• 콘크리트학회논문집
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• 제23권2호
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• pp.211-217
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• 2011

현행 설계기준식에 따르면 초고강도 콘크리트에서는 철근 인장 이음 길이보다 압축 이음 길이가 더 길어지는 현상이 발생된다. 압축 이음은 단부 지압이 존재하므로, 인장 이음보다 길 필요는 없다. 초고강도 콘크리트의 경제적 실용화를 위해 합리적인 압축 이음 강도의 평가가 필요하다. 이를 위해 설계강도 80, 100 MPa 콘크리트를 이용하여 횡보강근이 있는 압축 이음 실험을 수행하였다. 실험 결과 횡보강근량이 많을수록 압축 이음 강도는 향상되었다. 이음강도를 부착과 지압의 기여분으로 나눠서 분석하면, 이음 길이에 횡보강근량이 증가하여도 지압은 상승하지 않았다. 따라서 횡보강근 배근에 따른 압축 이음 강도의 상승은 부착 강도의 증가에 기인한 것으로 평가되었다. 실험 결과에서 분석된 이음 길이와 콘크리트 강도의 영향 특성을 고려하고, 기존 실험 자료를 통합하여 압축 이음 강도 평가식과 압축이음 길이 설계식을 제안하였다. 제안된 설계식을 이용하면 고강도 콘크리트 특성과 횡보강근 효과가 반영된 합리적인 압축 이음 길이가 산정되어 인장 이음 길이보다 길어지는 이상 현상을 해소할 수 있다.

• Parasites, Hosts and Diseases
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• 제51권6호
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• pp.645-650
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• 2013

A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at $81.5{\pm}0.2^{\circ}C$, $79.0{\pm}0.3^{\circ}C$, $76.8{\pm}0.1^{\circ}C$, and $79.9{\pm}0.1^{\circ}C$, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.

• 한국강구조학회 논문집
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• 제29권2호
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• pp.181-192
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• 2017

최근 경사단부강판과 고장력 볼트를 이용한 보 이음(Inclined end-plate beam splice) 공법이 개발되었다. 단부강판은 브래킷 단부에 용접되고 연결보는 고장력 볼트를 통해 이음시킨다. 기둥면에는 브래킷이 용접되고, 브래킷과 연결보 단부에 각각 경사단부강판과 고장력 볼트를 이용하여 이음 시킨다. 이 연구에서는 총 6개의 외부 보-기둥 모멘트접합부의 반복가력실험을 수행하였다. 실험변수는 단부강판 상세와 볼트 배열 상세이다. 모든 실험체는 AISC Design Guide 4에 따라 단부강판 및 볼트에 의한 모멘트 저항성능이 보 이음부 요구모멘트보다 크도록 설계되었다. 실험결과, 확장된 단부강판(Extended end-plate)을 사용한 보이음부의 경우 이음부 단부강판의 지레작용 및 볼트의 취성 파단 없이 중앙 보 모멘트가 단부 브래킷으로 효과적으로 전달되었다. 하지만, 보-기둥 접합부의 변형능력은 기둥면 보 플랜지 용접부의 취성파단으로 제한적이었다. 실험결과를 바탕으로, 기울어진 단부강판 이음부를 갖는 보-기둥 모멘트접합부의 내진설계를 위한 개선사항을 제안하였다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.287-294
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• 2011

Autophagy is a process of intracellular bulk protein degradation, in which the accumulated proteins and cytoplasmic organelles are degraded. It plays important roles in cellular homeostasis, apoptosis, and development, but its role during early embryo development remains contentious. Therefore, in the present study, we investigated the effects of 3-methyladenine (3-MA) on early embryonic development in pigs, we also investigated several indicators of developmental potential, including mitochondrial distribution, genes expressions (autophagy-, apoptosis- related genes), apoptosis and ER-stress, which are affected by 3-MA. After in vitro maturation and fertilization, presumptive pig embryos were cultured in PZM-3 medium supplemented with 3-MA for 2 days at $39^{\circ}C$ 5% $CO_2$ in air. Developmental competence to the blastocyst stage in the presence of 3-MA was gradually decreased according to increasing concentration. Thus, all further experiments were performed using 2 mM 3-MA. Blastocysts that developed in the 3-MA treated group decreased LC3-II intensity and expressions of autophagy related genes than those of the untreated control, resulting in down-regulates the autophagy. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 3-MA treated group compared with control ($6.0{\pm}1.0$ vs $3.3{\pm}0.6$, p<0.05). Also, the expression of the pro-apoptotic gene Bax increased in 3-MA treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. Mito Tracker Green FM staining showed that blastocysts derived from the 3-MA treated group had lower mitochondrial integrity than that of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. Then, the expression of the spliced form of pXBP-1 product (pXBP-1s) increased in 3-MA treated group, resulting increase of ER-stress. Taken together, these results indicate that inhibition of autophagy by 3-MA is closely associated with apoptosis and ER-stress during preimplantation periods of porcine embryos.

• 콘크리트학회논문집
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• 제27권1호
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• pp.29-35
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• 2015

최근 연구되고 있는 해상 초대형 부유 콘크리트 구조물의 제작은 현장 타설이 어렵다는 단점이 있으므로 프리캐스트 콘크리트로 제작한 모듈을 수상에서 인양 후 프리스트레스력으로 접합 및 제작하는 것을 목표로 한다. 그러나 해상 환경에서 발생하는 다양한 하중 및 예상되는 상부 구조물에 의한 하중을 고려하였을 때, 프리스트레스력으로 접합되는 콘크리트 모듈 간의 접합부의 안정적인 거동 및 성능이 요구된다. 이에 프리스트레스력이 적용되기 전 가접합에 수중용 에폭시를 이용한 접합 방법이 고려되고 있는데, 수중용 에폭시는 고점성의 재료로서 내부에 공극이 발생하기 쉽다. 이를 해결하기 위하여 마이크로 실리카를 혼입하여 공극을 감소시킨 수중용 에폭시를 이용하여 콘크리트 부유체의 접하 거동에 대한 평가가 요구된다. 그러므로 이번 연구에서는 마이크로 실리카를 혼입한 수중용 에폭시를 이용하여 가접합하고 프리스트레싱을 적용한 시험체를 제작하여 성능을 평가하였다. 마이크로 실리카를 혼입한 수중용 에폭시로 접합된 콘크리트 모듈은 프리스트레싱만을 적용한 시험체에 비하여 균열 발생 및 응력 집중 현상이 완화되었으며, 최대 하중 및 변위는 일체형 RC 시험체에 비하여 10% 미만의 감소율을 나타내어 모듈형 콘크리트 부유체의 수상 접합을 위한 안정적인 접합재료로 사용 가능할 것으로 보인다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권5호
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• pp.2153-2158
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• 2014

Beclin 1 is a key factor for initiation and regulation of autophagy, which is a cellular catabolic process involved in tumorigenesis. To investigate the role of alternative splicing of Beclin1 in the regulation of autophagy in leukemia cells, Beclin1 mRNA from 6 different types of cell lines and peripheral blood mononuclear cells from 2 healthy volunteers was reversely transcribed, subcloned, and screened for alternative splicing. New transcript variants were analyzed by DNA sequencing. A transcript variant of Beclin 1 gene carrying a deletion of exon 11, which encoded a C-terminal truncation of Beclin 1 isoform, was found. The alternative isoform was assessed by bioinformatics, immunoblotting and subcellular localization. The results showed that this variable transcript is generated by alternative 3' splicing, and its translational product displayed a reduced activity in induction of autophagy by starvation, indicating that the spliced isoform might function as a dominant negative modulator of autophagy. Our findings suggest that the alternative splicing of Beclin 1 might play important roles in leukemogenesis regulated by autophagy.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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• pp.1581-1590
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• 2018

Objective: The aim of this study was to clone alternative splicing isoforms of pig myoneurin (MYNN), predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time polymerase chain reaction (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST), and longissimus dorsi (LD). Results: The results showed that MYNN had two alternatively spliced isoforms, MYNN-1 (GenBank accession number: KY470829) and MYNN-2 (GenBank accession number: KY670835). MYNN-1 coding sequence (CDS) is composed of 1,830 bp encoding 609 AA, whereas MYNN-2 CDS is composed of 1,746 bp encoding 581 AA. MYNN-2 was 84 bp less than MYNN-1 and lacked the sixth exon. MYNN-2 was found to have one $C_2H_2$ type zinc finger protein domain less than MYNN-1. Two variants were ubiquitously expressed in all pig tissues, and there were significant differences in expression of different tissues (p<0.05; p<0.01). The expression of MYNN-1 was significantly higher than that of MYNN-2 in almost tissues (p<0.05; p<0.01), which testified that MYNN-1 is the main variant. The expression of two isoforms decreased gradually with increase of age in ST and CE of MS pig, whereas increased gradually in LW pig. In LD, the expression of two isoforms increased first and then decreased with increase of age in MS pig, and decreased gradually in LW pig. Conclusion: Two transcripts of pig MYNN were successfully cloned and MYNN-1 was main variant. MYNN was highly expressed in ST, CE, and LD, and their expression was regular. We speculated that MYNN plays important roles in digestion/absorption and skeletal muscle growth, whereas the specific mechanisms require further elucidation.

• Asian-Australasian Journal of Animal Sciences
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• 제28권6호
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• pp.870-875
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• 2015

The purpose of this study was to investigate the alternative splicing in equine cordon-bleu WH2 repeat protein-like 1 (COBLL1) gene that was identified in horse muscle and blood leukocytes, and to predict functional consequences of alternative splicing by bioinformatics analysis. In a previous study, RNA-seq analysis predicted the presence of alternative spliced isoforms of equine COBLL1, namely COBLL1a as a long form and COBLL1b as a short form. In this study, we validated two isoforms of COBLL1 transcripts in horse tissues by the real-time polymerase chain reaction, and cloned them for Sanger sequencing. The sequencing results showed that the alternative splicing occurs at exon 9. Prediction of protein structure of these isoforms revealed three putative phosphorylation sites at the amino acid sequences encoded in exon 9, which is deleted in COBLL1b. In expression analysis, it was found that COBLL1b was expressed ubiquitously and equivalently in all the analyzed tissues, whereas COBLL1a showed strong expression in kidney, spinal cord and lung, moderate expression in heart and skeletal muscle, and low expression in thyroid and colon. In muscle, both COBLL1a and COBLL1b expression decreased after exercise. It is assumed that the regulation of COBLL1 expression may be important for regulating glucose level or switching of energy source, possibly through an insulin signaling pathway, in muscle after exercise. Further study is warranted to reveal the functional importance of COBLL1 on athletic performance in race horses.

• Asian-Australasian Journal of Animal Sciences
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• 제30권10호
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• pp.1471-1477
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• 2017

Objective: Since athletic performance is a most importance trait in horses, most research focused on physiological and physical studies of horse athletic abilities. In contrast, the molecular analysis as well as the regulatory pathway studies remain insufficient for evaluation and prediction of horse athletic abilities. In our previous study, we identified AXL receptor tyrosine kinase (AXL) gene which was expressed as alternative spliced isoforms in skeletal muscle during exercise. In the present study, we validated two AXL alternative splicing transcripts (named as AXLa for long form and AXLb for short form) in equine skeletal muscle to gain insight(s) into the role of each alternative transcript during exercise. Methods: We validated two isoforms of AXL transcripts in horse tissues by reverse transcriptase polymerase chain reaction (RT-PCR), and then cloned the transcripts to confirm the alternative locus and its sequences. Additionally, we examined the expression patterns of AXLa and AXLb transcripts in horse tissues by quantitative RT-PCR (qRT-PCR). Results: Both of AXLa and AXLb transcripts were expressed in horse skeletal muscle and the expression levels were significantly increased after exercise. The sequencing analysis showed that there was an alternative splicing event at exon 11 between AXLa and AXLb transcripts. 3-dimentional (3D) prediction of the alternative protein structures revealed that the structural distance of the connective region between fibronectin type 3 (FN3) and immunoglobin (Ig) domain was different between two alternative isoforms. Conclusion: It is assumed that the expression patterns of AXLa and AXLb transcripts would be involved in regulation of exercise-induced stress in horse muscle possibly through an $NF-{\kappa}B$ signaling pathway. Further study is necessary to uncover biological function(s) and significance of the alternative splicing isoforms in race horse skeletal muscle.

• BMB Reports
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• 제39권6호
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• pp.677-685
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• 2006

Proteins that are unfolded or misfolded in the endoplasmic reticulum (ER) must be targeted for refolding or degradation to maintain the homeostasis of the ER. Derlin-1 was reportedly implicated in the retro-translocation of misfolded proteins from the ER to the cytosol for degradation. In this report, we showed that Derlin-1 was down-regulated in the endothelial cells derived from human hepatic cavernous hemangioma (CHEC) compared with other tested cells. Electron microscopy analysis showed that ER was aberrantly enlarged in CHEC cells, but not in other tested cells. When overexpressed, Derlin-1 induced the dilated ER to return normal size. This ER dynamic was associated with the activation of unfolded protein response (UPR). In CHEC cells where Derlin-1 was down-regulated, increased expression of the immunoglobulin heavy chain-binding protein (Bip) and UPR-specific splicing of X-box DNA-binding protein 1 (XBP1) mRNA were detected, as compared with that in other tested cells, indicating that UPR was activated. After Derlin-1 overexpression, the extent of UPR activation diminished, as evidenced by decreased expression of Bip, reduced amount of the spliced form of XBP1 ($XBP1_S$), and elevated expression of the unspliced form of XBP1 ($XBP1_U$). Taken together, these findings provide another example of a single protein being able to affect ER dynamic in mammalian cells, and an insight into the possible molecular mechanism(s).