• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.30-38
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• 2010

The contribution of a type II restriction-modification system (R-M system) to genome integrity and cell viability was investigated. We established experimental conditions that enabled the achievement of hemimethylated and unmethylated states for the specific bases of the recognition sequences of the host's DNA. To achieve this, we constructed the MboII R-M system containing only one (i.e., M2.MboII) out of two functional MboII methyltransferases found in Moraxella bovis. Using the incomplete R-M system, we were able to perturb the balance between methylation and restriction in an inducible manner. We demonstrate that upon the SOS-induced DNA repair in mitomycin C treated cells, restriction significantly reduces cell viability. Similar results for the well-studied wild-type EcoRI R-M system, expressed constitutively in Escherichia coli, were obtained. Our data provide further insights into the benefits and disadvantages of maintaining of a type II R-M system, highlighting its impact on host cell fitness.

• Animal cells and systems
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• 제11권2호
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• pp.175-182
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• 2007

The lysozyme II gene of cabbage butterfly Artogeia rapae was cloned from fat body of the larvae injected with E. coli and its nucleotide sequence was determined by the RACE-PCR. It has an open reading frame of 414 bp nucleotides corresponding to 138 amino acids including a signal sequence of 18 amino acids. The estimated molecular weight and the isoelectric point of the lysozyme II without the signal peptide were 13,649.38 Da and 9.11, respectively. The A. rapae lysozyme II (ARL II) showed the highest identity (81%) in the amino acid sequence to Manduca sexta lysozyme among other lepidopteran species. The two catalytic residues ($Glu^{32}$ and $Asp^{50}$) and the eight Cys residue motifs, which are highly conserved among other c-type lysozymes in invertebrates and vertebrates, are also completely conserved. A phylogenetic analysis based on amino acid sequences indicated that the ARL II was more closely related to M. sexta, Hyphantria cunea, Heliothis virescens, and Trichoplusia ni lysozymes. The ARL II gene was expressed in Spodoptera frugiperda 21 insect cells and the recombinant ARL II (rARL II) was purified from cell-conditioned media by cation exchange column chromatography and reverse phase FPLC. The purified rARL II was able to form a clear zone in lysoplate assay against Micrococcus luteus. The lytic activity was estimated to be 511.41 U/mg, 1.53 times higher than that of the chicken lysozyme. The optimum temperature for the lytic activity of the rARL II was $50^{\circ}C$, the temperature dependency of the absolute lytic activity of rARL II was higher than that of the chicken lysozyme at low temperatures under $65^{\circ}C$.

• International Journal of Reliability and Applications
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• 제7권1호
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• pp.41-53
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• 2006

The exact cause of the system's failure is often unknown in the masked system lifetime data. In such type of data, there are two observable quantities, namely (i) the systems time to failure and (ii) the set of systems components that contains the component, which might cause the system to fail. Our objective in this paper is to use the maximum likelihood procedure in the presence of masked data to make inference for the reliability of the system's components. We assume a multi-component series system where each component has a constant failure rate. Different cases that permit for closed form solutions of point estimates are considered. The results obtained in this paper generalize other published results.

• International Journal of Reliability and Applications
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• 제2권4호
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• pp.253-262
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• 2001

A k-out-of-n standby system is considered where all of its components are s-independent and classified either working or cold standby connected with imperfect switches. The probability density function of the life length for this system is established in closed form, when the underlying components have constant failure rates. Also the reliability function of the system is derived. Finally, the reliability functions for one, two and three out of four systems are deduced for perfect or imperfect switches and identical or non-identical constant failure rates for working and standby components.

• Steel and Composite Structures
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• 제15권5호
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• pp.539-566
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• 2013

This paper summarises the results of a numerical study on the non linear response of steel concentric braced frames under monotonic and cyclic loads, using force-based finite elements with section fibre discretisation. The first part of the study is addressed to analyse the single brace response. A parametric analysis was carried out and discussed to evaluate the accuracy of the model, examining the influence of the initial camber, the material modelling, the type of force-based element, the number of integration points and the number of fibers. The second part of the paper is concerned with the modelling issues of whole braced structures. The effectiveness of the modelling approach is verified against the nonlinear static and dynamic behaviour of different type of bracing configurations. The model sensitivity to brace-to-brace interaction and the capability of the model to mimic the response of complex bracing systems is analyzed. The influence of different approaches for modelling the inertia, the equivalent viscous damping and the brace hysteretic response on the overall structural response are also investigated. Finally, on the basis of the performed numerical study general modelling recommendations are proposed.

• Animal cells and systems
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• 제13권4호
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• pp.447-452
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• 2009

We cloned the complete complementary DNA (cDNA) of a Pacific oyster (Crassostrea gigas) cytochrome P450 (CYP450)-related protein using rapid amplification of cDNA ends (RACE). The cDNA included a 1470 bp open reading frame that began with the first ATG codon at position 103 bp and ended with a TAG stop codon at position 1573 bp (GenBank accession EF451959). The sequence had all major functional domains and characteristics of previously characterized CYP450 molecules, including the heme-binding region (FGVGRRRCVG) and putative arginine codon (R) integral to enzymatic function. An NCBI/GenBank database comparison to other CYP450 genes revealed that the deduced C. gigas CYP450 amino acid sequence is similar to that of mouse (Mus musculus) CYP450 2D/II (28%, accession AK078880), rabbit (Oryctolagus cuniculus) CYP450 2D/II (28%, AB008785), and white-tufted-ear marmoset (Callithrix jacchus) CYP450 2D (28%, AY082602). Thus, although the C. gigas CYP450 we cloned appears to belong to the 2D type of the CYP450 group, it has low similarity to this type. CYP450 mRNA expression increased over 6 h in C. gigas gills at $30^{\circ}C$ and $10^{\circ}C$, and then decreased, indicating that CYP450 plays an important role in C. gigas exposed to water temperature changes. This finding can be used as a physiological index for Pacific oysters exposed to changing water temperatures.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1737-1746
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• 2013

IbMYB1, a transcription factor (TF) for R2R3-type MYB TFs, is a key regulator of anthocyanin biosynthesis during storage of sweet potatoes. Anthocyanins provide important antioxidants of nutritional value to humans, and also protect plants from oxidative stress. This study aimed to increase transgenic potatoes' (Solanum tuberosum cv. LongShu No.3) tolerance to environmental stress and enhance their nutritional value. Transgenic potato plants expressing IbMYB1 genes under the control of an oxidative stress-inducible peroxidase (SWPA2) promoter (referred to as SM plants) were successfully generated through Agrobacterium-mediated transformation. Two representative transgenic SM5 and SM12 lines were evaluated for enhanced tolerance to salinity, UV-B rays, and drought conditions. Following treatment of 100 mM NaCl, seedlings of SM5 and SM12 lines showed less root damage and more shoot growth than control lines expressing only an empty vector. Transgenic potato plants in pots treated with 400 mM NaCl showed high amounts of secondary metabolites, including phenols, anthocyanins, and flavonoids, compared with control plants. After treatment of 400 mM NaCl, transgenic potato plants also showed high DDPH radical scavenging activity and high PS II photochemical efficiency compared with the control line. Furthermore, following treatment of NaCl, UV-B, and drought stress, the expression levels of IbMYB1 and several structural genes in the flavonoid biosynthesis such as CHS, DFR, and ANS in transgenic plants were found to be correlated with plant phenotype. The results suggest that enhanced IbMYB1 expression affects secondary metabolism, which leads to improved tolerance ability in transgenic potatoes.

• 대한핵의학회지
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• 제28권1호
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• pp.75-84
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• 1994

수신증 또는 요관수류수신증으로 이뇨 신장스캔을 시행한 80명의 환아중 이노신장스캔영화영상에서 10분 이상의 뚜렷한 지속적인 요관 영상을 보이는 12명의 16예의 신장요관계를 대상으로 요관 관심영역에 의한 이뇨요관그람 요관곡선과 신장 관심 영역에 의한 이뇨 레노그람 신장곡선의 배뇨 반감시간을 포함한 각각의 정량적 지표를 비교 분석하여 다음의 결과를 얻었다. 1) 이뇨 신장스캔영화영상에서 10분 이상의 뚜렷한 지속적 요관 영상을 보이는 경우인 스캔상 요관 이상(요관확장)의 진단률을 방사선학적 검사에 의한 결과와 비교하였을 때, 예민도와 특이도는 각각 88%와 99%이었다. 2) 이뇨 요관그람 요관곡선상 이뇨 요관배뇨 반감시간의 비폐쇄의 폐쇄에 대한 경계치를 10분 이하로 하였을 때 요관 폐쇄 여부를 판별한 예민도와 특이도는 각각 86% (7/8) 이었다. 3) 이뇨 요관그람 요관곡선상 이뇨 요관배뇨 반감시간(Ut1/2)과 이뇨 레노그람 신장곡선상 이뇨 신장배노 반감시간(Kt1/2)간에 유의한 상관 관계를 보였다(r=0,70: p<0.01). 결론적으로 소아의 폐쇄성 요로질환에서 이뇨 신장스캔상 10분 이상의 지속적 요관 영상을 보이는 경우 이뇨요관그람 요관곡선을 이뇨 레노그람 신장곡선과 함께 분석하여 요관 폐쇄의 유무와 정확한 부위를 평가하여 치료 방침을 결정하는 것이 필수적일 것으로 사료된다.