Alkaline protease-overproducing strains of Vibrio metschnikovii were developed by using the molecular evolution from the classical mutants V. metschnikovii L12-23, N4-8, and KS1. Each vapK (Vibrio alkaline protease K) was obtained from the genomic DNAs of mutants by PCR to carry out the DNA shuffling. The modified vapK-1 obtained by DNA shuffling was used again as a template for the error-prone PCR to make the vapK-2. Both genes were cloned in the plasmid pKF3 to construct the recombinant plasmids which have one or two copies of the modified genes. The recombinant plasmids were back-transformed to V. metschnikovii KS1 to construct recombinant V. metschnikovii that expresses the alkaline protease. About 3.9-fold more protease activity was measured in the strain which has the plasmid containing two copies of vapK-2 when compared to strain KS1. When compared to wild type V. metschnikovii RH530, 43-fold more activity was achieved. Comparison of amino acids among vapK, vapK-1, and vapK-2 revealed that the active sites was highly conserved and not changed. However, many amino acids except the active sites were changed. These results suggested that the changes in amino acids might play an important role in the increase of protease activity by allowing the easy access of substrate to active sites of the protease. The fermentation of alkaline protease from the V. metschnikovii KS1 harboring the plasmid that contains two copies of vapK-1 showed the possibility of this strain to be used as industrial producer.
Performance of an image retrieval system is usually very low because of the semantic gap between the low level feature and the high level concept in a query image. Semantically relevant images may exhibit very different visual characteristics, and may be scattered in several clusters. In this paper, we propose a content based image rertrieval approach which combines region based image retrieval and a new relevance feedback method using adaptive clustering together. Our main goal is finding semantically related clusters to narrow down the semantic gap. Our method consists of region based clustering processes and cluster-merging process. All segmented regions of relevant images are organized into semantically related hierarchical clusters, and clusters are merged by finding the number of the latent clusters. This method, in the cluster-merging process, applies r: using v principal components instead of classical Hotelling's $T_v^2$ [1] to find the unknown number of clusters and resolve the singularity problem in high dimensions and demonstrate that there is little difference between the performance of $T^2$ and that of $T_v^2$. Experiments have demonstrated that the proposed approach is effective in improving the performance of an image retrieval system.
Journal of the Korea Academia-Industrial cooperation Society
/
v.19
no.9
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pp.333-340
/
2018
Recently, free-form buildings have been designed with complex shapes due to digitization of the construction industry. Exterior and interior components of free-form buildings have free cross sections and curved shapes. Therefore, structural members with curvature are frequently seen. In the modeling and stability evaluation of these structures, commercial programs using classical finite element analysis are not able to perform rapid shape modeling, resulting in a decrease in productivity. Therefore, in this study, pre- and post-processing modules were developed using a prior study to rapidly model the surface of a free-form building and to automatically generate frame structures that make up the cladding. The developed modules use a subdivision algorithm with spline curves. This algorithm is used to automatically generate analytical elements from the configuration information of NURBS curves. In addition, the deformation after analysis can be viewed more realistically. The modules can quickly construct complex curved surfaces. An analysis model of the frame structure was also automatically generated. Therefore, the modules could contribute to the productivity improvement of free-form building design.
The estrogen receptor-related receptors (ERRs) are a group of nuclear receptor superfamily of transcription factors. ERRs and estrogen receptors (ERs) have overlapping affinities for coactivators and DNA binding sites, but differ markedly in ligand binding and activation. The three mammalian ERR genes have been implicated in diverse physiological processes ranging from placental development to maintenance of bone density, whereas the molecular diversity, function, and regulation of ERRs in non-mammalian species are not well understood. In the present study, to investigate the involvement of ERR in transcription and embryogenesis in marine invertebrates, a cDNA encoding ERR (SnERR) was cloned from the gonad in Strongylocentrotus nudus, by polymerase chain reaction (PCR). The amino acid sequence of SnERR showed high homology with that of S. purpuratus (91%). A phylogenetic tree clearly showed that SnERR is a member of the ERR family and clustered in echinodermata group as supported by a high bootstrap value. We examined gene expression of SnERR during embryonic development of S. nudus using real-time PCR. During the embryonic development, the mRNA of ERR was significantly high levels in early development stages (4~64 cell) and larval stages. The SnERR slightly activated transcription through the classical estrogen response elements (EREs) in the presence of genistein. In addition, peroxisome proliferator-activated receptor $\gamma$ coactivator (PGC)-$1\alpha$ knwon as a coactivator of ERR enhanced the snERR-mediated transactivation, suggesting that the PGC-$1\alpha$ is a coactivator of SnERR.
We explore the question of whether adenosine 5'-triphosphate (ATP) acts as an excitatory neurotransmitter in guinea-pig gastric smooth muscle. In an organ bath system, isometric force of the circular smooth muscle of guinea-pig gastric antrum was measured in the presence of atropine and guanethidine. Under electrical field stimulation (EFS) at high frequencies (>20 Hz), NO-mediated relaxation during EFS was followed by a strong contraction after the cessation of EFS (a 'rebound-contraction'). Exogenous ATP mimicked the rebound-contraction. A known $P_{2Y}-purinoceptor$ antagonist, reactive blue 2 (RB-2), blocked the rebound-contraction while selective desensitization of $P_{2Y}-purinoceptor$ with ${\alpha},{\beta}-MeATP$ did not affect it. ATP and 2-MeSATP induced smooth muscle contraction, which was effectively blocked by RB-2 and suramin, a nonselective $P_2-purinoceptor$ antagonist. Particularly, in the presence of RB-2, exogenous ATP and 2-MeSATP inhibited spontaneous phasic contractions, suggesting the existence of different populations of purinoceptors. Both the rebound-contraction and the agonist-induced contraction were not inhibited by indomethacin. The rank orders of agonists' potency were 2-MeSATP > ATP ${ge}$ UTP for contraction and ${\alpha},{\beta}-MeATP\;{\ge}\;{\beta},{\gamma}-MeATP$ for inhibition of the phasic contraction, that accord with the commonly accepted rank order of the classical $P_{2Y}-purinoceptor$ subtypes. Electrical activities of smooth muscles were only slightly influenced by ATP and 2-MeSATP, whereas ${\alpha},{\beta}-MeATP$ attenuated slow waves with membrane hyperpolarization. From the above results, it is suggested that ATP acts as an excitatory neurotransmitter, which mediates the rebound-contraction via $P_{2Y}-purinoceptor$ in guinea-pig gastric antrum.
For medical students and doctors, knowledge of the 3-dimensional (3D) structure of the heart is very important in diagnosis and treatment of the heart diseases. 2-dimensional (2D) tools (e.g. anatomy book) or classical 3D tools (e.g. plastic model) are not sufficient or understanding the complex structures of the heart. Moreover, it is not always guaranteed to dissect the heart of cadaver when it is necessary. To overcome this problem, virtual dissection systems of the heart have been developed. But these systems are not satisfactory since they are made of radiographs; they are not true 3D images; they can not be used to dissect freely; or they can only be operated on the workstation. It is also necessary to make the dissection systems incorporating the various races and tribes because of the organ's difference according to race and tribe. This study was intended to make the 3D image of the heart from a Korean cadaver, and to establish a virtual dissection system of the heart with a personal computer. The procedures or manufacturing this system were as follows. 1. The heart from a Korean adult cadaver was embedded with gelatin solution, and serially cross-sectioned at 1mm-thickness on a meat slicer. Pictures or 153 cross-sectioned specimens were inputted into the computer using a digital camera ($756{\times}504$ resolution, true color). 2. The alignment system was established by means of the language of IDL, and applied to align 2D images of the heart. In each of 2D images, closed curves lining clean and dirty blood pathways were drawn manually on the CorelDRAW program. 3. Using the language of IDL, the 3D image and the virtual dissection system of the heart were constructed. The virtual dissection system of the heart allowed or ree rotation, any-directional sectioning, and selected visualization of the heart's structure. This system is expected to become more advanced, and to be used widely through Internet or CD-title as an educational tool for medical students and doctors.
Suh, Bo Ik;Kim, Sang Woo;Chung, Ho Yun;Kim, Il Hwan;Yang, Jung Dug;Park, Jae Woo;Cho, Byoung Chae
Archives of Plastic Surgery
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v.34
no.3
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pp.279-284
/
2007
Purpose: The vein graft was considered as a useful conduit for nerve defect. But the problem is that it might be collapsed in long vein graft state. A new experimental model using vein graft filled with hyaluronic acid was considered. Methods: Thirty rats were used for the experimental animal. In group I, one side of the femoral nerve was exposed and a segment was removed about 15mm. The neural gap was connected with nerve graft. In group II, the nerve gap was connected with vein graft only. In group III, the nerve gap was connected with vein graft filled with hyaluronic acid. A walking track analysis was made periodically for 2 months and NCV(nerve conduction velocity) was executed at the end of the experiment. And morphologic studies were also done for all groups Results: In a walking track analysis, the toe-spread was widen and the foot-length was lengthened. The recovery of the toe-spread and foot length was checked 2 weeks interval, periodically for two months. The SFI (sciatic function index) was $-52.5{\pm}8.2$ in group I, $-68.1{\pm}4$ in group II, $-55.3{\pm}7.9$ in group III. In electrophysiological study, NCV(nerve conduction velocity) was $26.71{\pm}3.11m/s$ in group I, $17.94{\pm}4.35m/s$ in group II, $25.69{\pm}2.81m/s$ in group III. The functional recovery in group I and III was superior to that the group II statistically(p < 0.05) Under electromicroscopic study, the number of the myelinated axons were $1419.1{\pm}240$ in group I, $921.7{\pm}176.8$ in group II, $1322.2{\pm}318$ in group III. The number of the myelinated axons were much more in group I and III than group II statistically (p<0.05). Conclusion: This study suggested that the vein graft filled with hyaluronic acid is more effective than vein graft only for the conduit of the nerve gap. It was thought that the technique could be used in clinical cases with nerve defects as an alternative method to classical nerve grafts.
Lee, Seulki;Kim, Jung Sun;Kang, Sang-Ho;Sohn, Seong-Han;Won, So Youn
Journal of Plant Biotechnology
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v.43
no.1
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pp.12-20
/
2016
Advances in DNA sequencing technologies have contributed to revolutionary understanding of many fundamental biological processes. With unprecedented cost-effective and high-throughput sequencing, a single laboratory can afford to de novo sequence the whole genome for species of interest. In addition, population genetic studies have been remarkably accelerated by numerous molecular markers identified from unbiased genome-wide sequences of population samples. As sequencing technologies have evolved very rapidly, acquiring appropriate individual plants or populations is a major bottleneck in plant research considering the complex nature of plant genome, such as heterozygosity, repetitiveness, and polyploidy. This challenge could be overcome by the old but effective method known as haploid induction. Haploid plants containing half of their sporophytic chromosomes can be rapidly generated mainly by culturing gametophytic cells such as ovules or pollens. Subsequent chromosome doubling in haploid plants can generate stable doubled haploid (DH) with perfect homozygosity. Here, classical methodology to generate and identify haploid plants or DH are summarized. In addition, haploid induction by epigenetic regulation of centromeric histone is explained. Furthermore, the utilization of haploid plant in the genomics era is discussed in the aspect of genome sequencing project and population genetic studies.
As classical acute aortic dissection, atherosclerotic penetrating ulcers and intramural hematoma have different pathophysiology and natural history, treatment strategy should be different and, therefore, accurate differential diagnosis is necessary. However, these three aortic diseases may be indistinguishable by clinical observation and even by various diagnostic modalities such as cardiac echocardiography, CT and MRI. The patients was a 71-year-old female with chief complaints of anterior chest pain, nausea and vomiting which occurred suddenly 3 days before admission. CT angiographic with 3 dimensional reconstruction shows intramural hematoma in ascending aorta, aortic arch, descending thoracic aorta and right brachiocephalic trunk, heompericardium, and blood in mediastinum and both pleural cavities. The CT angiographic finding of focal out-bulging in the ascending thoracic aorta was diagnosed as penetrating atherosclerotic ulcer. The patient underwent emergency operation under a preoperative diagnosis of penetrating atherosclerotic ulcer with a sign of aortic rupture. In the intraoperative findings, however, intimal tear was seen in the anterior portion of the ascending aorta about 1cm below the brachiocephalic trunk and falselumen appeared after hematoma was removed from the layer of tunica media. We report a case of type A aortic dissection which mimicked clinical and diagnostic features of penetrating atherosclerotic ulcer.
Park, Nam-Gyu;Go, Hye-Jin;Kim, Gun-Do;Lee, Jong-Kyu;Kil, Sang-Hyeong;Lee, Byung-Woo
Journal of Life Science
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v.22
no.8
/
pp.1046-1051
/
2012
In order to investigate the changes in bioactive materials induced in goldfish brains by $^{18}F$ irradiation, the variations in the neurotransmitter levels in the whole brain were studied. The distance between the goldfish and 580 mCi of $^{18}F$ was about 4 cm, and the exposure lasted for 4 hrs. The absorption level calculated based on the distance, exposure time, and half-life of $^{18}F$ was approximately 2 Gy. After sacrifice by $^{18}F$ irradiation or untreated conditions, ten brains were dissected or immediately frozen, respectively. The tissues were extracted in acetic acid. After lyophilization, the samples were dissolved in distilled water and were further purified on a reverse-phase HPLC column. There were no differences in the intensities of the bioactive materials between $^{18}F$-exposed goldfish and control goldfish, while the only peak corresponded to 13 min, which indicated a significant increase in the irradiated brains. Our analysis has found that this compound is tryptophan. This result suggests that $^{18}F$ leads to changes in a classical neurotransmitter, tryptophan, in both the brains of control goldfish and goldfish contaminated by irradiation.
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