Hyo-Jung;Kang, Hui-Jung;YoonPark, Jung-Han;Kim, Jong-Sang
Nutrition Research and Practice
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v.4
no.2
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pp.93-98
/
2010
Our previous study demonstrated that methanolic extract of Chrysanthemum zawadskii Herbich var. latilobum Kitamura (Compositae) has the potential to induce detoxifying enzymes such as NAD(P)H:(quinone acceptor) oxidoreductase 1 (EC 1.6.99.2) (NQO1, QR) and glutathione S-transferase (GST). In this study we further fractionated methanolic extract of Chrysanthemum zawadskii and investigated the detoxifying enzyme-inducing potential of each fraction. The fraction (CZ-6) shown the highest QR-inducing activity was found to contain (+)-(3S,4S,5R,8S)-(E)-8-acetoxy-4-hydroxy-3-isovaleroyloxy-2-(hexa-2,4-diynyliden)-1,6-dioxaspiro [4,5] decane and increased QR enzyme activity in a dose-dependent manner. Furthermore, CZ-6 fraction caused a dose-dependent enhancement of luciferase activity in HepG2-C8 cells generated by stably transfecting antioxidant response element-luciferase gene construct, suggesting that it induces antioxidant/detoxifying enzymes through antioxidant response element (ARE)-mediated transcriptional activation of the relevant genes. Although CZ-6 fraction failed to induce hepatic QR in mice over the control, it restored QR activity suppressed by $CCl_4$ treatment to the control level. Hepatic injury induced by $CCl_4$ was also slightly protected by pretreatment with CZ-6. In conclusion, although CZ-6 fractionated from methanolic extract of Chrysanthemum zawadskii did not cause a significant QR induction in mice organs such as liver, kidney, and stomach, it showed protective effect from liver damage caused by $CCl_4$.
The cysteine and glycine rich protein 3 (CSRP3), apolipoprotein B mRNA editing enzyme catalytic polypeptide‐like 2(APOBEC2) and caveolin (CAV) gene family(CAV1, CAV2, CAV3) have been reported to play important roles for carcass and meat quality traits in pig, mouse, human and cattle. As an initial step, we investigated SNPs in these 5 genes among eight different cattle breeds. Eighteen primer pairs were designed from bovine sequence data of NCBI database to amplify the partial gene fragments. Sequencing results revealed 9 SNPs in the coding regions of three caveolin genes, 1 SNP in CSRP3 and 3 SNPs in APOBEC2 gene. All the identified SNPs were confirmed by PCR-RFLP. Also, 9 more intronic SNPs were detected in these genes. However, all identified mutations in the coding region do not change amino acid sequence. Allelic distributions were significantly different for 5 SNPs in CAV2, CAV3, CSRP3 and APOBEC2 genes among the eight different breeds. These results gave some clues about the polymorphisms of these genes among the cattle breeds and will be useful for further searches for identifying association between these SNPs and meat quality traits in cattle.
Purpose: The shape and location, the amount of the wound and the characteristics of the remaining tissues are known to influence wound contraction. The previous studies using small animals have not been an appropriate model because the wound healing mechanisms and skin structures are different from those of the human. The purpose of this study is to evaluate wound contraction according to the shape and location of the wound using a $Micropig^{(R)}$. Methods: Four $Micropigs^{(R)}$ (Medikinetics, Pyeongtaek, Korea) that were 10 months of age weighed 25 kg were used. Full thickness skin defects were made by clearing all the tissues above the fascial layer in the shape of square, a regular triangle and a circle of 9 $cm^2$ each on the back around the spine. Eight wounds were created on the back of each pig, 50 mm apart from each other. The randomly chosen wound shapes included 11 squares, 11 regular triangles, and 10 circles. Wound dressing was done every other day with polyurethane foam. The wound size was measured using a Visitrak $Digital^{(R)}$ (Smith & Nephew, Hull, UK) on every other day after surgery from day 2 to day 28. A biopsy was performed on day 3, and 1, 2, 3 and 4 weeks to investigate the degree of acute and chronic inflammation, the number of microvesssel and myofibroblast density using H & E stain and immunohistochemistry. The wound contraction rate was calculated to figure out the differences among each of the shapes and the locations. Results: The ultimate shape of the circle wound was oval, and that of the regular triangle and square were stellate. The maximum contraction rate was obtained on 8 to 10 days for all the shapes, which corresponds with the immunohistochemical finding that myofibroblast increases in the earlier 2 weeks whereas it decreases in the later 2 weeks. Epithelialization was seen in the wound margin on day 7 and afterwards. The final wound contraction rates were highest for the regular triangle shapes; however, there were no statistically significant differences. The wound contraction rates by locations showed statistically significant differences. The wound in the cephalic area presented more contractions than that of the wounds in the caudal area. Conclusion: The location of a wound is more important factor than the wound shape in wound contraction.
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.4
/
pp.520-526
/
2013
To determine the kimchi with the best quality and functional characteristics, we manufactured and compared recipes for Korean and Japanese kimchi made either Korean or Japanese baechu cabbages. All batches were fermented for 4 weeks at $5^{\circ}C$, and tested for pH, texture, microbial count, sensory evaluation, DPPH radical-scavenging activity, and cell proliferation (using the MTT assay on AGS human gastric cancer cells). By the third week of fermentation, Korean kimchi made with Korean baechu (KK) and Japanese kimchi made with Korean baechu (KJ) showed a higher acidity than Korean or Japanese kimchi made with Japanese baechu (JK and JJ, respectively). KK ranked highest in springiness, followed by KJ, JK, and JJ. Therefore, the texture of kimchi produced with Korean baechu was appears better than kimchi produced with Japanese baechu. This was confirmed in masticatory tests. Kimchi produced with Korean baechu (KK and KJ) showed lower total aerobic bacterial counts, while the total lactic acid bacterial counts were higher (p<0.05). In sensory evaluation test, KK received the highest overall acceptability score, while JJ earned the lowest score. In the DPPH assay for anti-oxidative activity, KK showed a 94% anti-oxidative effect, followed by KJ (92%), JK (91%), and JJ (88%) (p<0.05). In the MTT assay for analyzing the cell proliferation of AGS human gastric cancer cells, KK showed a 64% anticancer effect in vitro, followed by KJ (57%), JK (38%), and JJ (26%). Therefore, the anti-oxidative and anti-cancer functionalities of kimchi made with Korean baechu were higher than those made with Japanese baechu, regardless of the kimchi recipe applied. Overall, Korean baechu had important and superior effects on the quality and functionality of kimchi.
The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.
Pyrethroid insecticide have widely been used for agricultural sector and residential environments. To assess the exposure of insecticide which is absorbed through skin the analysis of urinary metabolite is essential. At present, the urinary 3-PBA was analyzed using liquid-phase extraction. But LPE have many limitations, such as long pre-treatment time and low recovery. So, this study was conducted to determine the optimum conditions for analysing 3-PBA in urine using solid phase extraction. Furthermore, this study intend to investigate the relation of concentrations of pyrethroid, deltamethrin in air and 3-PBA in urine. The optimum condition for hydrolysis was found to be done with hydrochloric acid for one hour. The recovery rates of 3-PBA were $84.6%{\pm}1.2%$, $54.8{\pm}0.9%$, $99.8{\pm}1.2%$ with XAD-2, XAD-7, XAD-16 using as the aborbents and acetone as eluents respectively. But acetonitrle and methanol gave low recovery rate and methyl cellosolve could not elute the compound. The amount of acetone for elution were 6mL, 9mL, 3mL for XAD-2, XAD-7, XAD-16 as absorbents respectively. The non-absorbed rates was $0.8{\pm}0.5%$, and $0.7{\pm}0.3%$ under XAD-16, mesh size 140-200, amount of resin 1.4g and the flow rate of eluent was 0.1mL/min. In the concentration process, we obtained 11 times higher concentration of material. The amounts of urinary 3-PBA were. The LODs of 3-PBA and deltamethrin were 0.004 mg/L, 0.038 mg/L, respectively. The further research of minute monitoring which include spray pattern, environmental condition is needed And more research about the relation between total pyrethroid exposure and urinary various metabolite are also necessary.
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