• Journal of Microbiology and Biotechnology
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• v.20 no.1
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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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• v.11 no.2
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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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• v.7 no.1
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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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• v.2 no.4
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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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• v.15 no.5
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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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• v.13 no.4
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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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• v.23 no.12
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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.

• The Korean Journal of Nuclear Medicine
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• v.28 no.1
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• pp.75-84
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• 1994

The need for assessment of ureteric function in the patient with an obviousely dilated ureter has increased particularly with the added spectrum of asymptomatic patients presenting with hydrone-phrosis and hydroureter on antenatal and perinatal ultrasound. To assess the influence of ureteral status on kidney washout during $^{99m}Tc$-DTPA diuretic renography, ureteral images were reviewed in 80 children referred for hydronephrosis. A scintigraphically abnormal ureter was defined as an intense and continuous image of > 10 min during diuretic renography. Out of them, a total of 16 nephroureteral systems in 12 children with scintigraphically abnormal ureter were analyzed. A diuretic washout index using response half time (t1/2) by linear fitting after lasix injection, was determined on renal (Kt1/2) and ureteral (Ut1/2) curves (diuretic renogram vs. diuretic ureterogram). Diuretic ureterogram curve patterns corresponding to normal (type I), obstructive (II) and non-obstructive (III) cases were described. Compared with X-ray data, diuretic renography was highly sensitive (88%) and specific (99%) for detecting any ureteral abnormality. Despite an obstructive Kt1/2 (>20 min), no patient with an abnormal ureter underwent therapy at the ureteropelvic junction because the hydronephrosis regressed after surgery at the lower level. Our data indicate that the abnormal ureter findings during diuretic renography have to be recognized before therapy for children with hydeonephrosis.