• Development and Reproduction
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• 제11권3호
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• pp.263-272
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• 2007

Contrast to mouse where its in vitro maturation rates are high without specific supplements or presence of the cumulus cells, there are some species, such as porcine, where its in vitro oocyte maturation rates are still very low. This comparative study was conducted to investigate the role of malate dehydrogenase(Mor2) during oocyte maturation by RNAi in the mouse and porcine. The Mor2 double-stranded RNA(dsRNA) was prepared speciesspecifically and microinjected into the cytoplasm of denuded germinal vesicle(GV) oocytes. Oocytes were cultured for 48 h(porcine) and 16 h(mouse) in M199 with 10% porcine follicular fluid, pyruvate, p-FSH, EGF, cystein, and estradiol-$17{\beta}$. We measured changes in oocyte morphology, maturation rates and mRNA levels after Mor2 RNAi. We confirmed gene sequence-specific knock down of Mor2 mRNA in both species after Mor2 RNAi. In contrast to our previous finding that mMor2 RNAi resulted in GV arrest in the mouse, we found that pMor2 RNAi resulted in MI arrest in denuded porcine oocytes(58%), but developed to MII(84.4%) in COCs. To determine whether this difference between mouse and porcine RNAi is due to differences in culture media, we cultured mouse oocytes in the M199 media for 16 h after mMor2 RNAi. Mouse oocytes were developed to MII stage(62%) and there was no statistical difference compared to that of non-injected(76.8%) and buffer-injected(73.3%) control groups. Therefore, we concluded that the mouse and porcine oocytes are having different metabolic systems in relation to malate dehydrogenase for oocyte maturation. This could be a basis for differences in maturation rates in vitro in two species. Further scrutinized studies on the metabolic pathways would led us in finding better culture system to improve oocyte maturation rates in vitro, especially in more challenging species like the porcine.

• Restorative Dentistry and Endodontics
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• 제26권4호
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• pp.263-272
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• 2001

The aim of this study was to investigate the influence of four different light curing modes on the marginal leakage of Class V composite resin restoration. Eighty extracted human premolars were used. Wedge-shaped class Y cavities were prepared on the buccal surface of the tooth with high-speed diamond bur without bevel. The cavities were positioned half of the cavity above and half beyond the cemento-enamel junction. The depth, height, and width of the cavity were 2 mm, 3 mm and 2 mm respectively. The specimens were divided into 4 groups of 20 teeth each. All the specimen cavities were treated with Prime & Bond$^{R}$ NT dental adhesive system (Dentsply DeTrey GmbH, Germany) according to the manufacturer's instructions and cured for 10 seconds except group VI which were cured for 3 seconds. All the cavities were restored with resin composite Spectrum$^{TM}$ TPH A2 (Dentsply DeTrey GmbH, Germany) in a bulk. Resin composites were light-cured under 4 different modes. A regular intensity group (600 mW/${cm}^2$, group I) was irradiated for 30 s, a low intensity group (300 mW/${cm}^2$, group II) for 60 s and a ultra-high intensity group (1930 mW/${cm}^2$, group IV) for 3 s. A pulse-delay group (group III) was irradiated with 400 mW/${cm}^2$ for 2 s followed by 800 mW/${cm}^2$ for 10 s after 5 minutes delay. The Spectrum$^{TM}$ 800 (Dentsply DeTrey GmbH, Germany) light-curing units were used for groups I, II and III and Apollo 95E (DMD, U.S.A.) was used for group IV. The composite resin specimens were finished and polished immediately after light curing except group III which were finished and polished during delaying time. Specimens were stored in a physiologic saline solution at 37$^{\circ}C$ for 24 hours. After thermocycling (500$\times$, 5-55$^{\circ}C$), all teeth were covered with nail varnish up to 0.5 mm from the margins of the restorations, immersed in 37$^{\circ}C$, 2% methylene blue solution for 24 hours, and rinsed with tap water for 24 hours. After embedding in clear resin, the specimens were sectioned with a water-cooled diamond saw (Isomet$^{TM}$, Buehler Co., Lake Bluff, IL, U.S.A.) along the longitudinal axis of the tooth so as to pass the center of the restorations. The cut surfaces were examined under a stereomicroscope (SZ-PT Olympus, Japan) at ${\times}$25 magnification, and the images were captured with a CCD camera (GP-KR222, Panasonic, Japan) and stored in a computer with Studio Grabber program. Dye penetration depth at the restoration/dentin and the restoration/enamel interfaces was measured as a rate of the entire depth of the restoration using a software (Scion image, Scion Corp., U.S.A.) The data were analysed statistically using One-way ANOVA and Tukey's method. The results were as follows : 1. Pulse-Delay group did not show any significant difference in dye penetration rate from other groups at enamel and dentin margins (p>0.05) 2. At dentin margin, ultra-high intensity group showed significantly higher dye penetration rate than both regular intensity group and low intensity group (p<0.05). 3. At enamel margin, there were no statistically significant difference among four groups (p>0.05). 4. Dentin margin showed significantly higher dye penetration rate than enamel margin in all groups (p<0.05).

• Journal of the Society of Cosmetic Scientists of Korea
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• 제30권2호
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• pp.263-278
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• 2004

Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1mg/mL UA or 0.1-1mg/mL ONA after tape stripping, and TEWL (transepidermal water loss) was measured. The recovery rate increased in those UA or ONA treated groups (0.1mg/mL UA and 0.5mg/mL ONA) at 6h more than 20% compared to vehicle treated group (p < 0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/mL per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from 1 week without TEWL alteration (p < 0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent (ONA=UA > vehicle). LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA > ONA > vehicle). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via PPAR Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either ONA (10${\mu}$M) or UA (10${\mu}$M) for 24 h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via PPAR Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

• Korean Journal of Soil Science and Fertilizer
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• 제21권3호
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• pp.254-263
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• 1988

Contents and distribution of amino acids in the humic acid and fulvic acid fractions of different types ($R_p$, B, A, P) were investigated. Extracted humic and fulvic acids were purified and analyzed. The results are summarized as the following: (1) Composition of Humus The total humus ($H_T$), amount of humic acid (a), amount of fulvic acid (b), and ${\Delta}logK$ all decrease in the order of $R_p$ > B > A > P type. The same trend was observed in the total nitrogen and carbon. (2) Contents and composition of amino acids in humic acids. 1) The total amounts of amino acids in the humic acid fraction of different types were in the following order for soils under coniferous forest trees: $R_p$ > B > A > P type, but for soils under deciduous forest trees the order was P > A > $R_p$ > B type. There were positive correlationships between total amino acids and total carbon and ${\Delta}logK$ for humic acids from soils under coniferous forest trees, but a negative correlationship was existed. between total amino acids and C/N ratios. No significant correlation was found for samples taken from soils under deciduous forest trees. 2) The ratios of one group of amino acids to the others were compared. The ratios of acidic amino acids were in the order of P > $R_p$ > B > A type. those of neutral amino acids followed the order of $R_p$ > B > A > P type and those of the basic amino acids were in the order of B > A >$R_p$ > P type for soils under coniferous forest trees. Contents of total amino acids were in the order of the neutral > the acidic > the basic amino acids. For the soils under deciduous forest trees the order of the ratio was different. Acidic amino acids followed the order of A > P > B > $R_p$ type, neutral ones followed the order of P > $R_p$ > A > B type, and the basic amino acids did the order of $$P{\geq_-}$$ A > B $$\geq_-$$ $-R_p$ type where the difference was very small. 3) In general aspartic aicd, glycine and glutamic acid were the major components in all samples. Histidine, tyrosine and methionine belonged to the group contained in a small amount. (3) Contents and composition of amino acids in fulvic acids. 1) The total amounts of amino acids of different types of fulvic acids were in the order of $R_p$ > B > P > A type regardless of origin of samples. There were positive correlationships observed between the toal amino acids and total carbon and ${\Delta}logK$ for soils under coniferous forest trees. For soils under deciduous forest trees, positive correlationships were observed among total amino aicds, total nitrogen, total humus ($H_T$), total humic aicd (a), and ${\Delta}logK$, but a negative correlationship existed between total amino acids and C/N ratio. 2) Thr ratio among acidic amino acids, neutral amino acids and basic amino acids of different types were $R_p$ > B > P > A type. In this respect there was no difference between the two soils. 3) In general glycine, aspartic acid, and alanine were the major constituents in all samples of different types, while tyrosine and methionine were contained in a small amount. Virtually no amount of arginine was measured.

• Journal of Animal Science and Technology
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• 제45권6호
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• pp.917-922
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• 2003

This study was conducted to investigate effects of farm, type of sow Landrace${\times}$Yorkshire, LY; Yorkshire${\times}$ Landrace, YL; Yorkshire${\times}$Yorkshire, YY and multi-cross bred sow, MBS), parity, farrowing year, farrowing season and mating method on reproductive traits such as total number born per litter, number of born alive per litter, number of weaned per litter, number of mummified per litter, number of stillbirth. The Reproductive Records of 3,387 litters from January, 1999 to September, 2002 were obtained from four pig farms managed by Electronic Data Processing(EDP) system. Reproductive performances for two types of F1 Sows(YL and LY) were estimated as 11.34${\pm}$0.266 and 11.57${\pm}$0.263 heads for total number of born per litter, 10.56${\pm}$0.216 and 11.81${\pm}$0.251 heads for number of born alive per litter, and 10.05${\pm}$0.131 and 9.96${\pm}$0.153 heads for number of weaned per litter, respectively. These records are significantly higher(P〈0.05) than those of YY and MBS. However, number of mummified per litter, number of stillborn per litter, number of dead by diarrhea per litter, number of dead by other reasons during lactating period per litter and estrus interval did not show significant difference between types of sows. There were more total number of born per litter and number of weaned per litter in year 2001 than other year. As year passed, number of dead by crush per litter, number of dead by diarrhea per litter and number of dead by other reasons during lactating period per litter reduced from 0.18${\pm}$0.023 to 0.07${\pm}$0.022 head, 0.12${\pm}$0.21 to 0.02${\pm}$0.020 head and 0.43${\pm}$0.041 to 0.22${\pm}$0.040 head, respectively and weaning rate increased from 0.94${\pm}$0.005% to 0.97${\pm}$0.005%. The total number of born per litter and number of born alive per litter were higher(P〈0.05) in the case of mating twice than mating once. The number of weaned per litter, number of mummified per litter and number of stillborn per litter were not significant(P〈0.05) between mating methods. Estrus interval was shorter on the occasion of twice artificial insemination(5.24${\pm}$0.153days) than twice natural mating(6.51${\pm}$0.466days).

• Asian-Australasian Journal of Animal Sciences
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• 제14권5호
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• pp.615-623
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• 2001

A study of mammary function in relation to glucose metabolism of first lactation Bali cows on grass-legume diets was carried out using 12 primiparous cows (initial BW $263.79{\pm}21.66kg$) for 16 weeks starting immediately post calving. The animals were randomly allocated into 4 dietary treatment groups R1, R2, R3 and R4, receiving from the last 2 months of pregnancy onwards, rations based on a mixture of locally available grass and legume feed ad libitum. On a DM basis R1 contained 70% elephant grass (PP, Penicetum purpureum) plus 30% Gliricidia sepia leaves (GS), R2 was 30% PP plus 25% GS supplemented with 55% Hibiscus tilliacius leaves (HT, defaunating effect), R3 and R4 were 22.5% PP+41.25% GS+11.25% HT+25% concentrate, with R4 supplemented with zinc-diacetate. TDN, CP and zinc contents of the diets were 58.2%, 12.05% and 18.3 mg/kg respectively for R1, 65.05%, 16.9% and 25.6 mg/kg respectively for R2, 66.03%, 16.71% and 29.02 mg/kg respectively for R3 and 66.03%, 16.71% and 60.47 mg/kg respectively for R4. Milk production and body weights were monitored, an energy and protein balance trial conducted, overall glucose kinetics parameters assessed, mammary blood flow (MBF) and metabolite arteriovenous differences (${\Delta}AVs$) measured to get uptake data and mammary performance relationships. Parameters of glucose kinetics at peak lactation or during dry condition were not affected by ration quality. Glucose pool size, space of distribution and flux increased by 61.77, 62.26 and 82.08%, respectively, during lactation compared to the dry period. Mean glucose flux of lactating Bali cows was $5.52mg/min.kgBW^{0.807}$ which resembles the range of values of temperate dairy cows. Calculation showed that glucose requirements for maintenance, milk lactose and fat-glycerol synthesis, and the formation of NADPH reached 461.69 g for a yield of 1 kg/d or equal to 320.62 mg/min, which was less than the average glucose flux of lactating Bali cows of 481.35 mg/min. Mammary blood flow (MBF) values ranged from 56 to 83 l/h for the different treatments and the ratio MBF per kg milk produced improved from av. 1540 l/kg for R1 to av. 967 l/kg for R4 treated cows. Mammary glucose uptake ranged from 6.27 to 12.03 g/h or 120 to 140 g/kg milk. Glucose uptake was mass-wise 2 to 4 times the amount secreted as lactose, which indicated values less than the calculated mammary glucose needs and that little lactose was synthesized. The excess glucose taken-up was used for other metabolic processes. Linear relationships between metabolite ${\Delta}AVs$ and arterial blood plasma concentration [A] showed that in Bali cows triglycerides (TG), phenylalanine (Phe) and tyrosine (Tyr) have high coefficients of determination, i.e. 0.77, 0.81 and 0.69, respectively. For glucose, the relationship is quadratic with an $R^2$ value of 0.49. It was concluded that lactose synthesis was inadequate, which led to a speculation that milk yield could be improved by increased lactose synthesis.

• The Journal of Engineering Geology
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• 제23권2호
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• pp.171-186
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• 2013

A drilling project was undertaken to characterize the geochemical relationship and the occurrence of radioactive materials at a test site among public-use groundwaters previously known to have high occurrence of uranium and radon-222 in the Daejeon area. A borehole (121 m deep) was drilled and core rocks mainly consist of two-mica granite, and associated with pegmatite and dykes of intermediate composition. The groundwater samples collected at six different depths in the borehole by a double-packed system showed the pH values ranging from neutral to alkaline (7.10-9.3), and electrical conductivity ranging from 263 to 443 ${\mu}S/cm$. The chemical composition of the borehole groundwaters was of the $Ca-HCO_3(SO_4+Cl)$ type. The uranium and Rn-222 contents in the groundwater were 109-1,020 ppb and 9,190-32,800 pCi/L, respectively. These levels exceed the regulation guidelines of US EPA. The zone of the highest groundwater uranium content occurred at depths of 45 to 55m. The groundwater chemistry in this zone (alkaline, oxidated, and high in bicarbonate) is favorable for the dissolution of uranium into groundwater. The dominant uranium complex in groundwater is likely to be $(UO_2CO_3)^0$ or $(UO_2HCO_3)^+$. Radon-222 content in groundwater shows an increasing trend with depth. The uranium and thorium contents in the core were 0.372-47.42 ppm and 0.388-11.22 ppm, respectively. These levels are higher values than those previously been reported in Korea. Microscopic observations and electron microprobe analysis(EPMA) revealed that the minerals containing U and Th are monazite, apatite, epidote, and feldspar. U and Th in these minerals are likely to substitute for major elements in crystal lattice.

• Journal of the Society of Cosmetic Scientists of Korea
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• 제33권4호
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• pp.263-267
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• 2007

Parabens are used in nearly all types of cosmetics and toiletries because they are formulated well and have broad spectrum of activity, interness, low costs and excellent chemical stability in relation to pH. 2-phenoxyethanol and chlorphenesin are common preservatives which are usually used in combination with parabens in cosmetics. Toxicity of parabens is generally low but application of parabens to damaged or broken skin has resulted in sensitization. Moreover, the possibility of their estrogenic potential, anesthetic effects and reproductive toxicity has been reported. Consequently there are some regulations in use of parabens. And the maximum permitted concentrations of chlorphenesin and 2-phenoxyethanol in cosmetic products are authorized by the same reasons. So it is important to control and estimate the amount of parabens in products. In this article, we proposed a valid method for the simultaneous determination of 8 preservatives including parabens in a short time using ultra performance liquid $chromatography^{TM}\;(UPLC^{TM})$. Separation of eight components was achieved in less than 10 min and resolutions were reasonable (USP resolution ${\geqq}\;2$). And limit of detection and quantification were evaluated. The method was suitably validated for specificity, linearity, precision (repeatability, intermediate precision) and accuracy for assay (recovery) based on International conference on harmonisation (ICH) guideline. The method was applicable to analysis of preservatives in cosmetic products.

• Food Science and Preservation
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• 제22권4호
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• pp.520-527
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• 2015

This study investigated the physiological activities of garlic extracts frozen at -20, -80, and $-196^{\circ}C$. To determine the optimum freezing temperature for maintaining garlic's physiological activity, antioxidant and antibacterial activities were investigated. The antioxidant activities were determined by DPPH radical scavenging ability, SOD-like activity, nitrite-scavenging ability, and reducing power. Total phenolic compounds and flavonoids of garlic extract frozen at $4^{\circ}C$ (control) were measured as $6.91{\pm}0.69{\mu}gGAE/g$ and $0.315{\pm}0.017{\mu}gQE/g$, respectively. Although the content of total phenolic compounds was not affected by the different freezing temperatures, the flavonoids contents of garlic extract frozen at $-20^{\circ}C$ were slightly decreased. The DPPH radical scavenging ability of garlic extracts (2 mg/mL) frozen at $-80^{\circ}C$ was 61%, whereas those frozen at $-20^{\circ}C$ and $-196^{\circ}C$ were 51%. SOD-like activity was slightly increased by freezing. However, the nitrite scavenging ability (18% at pH 3.0) and reducing power (OD700=1.6) were not affected by freezing temperatures. Antimicrobial activities did not show significant differences depending on freezing temperatures. Taken together, the physiological activities of the frozen garlic extracts were not significantly changed by the freezing temperatures; however, the antioxidant and antibacterial effects of the phenolic compounds and flavonoids were maintained at $-80^{\circ}C$. These results suggest that $-80^{\circ}C$ frozen garlic could maintain a higher quality than the conventional freezing method ($-20^{\circ}C$) without loss of physiological activities during the storage.

• Journal of fish pathology
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• 제22권3호
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• pp.263-273
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• 2009

There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14${^{\circ}C}$ and 21${^{\circ}C}$) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to $2.0{\times}10^{7}$ live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21${^{\circ}C}$ and 14${^{\circ}C}$ water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.