• Title/Summary/Keyword: Outer membrane

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Development of an Improved Endotoxin Detection Method Using Centrifugation (원심분리법을 이용한 혈청 내 내독소의 개선된 측정방법 연구)

  • Choi, Hyeong Jwa;Lim, Yoo Jung;Lee, Eun Hee;Park, Jin Yeon;Prabagar, Miglena G.;Park, Hyung Soon;Kang, Young Sun
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.242-248
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    • 2013
  • Endotoxins are part of the outer membrane of the cell wall of gram-negative bacteria and are continuously released during bacterial growth. Endotoxins typically induce severe sepsis and septic shock, which cause more than 50% of mortalities. Endotoxins are easily measured in the serum by the limulus amebocyte lysate (LAL) test. However, a nonspecific result is obtained, because the high concentration of serum proteins disturbs the enzyme reaction of the LAL test. In order to solve this problem, the LAL test was performed in this study after the centrifugation of the boiled serum samples to remove the impurities. As a result, among the various conditions examined, endotoxin measurement with the LAL test was the most accurate and repeatable after centrifugation of the boiled serum at $100^{\circ}C$. Moreover, the endotoxin was accurately and repeatedly measured from the prepared sera of mice that had been administered an intraperitoneal injection of purified lipopolysaccharides (LPS) or E. coli. Therefore, the application of centrifugation to remove impurities from boiled serum gives an accurate measurement of endotoxins in the sera of normal subjects or patients, and this will lead to the improved diagnosis and prevention of diseases caused by endotoxins. In addition, the centrifugation of boiled serum samples should be considered and included in the development of endotoxin test kits.

Biophysical Studies Reveal Key Interactions between Papiliocin-Derived PapN and Lipopolysaccharide in Gram-Negative Bacteria

  • Durai, Prasannavenkatesh;Lee, Yeongjoon;Kim, Jieun;Jeon, Dasom;Kim, Yangmee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.671-678
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    • 2018
  • Papiliocin, isolated from the swallowtail butterfly (Papilio xuthus), is an antimicrobial peptide with high selectivity against gram-negative bacteria. We previously showed that the N-terminal helix of papiliocin (PapN) plays a key role in the antibacterial and anti-inflammatory activity of papiliocin. In this study, we measured the selectivity of PapN against multidrug-resistant gram-negative bacteria, as well as its anti-inflammatory activity. Interactions between Trp2 of PapN and lipopolysaccharide (LPS), which is a major component of the outer membrane of gram-negative bacteria, were studied using the Trp fluorescence blue shift and quenching in LPS micelles. Furthermore, using circular dichroism, we investigated the interactions between PapN and LPS, showing that LPS plays critical roles in peptide folding. Our results demonstrated that Trp2 in PapN was buried deep in the negatively charged LPS, and Trp2 induced the ${\alpha}$-helical structure of PapN. Importantly, docking studies determined that predominant electrostatic interactions of positively charged arginine residues in PapN with phosphate head groups of LPS were key factors for binding. Similarly, hydrophobic interactions by aromatic residues of PapN with fatty acid chains in LPS were also significant for binding. These results may facilitate the development of peptide antibiotics with anti-inflammatory activity.

Brucella melitensis omp31 Mutant Is Attenuated and Confers Protection Against Virulent Brucella melitensis Challenge in BALB/c Mice

  • Verdiguel-Fernandez, L;Oropeza-Navarro, R;Ortiz, Adolfo;Robles-Pesina, MG;Ramirez-Lezama, J;Castaneda-Ramirez, A;Verdugo-Rodriguez, A
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.497-504
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    • 2020
  • For control of brucellosis in small ruminants, attenuated B. melitensis Rev1 is used but it can be virulent for animals and human. Based on these aspects, it is essential to identify potential immunogens to avoid these problems in prevention of brucellosis. The majority of OMPs in the Omp25/31 family have been studied because these proteins are relevant in maintaining the integrity of the outer membrane but their implication in the virulence of the different species of this genus is not clearly described. Therefore, in this work we studied the role of Omp31 on virulence by determining the residual virulence and detecting lesions in spleen and testis of mice inoculated with the B. melitensis LVM31 mutant strain. In addition, we evaluated the conferred protection in mice immunized with the mutant strain against the challenge with the B. melitensis Bm133 virulent strain. Our results showed that the mutation of omp31 caused a decrease in splenic colonization without generating apparent lesions or histopathological changes apparent in both organs in comparison with the control strains and that the mutant strain conferred similar protection as the B. melitensis Rev1 vaccine strain against the challenge with B. melitensis Bm133 virulent strain. These results allow us to conclude that Omp31 plays an important role on the virulence of B. melitensis in the murine model, and due to the attenuation shown by the strain, it could be considered a vaccine candidate for the prevention of goat brucellosis.

Changes of Morphology and Morphometric Characteristics of the Oocyte during Oogenesis of the Abalone Haliotis discus hannai (북방전복 Haliotis discus hannai 난자형성과정 동안 난모세포의 형태 및 계측형질 변화)

  • Ju, Sun Mi;Lee, Jung Sick
    • The Korean Journal of Malacology
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    • v.32 no.1
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    • pp.1-7
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    • 2016
  • The developmental stage of germ cells during oogenesis can be categorized into six stages with histological features: (1) oogonium, (2) previtellogenic oocyte, (3) initial vitellogenic oocyte, (4) early active vitellogenic oocyte, (5) late active vitellogenic oocyte and (6) ripe oocyte. The size of oocyte, nucleus and nucleolus illustrated the increase tendency but size ratio of nucleolus to nucleus was decreased during oogenesis. During oogenesis the stainability in the cytoplasm of oocyte changes from basophilic to eosinophilic in H-E stain. And egg stalk and outer jelly membrane was developed in the oocyte. These histological changes are seemed to be yolk accumulation in the oocyte and preparation process for spawning.

Intracapsular Chondroma of the Knee - A case report - (슬관절에 생긴 연골종 - 증례보고 -)

  • Ji, Jong-Hoon;Kim, Young-Yul;Kim, Jin-Young;Shafi, Mohamed;Kim, Ji-Chang;Lee, Sang-Wook;Kim, Weon-Yoo
    • Journal of the Korean Arthroscopy Society
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    • v.8 no.1
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    • pp.54-57
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    • 2004
  • Intracapsular chondromas of the knee have been reported rarely, so the authors report one case about its clinical and histologic findings. This is the case about 38-year-old man who feel painful swelling of the knee joint and the tenderness on the anterior aspect of knee. After arthroscopic excision, grossly there was a 2.5${\times}$l.5${\times}$1cm sized white colored firm mass which had central brownish lesion. Microscopically it was confirmed to chondroma which was consisted of hyaline cartilage and outer thinly synovial membrane.

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Protective Immunity of 56-kDa Type-Specific Antigen of Orientia tsutsugamushi Causing Scrub Typhus

  • Choi, Sangho;Jeong, Hang Jin;Ju, Young Ran;Gill, Byoungchul;Hwang, Kyu-Jam;Lee, Jeongmin
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1728-1735
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    • 2014
  • Scrub typhus, caused by infection with Orientia tsutsugamushi, is a mite-borne zoonotic disease endemic to the Asian-Pacific region. In Korea, the incidence of this disease has increased with climate changes, and over 10,000 cases of infection were reported in 2013. Although this infection is treatable with antibiotics such as doxycycline and azithromycin, an effective prophylactic vaccine against O. tsutsugamushi would be more desirable for preventing scrub typhus in endemic areas. In this study, we investigated the 56-kDa type-specific antigen (TSA56), which is a major outer membrane protein of O. tsutsugamushi, as a vaccine candidate. Intranasal immunization of recombinant TSA56 (rec56) induced a higher level of TSA56-specific IgG than that induced by intramuscular immunization of tsa56-expressing DNA (p56). Both types of immunization induced a cell-mediated immune response to TSA56, as demonstrated by the splenic cell proliferation assay. Mice immunized with p56, followed by rec56 plus heat-labile enterotoxin B subunit from E. coli, had a stronger protection from a homologous challenge with the O. tsutsugamushi Boryong strain than with other combinations. Our preliminary results suggest that an effective human vaccine for scrub typhus can include either recombinant TSA56 protein or tsa56-expressing DNA, and provide the basis for further studies to optimize vaccine performance using additional antigens or different adjuvants.

Increased Immunogenicity and Protective Efficacy of a P. aeruginosa Vaccine in Mice Using an Alum and De-O-Acylated Lipooligosaccharide Adjuvant System

  • Ryu, Ji In;Wui, Seo Ri;Ko, Ara;Do, Hien Thi Thu;Lee, Yeon Jeong;Kim, Hark Jun;Rhee, Inmoo;Park, Shin Ae;Kim, Kwang Sung;Cho, Yang Je;Lee, Na Gyong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1539-1548
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    • 2017
  • Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that commonly causes fatal infections in cystic fibrosis and burn patients as well as in patients who are hospitalized or have impaired immune systems. P. aeruginosa infections are difficult to treat owing to the high resistance of the pathogen to conventional antibiotics. Despite several efforts, no effective prophylactic vaccines against P. aeruginosa are currently available. In this study, we investigated the activity of the CIA06 adjuvant system, which is composed of alum and de-O-acylated lipooligosaccharide, on a P. aeruginosa outer membrane protein (OMP) antigen vaccine in mice. The results indicated that CIA06 significantly increased the antigen-specific IgG titers and opsonophagocytic activity of immune sera against P. aeruginosa. In addition, the antibodies induced by the CIA06-adjuvanted vaccine exhibited higher cross-reactivity with heterologous P. aeruginosa strains. Finally, mice immunized with the CIA06-adjuvanted vaccine were effectively protected from lethal P. aeruginosa challenge. Based on these data, we suggest that the CIA06 adjuvant system might be used to promote the immunogenicity and protective efficacy of the P. aeruginosa OMP vaccine.

Speech Signal Processing using Pitch Synchronous Multi-Spectra and DSP System Design in Cochlear Implant (피치동기 다중 스펙트럼을 이용한 청각보철장치의 음성신호처리 및 DSP 시스템 설계)

  • Shin, J. I.;Park, S. J.;Shin, D. K.;Lee, J. H.;Park, S. H.
    • Journal of Biomedical Engineering Research
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    • v.20 no.4
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    • pp.495-502
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    • 1999
  • We propose efficient speech signal processing algorithms and a system for cochlear implant in this paper. The outer and the middle car which perform amplifying, lowpass filtering and AGC, are modeled by an analog system, and the inner ear acting as a time-delayed multi filter and the transducer is implemented by the DSP circuit which enables real-time processing. Especially, the basilar membrane characteristic of the inner ear is modeled by a nonlinear filter bank, and then tonotopy and periodicity of the auditory system is satisfied by using a pitch-synchronous multi-spectra(PSMS) method. Moreover, most of the speech processing is performed by S/W so the system can be easily modified. And as our program is written in C-language, it can be easily transplanted to the system using other processors.

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Characteristics of Cytophaga columnaris isolated from rainbow trout (Oncorhynchus mykiss), goldfish(Carassius auratus), and ayu(Plecoglossus altivels) in Korea (국내에서 양식되는 무지개송어(Oncorhynchus mykiss), 금붕어(Carassius auratus), 은어(Plecoglossus altivelis) 로부터 분리된 Cytophaga columnaris의 특성)

  • Park, Se-chang;Heo, Gang-joon
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.371-378
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    • 1998
  • Five strains of gram-negative and yellow-pigmented bacteria were recently isolated from diseased freshwater fishes in Korea. All isolates were confirmed as a known fish pathogen of columnaris disease, Cytophaga columnaris based on their colonial and cellular morphology, and on physiological, biochemical and antigenic characteristics. Although the isolates were from different fish species, their characteristics of them were very similar to those of the reference strains of C columnaris (NCMB $2248^T$ and EK 28). Also, profiles of OMPs of Korean isolates were similar to those of the reference strain, C columnaris NCMB $2248^T$ when analyzed by SDS-PAGE.

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Changes of the Osmolarity in Embryonic Body Fluid and the Maternal Ovarian Tissue of the Viviparous Teleost, Ditrema temmincki, during the Gestation Period (태생 경골어류, 망상어(Ditrema temmincki)의 임신기 동안 체내자어의 체액삼투압 및 모체 난소조직상의 변화)

  • LEE Jung Sick;CHIN Pyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.2
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    • pp.157-164
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    • 1996
  • Dry weight and body fluid osmolarity of embryo, and maternal ovarian fluid and the ovarian inner tissue of the viviparous teleost, Ditrema temmincki, changed considerably during the gestation period. After the complete absorption of the egg yolk, average dry weight of the embryo increased to 373.76 mg, and the range of total length (TL) was from 6.0 to 60.0 mm. Osmolarity of the embryonic body fluid was 796,8 mOsmol/kg with TL 64.0 mm right before parturition. Ovarian outer membrane started to swelling clearly after fertilization, and maximized in March. The swelling of ovigerous folds was maximized in late April. Expansion of blood vessels and increase of hemocytes reached to their maximum right before parturition. The results of this study indicated that these changes are related to the nutritional and environmental adaptation of both the embryo and the maternal body during the gestation period.

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