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Effects of the Changes in Ca-current and Intracellular Ca-concentration on the Contraction and Action Potential Staircase (수축과 활동전압의 Staircase 현상에 대한 Ca-전류 및 세포내 $Ca^{2+}$ 농도 변화의 영향)

  • Park, Choon-Ok;So, In-Suk;Ho, Won-Kyung;Earm, Yung-E;Kim, Woo-Gyeum
    • The Korean Journal of Physiology
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    • v.23 no.2
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    • pp.301-312
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    • 1989
  • It well known that the magnitude of contraction and the shape of action potential depend upon the stimulation frequency and the duration of resting period (positive and negative staircase). Although the underlying mechanism of the staircase phenomenon is not fully understood, it has been suggested that staircase could be related to the intracllular $Ca^{2+}$ concentration. In order to elucidate the role of intracellular $Ca^{2+}$ on the contraction and action potential staircases, we examined the effects of 1 mM 4-aminopyridine (4-AP), 0.5 uM verapamil, 1 uM ryanodine, or reduction of extracellular Na concentration to 30% $(substituted\;by\;equimolar\;Li^+)$ in small atrial strips of the rabbit $(3{\times}10\;mm)$. The results obitained were as follows; 1) When the stimulation frequency was increased from 0.1 Hz to 2 Hz, positive staircase of the contraction and elevation of plateau level in action potential were found in control and the conditions of Na reduction and treatments of 4-AP, verapamil and ryanodine. 2) When stimulation frequency returned to 0.1 Hz from 1 min rest just after 2 Hz stimulation fer 1 min, the magnitudes of initial few contractions were larger than that of steady state contraction (post-rest potentiation) except, ryanodine or Na-reduction groups. 3) Negative staircase of contraction was developed in control and 4-AP group at post-rest 0.1 Hz stimulation and the plateau level of the action potential was decreased at the same time. But the reduction of contraction or the plateau level was much smaller in 4-AP group and than in control. From the above results it can be concluded that contraction and action potential staircase is dependent upon transmembrane $Ca^{2+}-current\;and\;Ca^{2+}$release from the SR.

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A Color-Reaction-Based Biochip Detection Assay for RIF and INH Resistance of Clinical Mycobacterial Specimens

  • Xue, Wenfei;Peng, Jingfu;Yu, Xiaoli;Zhang, Shulin;Zhou, Boping;Jiang, Danqing;Chen, Jianbo;Ding, Bingbing;Zhu, Bin;Li, Yao
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.180-189
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    • 2016
  • The widespread occurrence of drug-resistant Mycobacterium tuberculosis places importance on the detection of TB (tuberculosis) drug susceptibility. Conventional drug susceptibility testing (DST) is a lengthy process. We developed a rapid enzymatic color-reaction-based biochip assay. The process included asymmetric multiplex PCR/templex PCR, biochip hybridization, and an enzymatic color reaction, with specific software for data operating. Templex PCR (tem-PCR) was applied to avoid interference between different primers in conventional multiplex-PCR. We applied this assay to 276 clinical specimens (including 27 sputum, 4 alveolar lavage fluid, 2 pleural effusion, and 243 culture isolate specimens; 40 of the 276 were non-tuberculosis mycobacteria specimens and 236 were M. tuberculosis specimens). The testing process took 4.5 h. A sensitivity of 50 copies per PCR was achieved, while the sensitivity was 500 copies per PCR when tem-PCR was used. Allele sequences could be detected in mixed samples at a proportion of 10%. Detection results showed a concordance rate of 97.46% (230/236) in rifampicin resistance detection (sensitivity 95.40%, specificity 98.66%) and 96.19% (227/236) in isoniazid (sensitivity 93.59%, specificity 97.47%) detection with those of DST assay. Concordance rates of testing results for sputum, alveolar lavage fluid, and pleural effusion specimens were 100%. The assay provides a potential choice for TB diagnosis and treatment.

Validation of Reference Genes for Quantitative Real-Time PCR in Bovine PBMCs Transformed and Non-transformed by Theileria annulata

  • Zhao, Hongxi;Liu, Junlong;Li, Youquan;Yang, Congshan;Zhao, Shuaiyang;Liu, Juan;Liu, Aihong;Liu, Guangyuan;Yin, Hong;Guan, Guiquan;Luo, Jianxun
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.39-46
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    • 2016
  • Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (${\beta}-actin$), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.

Effects of Bacillus subtilis KN-42 on Growth Performance, Diarrhea and Faecal Bacterial Flora of Weaned Piglets

  • Hu, Yuanliang;Dun, Yaohao;Li, Shenao;Zhao, Shumiao;Peng, Nan;Liang, Yunxiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.8
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    • pp.1131-1140
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    • 2014
  • This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight $7.14{\pm}0.63$ kg) weaned at $26{\pm}2$ days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with $2{\times}10^9$ (L), $4{\times}10^9$ (M) and $20{\times}10^9$ (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p<0.05), and all probiotics and antibiotics groups had a lower diarrhea index than group NC (p<0.05). The 16S rDNA gene-based methods were used to analyze faecal bacterial flora on day 28 of experiment. The result of denaturing gradient gel electrophoresis analysis showed that supplementation of B. subtilis KN-42 to the diet changed the bacterial communities, with a higher bacterial diversity and band number in group M than in the other four groups. Real-time polymerase chain reaction analysis showed that the relative number of Lactobacillus were higher in groups PC and H than in group NC (p<0.05), and the supplemented B. subtilis KN-42 to the diet also reduced the relative number of E. coli (p<0.05). These results suggest that dietary addition of B. subtilis KN-42 can improve the growth performance and gastrointestinal health of piglets.

Evaluation of Salt-Tolerance Plant for Improving Saline Soil of Reclaimed Land (간척지 토양개량을 위한 내염성 식물의 활용성 평가)

  • Lee, Kyeong-Bo;Kang, Jong-Gook;Li, Jumei;Lee, Deog-Bae;Park, Chan-Won;Kim, Jae-Duk
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.3
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    • pp.173-180
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    • 2007
  • Reclaimed tidal area is a great agricultural resource in the world. Improvement and utilization of reclaimed soil is an important measure for expanding land resource. This study was conducted to evaluate relative salt-tolerance of plants and its effect for improving saline soil. Eighteen tolerance plants were selected from China, Pakistan and Korea the climate of which is different. The emergence of different varieties in reclaimed soil was in order as FL478>Barnyard grass>Pokkali>Atriplex>Sesbania>Rumex>Alfalfa>Tall Fescue>Ryegrass>Sudan grass. Four varieties, Barnyard grass, Sesbania, Atriplex and Limonium were selected for soil improvement in reclaimed land. Cultivation of Sesbania, Barnyard grass and Atriplex were good to soil physico-chemical quality. Also these plants increased soil organic matter contents and reduced soil salt concentration. Organic matter contents of cultivated soils of Sesbania, Barnyard grass and Atriplex were $4.10g\;kg^{-1}$, $4.60g\;kg^{-1}$ and $2.81g\;kg^{-1}$ respectively. On the other hand organic matter content of uncultivated soils was $2.65g\;kg^{-1}$. As Sesbania and Barnyard grass were applied to cultivated soil like green manure, bulk density improved from $1.42Mg\;m^{-3}$ to $1.39Mg\;m^{-3}$.

Human Intersectin 2 (ITSN2) binds to Eps8 protein and enhances its degradation

  • Ding, Xiaofeng;Yang, Zijian;Zhou, Fangliang;Hu, Xiang;Zhou, Chang;Luo, Chang;He, Zhicheng;Liu, Qian;Li, Hong;Yan, Feng;Wang, Fangmei;Xiang, Shuanglin;Zhang, Jian
    • BMB Reports
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    • v.45 no.3
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    • pp.183-188
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    • 2012
  • Participates in actin remodeling through Rac and receptor endocytosis via Rab5. Here, we used yeast two-hybrid system with Eps8 as bait to screen a human brain cDNA library. ITSN2 was identified as the novel binding factor of Eps8. The interaction between ITSN2 and Eps8 was demonstrated by the in vivo co-immunoprecipitation and colocalization assays and the in vitro GST pull-down assays. Furthermore, we mapped the interaction domains to the region between amino acids 260-306 of Eps8 and the coiled-coil domain of ITSN2. In addition, protein stability assays and immunofluorescence analysis showed ITSN2 overexpression induced the degradation of Eps8 proteins, which was markedly alleviated with the lysosome inhibitor NH4Cl treatment. Taken together, our results suggested ITSN2 interacts with Eps8 and stimulates the degradation of Eps8 proteins.

Association of SNP Haplotypes at the Myostatin Gene with Muscular Hypertrophy in Sheep

  • Gan, S.Q.;Du, Z.;Liu, S.R.;Yang, Y.L.;Shen, M.;Wang, X.H.;Yin, J.L.;Hu, X.X.;Fei, J.;Fan, J.J.;Wang, J.H.;He, Q.H.;Zhang, Y.S.;Li, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.7
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    • pp.928-935
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    • 2008
  • The myostatin gene of seven important meat (Beltex (Australia), Beltex$\times$Huyang (F1), Meat and Multi-Prolific Chinese Merino Fine Wool, Meat Chinese Merino Fine Wool and Dorper (South Africa)) and non-meat (Huyang and Kazak) sheep breeds was analyzed to study the genetic basis of muscular hypertrophy (double muscling) phenotype in sheep. SNPs, four in regulatory regions and several in the introns in the myostatin gene, were identified, and the former four SNPs were used for further studies. Twelve haplotypes were predicted by PHASE program, of which four main haplotypes (1, 3, 7, 9) were present in 90% of the 364 sheep in the study. Haplotypes 1-4 were mainly present in meat breeds while haplotypes 7 and 9 dominated the non-meat breeds. The association between haplotypes and average daily gain (ADG) was analyzed among 116 sheep with production data, Haplo2 (CGAA) and Haplo8 (TGAA) were identified to have significant (p<0.05) effect on ADG by the model (JMP5.1 software) taking into account the effects of breed, family background, haplotype, birth weight and sex. ADG of these haplotype groups also correlated well (r = 0.82) with hypertrophic phenotype scores. In conclusion, the mutations -956 (T$\rightarrow$C), -41 (C$\rightarrow$A) and 6223 (G$\rightarrow$A) involved in Haplo2 and 8 may be associated with the double-muscling trait by influencing myostatin function and be suitable markers in selecting meat sheep.

Novel Nucleotide Variations, Haplotypes Structure and Associations with Growth Related Traits of Goat AT Motif-Binding Factor (ATBF1) Gene

  • Zhang, Xiaoyan;Wu, Xianfeng;Jia, Wenchao;Pan, Chuanying;Li, Xiangcheng;Lei, Chuzhao;Chen, Hong;Lan, Xianyong
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1394-1406
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    • 2015
  • The AT motif-binding factor (ATBF1) not only interacts with protein inhibitor of activated signal transducer and activator of transcription 3 (STAT3) (PIAS3) to suppress STAT3 signaling regulating embryo early development and cell differentiation, but is required for early activation of the pituitary specific transcription factor 1 (Pit1) gene (also known as POU1F1) critically affecting mammalian growth and development. The goal of this study was to detect novel nucleotide variations and haplotypes structure of the ATBF1 gene, as well as to test their associations with growth-related traits in goats. Herein, a total of seven novel single nucleotide polymorphisms (SNPs) (SNP 1-7) within this gene were found in two well-known Chinese native goat breeds. Haplotypes structure analysis demonstrated that there were four haplotypes in Hainan black goat while seventeen haplotypes in Xinong Saanen dairy goat, and both breeds only shared one haplotype (hap1). Association testing revealed that the SNP2, SNP5, SNP6, and SNP7 loci were also found to significantly associate with growth-related traits in goats, respectively. Moreover, one diplotype in Xinong Saanen dairy goats significantly linked to growth related traits. These preliminary findings not only would extend the spectrum of genetic variations of the goat ATBF1 gene, but also would contribute to implementing marker-assisted selection in genetics and breeding in goats.

A Study on the Application of Universal Design in the Public Cultural Space -Focused on the Museums and Art Museums in Busan City- (공공 문화공간의 유니버설디자인 적용 현황 연구 -부산시 박물관 및 미술관을 중심으로-)

  • Wang, Dan;Yoon, Ji-Young;Tian, Hui;Li, Jian-Hua
    • The Journal of the Korea Contents Association
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    • v.20 no.11
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    • pp.58-76
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    • 2020
  • This study takes cultural space facilities as the research object, trying to grasp the status quo of universal design. By selecting Busan Museum, Busan Museum of Art, and Busan Museum of Modern Art, masters the applicability and characteristics of universal design, and gets the improvements. First, through literature review, the concepts of universal design and evaluation elements are considered. Secondly, obtains five evaluation elements by analyzing previous research. Thirdly, a field survey of various cultural spaces were conducted. gains the relevant status and problems of universal design. According to the results of the detailed evaluation project of cultural space also shuowed that the Busan Museum of Modern Art, Busan Museum, and Busan Museum of Modern Art followed in order, while the average applicability of the universal design of the three cultural spaces accounts for 53.6%. In addition, the results of the average evaluation of the facilities show that the additional facilities are the best, in turn internal facilities, sanitation facilities, other facilities, and guidelines facilities. In the future, it is deemed necessary to conduct regular surveys on universal design and renovate related facilities for public cultural facilities.

Influence of Expression Plasmid of Connective Tissue Growth Factor and Tissue Inhibitor of Metalloproteinase-1 shRNA on Hepatic Precancerous Fibrosis in Rats

  • Zhang, Qun;Shu, Fu-li;Jiang, Yu-Feng;Huang, Xin-En
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7205-7210
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    • 2015
  • Background: In this study, influence caused by expression plasmids of connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) short hairpin RNA (shRNA) on mRNA expression of CTGF,TIMP-1,procol-${\alpha}1$ and PCIII in hepatic tissue with hepatic fibrosis, a precancerous condition, in rats is analyzed. Materials and Methods: To screen and construct shRNA expression plasimid which effectively interferes RNA targets of CTGF and TIMP-1 in rats. 50 cleaning Wistar male rats are allocated randomly at 5 different groups after precancerous fibrosis models and then injection of shRNA expression plasimids. Plasmid psiRNA-GFP-Com (CTGF and TIMP-1 included), psiRNA-GFP-CTGF, psiRNA-GFP-TIMP-1 and psiRNA-DUO-GFPzeo of blank plasmid are injected at group A, B, C and D, respectively, and as model control group that none plasimid is injected at group E. In 2 weeks after last injection, to hepatic tissue at different groups, protein expression of CTGF, TIMP-1, procol-${\alpha}1$ and PC III is tested by immunohistochemical method and,mRNA expression of CTGF,TIMP-1,procol-${\alpha}1$ and PCIII is measured by real-time PCR. One-way ANOVA is used to comparison between-groups. Results: Compared with model group, there is no obvious difference of mRNA expression among CTGF,TIMP-1,procol-${\alpha}1$, PC III and of protein expression among CTGF, TIMP-1, procol-${\alpha}1$, PC III in hepatic tissue at group injected with blank plasmid. Expression quantity of mRNA of CTGF, TIMP-1, procol-${\alpha}1$ and PCIII at group A, B and C decreases, protein expression of CTGF, TIMP-1, procol-${\alpha}1$, PC III in hepatic tissue is lower, where the inhibition of combination RNA interference group (group A) on procol-${\alpha}1$ mRNA transcription and procol-${\alpha}1$ protein expression is superior to that of single interference group (group B and C) (P<0.01 or P<0.05). Conclusions: RNA interference on CTGF and/or TIMP-1 is obviously a inhibiting factor for mRNA and protein expression of CTGF, TIMP-1, procol-${\alpha}1$ and PCIII. Combination RNA interference on genes of CTGF and TIMP-1 is superior to that of single RNA interference, and this could be a contribution for prevention of precancerous condition.