Backgrounds: This study investigated whether or not a polymorphism in the gene encoding the surfactant protein A(SP-A) has any bearing on the individual susceptibility to the development of chronic obstructive pulmonary disease(COPD) in a genetically homogenous Korean population. Methods: The genotypes of 19 COPD patients and 20 healthy neonates as controls were tested using a polymerase chain reaction followed by restriction fragment length polymorphism analysis for the SP-A gene. Results: The specific frequencies of the 6A2 and 6A18 alleles of SP-A1 and the 1A2 allele of SP-A2 were much higher in the COPD group than control group (p<0.05). However, the frequencies of the 6A3 and 6A4 alleles of SP-A1 and the 1A0 allele of SP-A2 in the COPD group were significantly lower than the control group. In the COPD group, the frequencies of the +50 locus genotypes GG of SP-A1 and the +9 locus genotypes CC of SP-A2 were 85.0% and 60.6%, respectively, and 19.7% and 24.8% in the control group, respectively. The frequencies of the polymorphic genotypes or alleles showed a statistically significant difference between the COPD group and the control group (P<0.05). Conclusion: A genetic polymorphism in SP-A is associated with the development of COPD in the Korean population.
Seo, Eun-Sun;Chae, Soo-Chul;Kho, Eun-Gyeong;Lee, Jong-Bin
Korean Journal of Environmental Biology
/
v.27
no.1
/
pp.66-72
/
2009
Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.
This study was carried out to develop a novel hydroponic medium far fruit vegetable crops by using waste synthetic fibers. In physical analysis of the synthetic fiber medium (SFM), the bulk density and percent solid phase were lower, while the porosity and water content were greater in comparison with the rockwool slab. The SFM had pH of 6.5 and EC of $0.03dS{\cdot}m^{-1}$ both of which are similar to those of the rockwool slab. The CEC of 0.39me/100mL of the SFM was lower than compared with 3.29me/100mL of the rockwool slab. However, concentrations K, Ca, Mg and Na were slightly higher in the SFM than those in the rockwool slab. The 'Momotaro' tomato crop in the SFM gave comparable plant height, stem diameter, days to first flowering, fruit weight and percent marketable yield as the rockwool slab. In the SFM and in the rockwool slab, mean fiuit weight were 182g and 181g, percent marketable yield were $93.8\%$ and $92.0\%$, respectively. The marketable yield per 10a in the SFM was 12,799 kg, which was $97\%$ of that in the rockwool slab. Growth parameters such as leaf length and width, leaf number, stem diameter and chlorophyll content of an exportable cucumber crop grown in the SFM and the rockwool slab were not different. Fruit weight was greater in the rockwool slab, while percent marketable yield was greater in the SFM. The marketable fruit yield per 10a of 5,062kg in the SFM was $2\%$ greater than that in the rockwool slab. $NO_3$ concentration in nutrient solution during the crop cultivation was higher in the SFM than in the rockwool slab, while concentrations $NH_4$, K, Ca, Mg and $SO_4$ were not different between the two media.
Preferential flow has recently been the subject of increasing interest because these phenomena contribute to solute transport in soils. Commonly, preferential flow paths are associated with macropores or highly structured soils. We presented an analysis of the measured breakthrough curves (BTCs) of $Cl^-$ and $Cu^{2+}$ ions to test the occurrence of preferential flow in soils using miscible displacement technique under steady flow conditions. We also analyzed soil water retention curves and from this curves induced cumulative pore size distribution of undisturbed soils, which sampled from Ap1, B1, and C horizons of Songjeong series soils (the fine loamy, mesic family of Typic Hapludults). In this study, miscible displacement experiment on C horizon was excluded, because it is structureless sandy loam with saturated hydraulic conductivity of $5.2cmhr^{-1}$. The saturated hydraulic conductivity of Ap1 horizon was $2.0cmhr^{-1}$, which was about 7 times higher than that of B1 horizon ($0.27cm hr^{-1}$). Cumulative pore size distribution predicted that Ap1 horizon had more macropores (pore diameter larger than $49{\mu}m$, equivalent to -6 kpa of soil matric potential) than B1 horizon. The hydrodynamic dispersion coefficient from chloride BTCs was estimated as $1.3cm^2hr^{-1}$ for B1 and $34cm^2hr^{-1}$ for Ap1 horizon. However the retardation factors of B1 and Ap1 horizon were significantly different, i.e. 1 and 0.6, respectively, which means that there was distinct partition between mobile water and immobile phase in Ap1 horizon. The copper retardation effect of Ap1 horizon was less than that of B1 horizon, even though cation exchange capacity of Ap1 horizon was higher than that of B1 horizon. Thus, breakthrough curves of $Cl^-$ and $Cu^{2+}$ obviously showed the probability that preferential flow would occur in Ap1 horizon.
Prupose : The genes involved on the suppression or radiation-induced apoptosis by genistein in K562 leukemia cell line was investigated. Materials and methods : K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For X-ray irradiation and drug treatment, cultures were prepared at $2\times10^5\;cells/mL$. The cells were irradiated with 10 Gy (Clinac 1800C, Varian, USA), Stock solutions of herbimycin A (HMA, Calbiochem, UK) and genistein (Calbiochem, UK) were prepared in dimethylsulfoxide (DMSO, Sigma, UK). After incubation at $37^{\circ}C$ for 24 h, PCR-select cDNA subtractive hybridization, dot hybridization, DNA sequencing and Northern hybridization were examined. Results : Smad6 gene was identified from the differentially expressed genes in K562 cells incubated with genistein which had been selected by PCR-select cDNA subtractive hybridization. The mRNA expression of Smad6 in K562 cells incubated with genistein was also higher than control group by Northern hybridization analysis. Conclusion : We have shown that Smad6 involved on the suppression of radiation-induced apoptosis by genistein in K562 leukemia cell line. It is plausible that the relationship between Smad6 and the suppression of radiation-induced apoptosis is essential for treatment development based on molecular targeting designed to modify radiation-induced apoptosis.
Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.
Kim, Eun-Ji;Park, Hee-Sook;Lim, Soon-Sung;Kim, Jong-Sang;Shin, Hyun-Kyung;Yoon, Jung-Han
Korean Journal of Food Science and Technology
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v.40
no.2
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pp.207-214
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2008
In Asia Saussurea lappa (SL) has been used as a traditional herbal medicine to treat abdominal pain and tenesmus. Recently, in vitro cell culture studies have shown that SL has anti-ulcer, anti-inflammatory, and anti-tumor properties. To explore its potential chemopreventive and chemotherapeutic effects in colon cancer, we examined whether the hexane extract of SL (HESL) could inhibit the growth of HT-29 human colon cancer cells, and investigated the mechanisms for this effect. The cells were cultured with various concentrations (0-5 ${\mu}g/mL$) of HESL. The results indicated that HESL markedly decreased the numbers of viable HT-29 cells; whereas at the concentration of 5 ${\mu}g/mL$, HESL slightly decreased the viable cell numbers of CCD 1108Sk human skin normal fibroblasts at 72 hr. HESL substantially increased the numbers of cells in the sub G1 phase, and dose-dependently increased apoptotic cell numbers. Western blot analysis of the total cell lysates revealed that HESL increased Bax protein levels, but did not affect Bcl-2 levels. HESL induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3. This study demonstrated that HESL inhibits cell growth and induces apoptosis in HT-29 cells, which may be mediated by its ability to increase Bax levels and activate the caspase pathway. These findings may lead to the development of new therapeutic strategies for colon cancer treatment.
This study compared and analyzed the effect of income-redistribution, collecting data on the basis of the estimated details of insurance contribution and individual money wage lists for each one year before and after the combination of medical insurance program for industrial workers, by systematic sampling, extracting 4,160 families(14,764 people) among people applied to medical insurance program for self employees in Taegu City on the basis of Oct. 1st in 1998 with 227 associations of medical insurance program for self employees and medical insurance program for government employees and private school teachers combined, comparing the effect of income redistribution of before and after the combination of medical insurance program for self employees. The insurance contribution by household after the combination of medical insurance program for self employees showed the increase rate of average 20.9%, among them households of 68.8% increased and 31.2% decreased. The effect of income-redistribution was more positive because the degree of inequality was more deepened from 0.64 of the before-combination to 0.45 of the after-one in decile distribution ratio, from 0.26 to 0.34 in Gini -coefficient. Decile distribution ratio on the basis of insurance benefits by household was from 0.09 in the before-combination to 0.14 in the after-one, Gini-coefficient from 0.16 in the before-combination to 0.57 in the after-one was a little lowered. And decile distribution ratio of insurance benefits on the basis of insurance contribution was higher from 1.08 in the before-combination to 1.23 in the after-one, concentration index was a little lowered from 0.14 to 0.11, the effect of income-redistribution was improved in the phase of insurance benefits. The income-transfer rate of medical insurance program for self employees (the occupied rate of insurance benefits/ the occupied rate of insurance contribution) showed a lower trend in all of the before and after-combination towards upper classes, it was known that the income-transfer rate was higher from 1st degree to 7th degree in the after-combination in comparison with the before-one, but the effect of income¬redistribution was high because the income-transfer rate was lowered from 8th degree to 10th degree. The rate of medical insurance benefits (insurance benefits/ insurance contribution) increased from 0.79 in the before-combination to 1.07 in the after-one, and showed over 1.0 under 3th degree before the combination, but all of it was higher than 1.0 under 7th degree after the combination, the after-combination was more improved than the before-one in view of the rate of insurance benefits. As the result of above, on the basis of Oct. 1st in 1998 that 227 associations of medical insurance program for self employees was combined into one, we could say that the equality of imposing medical insurance contribution was more re-considered in the after-combination than in the before-one. But this study analyzed with classes divided, anyway, on the basis of insurance contribution, we have limit in explaining the correct effect of income-redistribution, because it was not analyzed according to classes of income, though it helps to analogize the effect of income-redistribution. So there must be analysis about the effect of income-redistribution, on the basis of the system, building up the system to grasp the correct income of the insureds of medical insurance program for self employees.
Ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin in chicken muscle were seperated by liquid extraction and determined with high performance liquid chromatography (HPLC) with fluorescence detector. Analysis was carried out using following conditions; Cl8 column (250${\times}$4.6 mm i.d. 5 ${\mu}{\textrm}{m}$ particle size), mobile phase composed of D.W. (containing 0.4% triethylamine and phospholic acid): methanol : acetonitrile (800:100:100, v/v/v), isocratic pump at a flow rate of 1.0 $m\ell$/min and 50 ${mu}ell$ of injection volume, fluorescence detector with EX278 nm/EM.456 nm. The calibration curves of four fluoroquinolones showed linearity (${\gamma}$$^2$$\geq$0.999) at concenration range of 0.025-0.6 $\mu\textrm{g}$/ml. The recoveries in fortified chicken muscle represented more than 80% with low coefficient of variation (〈10%) for concentration range of four fluoroquinolones. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin were 23.5, 3.4, 3.0 and 2.5 ng/g in chicken muscle, respectively. We also monitored fluoroquinolones residue in muscle of chickens (broiler 1:227, Korean native chicken 219, laying chicken 77) using EEC-4-plate screening and HPLC conformation methods. Ten(broiler 5, Korean native chicken 5) out of the fifteen samples which were positively detected by EEC-plate screening method from 1,523 chicken meat were confirmed with ciprofloxacin and enrofloxacin by HPLC. The ranges of residual concentration were 0-0.12 ppm for ciprofloxacin and 0.01-6.79 ppm for enrofloxacin. In conclusion, our method could be applied effectively to determine four fluoroquinolones residues in chicken meat, and further survey for fluoroquinolones residue in chicken meat are needed for more effective control of fluoroquinolones used in livestock.
The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.
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