• Title/Summary/Keyword: Strain sensing

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Sensing the Stress: the Role of the Stress-activated p38/Hog1 MAPK Signalling Pathway in Human Pathogenic Fungus Cryptococcus neoformans

  • Bahn, Yong-Sun;Heitman, Joseph
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2007.05a
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    • pp.120-122
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    • 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.

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Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1 (Methylophaga aminosulfidovorans SKI bfmo 오페론의 클로닝 및 구조 분석)

  • Lim Hyun Sook;Goo Jae Whan;Kim Lee Hyun;Kim Si Wouk;Cho Eun Hee
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.1-7
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    • 2005
  • Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP) can utilize trimethylamine as a sole carbon, nitrogen, and energy source. The bacterial flavin-containing monooxygenase (bFMO) gene was identified in the strain and the recombinant enzyme expressed in E. coli oxidized trimethylamine. To study the function and regulation of the bfmo, over 8,000 nucleotide sequences of the neighboring regions including the bfmo were determined. Three open reading frames proceeding to the bfmo gene encoded analogues to highly conserved nitrate/nitrite sensing two-component system regulators and a methyl accepting protein. Two small open reading frames just downstream of the bfmo gene showed no similar proteins of known functions but the sequences were conserved among other bacteria. Reverse transcription-polymerase chain reaction analysis showed that the six putative genes consisted of three transcription units. The three regulatory genes located upstream of the bfmo gene formed two separate transcription units. The bfmo and the two downstream genes were transcribed from a single promoter.

Serviceability Assessment of a K-AGT Test Bed Bridge Using FBG Sensors (광섬유 센서를 이용한 경량전철 교량의 사용성 평가)

  • Kang, Dong-Hoon;Chung, Won-Seok;Kim, Hyun-Min;Yeo, In-Ho
    • Journal of the Korean Society for Nondestructive Testing
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    • v.27 no.4
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    • pp.305-312
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    • 2007
  • Among many types of light rail transits (LRT), the rubber-tired automated guide-way transit (AGT) is prevalent in many countries due to its advantages such as good acceleration/deceleration performance, high climb capacity, and reduction of noise and vibration. However, AGT is generally powered by high-voltage electric power feeding system and it may cause electromagnetic interference (EMI) to measurement sensors. The fiber optic sensor system is free from EMI and has been successfully applied in many applications of civil engineering. Especially, fiber Bragg grating (FBG) sensors are the most widely used because of their excellent multiplexing capabilities. This paper investigates a prestressed concrete girder bridge in the Korean AGT test track using FBG based sensors to monitor the dynamic response at various vehicle speeds. The serviceability requirements provided in the specification are also compared against the measured results. The results show that the measured data from FBG based sensors are free from EMI though electric sensors are not, especially in the case of electric strain gauge. It is expected that the FBG sensing system can be effectively applied to the LRT railway bridges that suffered from EMI.

Non-contact Stress Measurement in Steel Member of PSC Box Bridge Using Raman Spectroscopy (라만 형광 분광법을 이용한 PSC 박스교 인장케이블 응력측정방법 연구)

  • Kim, Jongwoo;Kim, Namgyu
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.23 no.2
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    • pp.130-134
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    • 2019
  • In this paper, a laser-based non-contact load cell is newly developed for measuring forces in prestressed concrete tendons. First, alumina particles have been sprayed onto an empty load cell which has no strain gauges on it, and the layer has been used as a passive stress sensor. Then, the spectral shifts in fluorescence spectroscopy have been measured using a laser-based spectroscopic system under various force levels, and it has been found that the relation of applied force and spectral shift is linear in a lab-scale test. To validate the field applicability of the customized load cell, a full-scale prestressed concrete specimen has been constructed in a yard. During the field test, it was, however, found that the coating surface has irregular stress distribution. Therefore, the location of a probe has to be fixed onto the customized load cell for using the coating layer as a passive stress sensor. So, a prototype customized load cell has been manufactured, which consists of a probe mount on its casing. Then, by performing lab-scale uniaxial compression tests with the prototype load cell, a linear relation between compression stress and spectrum shift at a specific point where laser light had been illuminated has been detected. Thus, it has a high possibility to use the prototype load cell as a force sensor of prestressed concrete tendons.

OmpR Is Essential for Growth and Expression of Virulence-related Genes in the Fish Pathogen Edwardsiella piscicida (어류 병원체 Edwardsiella piscicida의 OmpR은 생육과 병원성과 관련된 유전자의 발현에 필수적)

  • Ray, Durga;Kim, Yeon Ha;Choe, unjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.31 no.1
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    • pp.28-36
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    • 2021
  • Edwardsiella piscicida is a significant cause of hemorrhagic septicemia in fish and gastrointestinal infections in humans. Survival bacteria require specialized mechanisms to adapt to environmental fluctuations. Hence, to understand the mechanism through which E. piscicida senses and responds to environmental osmolarity changes, we determined the protein expression profile and physiological properties under various salinity conditions in this study. The OmpR protein is a part of the Env-ZOmpR two-component system that has been implicated in sensing salt stress in bacteria. However, the physiological role played by this protein in E. piscicida remains to be elucidated. Therefore, in this work, the function of the OmpR protein in response to salt stress was investigated. Phenotypic analysis revealed that, in the mutant, three of the biochemical phenotypes were different from the wild type, including, citrate utilization, hydrogen sulfide, and indole production. Introduction of the plasmid containing the entire ompR gene to the mutant strain returned it to its parental phenotype. The retarded growth rate also partially recovered. Furthermore, in our studies, OmpR was not found to be related to cell motility. Taken together, our results from the mutational analysis, the growth assay, MALDI-TOF MS, qRT-PCR, and the phenotype studies suggest that the OmpR of E. piscicida is implicated in osmoregulation, growth, expression of porins (ETAE_1826), virulence-related genes (EseC, EseD and EvpC), and certain genes of unknown function (ETAE_1540 and ETAE_2706).

Analysis of whole genome sequencing and virulence factors of Vibrio vulnificus 1908-10 isolated from sea water at Gadeok island coast

  • Hee-kyung Oh;Nameun Kim;Do-Hyung Kim;Hye-Young Shin;Eun-Woo Lee;Sung-Hwan Eom;Young-Mog Kim
    • Fisheries and Aquatic Sciences
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    • v.26 no.9
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    • pp.558-568
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    • 2023
  • Vibrio vulnificus is an aquatic bacterium causing septicemia and wound infection in humans. To understand this pathogen at the genomic level, it was performed whole genome sequencing of a cefoxitin-resistant strain, V. vulnificus 1908-10 possessing virulence-related genes (vvhA, viuB, and vcgC) isolated from Gadeok island coastal seawater in South Korea. The genome of V. vulnificus 1908-10 consisted of two circular contigs and no plasmid. The total genome size was estimated to be 5,018,425 bp with a guanine-cytosine (GC) content of 46.9%. We found 119 tRNA and 34 rRNA genes respectively in the genome, along with 4,352 predicted protein sequences. Virulence factor (VF) analysis further revealed that V. vulnificus 1908-10 possess various virulence genes in classes of adherence, antiphagocytosis, chemotaxis and motility, iron uptake, quorum sensing, secretion system, and toxin. In the comparison of the presence/absence of virulence genes, V. vulnificus 1908-10 had fur, hlyU, luxS, ompU, pilA, pilF, rtxA, rtxC, and vvhA. Of the 30 V. vulnificus comparative strains, 80% of the C-genotype strains have all of these genes, whereas 40% of the E-genotype strains have all of them. In particular, pilA were identified in 80% of the C-type strains and 40% of the E-type strains, showing more difference than other genes. Therefore, V. vulnificus 1908-10 had similar VF characteristics to those of type C strains. Multifunctional-autoprocessing repeats-in-toxin (MARTX) toxin of V. vulnificus 1908-10 contained 8 A-type repeats (GXXGXXXXXG), 25 B.1-type repeats (TXVGXGXX), 18 B2-type repeats (GGXGXDXXX), and 7 C-type repeats (GGXGXDXXX). The National Center for Biotechnology Information (NCBI) Basic Local Alignment Search Tool (BLAST) showed that the RtxA protein of V. vulnificus 1908-10 had the effector domain in the order of cross-liking domain (ACD)-C58_PaToxP-like domain- α/β hydrolase-C58_PaToxP-like domain.

Analysis of Tidal Deflection and Ice Properties of Ross Ice Shelf, Antarctica, by using DDInSAR Imagery (DDInSAR 영상을 이용한 남극 로스 빙붕의 조위변형과 물성 분석)

  • Han, Soojeong;Han, Hyangsun;Lee, Hoonyol
    • Korean Journal of Remote Sensing
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    • v.35 no.6_1
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    • pp.933-944
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    • 2019
  • This study analyzes the tide deformation of land boundary regions on the east (Region A) and west (Region B) sides of the Ross Ice Shelf in Antarctica using Double-Differential Interferometric Synthetic Aperture Radar (DDInSAR). A total of seven Sentinel-1A SAR images acquired in 2015-2016 were used to estimate the accuracy of tide prediction model and Young's modulus of ice shelf. First, we compared the Ross Sea Height-based Tidal Inverse (Ross_Inv) model, which is a representative tide prediction model for the Antarctic Ross Sea, with the tide deformation of the ice shelf extracted from the DDInSAR image. The accuracy was analyzed as 3.86 cm in the east region of Ross Ice Shelf and it was confirmed that the inverse barometric pressure effect must be corrected in the tide model. However, in the east, it is confirmed that the tide model may be inaccurate because a large error occurs even after correction of the atmospheric effect. In addition, the Young's modulus of the ice was calculated on the basis of the one-dimensional elastic beam model showing the correlation between the width of the hinge zone where the tide strain occurs and the ice thickness. For this purpose, the grounding line is defined as the line where the displacement caused by the tide appears in the DDInSAR image, and the hinge line is defined as the line to have the local maximum/minimum deformation, and the hinge zone as the area between the two lines. According to the one-dimensional elastic beam model assuming a semi-infinite plane, the width of the hinge region is directly proportional to the 0.75 power of the ice thickness. The width of the hinge zone was measured in the area where the ground line and the hinge line were close to the straight line shown in DDInSAR. The linear regression analysis with the 0.75 power of BEDMAP2 ice thickness estimated the Young's modulus of 1.77±0.73 GPa in the east and west of the Ross Ice Shelf. In this way, more accurate Young's modulus can be estimated by accumulating Sentinel-1 images in the future.