Lysin motif (LysM) proteins are reported to be necessary for the virulence and immune response suppression in many herbaceous plant pathogens, while far less is documented in woody plant pathogens. In this study, we preliminarily characterized the molecular function of a LysM protein LtLysM1 in woody plant pathogen Lasiodiplodia theobromae. Transcriptional profiles revealed that LtLysM1 is highly expressed at infectious stages, especially at 36 and 48 hours post inoculation. Amino acid sequence analyses revealed that LtLysM1 was a putative glycoprotein with 10 predicted N-glycosylation sites and one LysM domain. Pathogenicity tests showed that overexpressed transformants of LtLysM1 displayed increased virulence on grapevine shoots in comparison with that of wild type CSS-01s, and RNAi transformants of LtLysM1 exhibited significantly decreased lesion length when compared with that of wild type CSS-01s. Moreover, LtLysM1 was confirmed to be a secreted protein by a yeast signal peptide trap assay. Transient expression in Nicotiana benthamiana together with protein immunoblotting confirmed that LtLysM1 was an N-glycosylated protein. In contrast to previously reported LysM protein Slp1 and OsCEBiP, LtLysM1 molecule did not interact with itself based on yeast two hybrid and co-immunoprecipitation assays. These results indicate that LtLysM1 is a secreted protein and functions as a critical virulence factor during the disease symptom development in woody plants.
Hong, Jin Ho;Lee, Ha Young;Kang, Young Hye;Lim, Myung Kwan;Kim, Yeo Ju;Cho, Soon Gu;Kim, Mi Young
Investigative Magnetic Resonance Imaging
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v.20
no.1
/
pp.44-52
/
2016
Purpose: To quantitatively and qualitatively compare fat-suppressed MRI quality using iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) with that using frequency selective fat-suppression (FSFS) T2- and postcontrast T1-weighted fast spin-echo images of the head and neck at 3T. Materials and Methods: The study was approved by our Institutional Review Board. Prospective MR image analysis was performed in 36 individuals at a single-center. Axial fat suppressed T2- and postcontrast T1-weighted images with IDEAL and FSFS were compared. Visual assessment was performed by two independent readers with respect to; 1) metallic artifacts around oral cavity, 2) susceptibility artifacts around upper airway, paranasal sinus, and head-neck junction, 3) homogeneity of fat suppression, 4) image sharpness, 5) tissue contrast of pathologies and lymph nodes. The signal-to-noise ratios (SNR) for each image sequence were assessed. Results: Both IDEAL fat suppressed T2- and T1-weighted images significantly reduced artifacts around airway, paranasal sinus, and head-neck junction, and significantly improved homogeneous fat suppression in compared to those using FSFS (P < 0.05 for all). IDEAL significantly decreased artifacts around oral cavity on T2-weighted images (P < 0.05, respectively) and improved sharpness, lesion-to-tissue, and lymph node-to-tissue contrast on T1-weighted images (P < 0.05 for all). The mean SNRs were significantly improved on both T1- and T2-weighted IDEAL images (P < 0.05 for all). Conclusion: IDEAL technique improves image quality in the head and neck by reducing artifacts with homogeneous fat suppression, while maintaining a high SNR.
Journal of the Korean Institute of Telematics and Electronics S
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v.36S
no.8
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pp.8-16
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1999
In this paper, the performance of a hybrid DS/SFH-SS(direct-sequence/slow-frequency-hopped spread-spectrum) system with coherent BPSK modulation over Nakagami fading channel in the presence of multiple tone jamming is analyzed. Because the Nakagami m-distribution can describe not only Rayleigh fading but also more general fluctuations involving a specular component by adjusting the value of the fading index m. It is known that for m=1 corresponds to Rayleigh fading, for $1/2{\le}m{\le}1$ corresponds to the worst case fading condition, for m>1 corresponds to Rician fading, and for $m{\to}{\infty}$ corresponds to the nonfading condition. The bit error probability is derived over Nakagami model and numerical evaluations are presented for some combinations of system parameters. The results show that as m increases, the bit error probability is better. Also, at a low JSR(jamming-to-signal power ratio), a pure DS-SS system can achieve lower bit error probability than a hybrid DS/SFH-SS system. But at a high JSR, a hybrid DS/SFH-SS system is shown to be superior to a pure DS-SS system. Therefore, it is demonstrated that without increasing the total system bandwidth, the performance of a hybrid DS/SFH-SS is superior to that of a pure DS-SS system in the presence of multiple tone jamming.
The expression and secretion of human lactoferrin (hLf) in Sacclnromyces diastaticus were performed. 1. For the secretion of hLf in yeast, recombinant plasmid pYEGLf was constructed using promoter, secretion signal sequence of glucoamylase I gene (STA1) and transcriptional terminator of GAL7 gene. 2. Each correct recombinant plasmid was selected by mini-preparation of plasmid DNA from E coli transformant and restriction enzyme digestion analysis. The selected plasmids, pYEGLf, were transformed into S. diastaticus YIY345 as a expression host, respectively. 3. Western blot analysis using rabbit anti-hLf was carried out to identify expressed hLf. Positive signals were shown in culture supernatant of pYEGLf transformant. 4. About $100{\mu}g-1mg$ of concentrated culture supernatant of positive clone were loaded on paper disc and tested for the antimicrobial activity against E coli. However, no activity was observed. We concluded that this fact results from low concentration of hLf secreted from yeast, compared with the fact that MIC of hLf is as high as $3mg/m{\ell}$. Therefore, the purification of secreted hLf may be require to investigate the antimicrobial activity. From this study, the feasibility of low-cost production of sufficient quantities of human lactofferin for nutritional and therapeutical applications were suggested.
Kim, Dae Yeon;Hong, Min Jeong;Jung, Woo Joo;Seo, Yong Weon
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
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pp.140-140
/
2017
Nutritious and functional foods from crop have received great attention in recent years. Colored-grain wheat contains high phenolic compound and a large number of flavonoid. The anthocyanin and polyphenolic synthesis and accumulation is generally stimulated in response to biotic or abiotic stresses. Here, we analyzed genome wide transcripts in seedling of colored-grain wheat response to ABA and PEG treatment. About 900 and 1500 transcripts (p-value < 0.05) from ABA and PEG treatment were aligned to IWGSC1+popseq DB which is composed of over 110,000 transcripts including 100,934 coding genes. NR protein sequences of Poaceae from NCBI and protein sequence of transcription factors originated from 83 species in plant transcription factor database v3.0 were used for annotation of putative transcripts. Gene ontology analysis were conducted and KEGG mapping was performed to show expression pattern of biosynthesis genes related in flavonoid, isoflavonoid, flavons and anthocyanin biopathway. DroughtDB (http://pgsb.helmholtz-muenchen.de/droughtdb/) was used for detection of DEGs to explain that physiological and molecular drought avoidance by drought tolerance mechanisms. Drought response pathway, such as ABA signaling, water and ion channels, detoxification signaling, enzymes of osmolyte biosynthesis, phospholipid metabolism, signal transduction, and transcription factors related DEGs were selected to explain response mechanism under water deficit condition. Anthocyanin, phenol compound, and DPPH radical scavenging activity were measured and antioxidant activity enzyme assays were conducted to show biochemical adaptation under water deficit condition. Several MYB and bHLH transcription factors were up-regulated in both ABA and PEG treated condition, which means highly expressed MYB and bHLH transcription factors enhanced the expression of genes related in the biosynthesis pathways of flavonoids, such as anthocyanin and dihydroflavonols in colored wheat seedlings. Subsequently, the accumulation of total anthocyanin and phenol contents were observed in colored wheat seedlings, and antioxidant capacity was promoted by upregulation of genes involved in maintaining redox state and activation of antioxidant scavengers, such as CAT, APX, POD, and SOD in colored wheat seedlings under water deficit condition. This work may provide valuable and basic information for further investigation of the molecular responses of colored-grain wheat to water deficit stress and for further gene-based studies.
Transactions of the Korean Society of Mechanical Engineers A
/
v.37
no.9
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pp.1093-1098
/
2013
A single-crystal sapphire is used as a transparent bulletproof window material; however, few studies have investigated the dynamic behavior and fracture properties under high-speed impact. High-speed and high-resolution sequential images are required to study the interaction of the bullet with the brittle ceramic materials. In this study, a device is developed to capture the sequence of high-speed impact/penetration phenomena. This system consists of a speed measurement device, a microprocessor-based camera controller, and multiple CCD cameras. By using a linear array sensor, the speed-measuring device can measure a small (diameter: up to 1 2 mm) and fast (speed: up to Mach 3) bullet. Once a bullet is launched, it passes through the speed measurement device where its time and speed is recorded, and then, the camera controller computes the exact time of arrival to the target during flight. Then, it sends the trigger signal to the cameras and flashes with a specific delay to capture the impact images sequentially. It is almost impossible to capture high-speed images without the estimation of the time of arrival. We were able to capture high-speed images using the new system with precise accuracy.
Kim, Sung Chan;Kang, Seung Ha;Choi, Eun Young;Hong, Yeon Hee;Bok, Jin Duck;Kim, Jae Yeong;Lee, Sang Suk;Choi, Yun Jaie;Choi, In Soon;Cho, Kwang Keun
Asian-Australasian Journal of Animal Sciences
/
v.29
no.1
/
pp.126-133
/
2016
A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-${\beta}$-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) $DH5{\alpha}$. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli $DH5{\alpha}$ harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine.Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was $55^{\circ}C$, but it retained over 90% of maximum activity in a broad temperature range ($40^{\circ}C$ to $60^{\circ}C$). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, $K_m$ and $V_{max}$ of rEG1 were 0.39% CMC and 143 U/mg, respectively.
Authors investigated neuronal changes of local cellular metabolism in the cerebral lesions of Parkinsonian symptomatic side between before and after stereotactic neurosurgery by follow-up 1H magnetic resonance spectroscopy (MRS). Patients with Parkinson's disease (PD) (n = 15) and age-matched normal controls (n = 15) underwen MRS examinations using a stimulated echo acquisition mode (STEAM) pulse sequence that provided 2${\times}$2${\times}$2 ㎤ (8ml) volume of interest in the regions of substantia nigra, thalamus, and lentiform nucleus. Spectral parameters were 20 ms TE, 2000 ms TR, 128 averages,2500 Hz spectral width, and 2048 data points. Raw data were processed by the SAGE data analysis package (GE Medical Systems). Peak areas of N-acetylaspartate (NAA), creatine (Cr), choline-containing compounds (Cho), inositols (Ins), and the sum (Glx) of glutamate and GABA were calculated by means of fitting the spectrum to a summation of Lorentzian curves using Marquardt algorithm. After blindly processed, we evaluated neuronal alterations of observable metabolite ratios between before and after stereotactic neurosurgery using Pearson product-moment analysis (SPSS, Ver. 6.0). A significant reduction of NAA/Cho ratio was observed in the cerebral lesion in substantia nigra of PD patient related to the symptomatic side after neurosurgery (P : 0.03). In thalamus, NAA/Cho ratio was also significantly decreased in the cerebral lesion including the electrode-surgical region (P : 0.03). A significant reduction of NAA/Cho ratio in lentiform nucleus was not oberved, but tended toward significant reduction after neurosurgery (P = 0.08). In particular, remarkable lactate signal was noted from the surgical thalamic lesions of 6 among 8 patients and internal segments of globus pallidus of 6 among 7 patients, respectively. Significant metabolic alterations of NAA/Cho ratio might reflect functional changes of neuropathological processes in the lesion of substantia nigra, thalamus, and lentiform nucleus, and could be a valuable finding fur evaluation of Parkinson's disease after neurosurgery. Increase of lactate signals, being remarkable in surgical lesions, could be consistent with a common consequence of neurosurgical necrosis. Thus, IH MRS could be a useful modality to evaluate the diagnostic and prognostic implications fur Parkinsons disease after functional neurosurgery.
Proceedings of the Microbiological Society of Korea Conference
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2007.05a
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pp.120-122
/
2007
All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.
Consumers changed to the subject to produce a digital contents as data transmission technique is advanced and a digital machine is diffused variously. Users are interested greatly in a user creative movie (UCM) production among various online contents. The UCM production method which uses the music and picture is the method that users make the UCM more easily. However, the UCM production service has the problem that any association does not exist in the music and picture and that the picture changes according to fixed time interval without the relation at a music rhythm. To solve this problem, we propose the UCM production method which uses a music analysis and picture analysis in the paper. A music analysis finds a picture change time according to the rhythm and a picture analysis finds the association of the picture. A music analysis finds strong parts of the sound which uses Root-Mean-Square (RMS). And a picture analysis classifies the picture as a scenery picture and people picture which uses structure simplicity of the picture(SSP) and face region detection. A picture analysis got correct result of 86.4% in the experiment and we can finds the association at each picture and arranges the sequence which the picture appears. Therefore, if we use a music and picture analysis at the UCM production, users may make natural and efficient movie.
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