• Asian-Australasian Journal of Animal Sciences
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• 제20권1호
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• pp.36-40
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• 2007

Proteins and lipids not only provide a source of energy to the cell, but also play vital roles in modifying the physical properties and function of the biological membranes. In the present study, we investigated the biochemical constituents, viz. proteins and lipids, in growing oocytes of goat antral follicles during summer and winter seasons. Goat genitalia in phosphate buffered saline (pH 7.4) were brought to the laboratory within one hour of slaughter under aseptic conditions at $37^{\circ}C$. Oocytes were aspirated from normal small (<3 mm in diameter) and large (>3 mm) follicles and pooled for biochemical estimations. A significant increase in the amount of protein and lipid was observed with the growth of the oocyte. The amount of protein varied non-significantly with the season, while the amount of lipid varied significantly. The amounts of phospholipid, cholesterol, free fatty acid, and triglyceride increased with the growth of the oocyte, but no significant effect of season in these constituents was observed. Lysolecithin, sphingomyelin, and sterols were the polar lipids identified in both oocytes prepared from small follicles (small oocytes) as well as large follicles (large oocytes). In addition, the small oocytes also contained phosphatidyl serine, while large oocytes contained phosphatidyl glycerol phosphate and phosphatidyl inositol. Among non-polar lipids, triglycerides and long chain alcohols appear only in small oocytes and not in large oocytes. Monoglycerides, 1,2-diglycerides, 1,3-diglycerides and o-dialkyl glycerol ethers, fatty acids, fatty acid methyl esters, and wax esters were identified in both small and large oocytes. Information on biochemical composition of growing oocytes is relevant to oocyte and embryo competence, culture and cryopreservation.

• 대한화장품학회지
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• 제27권1호
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• pp.17-27
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• 2001

A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

• 전기화학회지
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• 제4권3호
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• pp.109-112
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• 2001

활성화된 탄소반죽전극 표면을 N피드록시숙신이미드(NHS)층으로 수식한 후, 이 전극을 이용하여 square-wave voltammetry방법으로 과량의 아스코빅산 존재 하에서 도파민을 측정하였다. 수식된 전극의 특성은 도파민과 아스코빅산 혼합용액에서 순한전압전류법을 이용하여 조사하였다. 도파민과 아스코빅산의 산화 피크의 분리는 시료용액의 pH에 큰 영향을 받았으며, pH 4.0에서 최대의 피크분리(172mv)를 보였다. 따라서 도파민을 정량하기 위한 square-wave voltammeoy는 140 mM NaCl을 포함하는 100 mM phosphate buffered saline (PBS)의 pH 4.0 조건에서 수행하였다. NHS로 수식된 전극은 0.2mM 아스코빅 산의 존재 하에서 도파민의 농도 $5.0\times10^{-2}$까지 검출한계와 감응기울기 $6.1{\mu}A/{\mu}M$의 감도를 나타내었다. 반면 수식되지 않은 전극은 $1.0{\mu}M$의 검출한계와 $0.93{\mu}A/{\mu}M$ 기울기를 나타내어 표면에 수식된 N-히드록시숙신이미드가 도파민의 감응을 촉진함을 보여주었다.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.80-88
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• 2002

Recent studies demonstrate that collagen IV selectively pro-motes the repair of physiological processes in sublethally injured renal proximal tubular ceils (RPTC). We sought to further define the mechanisms of cell repair by measuring the effects of toxicant injury and stimulation of repair by L-ascorbic acid-2-phosphate (AscP), exogenous collagen IV, or function-stimulating integrin antibodies on the expression and subcellular localization of collagen-binding integrins (CBI) in RPTC. Expression of CBI subunits ${\alpha}_1$, ${\alpha}_2$, and ${\beta}_1$ in RPTC was not altered on day 1 after sublethal injury by S-(1,2-dichlorovinyl)-L-cysteine (DCVC). On day 6, expression of ${\alpha}_1$ and ${\beta}_1$ subunits remained unchanged, whereas a 2.2-fold increase in ${\alpha}_2$ expression was evident in injured RPTC. CBI localization in control RPTC was limited exclusively to the basal membrane. On day 1 after injury, RPTC exhibited a marked inhibition of active $Na^+$ transport and a loss of cell polarity characterized by a decrease in basal CBI localization and the appearance of CBI on the apical membrane. On day 6 after injury, RPTC still exhibited marked inhibition of active $Na^+$ transport and localization of CBI to the apical membrane. However, DCVC-injured RPTC cultured in pharmacological concentrations of AscP (500 ${\mu}$M)or exogenous collagen IV (50 ${\mu}$g/ml) exhibited an increase inactive $Na^+$ transport, relocalization of CBI to the basal membrane, and the disappearance of CBI from the apical membrane on day 6. Function-stimulating antibodies to CBI ${\beta}_1$ did not promote basal relocalization of CBI despite stimulating the repair of $Na^+$/$K^+$-ATPase activity on day 6 after injury. These data demonstrate that DCVC disrupts integrin localization and that physiological repair stimulated by AscP or collagen IV is associated with the basal relocalization of CBI in DCVC-injured RPTC. These data also suggest that CBI-mediated repair of physiological functions may occur independently of integrin relocalization.

• 한국연초학회지
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• 제27권1호
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• pp.51-58
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• 2005

The objective of this study was to evaluate the effect of cigarette filter on in vitro cytotoxicity of cigarette mainstream smoke from the cigarette. In this work, we used 3 types of cigarettes included non-filtered 2R4F cigarette, cellulose acetate-filtered 2R4F cigarette, and carbon dual-filtered 2R4F cigarette which was made from original 2R4F by replacing with an acetate filter containing carbon. The cytotoxicity of both the cigarette smoke condensate (CSC), which was collected in Cambridge filter pad, and the gas/vapor phase (GVP), which was bubbled through in phosphate-buffered saline in a gas-washing bottle, was determined using a neutral red uptake assay with CHO-K1 cells. With regard to cytotoxicity when calculated on an equal puff basis, the cytotoxicity of CSC from the filtered cigarettes was lower than that of the non filtered cigarette. Also, $EC_{50}$ vlaue of GVP from carbon filter cigarette was 40.9 puff/L, indicating the cytotoxicity to be $20\%$ lower than that of the CA filter cigarette. The cytotoxicity of the GVP was correlated to the several vapor phase components (formaldehyde, acetaldehyde, acetone, acrolein, crotonaldehyde and MEK). In conclusion, carbon filter, which significantly reduced the amount of carbonyl compounds in mainstream cigarette smoke, results in significant reductions in the cytotoxicity potential of the smoke.

• 한방안이비인후피부과학회지
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• 제27권3호
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• pp.84-95
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• 2014

Objective : Ephedra sinica (ES) has has been used as remedy of allergic diseases for a long time in Korea. In the present study, we investigated the anti-allergic effects of ES on experimental allergic asthma mouse model using ovalbumin (OVA). Methods : BALB/c mice were sensitized and challenged with 100 ug of OVA and 1 mg of aluminum potassium sulfate of 0.2 ml phosphate-buffered saline(PBS) intraperitoneally on day 1 and 15. mice were challenged on 3 consecutive days with 5% OVA and AHR was assessed 24 h after the last challenge. we examined the lung histology, airway hyper sensitivity, total inflammatory cell count in bronchoaveloar lavage fluid(BALF), Th2-associated cytokines production and IgE production. Results : ES potently inhibited the lung damage and the development of Penh. ES also reduced the number of BAL cells during OVA-induced allergic asthma. Furthermore, ES inhibited cytokines production such as IL-4, IL-13 productions, and IgE level of serum. Conclusion : These results suggest that ES may inhibit the production of IL-4, IL-13, IgE and infiltration of inflammatory cell and be beneficial oriental medicine for allergic asthma.

• 한방안이비인후피부과학회지
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• 제27권3호
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• pp.96-105
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• 2014

Objective : Glycyrrhiza uralensis Fisch (GUF) has been used as remedy of allergic diseases for a long time in Korea. In the present study, we investigated the anti-allergic effects of GUF on experimental allergic asthma mouse model using ovalbumin (OVA). Methods : BALB/c mice were sensitized and challenged with 100 ug of OVA and 1 mg of aluminum potassium sulfate of 0.2 ml phosphate-buffered saline (PBS) intraperitoneally on day 1 and 15. Mice were challenged on 3 consecutive days with 5% OVA and AHR was assessed 24 hrs after the last challenge. We examined total inflammatory cell number in bronchoaveloar lavage fluid (BALF), Th2-associated cytokine productions and lung histology. Results : GUF potently inhibited the development of airway hypersensitivity and also reduced the number of BAL cells during OVA-induced allergic asthma. GUF also inhibited cytokine productions such as IL-4, IL-13 in lung tissue. Furthermore, GUF treatment inhibited allergic airway inflammation. Conclusion : These results suggest that GUF may inhibit the production of IL-4, IL-13 and infiltration of inflammatory cell and be beneficial oriental medicine for allergic asthma.

• 농업과학연구
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• 제43권5호
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• pp.788-793
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• 2016

Post-weaning diarrhea (PWD), mostly caused by enterotoxigenic Escherichia coli (ETEC), remains to be a major source of economic loss in swine industry. The use of the ETEC-oral challenge model is often applied to mimic unsanitary commercial swine farm conditions where pathogens and unknown complex microbes exist and can cause severe infections in pigs. The purpose of this study was (1) to estimate ETEC density using spectrophotometric computation, (2) to determine survivability of ETEC after storing at $-20^{\circ}C$ for 7 days, and (3) to evaluate survivability of ETEC after blending with diluted sweeteners (0, 5, 10, 20, and 40% sucrose in phosphate buffered saline [PBS]). Cell density was quantified using UV-VIS spectrophotometer and counting ETEC colony forming units (cfu) at 0, 30, 60, 90, 120, 150, 180, 210, and 240 min. The established linear equation ($y=0.0031x^2-0.0079x+0.0043$ and $y=0.0046x^2-0.0151x+0.0113$) was used for robust quantification of each ETEC cell density. ETEC stored at $-20^{\circ}C$ showed 108 cfu/mL after thawing and incubation. When ETEC was blended with sweeteners (20 and 40%), survival of ETEC was decreased by 58 and 54% in 5 min post blending. However, addition of 20% of sweetener resulted in a higher survivability than those with other media concentrations. Therefore, the use of ETEC-oral challenge model would be possible as a stable method if we could confirm the appropriate medium that increases survivability of ETEC in weaner pigs.

• 한국수정란이식학회지
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• 제31권3호
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• pp.185-190
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• 2016

During the ovary preservation in low temperature, the cumulus oocyte complexes(COCs) lose their developmental competences after in vitro fertilization. We used phosphate-buffered saline (PBS) as a basic solutions of at various temperatures (25, 15 or $5^{\circ}C$) and supplemented them with 1mM glucose and 0.5mM glutamine as a source of carbohydrate metabolites. After recovery of COCs and in vitro fertilization, a significantly higher number of oocytes developed into blastocysts. The developmental competence of embryos that were originated from ovaries preserved at $15^{\circ}C$ was increased compared to those of 25 or $5^{\circ}C$. The maturation rate of oocytes was not differed between 24 and 36 h at $15^{\circ}C$ but showed lower than control group (71% versus 78%). In vitro-fertilized oocytes from ovaries stored at $25^{\circ}C$ for 24 h or at $5^{\circ}C$ for 24 h had a significantly decreased developmental potentials, but at $15^{\circ}C$ did not (27% versus 29% of blastocysts to develop into day 8). With these results, bovine ovaries can be preserved at $15^{\circ}C$ for 36 h without decreasing developmental capacity of in vitro-fertilized oocyte at least to the blastocyst stage. This information provides valuable information of preserving ovaries for embryo transfer or in vitro embryo production.

• International Journal of Oral Biology
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• 제37권1호
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• pp.9-16
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• 2012

The aim of this study was to evaluate surface characteristics and biological properties of the dentin -derived hydroxyapatite (HA) coating on titanium substrate. Dentinderived HA was obtained from extracted human teeth using a calcination method at $850^{\circ}C$. The commercially pure titanium (cp-Ti, ASTM Grade II) was used as a metallic substrate and a radio frequency magnetron sputtering method was employed as a coating method. Scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX) were utilized to investigate the coating aspects and composition. Atomic forced microscopy (AFM) and a surface profiler were used to assess the surface morphology and roughness. Corrosion tests were performed in phosphate-buffered saline at a $36.5{\pm}1^{\circ}C$ in order to determine the corrosion behavior of the uncoated and coated specimens. The biocompatibility of dentin-derived HA coated specimens with fetal rat calvarial cells and human gingival fibroblasts was assessed by SEM and cell proliferation analysis. The results showed that the dentin-derived HA coatings appeared to cover thinly and homogeneously the surfaces without changing of the titanium substrate. The EDX analysis of this the coating surface indicated the presence of Ca and P elements. The mean surface roughness of cp-Ti and dentin-derived coating specimens was $0.27{\mu}m$ and, $1.7{\mu}m$, respectively. Corrosion tests indicated a stable passive film of the dentin-derived HA coating specimens. SEM observations of fetal rat calvarial cells and human fibroblast cells on coated surfaces showed that the cells proliferated and developed a network of dense interconnections. The cells on all specimens proliferated actively within the culture period, showing good cell viability. At day 1 and 3, dentin-derived coating specimens showed 89% and 93% cell viability, respectively, when normalized to cp-Ti specimens. These results suggest that dentin-derived HA coating using the RF magnetron sputtering method has good surface characteristics and biocompatibility.