• Journal of Life Science
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• v.20 no.1
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• pp.60-65
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• 2010

Since organic sulfur compounds respond to GC/ECD sensitively, they interfere with quantitative separation of pesticides during residual pesticide analysis of Allium species. In this study, it was intended to develop a rapid and simple method for pesticide multi-residues analysis through clean-up and interferences by a solid-phase extraction (SPE). An SPE method employing silver nitrate impregnated Florosil cartridge was developed and evaluated for the elimination of sulfur compounds from the test solution of Allium species during pesticide residues analysis. The silver nitrate impregnated Florosil cartridge was prepared by efflux of 3 ml of 20% silver nitrate solution through Florosil cartridge (1 g packing, 6 ml tube). The extracts equivalent to 2, 4 6, and 10 g of each sample were loaded onto the cartridge and allowed to exude, and then the exudations were analyzed by GC/ECD. More than 95% of sulfur compounds were removed from the loaded extracts equivalent, up to 6 g in onion, 4 g in spring onion and 4 g in shallot, respectively. 40 pesticides were spiked in the Allium species and loaded onto the cartridge to determine the recoveries; from this, the recoveries of 34 pesticides were within 70~120%.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.148-151
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• 2010

The aim of this study was to investigate the erythromycin resistance patterns of Enterococci sp. present in cow milk. A total 110 erythromycin resistant Enterococci were isolated from milk samples; E. faecalis (n=101), E. avium (n=7), and E. durans (n=2). The minimum inhibitory concentration of erythromycin against 110 Enterococci were determined. The 66.3% of Entercocci (n=73) showed high level resistance (${\geq}64$ mg/ml). Among 110 isolates, 86.3% (n=95) showed $cMLS_B$ phenotype and 13.6% (n=15) showed $iMLS_B$ The aim of this study was to investigate the erythromycin resistance patterns of Enterococci sp. present in cow milk. A total 110 erythromycin resistant Enterococci were isolated from milk samples; E. faecalis (n=101), E. avium (n=7), and E. durans (n=2). The minimum inhibitory concentration of erythromycin against 110 Enterococci were determined. The 66.3% of Entercocci (n=73) showed high level resistance (${\geq}64$ mg/ml). Among 110 isolates, 86.3% (n=95) showed $cMLS_B$ phenotype and 13.6% (n=15) showed $iMLS_B$ phenotype. All of isolates have erm(B) determinant, 75.45% (n=83) have mef(A) an efflux system determinant. The majority of Enetrcococci isolated from raw milk samples in northern area of Gyeonggi-Do showed high level of resistance to erythromycin.

• Korean Journal of Veterinary Research
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• v.40 no.2
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• pp.267-274
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• 2000

Since intracellular free $Mg^{2+}$ ($[Mg^{2+}]_i$) appears to be tightly regulated following cellular energy depletion, we hypothesized that the increase in $[Mg^{2+}]_i$ would result in $Mg^{2+}$ extrusion into circulation. Extracellualr $Mg^{2+}$ contents ($[Mg^{2+}]_o$) were measured in rat erythrocytes, the perfused heart and liver, and plasma in the anesthetized rat. Animals were injected intraperitoneally with sodium nitrite ($NaNO_2$) and plasma $Mg^{2+}$ was measured after the injection and then 10 and 20 minutes later. An increase in circulating (plasma) $Mg^{2+}$ ($[Mg^{2+}]_c$) and methemoglobin was observed in animals injected with $NaNO_2$ (30 mg/Kg). The time course of the effects demonstrated that $[Mg^{2+}]_c$ and methemoglobin continued to increase 10 minutes after the $NaNO_2$ injection. Under these conditions, there was a sustained increase in $[Mg^{2+}]_c$, but not in methemoglobin, which was inhibited by pretreatment with potassium cyanide (KCN, 4 mg/Kg), indicating that an increase in $[Mg^{2+}]_c$ was accompanied by ATP depletion. Injection of rotenone (0.9 mg/Kg) or 2,4-dinitrophenol (15 mg/Kg) also induced an increase in $[Mg^{2+}]_c$. Reduced respiration rate from 100/min to 10/min during 30 minutes also caused a time-dependent rise in $[Mg^{2+}]_c$. These increase in $[Mg^{2+}]_c$ were inhibited by pretreatment with KCN. In addition, ATP depletion by $NaNO_2$ or KCN sustainedly increased the $[Mg^{2+}]_o$ in rat erythrocytes. $Mg^{2+}$ efflux was stimulated by KCN in the perfused heart and liver, but not by $NaNO_2$. These results suggest that the activation of $Mg^{2+}$ effluxes into the circulation is directly dependent on the ATP depletion-induced increase in $[Mg^{2+}]_i$ and heart, liver and erythrocytes have a major pool of $Mg^{2+}$ that can be mobilized upon cellular energy state.

• Archives of Pharmacal Research
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• v.29 no.10
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• pp.866-873
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• 2006

When patients with cancers are treated with chemotherapeutic agents a long time, some of the cancer cells develop the multidrug resistance (MDR) phenotype. MDR cancer cells are characterized by the overexpression of multidrug resistance1 (MDR1) gene which encodes P-glycoprotein (Pgp), a surface protein of tumor cells that functions to produce an excessive efflux and thereby an insufficient intracellular concentration of chemotherapeutic agents. A variety of studies have sought potent MDR modulators to decrease MDR1 gene expression in cancer cells. Our previous study has shown that curcumin exhibits characteristics of a MDR modulator in KB-V1 multidrug-resistant cells. The aim of this study was to further investigate the effect of curcumin on MDR1 gene expression in patient leukemic cells. The leukemic cells were collected from 78 childhood leukemia patients admitted at Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand, in the period from July 2003 to February 2005. There were 61 cases of acute lymphoblastic leukemia (ALL), 14 cases of acute myeloblastic leukemia (AML), and 3 cases of chronic myelocytic leukemia (CML). There were 47 males and 31 females ranging from 1 to 15 years old. Bone marrows were collected. The leukemic cells were separated and cultured in the presence or absence of $10{\mu}M$ curcumin for 48 hours. MDR1 mRNA levels were determined by RT-PCR. It was found that curcumin reduced MDR1 gene expression in the cells from 33 patients (42%). Curcumin affected the MDR1 gene expression in 5 of 11 relapsed cases (45%), 10 of 26 cases of drug maintenance (38%), 7 of 18 cases of completed treatment (39%), and 11 of 23 cases of new patients (48%). The expression levels of MDR1 gene in leukemic patient cells as compared to that of KB-V1 cells were classified as low level (1-20%) in 5 of 20 cases (25%), medium level (21-60%) in 14 of 32 cases (44%), and high level (61-100%) in 14 of 20 cases (70%). In summary, curcumin decreased MDR1 mRNA level in patient leukemic cells, especially in high level of MDR1 gene groups. Thus, curcumin treatment may provide a lead for clinical treatment of leukemia patients in the future.

• Journal of Ginseng Research
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• v.26 no.1
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• pp.1-5
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• 2002

It has been well known that Korea red ginseng has an antihypertensive effect. The antihypertensive effect may be due to its ability to change the peripheral resistance. Change of vascular tone in the resistance-sized artery contribute to the peripheral resistance, thereby regulate the blood pressure. Therefore, we investigated to clarify the vasorelaxing mechanism induced by crude saponin of Korea red ginseng in the resistance-sized mesenteric artery of rats. The resistance-sized mesenteric artery was isolated and cut into a ring. The ring segment was immersed in HEPES-buffered solution and its isometric tension was measured using myograph force-displacement transducer. Crude saponin of ginseng relaxed the mesenmetric arterial rings precontracted with norepinephrine (3$\mu$M) in dose-dependent manner (0.01 mg/㎖ -1 mg/㎖. The relaxation by crude saponin was smaller in endothelium-intact preparation than that in endothelium-denuded preparation. The contraction induced by A23187 or phorbol 12,13-dibutyrate was not affected by crude saponin of ginseng. The vasorelaxing effect of crude saponin of ginseng was significantly attenuated by the increase of the extracellular K$\^$+/ concentration. Crude saponin-induced vasorelaxation was not affected by tetraethylammonium (1 mM), glybenclamide (10$\mu$M), and 4-aminopyridine (0.1 mM) in these preparations. Ba$\^$2+/(10$\mu$M ∼100$\mu$M) markedly reduced the crude saponin-induced vasorelakation dose-dependently. From the above results, we suggest that crude saponin of ginseng may stimulate K$\^$+/ efflux and hyperpolarize the membrane, thereby cause the vasorelaxation in the resistance-sized mesenteric artery of rats.

• Proceedings of the Korean Biophysical Society Conference
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• 1999.06a
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• pp.61-62
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• 1999

;The mechanisms inducing hypertension are actively investigated and are still challenging topics. Basically hypertension must be caused by the disorder of $Ca^{2+}$ metabolism in vascular smooth muscle, such as the increase of $Ca^{2+}$ influx, the decrease of ci+ efflux, or the change of sensitivity of contractile protein etc. The one of cause of the increase of ci+ influx may be the change of ci+ channel activity. Even though the relationships of ci+ channel activity and hypertension were studied using various hypertension models, still it is not clear how much change of $Ca^{2+}$ channel activity in diabetes mellitus (DM) induced hypertension is occurred. We induced DM hypertension in SD rat and compared the $Ca^{2+}$ channel activity with age-matched normotensive SD rat. For inducing DM hypertension, left kidney was removed with 200 gm rat and, after 1 month, 60 mg/kg of streptozotocin was injected into peritoneal space to induce diabetes mellitus. Usually after 4-6 weeks, hypertension was fully induced. For isolating vascular smooth muscle cells (VSMC), we used mesenteric arteriole (3rd - 4th branch of mesenteric artery) of which diameter is below 150 urn. VSMCs were isolated enzymatically. $Ca^{2+}$ current was measured using whole cell patch clamp technique. All experiments were performed at $37^{\circ}C$. The cell membrane area of VSMC of DM hypertensive rat is larger than that of control VSMC($36.6{\pm}3.64{\;}pF{\;}vs{\;}22.4{\pm}1.29{\;}pF, {\;}mean{\pm}S.E.$) When we compared the current amplitude, the $Ca^{2+}$ current amplitude in VSMC of DM hypertensive rat is much larger than that in VSMC of normotensive age-matched rat. After $Ca^{2+}$ current amplitude was normalized by cell membrane area, the current amplitude in DM hypertension is increased to $249.1{\pm}15.9{\;}%{\;}(mean{\pm}S.E.M)$, which means the ;absolute current amplitude is about 4 times larger in DM hypertension. When we compared the steady state activation and inactivation. there were no noticeable differences. From these results. one of cause of the DM hypertension is due to the increase of $Ca^{2+}$ current amplitude. But it need further study why the $Ca^{2+}$ current is so large in VSMC of DM hypertension and how much $Ca^{2+}$ influx through $Ca^{2+}$ channel contribute to the increase of intracellular $Ca^{2+}$ and eventually contribute to development of hypertension.ypertension.

• YAKHAK HOEJI
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• v.49 no.2
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• pp.185-191
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• 2005

The absorption of a P-gp substrate, rhodamine 123, from a liposomal dosage form was investigated across Caco-2 cell monolayers, rat intestines and rat intestinal Peyer's patches in Ussing chamber, Rhodamine 123 was incorporated into liposomes according to the standard evaporation method, which led to a production of liposomes with a mean diameter of 71.3 nm. The permeability (Papp of rhodamine 123 from a water solution across the monolayer was $2.45{\times}10^{-6}$ cm/s for $A{\leftrightarrow}B$ (apical to basal) and $14.0{\times}10^{-6}$ cm/s for $B{\leftrightarrow}A$ (basal to apical) directions, consistent with the fact that rhodamine 123 is one of the P-gp substrates. The transport of rhodamine 123 from the liposomal dosage form was much lower for both directions compared to the solution of rhodamine 123. The transport of rhodamine 123 across the rat intestine was also significantly decreased for both directions, I.e., influx and efflux, by the liposomal incorporation of the compound. The transport of rhodamine 123 across the Peyer's patch was substantially reduced by liposomal incorporation. No difference was found in the transport between the Peyer's patch and non-Peyer's patch. These observations suggest that the contribution of transport via Peyer's patches in the uptake of liposomes may be minimal, especially for rapidly absorbed compounds like rhodamine 123. Therefore, the increased absorption of P-gp substrates does not appear to be feasible by incorporating the compounds in liposomes, due to negligible involvement of Peyer's patches in the uptake of particulate dosage forms like liposomes. Liposomes may rather represent a sustained release dosage form of incorporated compounds.

• Korean Journal of Food Science and Technology
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• v.32 no.1
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• pp.218-224
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• 2000

The antimicrobial activities of chitosan oligosaccharide(chitohexaose) and two types of chitosans M.W.(10,000 and M.W. 100,000) were examined against Escherichia coli O157 : H7(ATCC 43894), Staphylococcus aureus(ATCC 144458) and Candida albicans(KFRI 432). Chitosan with molecular weight of 10,000 showed the strongest antimicrobial activities to E. coil O157 : H7 and S. aureus, whereas chitohexaose acted most strongly against C. albicans. The most effective concentration of chitosan was measured to be 0.1 mg/mL for E. coil O157 : H7 and S. aureus, and that of chitohexaose to be 1 mg/mL for C. albicans. Antimicrobial activities of chitosans and chitohexaose were maintained for 60 min after their treatment. They were found to induce leakage of intracellular proteins and nucleic acids from treated microorganisms. The efflux determined by assaying the ${\beta}-galactosidase$ leaked from the lactose-induced E. coli O157 : H7 cells was observed to reach the highest level within 60 min after treatment with the antimicrobial agents and chitosan with 10,000 molecular weight gave the highest ${\beta}-galactosidase$ activity. Therefore, it is supposed that the antimicrobial activity of chitosan with its unique polycationic nature might be caused by its binding to anionic component(s) of the cell envelope and thereby inhibiting the membrane metabolism and/or leaking intracellular materials.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.249-255
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• 2014

Programmed cell death or apoptosis is associated with changes in $K^+$ concentration in many cell types. Recent studies have demonstrated that two-pore domain $K^+$ ($K_{2P}$) channels are involved in mouse embryonic development and apoptotic volume decrease of mammalian cells. In cerebellar granule neurons that normally undergo apoptosis during the early developmental stage, TASK-1 and TASK-3, members of $K_{2P}$ channels, were found to be critical for cell death. This study was performed to identify the role of $K^+$ channels in the $H_2O_2$-induced or cryo-induced cell death of mouse and bovine embryos. Mouse and bovine two-cell stage embryos (2-cells) exposed to $H_2O_2$ for 4 h suffered from apoptosis. The 2-cells showed positive TUNEL staining. Treatment with high concentration of KCl (25mM) inhibited $H_2O_2$-induced apoptosis of 2-cells by 19%. Cryo-induced death in bovine blastocysts showed positive TUNEL staining only in the cells near the plasma membrane. Cryoprotectant supplemented with 25 mM KCl reduced apoptosis slightly compared to cryoprotectant supplemented with 5 mM KCl. However, the combination of antioxidants (${\beta}$-mercaptoethanol) with 25 mM KCl significantly decreased the rate of $H_2O_2$-induced and cryo-induced apoptosis compared to treatments with only antioxidants or 25 mM KCl. These results show that blockage of $K^+$ channel efflux for a short-time reduces $H_2O_2$- and cryo-induced apoptosis in mouse and bovine embryos. Our findings suggest that apoptosis in mouse and bovine embryos might be controlled by modulation of $K^+$ channels which are highly expressed in a given cell type.

• Korean Journal of Ecology and Environment
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• v.42 no.2
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• pp.242-252
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• 2009

The purpose of this study was to develop a Multi-metric Water Quality Assessment (MWQA) model and apply it to dataset sampled from Paldang and Daechung reservoir in 2008. The various water dataset used to this study included 5 year data sets (2003${\sim}$2007) in Korean reservoirs which were obtained from the Ministry of Environment, Korea. In this study, suggested MWQA model has 4 metrics that were composed of 4 parameters such as chemical, physical, biological, and hydrological variables. And, each of the variables attributed total phosphorus (TP) concentration in water, secchi depth (SD) measure in water, chlorophyll-${\alpha}$(Chl-${\alpha}$) concentration in water and the ratio of inflow of water into lakes and efflux of water from lakes, input/output (I/O). First, we established the criteria for trophic boundaries. The boundary between oligotrophic and mesotrophic categories was defined by the lower third of the cumulative distribution of the values. The mesotrophic-eutrophic boundary was defined by the upper third of the distribution. Second, each metric was given by a point-oligo=1, meso=3, eu=5. And then, obtained total score from each metric was divided 5 grade-Excellent, Good, Fair, Poor, and Very poor. As the results of applying the proposed MWQA model, the Paldang reservoir obtained "Fair" or "Poor" grade and Daechung reservoir obtained "Excellent" or "Good" grade. The suggested MWQA model through these procedures will enable to manage efficiently the reservoir. And, more studies such as metric numbers and attributes should be done for the accurate application of the new model.