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Variation of Nephrotoxicity Biomarkers by Urinary Storage Condition in Rats

  • Lee, Jung-Min;Han, Young-Hwan;Choi, Su-Jeong;Park, Ju-Seong;Jang, Jeong-Jun;Bae, Re-Ji-Na;Lee, Mi Ju;Kim, Myoung Jun;Lee, Yong-Hoon;Kim, Duyeol;Lee, Hye-Young;Park, Sun-Hee;Park, Cheol-Beom;Kang, Jin Seok;Kang, Jong-Koo
    • Toxicological Research
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    • v.30 no.4
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    • pp.305-309
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    • 2014
  • Recently, there has been an increase in the use of several nephrotoxicity biomarkers in preclinical experiments. In addition, it has been indicated that the result may have been influenced by secondary factors, such as sample storage condition or storage period. In this study, we have assessed the variation in urinary nephrotoxicity biomarkers as a result of urine storage conditions and storage period of the urine. Urine was sampled from specific pathogen-free Sprague-Dawley rats (19 weeks old), which were housed individually in hanged stainless steel wire mesh cages. Urine was stored at $20^{\circ}C$, at $4^{\circ}C$, or at $-70^{\circ}C$ after sampling. The levels of the biomarkers such as beta-2 microglobulin (B2M), cystatin-C (Cys-C), N-acetyl-${\beta}$-D-glucosaminidase (NAG), micro albumin (MA), micro protein (MP) were measured at 6, 24, 48 and 144 hr after sampling. The B2M level was significantly decreased at 6, 24, 48, and 144 hr compared to 0 hr at $-70^{\circ}C$ (p < 0.05, p < 0.01, p < 0.05, and p < 0.05, respectively) and 24 and 144 hr at $20^{\circ}C$ (p < 0.01, p < 0.01, respectively). The Cys-C level was significantly decreased at 144 hr compared to 0 hr at $4^{\circ}C$ (p < 0.01), at $20^{\circ}C$ (p < 0.05) and at $70^{\circ}C$ (p < 0.01). MP and MA levels were not different for 144 hr in all storage conditions. Taken together, B2M and Cys-C levels were modulated by storage temperature and period. For the enhancement of test accuracy, it is suggested that strict protocols be established for samples to minimize the effects of the storage conditions on the detected levels of biomarkers.

Comparison of International Guidelines of Dermal Absorption Tests Used in Pesticides Exposure Assessment for Operators

  • So, Jaehwan;Ahn, Junyoung;Lee, Tae-Hee;Park, Kyung-Hun;Paik, Min-Kyoung;Jeong, Mihye;Cho, Myung-Haing;Jeong, Sang-Hee
    • Toxicological Research
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    • v.30 no.4
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    • pp.251-260
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    • 2014
  • The number of farmers who have suffered from non-fatal acute pesticide poisoning has been reported to vary from 5.7% to 86.7% in South Korea since 1975. Absorption through the skin is the main route of exposure to pesticides for farmers who operate with them. Several in vitro tests using the skins of humans or animal and in vivo tests using laboratory animals are introduced for the assessment of human dermal absorption level of pesticides. The objective of this study is to evaluate and compare international guidelines and strategies of dermal absorption assessments and to propose unique approaches for applications into pesticide registration process in our situation. Until present in our situation, pesticide exposure level to operator is determined just using default value of 10 as for skin absorption ratio because of data shortage. Dermal absorption tests are requested to get exposure level of pesticides and to ultimately know the safety of pesticides for operators through the comparison with the value of AOEL. When the exposure level is higher than AOEL, the pesticide cannot be approved. We reviewed the skin absorption test guidelines recommended by OECD, EFSA and EPA. The EPA recommends assessment of skin absorption of pesticides for humans through the TPA which includes all the results of in vitro human and animal and animal in vivo skin absorption studies. OECD and EFSA, employ a tiered approach, which the requirement of further study depends on the results of the former stage study. OECD guidelines accept the analysis of pesticide level absorbed through skin without radioisotope when the recovery using the non-labeled method is within 80~120%. Various factors are reviewed in this study, including the origin of skin (gender, animal species and sites of skin), thickness, temperature and, etc., which can influence the integrity of results.

Error-Prone and Error-Free Translesion DNA Synthesis over Site-Specifically Created DNA Adducts of Aryl Hydrocarbons (3-Nitrobenzanthrone and 4-Aminobiphenyl)

  • Yagi, kashi;Fujikawa, Yoshihiro;Sawai, Tomoko;Takamura-Enya, Takeji;Ito-Harashima, Sayoko;Kawanishi, Masanobu
    • Toxicological Research
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    • v.33 no.4
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    • pp.265-272
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    • 2017
  • Aryl hydrocarbons such as 3-nitrobenzanthrone (NBA), 4-aminobiphenyl (ABP), acetylaminofluorene (AAF), benzo(a)pyrene (BaP), and 1-nitropyrene (NP) form bulky DNA adducts when absorbed by mammalian cells. These chemicals are metabolically activated to reactive forms in mammalian cells and preferentially get attached covalently to the $N^2$ or C8 positions of guanine or the $N^6$ position of adenine. The proportion of $N^2$ and C8 guanine adducts in DNA differs among chemicals. Although these adducts block DNA replication, cells have a mechanism allowing to continue replication by bypassing these adducts: translesion DNA synthesis (TLS). TLS is performed by translesion DNA polymerases-Pol ${\eta}$, ${\kappa}$, ${\iota}$, and ${\zeta}$ and Rev1-in an error-free or error-prone manner. Regarding the NBA adducts, namely, 2-(2'-deoxyguanosin-$N^2$-yl)-3-aminobenzanthrone (dG-$N^2$-ABA) and N-(2'-deoxyguanosin-8-yl)-3-aminobenzanthrone (dG-C8-ABA), dG-$N^2$-ABA is produced more often than dG-C8-ABA, whereas dG-C8-ABA blocks DNA replication more strongly than dG-$N^2$-ABA. dG-$N^2$-ABA allows for a less error-prone bypass than dG-C8-ABA does. Pol ${\eta}$ and ${\kappa}$ are stronger contributors to TLS over dG-C8-ABA, and Pol ${\kappa}$ bypasses dG-C8-ABA in an error-prone manner. TLS efficiency and error-proneness are affected by the sequences surrounding the adduct, as demonstrated in our previous study on an ABP adduct, N-(2'-deoxyguanosine-8-yl)-4-aminobiphenyl (dG-C8-ABP). Elucidation of the general mechanisms determining efficiency, error-proneness, and the polymerases involved in TLS over various adducts is the next step in the research on TLS. These TLS studies will clarify the mechanisms underlying aryl hydrocarbon mutagenesis and carcinogenesis in more detail.

Quality and Yield Characteristics of Potato (Solanum tuberosum L.) Grown at Paddy Field in Spring Season

  • Im, Ju Sung;Cho, Ji Hong;Chang, Dong Chil;Jin, Yong Ik;Park, Young Eun;Chun, Chung Gi;Kim, Dong Un;Yu, Hong Seob;Lee, Jong Nam;Kim, Myung Jun
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.24-31
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    • 2013
  • This study was conducted to determine the characteristics of quality and yield in potatoes grown at paddy field before rice transplantation during the spring season. Three potato cultivars ('Jowon', 'Haryeong', and 'Goun') were grown in Gangneung (asl 5 m) and Seocheon (asl 20 m). In both locations, weather condition belonged to the fourth zone (spring cropping) in potato production location's distribution of Korea. Daily mean soil temperature in both the locations was $0.2-0.6^{\circ}C$ lower than air temperature, while soil moisture was adequate level to potato growth in spite of spring drought. TR ratio was not affected by location, but by cultivar. Specific gravity, starch content, dry matter rate, and yield were significantly influenced by location and by cultivar. There was no difference in total tuber number by location, however there was a large gap in marketable tuber yield according to locations and cultivars. There were high negative relationships between yield and main qualities such as dry matter rate and starch content, while high positive correlation was observed between main qualities. It was possible to produce potato before rice transplanting at drained paddy fields located in representative two locations of potato spring cropping and their characteristics in growth and quality were similar to those generally well known in upland cultivation. Paddy field was thought to be more favorable than upland in terms of available soil moisture supply against spring drought. Further research, however, was needed to increase soil temperature and also preliminary review on proper cultivar according to location seemed to be needed for high yield.

Simulation Study of a Large Area CMOS Image Sensor for X-ray DR Detector with Separate ROICs (센서-회로 분리형 엑스선 DR 검출기를 위한 대면적 CMOS 영상센서 모사 연구)

  • Kim, Myung Soo;Kim, Hyoungtak;Kang, Dong-uk;Yoo, Hyun Jun;Cho, Minsik;Lee, Dae Hee;Bae, Jun Hyung;Kim, Jongyul;Kim, Hyunduk;Cho, Gyuseong
    • Journal of Radiation Industry
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    • v.6 no.1
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    • pp.31-40
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    • 2012
  • There are two methods to fabricate the readout electronic to a large-area CMOS image sensor (LACIS). One is to design and manufacture the sensor part and signal processing electronics in a single chip and the other is to integrate both parts with bump bonding or wire bonding after manufacturing both parts separately. The latter method has an advantage of the high yield because the optimized and specialized fabrication process can be chosen in designing and manufacturing each part. In this paper, LACIS chip, that is optimized design for the latter method of fabrication, is presented. The LACIS chip consists of a 3-TR pixel photodiode array, row driver (or called as a gate driver) circuit, and bonding pads to the external readout ICs. Among 4 types of the photodiode structure available in a standard CMOS process, $N_{photo}/P_{epi}$ type photodiode showed the highest quantum efficiency in the simulation study, though it requires one additional mask to control the doping concentration of $N_{photo}$ layer. The optimized channel widths and lengths of 3 pixel transistors are also determined by simulation. The select transistor is not significantly affected by channel length and width. But source follower transistor is strongly influenced by length and width. In row driver, to reduce signal time delay by high capacitance at output node, three stage inverter drivers are used. And channel width of the inverter driver increases gradually in each step. The sensor has very long metal wire that is about 170 mm. The repeater consisted of inverters is applied proper amount of pixel rows. It can help to reduce the long metal-line delay.

Die Übersicht des rechtliche Struktur über die Rechtsschutzversicherung (법무비용보험의 법적 구조의 개관 - 독일 입법례를 중심으로 -)

  • Kim, Eun-Kyung
    • Journal of Legislation Research
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    • no.44
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    • pp.315-342
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    • 2013
  • Die Rechtsschutzversicherung ist als eine echte Schadenversicherung eine neue Versicherung, die $Spezialit{\ddot{a}}t$ des versichertes Risikos hat. Das bedeutet, dass der Rechtschutzversicherer nur bestimmte vertraglich vereinbarte Risiken $tr{\ddot{a}}gt$. Diese Risiken sind in den Allgemeinen Bedingungen $f{\ddot{u}}r$ die Rechtsschutzversicheurng (ARB) im Zusammenhang mit Versicherungsvertragsgesetz (VVG) ${\S}$ 125 exact beschrieben. VVG ${\S}$ 125 spricht allegemein von Leistung des Versicherers im vereinbarten versicherten Umfang. Entscheidend ist also die Vereinbarung in den ARB, welche in ${\S}$ 1 die Kostentragung $f{\ddot{u}}r$ den Versicherten als Hauptleistung des Versicherers beschrieben ist. Also in den VVG ${\S}{\S}$ 125 bis 129 gibt es keine Definition ${\ddot{u}}ber$ die Sparte der Rechtsschutzversicherung, jedoch mindestens es $w{\ddot{a}}re$ $m{\ddot{o}}glich$, diese Sparte zu definieren. Um die $k{\ddot{u}}nftige$ Produktentwicklung nicht zu hindern, $enth{\ddot{a}}lt$ die Vorschrift keine gesetzliche Definition der Rechtsschutzversicherung nach Angabe der amtlichen $Begr{\ddot{u}}ndung$. Weil in Korea die Rechtsschutzversicherung relativ neu in Versicherungsmarket ist, sind daher VVG ${\S}{\S}$ 125 ein gutes gesetzgeberisches Vorbild, um pragmatisch und auch dazu rechtswissenschaftlich zu diskutieren und diese fsetzustellen. Im Schritt von Ausdehnungen der juristischen Dienstleistung $w{\ddot{a}}re$ es $n{\ddot{o}}tig$, zu betrachten, wie Leistungsumfang des Versicherers in der Rechtsschutzversicherung erweitert werden kann. And noch dazu ist die Informationspflicht des Versicherers in Hinsicht auf Versicherungsunfall und Leistungsumfang noch weitert zu ${\ddot{u}}berlegen$, weil diese Sparte der Rechtsschutzversicherung noch professioneller als die anderen Versicherungsbereiche ist.

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

  • Cha, Hye Jin;Song, Yun Jeong;Lee, Da Eun;Kim, Young-Hoon;Shin, Jisoon;Jang, Choon-Gon;Suh, Soo Kyung;Kim, Sung Jin;Yun, Jaesuk
    • Toxicological Research
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    • v.35 no.1
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    • pp.37-44
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    • 2019
  • A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor ($CB_1$) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified $CB_1$ were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid ${\Delta}^9$-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to $CB_1$ were determined to be (from highest to lowest) $9.52{\times}10^{-3}M$ (JWH-210), $6.54{\times}10^{-12}M$ (JWH-250), $1.56{\times}10^{-11}M$ (${\Delta}^9$-tetrahydrocannabinol), $2.75{\times}10^{-11}M$ (RCS-4), and $6.80{\times}10^{-11}M$ (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.

Inhalation of Bacterial Cellulose Nanofibrils Triggers an Inflammatory Response and Changes Lung Tissue Morphology of Mice

  • Silva-Carvalho, Ricardo;Silva, Joao P.;Ferreirinha, Pedro;Leitao, Alexandre F.;Andrade, Fabia K.;da Costa, Rui M. Gil;Cristelo, Cecilia;Rosa, Morsyleide F.;Vilanova, Manuel;Gama, F. Miguel
    • Toxicological Research
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    • v.35 no.1
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    • pp.45-63
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    • 2019
  • In view of the growing industrial use of Bacterial cellulose (BC), and taking into account that it might become airborne and be inhaled after industrial processing, assessing its potential pulmonary toxic effects assumes high relevance. In this work, the murine model was used to assess the effects of exposure to respirable BC nanofibrils (nBC), obtained by disintegration of BC produced by Komagataeibacter hansenii. Murine bone marrow-derived macrophages ($BMM{\Phi}$) were treated with different doses of nBC (0.02 and 0.2 mg/mL, respectively 1 and $10{\mu}g$ of fibrils) in absence or presence of 0.2% Carboxymethyl Cellulose (nBCMC). Furthermore, mice were instilled intratracheally with nBC or nBCMC at different concentrations and at different time-points and analyzed up to 6 months after treatments. Microcrystaline $Avicel-plus^{(R)}$ CM 2159, a plant-derived cellulose, was used for comparison. Markers of cellular damage (lactate dehydrogenase release and total protein) and oxidative stress (hydrogen peroxidase, reduced glutathione, lipid peroxidation and glutathione peroxidase activity) as well presence of inflammatory cells were evaluated in brochoalveolar lavage (BAL) fluids. Histological analysis of lungs, heart and liver tissues was also performed. BAL analysis showed that exposure to nBCMC or CMC did not induce major alterations in the assessed markers of cell damage, oxidative stress or inflammatory cell numbers in BAL fluid over time, even following cumulative treatments. $Avicel-plus^{(R)}$ CM 2159 significantly increased LDH release, detected 3 months after 4 weekly administrations. However, histological results revealed a chronic inflammatory response and tissue alterations, being hypertrophy of pulmonary arteries (observed 3 months after nBCMC treatment) of particular concern. These histological alterations remained after 6 months in animals treated with nBC, possibly due to foreign body reaction and the organism's inability to remove the fibers. Overall, despite being a safe and biocompatible biomaterial, BC-derived nanofibrils inhalation may lead to lung pathology and pose significant health risks.

Genetic diversity and phylogenetic relationship analyzed by microsatellite markers in eight Indonesian local duck populations

  • Hariyono, Dwi Nur Happy;Maharani, Dyah;Cho, Sunghyun;Manjula, Prabuddha;Seo, Dongwon;Choi, Nuri;Sidadolog, Jafendi Hasoloan Purba;Lee, Jun-Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.1
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    • pp.31-37
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    • 2019
  • Objective: At least eight local duck breeds have been recognized and documented as national germplasm of Indonesia so far. It is necessary to genetically characterize the local duck breeds for aiding conservation and future improvement strategies. Thus, this study was carried out to assess genetic diversity and phylogenetic relationship of eight local duck populations of Indonesia using microsatellite markers. Methods: In total, 240 individuals (30 individuals each population) from Alabio (AL), Bayang (BY), Magelang (MG), Mojosari (MJ), Pegagan (PG), Pitalah (PT), Rambon (RM), and Turi (TR) duck populations were genotyped using 22 microsatellite markers. Results: The results showed a moderate level of genetic diversity among populations, with a total of 153 alleles detected over all loci and populations, ranging from 3 to 22 alleles per locus. Observed (Ho) and expected heterozygosity (He), as well as polymorphism information content over all loci and populations were 0.440, 0.566, and 0.513, respectively. Heterozygote deficiency in the overall populations ($F_{IT}=0.237$), was partly due to the heterozygote deficiency within populations ($F_{IS}=0.114$) and moderate level of genetic differentiation among populations ($F_{ST}=0.137$). The most diverse population was MG (He = 0.545) and the least diverse population was AL (He = 0.368). The majority of populations were relatively in heterozygote deficiency (except AL), due to inbreeding. The genetic distances, phylogenetic trees, and principal coordinates analysis concluded that the populations can be grouped into two major clusters, resulting AL, MG, and MJ in one cluster separated from the remaining populations. Conclusion: The present study revealed a considerable genetic diversity of studied populations and thus, proper management strategies should be applied to preserve genetic diversity and prevent loss of alleles.

Colorimetric Analysis of Preformed Zirconia Anterior Crowns for Esthetic Restoration (심미수복용 기성 지르코니아 크라운의 색조 평가)

  • Lee, Changkeun;Park, Kibong;Yang, Yeonmi;Lee, Daewoo;Kim, Jaegon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.46 no.3
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    • pp.318-327
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    • 2019
  • The purpose of this study was to identify which combination of zirconia crowns and cements is most similar in color to the maxillary primary incisors by varying the color of zirconia crowns, crown thickness, and shade of cements. Prefabricated zirconia crowns in 3 shades and crowns fabricated using 6 types of zirconia blocks were used in this study. These were filled with A2-shade or translucent-shade resin cement and the $L^*$, $a^*$, and $b^*$ values were calculated using a spectrophotometer. The color differences between the natural teeth and the zirconia crowns were assessed. The shade of the final restoration was more similar to that of the natural teeth using A2-shade than translucent-shade resin cement. Application of A2-shade cement to a 0.5-mm-thick crown fabricated from a smile series 2 zirconia block resulted in the color most similar to that of the natural teeth. A2-shade resin cement is recommended for zirconia crown restoration in anterior primary teeth compared to TR-shade resin cement for more esthetic restoration. Since restorations with Nu-smile zirconia crowns were not esthetically favorable in terms of shade, improvement of the shade characteristics of the product or development of a new kind of zirconia crown is required.