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Effect of Calmodulin on Ginseng Saponin-Induced $Ca^{2+}$-Activated $Cl^{-}$ Channel Activation in Xenopus laevis Oocytes

  • Lee Jun-Ho;Jeong Sang-Min;Lee Byung-Hwan;Kim Jong-Hoon;Ko Sung-Ryong;Kim Seung-Hwan;Lee Sang-Mok;Nah Seung-Yeol
    • Archives of Pharmacal Research
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    • v.28 no.4
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    • pp.413-420
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    • 2005
  • We previously demonstrated the ability of ginseng saponins (active ingredients of Panax ginseng) to enhance $Ca^{2+}$-activated $Cl^{-}$ current. The mechanism for this ginseng saponin-induced enhancement was proposed to be the release of $Ca^{2+}$ from $IP_{3}-sensitive$ intracellular stores through the activation of PTX-insensitive $G\alpha_{q/11}$ proteins and PLC pathway. Recent studies have shown that calmodulin (CaM) regulates $IP_{3}$ receptor-mediated $Ca^{2+}$ release in both $Ca^{2+}-dependent$ and -independent manner. In the present study, we have investigated the effects of CaM on ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current responses in Xenopus oocytes. Intraoocyte injection of CaM inhibited ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement, whereas co-injection of calmidazolium, a CaM antagonist, with CaM blocked CaM action. The inhibitory effect of CaM on ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement was dose- and time-dependent, with an $IC_{50} of 14.9\pm3.5 {\mu}M$. The inhibitory effect of CaM on saponin's activity was maximal after 6 h of intraoocyte injection of CaM, and after 48 h the activity of saponin recovered to control level. The half-recovery time was calculated to be $16.7\pm4.3 h$. Intraoocyte injection of CaM inhibited $Ca^{2+}$-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement and also attenuated $IP_{3}$-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement. $Ca^{2+}$/CaM kinase II inhibitor did not inhibit CaM-caused attenuation of ginseng saponin-induced $Ca^{2+}$-activated $Cl^{-}$ current enhancement. These results suggest that CaM regulates ginseng saponin effect on $Ca^{2+}$-activated $Cl^{-}$ current enhancement via $Ca^{2+}$-independent manner.

Fabrication of FBAR (SMR) using Reflector (반사층을 이용한 FBAR(SMR)의 제조)

  • Lee, Jae-Bin;Kwak, Sang-Hyon;Kim, Hyeong-Joon;Park, Hee-Dae;Kim, Young-Sik
    • Korean Journal of Materials Research
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    • v.9 no.12
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    • pp.1263-1269
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    • 1999
  • An FBAR(Solidly Mounted Resonator) was fabricated using reflector layers which prohibit the penetration of bulk acoustic wave into substrate. The SMR consisted of top and bottom electrodes(Al films), a piezoelectric layer (ZnO film), reflector layers(W/$Si_2$ films) and Si substrate. The electrodes were deposited by dc sputtering. The piezoelectric layer and the reflector layers were deposited by rf magnetron sputtering. The control of crystallinity, microstructures and electric properties of each layer was essential for attaining the optimum FBAR characteristics. Under the best deposition conditions for FBAR devices, the ZnO films had highly c-axis preferred orientation(${\sigma}=2.17^{\circ}$), resistivity of $10^4\;{\omega}cm$, and surface roughness of 10.6 ${\AA}$. On the other hand, the surface roughness of W and $Si_2$ films was 16 ${\AA}$ and 33 ${\AA}$, respectively, and the resistivity of Al film was $5.1{\times}10^{-6}\;{\Omega}cm$. The SMR devices were fabricated by the conventional semiconductor processes. In the resonance conditions of the SMR, the series resonance frequency (fs) and the parallel resonance frequency(fp) were 1.244 GHz and 1.251 GHz, respectively and the quality factor(Q) was 1200.

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An Investigation into Oral Medicine Inpatients by Systemic Disease (전신질환으로 입원한 환자의 구강내과질환 발현실태)

  • Oh, Byung-Sub;Chun, Yang-Hyun;Hong, Jung-Pyo;Auh, Q-Schick
    • Journal of Oral Medicine and Pain
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    • v.34 no.2
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    • pp.123-132
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    • 2009
  • Purpose : To investigate the actual conditions of diagnosis and treatment of oral medicine inpatient with systemic disease. Methods : A total of 54 oral medicine subjects, inpatient due to systemic disease for diagnosis and treatment of oral disease was requested to answer the medical history and dental treatment record. Results : The ratio of gender is composed of male 44% and female 56%, the distribution of age is the order of the 50-59 group 37%, the 60-69 group 26%, the 40-49 group 22%. Systemic disease is composed of Endocrine, nutritional and metabolic diseases 36%, Diseases of the circulatory system 36%, Diseases of the nervous system 10%. Chief complain of oral disease is composed of routine check for oral health 26%, craniomandibular disorders 18%, soft tissue problem 18%. Oral disease is composed of Diseases of salivary glands 32%, Gingivitis and periodontal diseases 23%, Dentofacial anomalies 16% Conclusion : These findings indicate that oral medicine inpatient due to the systemic disease is significantly correlated to the oral disease. The patients of oral disease interrelationship between inpatient and outpatient of systemic disease should be validated by future research.

A Study on Microstructure and Mechanical Properties of TiC/Steel Composites Fabricated by Powder Metallurgy Process (분말야금공정으로 제조된 TiC/steel 금속복합재료의 미세조직 및 기계적 물성 연구)

  • Lee, Jihye;Cho, Seungchan;Kwon, Hansang;Lee, Sang-Kwan;Lee, Sang-Bok;Kim, Daeha;Kim, Junghwan
    • Composites Research
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    • v.34 no.5
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    • pp.311-316
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    • 2021
  • In this study, TiC/steel metal matrix composites were fabricated by powder metallurgy process using Fealloy powders with 3 wt.% Cr and 10 wt.% Cr, respectively, as matrix material. Subsequently, the composite samples were heat treated by the annealing and quenching-tempering(Q-T), respectively, to understand the effect of heat treatment on the mechanical properties of the composites. The correlation between microstructure and structural strength depending on the chromium content and the heat treatment conditions was studied through tensile, compressive, and transverse rupture test and microstructural analysis. In the case of TiC/steel composite containing 10 wt.% Cr, the tensile strength and transverse rupture strength at room temperature were significantly lowered by the influence of coarse chromium carbide formed at the TiC/steel interface. On the other hand, both TiC/steel composites containing 3 wt.% Cr and 10 wt.% Cr showed much higher compressive strength of about 4 GP after quenching-tempering compared to the annealed specimens regardless of the presence of the chromium carbide.

Persistence Study of Thiamethoxam and Its Metabolite in Kiwifruit for Establishment of Import Tolerance

  • Il Kyu Cho;Gyeong Hwan Lee;Woo Young Cho;Yun-Su Jeong;Danbi Kim;Kil Yong Kim;Gi-Woo Hyoung;Chul Hong Kim
    • Korean Journal of Environmental Agriculture
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    • v.41 no.4
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    • pp.355-364
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    • 2022
  • BACKGROUND: Pre-harvest interval and decline pattern of thiamethoxam were determined in kiwifruit using liquid chromatography-tandem mass spectrometry (LCMS/MS). The study was carried out to propose import tolerance using OECD maximum residue limit (MRL) calculator for the export promotion of kiwifruit to Taiwan. METHODS AND RESULTS: The thiamethoxam residue in kiwifruit was determined by using the LC-TriQ-MS/MS with the analytical process to set up the import tolerance under greenhouse conditions for Taiwan. Excellent linearity was observed for all of the analytes with a determination coefficient (R2)≥0.99. The limit of quantification was determined to be 0.01 mg/kg for both thiamethoxam and clothianidin in kiwifruit. Linearity was determined from the co-efficient of determinants (R2) obtained from the seven-point calibration curve. The standard calibration curve showed as follows; 1) Site 1 (Gimje): y = 944,406X + 1,583 (R2=0.9995), 2) Site 2 (Goheung): y = 1,356,205X + 934 (R2=0.9983), and 3) Site 3 (Jangheung): y = 1,239,937X - 3,090 (R2=0.9908). The residue of thiamethoxam in the kiwifruit for three decline trials showed the range of 0.35 to 0.56 mg/kg in site 1 (Gimje), 0.24 to 0.55 mg/kg in site 2 (Goheung), and 0.28 to 0.42 mg/kg in site 3 (Jangheung), respectively. However, clothianidin was not detected in all of the treatments. The maximum residual amounts (decline) in the samples, sprayed according to the safe-use standard for thiamethoxam 10% WG in kiwifruit (30 days before harvest, 3 sprays every 7 days) were 0.56 mg/kg in site 1, 0.55 mg/kg in site 2, and 0.42 mg/kg in site 3, respectively. CONCLUSION(S): The import tolerance (IT) of thiamethoxam for kiwifruit may be proposed to be 0.9 mg/kg by using the OECD MRL calculator.

Scavenging Capacities of DPPH and ABTS Free Radicals and Anti-inflammatory Activities of Ethanol Extracts and their Fractions from Sophora tonkinensis

  • Eun Sun Moon;Ji Yoon Lee;Seongdae Kim;Chang Won Choi
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.46-46
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    • 2021
  • The first purpose of this study was to evaluate the scavenging capacity (SC) of DPPH and ABTS free radicals for ethanol extract (STR-E) and its active fractions from Sophora tonkinensis root (STR). Four different fractions from STR-E were prepared by using different types of solvents such as chloroform (STR-E-C), ethyl acetate (STR-E-EA), n-butanol (STR-E-B), and water (STR-E-W). STR-E-C showed the highest value of total phenolic content, while STR-E showed the highest value of total flavonoid and terpenoid content. In STR-E and its four fractions, STR-E-EA showed the strongest SC with the lowest SC50 values of the DPPH radicals and ABTS radicals. The second purpose of this study was to evaluate anti-inflammatory activity in the lipopolysaccharide (LPS)-induced RAW 264.7 macrophages treated with STR-E, STR-E-C, and STR-E-EA, respectively. No cytotoxic effect to RAW 264.7 cells was observed at 20 ~ 25 ㎍/ml of STR-E, 10 ㎍/ml of STR-E-C, and 5 ㎍/ml of the STR-E-EA, presenting cell viability values close to that of the untreated control (100%). STR-E, STR-E-C, and STR-E-EA significantly suppressed the LPS-induced nitric oxide (NO) in a dose-dependent manner. Results of reverse-transcription (RT)-qPCR analysis showed that the peak mRNA levels of IL-1β, TNF-α, iNOS, IL-6, and IL-10 were observed in the LPS-stimulated macrophages at 4 h, 2 h, 12 h, 12 h, and 12 h, respectively. The peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 were significantly reduced in the LPS-stimulated macrophages co-treated with 20 ㎍/ml and 25 ㎍/ml of STR-E, respectively. In the case of IL-10, its peak mRNA level slightly increased without statistical significance. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 10 ㎍/ml and 20 ㎍/ml of STR-E-C, respectively. In contrast, the peak mRNA level of IL-10 significantly increased at 8 h. Compared with the LPS-stimulated macrophages, the peak mRNA levels of IL-1β, TNF-α, iNOS, and IL-6 reduced in the LPS-stimulated macrophages co-treated with 5 ㎍/ml and 10 ㎍/ml of STR-E-EA, respectively. In contrast, the peak mRNA level of IL-10 increased at 4 h. Taken together, our data indicated that STR-E, STR-E-C, and STR-E-EA activate macrophages to secrete both pro-inflammatory and anti-inflammatory cytokines.

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Transcriptome sequencing reveals non-coding RNAs respond to porcine reproductive and respiratory syndrome virus and Haemophilus parasuis co-infection in Kele piglets

  • Jing Zhang;Chunping Zhao;Min Yao;Jing Qi;Ya Tan;Kaizhi Shi;Jing Wang;Sixuan Zhou;Zhixin Li
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.663-681
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    • 2024
  • Co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and Haemophilus parasuis (HPS) has severely restricted the healthy development of pig breeding. Exploring disease resistance of non-coding RNAs in pigs co-infected with PRRSV and HPS is therefore critical to complement and elucidate the molecular mechanisms of disease resistance in Kele piglets and to innovate the use of local pig germplasm resources in China. RNA-seq of lungs from Kele piglets with single-infection of PRRSV or HPS and co-infection of both pathogens was performed. Two hundred and twenty-five differentially expressed long non-coding RNAs (DElncRNAs) and 30 DEmicroRNAs (DEmiRNAs) were identified and characterized in the PRRSV and HPS co-infection (PRRSV-HPS) group. Compared with the single-infection groups, 146 unique DElncRNAs, 17 unique DEmiRNAs, and 206 target differentially expressed genes (DEGs) were identified in the PRRSV-HPS group. The expression patterns of 20 DEmiRNAs and DElncRNAs confirmed by real-time quantitative polymerase chain reaction (RT-qPCR) were consistent with those determined by high-throughput sequencing. In the PRRSV-HPS group, the target DEGs were enriched in eight immune Gene Ontology terms relating to two unique DEmiRNAs and 16 DElncRNAs, and the unique target DEGs participated the host immune response to pathogens infection by affecting 15 immune-related Kyoto Encyclopedia of Genes and Genomes enrichment pathways. Notably, competitive endogenous RNA (ceRNA) networks of different groups were constructed, and the ssc-miR-671-5p miRNA was validated as a potential regulatory factor to regulate DTX4 and AEBP1 genes to achieve innate antiviral effects and inhibit pulmonary fibrosis by dual-luciferase reporter assays. These results provided insight into further study on the molecular mechanisms of resistance to PRRSV and HPS co-infection in Kele piglets.

Breast Cancer Association Studies in a Han Chinese Population using 10 European-ancestry-associated Breast Cancer Susceptibility SNPs

  • Guan, Yan-Ping;Yang, Xue-Xi;Yao, Guang-Yu;Qiu, Fei;Chen, Jun;Chen, Lu-Jia;Ye, Chang-Sheng;Li, Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.85-91
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    • 2014
  • Background: Genome-wide association studies (GWAS) have identified various genetic susceptibility loci for breast cancer based mainly on European-ancestry populations. Differing linkage disequilibrium patterns exist between European and Asian populations. Methods: Ten SNPs (rs2075555 in COL1A1, rs12652447 in FBXL17, rs10941679 in 5p12/MRPS30, rs11878583 in ZNF577, rs7166081 in SMAD3, rs16917302 in ZNF365, rs311499 in 20q13.3, rs1045485 in CASP8, rs12964873 in CDH1 and rs8170 in 19p13.1) were here genotyped in 1009 Chinese females (487 patients with breast cancer and 522 control subjects) using the Sequenom MassARRAY iPLEX platform. Association analysis based on unconditional logistic regression was carried out to determine the odds ratio (OR) and 95% confidence interval (95% CI) for each SNP. Stratification analyses were carried out based on the estrogen receptor (ER) and progesterone receptor (PR) status. Results: Among the 10 SNPs, rs10941679 showed significant association with breast cancer when differences between the case and control groups in this Han Chinese population were compared (30.09% GG, 45.4% GA and 23.7% AA; P = 0.012). Four SNPs (rs311499, rs1045485, rs12964873 and rs8170) showed no polymorphisms in our study. The remaining five SNPs showed no association with breast cancer in the present population. Immunohistochemical tests showed that rs2075555 was associated with ER status; the AA genotype showed greater association with ER negative than ER positive (OR = 0.54, 95% CI, 0.29-0.99; P = 0.046). AA of rs7166081 was also associated with ER status, but showed a greater association with ER positive than negative (OR = 1.59, 95% CI = 1.04-2.44; P = 0.031). However, no significant associations were found among the SNPs and PR status. Conclusion: In this study using a Han Chinese population, rs10941679 was the only SNP associated with breast cancer risk, indicating a difference between European and Chinese populations in susceptibility loci. Therefore, confirmation studies are necessary before utilization of these loci in Chinese.

Genetic Status of ESR Locus and Other Unidentified Genes As sociated with Litter Size in Chinese Indigenous Tongcheng Pig Breed after a Long Time Selection

  • Zhu, M.J.;Yu, M.;Liu, B.;Zhu, Z.Z.;Xiong, T.A.;Fan, B.;Xu, S.P.;Du, Y.Q.;Peng, Z.Z.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.598-602
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    • 2004
  • The Tongcheng pig breed is a famous Chinese indigenous breed. The Ministry of Agriculture of China has filed it as 1 of 19 national key conservation breeds selected from more than 100 Chinese indigenous pig breeds in 2000. In order to improve the reproductive performance, it has been intensively selected to increase the litter size for about 10 years. The population randomly sampled from conservation nucleus of eight families in the Tongcheng pigs was genotyped for identification of their estrogen receptor locus polymorphisms with the PCR-RFLPs method. Only AB heterozygotes and BB homozygotes were detected, and $X^2$ test demonstrated that the locus was in disequilibrium at a significant level (p<0.05). In the present paper, the litter sizes in different parities were regarded as different traits. Holistic status of other unspecific and unidentified genes was estimated by using the statistical methods. Coefficients of kurtosis and skewness showed that the litter size still presented segregating characteristic in the 2nd, 5th, 7th, 8th and 9th parities. Analysis of homogeneity of variance between families confirmed the results for the 5th, 7th and 8th parities. The heritability of litter size for the 1st to 10th parities was estimated with paternal half-sib model and individual estimated breeding values (EBVs) were evaluated by a single trait animal model as well. We found that the averages of EBVs for litter size in each parity did not differ significantly between genotypes, despite the significant difference for original phenotype records in the 3rd, 4th and 5th parities (p<0.05 or p<0.01). The results may be explained by the deduction that the polymorphisms of ESR locus are no longer the important genetic base of litter size variation when the frequency of allele B accumulated in the experience of selection procedure, and further conferring that there exist special genes associated with litter size in the recent Tongcheng pigs population can be made.

The Influence of Water Temperature and Body Weight on Metabolic Rate of Olive Flounder Paralichthys olivaceus (넙치 Paralichthys olivaceus 대사율에 미치는 수온과 체중의 영향)

  • Oh, Sung-Yong;Jang, Yo-Soon;Park, Heung-Sik;Choi, Young-Ung;Kim, Chong-Kwan
    • Ocean and Polar Research
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    • v.34 no.1
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    • pp.93-99
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    • 2012
  • The effect of water temperature and body weight on oxygen consumption by the fasted olive flounder Paralichthys olivaceus was investigated in order to assess the metabolic rate of this species under different conditions. The oxygen consumption rate (OCR) was measured at three different water temperatures (15, 20 and $25^{\circ}C$) and two different body weights [$9.1{\pm}1.2$ g (mean${\pm}$SD) for the juvenile group and $266.4{\pm}29.3$ g for the immature group] at an interval of 5 minutes for 24 hours using a closed flow-through respirometer. For each treatment condition, three replicates were set up and 135 fish in the juvenile group and 18 fish in the immature group were used. The OCRs exhibited a linear increase described by OCR=-82.06+28.30T ($r^2$=0.96, p<0.001) in the juvenile group and OCR=-52.52+14.73T ($r^2$=0.97, p<0.001) in the immature group. The OCRs decreased with increasing body weights at a given water temperature (p<0.001). The metabolic rate was related to the body weight of the fish as a power function with a weight exponent of between 0.77 and 0.82. $Q_{10}$ values ranged 1.67~2.28 when the temperature was between 15 and $20^{\circ}C$, 1.57~1.93 when the temperature was between 20 and $250^{\circ}C$, and 1.79~1.89 when the temperature was between 15 and $250^{\circ}C$. The energy expenditure by respiration increased with increasing water temperature and decreasing body weight (p<0.001). The mean energy loss rates at 15, 20 and $25^{\circ}C$ were 115.9, 149.8 and 208.2 kJ $kg^{-1}d^{-1}$ in the juvenile groups and 53.8, 81.2 and 101.9 kJ $kg^{-1}d^{-1}$ in the immature groups.