• Title/Summary/Keyword: Antibodies to $(Na^+,\

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"A La Recherche" of Functions for the Spore Protein SASP-E from Bacillus subtilis

  • Ruzal, Sandra M.;Bustos, Patricia L.;Sanchez-Rivas, Carmen
    • Journal of Microbiology and Biotechnology
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    • v.23 no.1
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    • pp.15-21
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    • 2013
  • We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

Delayed presentation of aggravation of thyrotoxicosis after radioactive iodine therapy at Graves disease (그레이브스병에서 방사성요오드 치료 후 발생한 갑상샘항진증 악화의 지연된 발현)

  • Lee, Ji-Hyun;Na, Hyun-Jin;Park, Jin-Woo;Lee, Cheol-Ho;Han, Hyun-Jeong;Kim, Tae-Ho;Kim, Se-Hwa
    • Journal of Yeungnam Medical Science
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    • v.31 no.2
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    • pp.148-151
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    • 2014
  • Radioactive iodine (RAI) therapy is widely used for the treatment of Graves disease. After RAI therapy, 44% become hypothyroid and up to 28% remain hyperthyroid. The development of thyrotoxicosis after RAI therapy is believed to be mediated by 2 different mechanisms: a transient increased release of thyroid hormone due to radiation thyroiditis and the rare development of Graves disease due to the formation of antibodies to the thyroid-associated antigens released from the damaged follicular cells. A 55-year-old woman was hospitalized with severe headache, weight loss, and palpitation. She received a dose of 7 mCi of RAI (I-131) about 6 weeks earlier. Thyroid function test showed 7.98 ng/dL free T4, >8 ng/mL T3, < $0.08{\mu}IU/L$ thyroid stimulating hormone, and high titer thyroid stimulating immunoglobulin (TSI) (85.8 IU/L). She improved with propylthiouracil, propranolol, and steroid treatment. The TSI, however, was persistently elevated for 11 months.

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.

A Case of Hereditary Spherocytosis with Hemolytic Anemia due to Mycoplasma pneumonia (마이코플라즈마 폐렴에 의해 용혈성 빈혈이 발현된 유전성 구상 적혈구증 1례)

  • Na, Hye-Yeon;Shin, Seon-Hee;Lee, Kyu-Man;Kim, Kwang-Nam
    • Pediatric Infection and Vaccine
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    • v.16 no.2
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    • pp.215-219
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    • 2009
  • Mycoplasma pneumoniae is a common cause of community-acquired pneumonia in children, with a peak incidence at 5-14 years. Extrapulmonary manifestations occur in 20-25% of patients with M. pneumoniae infection. Most auto-antibodies that cause immune hemolytic anemia in humans are cold agglutinins. The formation of cold agglutinins is frequently observed during M. pneumoniae infections, and cold agglutinin disease usually occurs during M. pneumoniae infections. Nevertheless, severe hemolysis is exceptional. If a patient has any underlying disease related to hemolysis, it is possible to accelerate hemolysis. Hereditary spherocytosis is a common cause of hereditary hemolytic anemia resulting from red blood cell membrane defects. Hemolysis of red cells may result from corpuscular abnormalities or extracorpuscular abnormalities, such as immune or non-immune mechanisms. We report a case of hereditary spherocytosis associated with severe hemolytic anemia due to Mycoplasma pneumonia.

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Relationship of Antibodies in Egg Yolk and Serum against Major Antigens of Bacterial Agents in Porcine Atrophic Rhinitis, Pneumonic Pasteurellosis and Pleuropneumonia (돼지 위축성 비염, 파스튜렐라성 폐렴 및 흉막폐렴 원인균의 주요 항원에 대한 IgG 와 IgY 의 상관 관계 분석)

  • Shin, Na-Ri;Kim, Jong-Man;Yoo, Han-Sang
    • Korean Journal of Veterinary Research
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    • v.42 no.3
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    • pp.371-376
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    • 2002
  • Swine respiratory diseases have induced severe economic lasses in swine industry worldwide. Therefore, several methods have been made and applied to prevent and control the diseases. However, these methods still have a problem and also induce side effects. Recently, the use of egg yolk antibody was introduced to control and prevent the diseases as one of new trials. As a study of using egg yolk antibody, antibody titers against several different antigens of major pathogens in swine respiratory diseases were compared in egg yolk and serum of hens immunized with those antigens. The titers were measured by ELISA using the antigens as coating antigens. The relationship in antibody titers between egg yolk and serum were identified by analysis of variance for linear regression. Almost of antigens used in this study showed the high relationship in antibody titers between egg yolk and serum (r = 0.87 ~ 0.93) even though the relationship in antibody titers against P. multocida A:3 IROMP was slightly low (r = 0.74)(P<0.01). These results indicated that antibody titer in egg yolk could be useful to predict the titer in serum of chicken.

Production of Monoclonal Antibody to Newcastle Disease Virus and its Diagnostic Use (뉴켓슬병 바이러스에 대한 단크론성 항체생산 및 진단에 이용)

  • Chung Ok Choi;Chung Gil Lee;Sung Man Cho;Jin Soo Na;Soo Hwan An;Joon Hun Kwon
    • Korean Journal of Poultry Science
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    • v.15 no.3
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    • pp.207-210
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    • 1988
  • A total of 3 hybridoma clones producting menoclonal antibody (MCA) against Newcastle disease virus(NDV) was raised by cell fusion method. The MCAs did not cross react against other avian or mammalian viruses tested. However, these antibodies reacted with all strains of velogenic and lentogenic NDVs tested indicating that they are unable to discriminate the possible antigenic differences among NDVs. All. the MCAs were classified as IgG type and did not show neutralizing and hemagglutination inhibition (HAI) activity except one clone which has low HAI activity. One of these MCA raised in mouse ascites revealed the titer of $10^6$ by indirect immunofluorescent antibody (IFA) test Using the MCA, virulent NDV could easily be detected from tracheal and conjunctival smears made 2 to 3 days after experimental infection.

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Identification of a Technique Optimized for the Isolation of Spermatogonial Stem Cells from Mouse Testes

  • Han, Na Rae;Park, Hye Jin;Lee, Hyun;Yun, Jung Im;Choi, Kimyung;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.327-336
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    • 2018
  • To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

Hard tissue formation after direct pulp capping with osteostatin and MTA in vivo

  • Ji-Hye Yoon;Sung-Hyeon Choi ;Jeong-Tae Koh ;Bin-Na Lee ;Hoon-Sang Chang;In-Nam Hwang; Won-Mann Oh;Yun-Chan Hwang
    • Restorative Dentistry and Endodontics
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    • v.46 no.2
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    • pp.17.1-17.9
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    • 2021
  • Objectives: In recent in vitro study, it was reported that osteostatin (OST) has an odontogenic effect and synergistic effect with mineral trioxide aggregate (MTA) in human dental pulp cells. Therefore, the aim of this study was to evaluate whether OST has a synergistic effect with MTA on hard tissue formation in vivo. Materials and Methods: Thirty-two maxillary molars of Spraque-Dawley rats were used in this study. An occlusal cavity was prepared and the exposed pulps were randomly divided into 3 groups: group 1 (control; ProRoot MTA), group 2 (OST 100 μM + ProRoot MTA), group 3 (OST 10 mM + ProRoot MTA). Exposed pulps were capped with each material and cavities were restored with resin modified glass ionomer. The animals were sacrificed after 4 weeks. All harvested teeth were scanned with micro-computed tomography (CT). The samples were prepared and hard tissue formation was evaluated histologically. For immunohistochemical analysis, the specimens were sectioned and incubated with primary antibodies against dentin sialoprotein (DSP). Results: In the micro-CT analysis, it is revealed that OST with ProRoot MTA groups showed more mineralized bridge than the control (p < 0.05). In the H&E staining, it is showed that more quantity of the mineralized dentin bridge was formed in the OST with ProRoot MTA group compared to the control (p < 0.05). In all groups, DSP was expressed in newly formed reparative dentin area. Conclusions: OST can be a supplementary pulp capping material when used with MTA to make synergistic effect in hard tissue formation.

Expression of CYP2A6, CYP2D6 and CYP4A11 Polymorphisms in COS7 Mammalian Cell Line

  • Lee, Hye-Ja;Park, Mi-Kyung;Park, Young-Ran;Kim, Dong-Hak;Yun, Chul-Ho;Chun, Young-Jin;Shin, Hee-Jung;Na, Han-Sung;Chung, Myeon-Woo;Lee, Chang-Hoon
    • Toxicological Research
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    • v.27 no.1
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    • pp.25-29
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    • 2011
  • The cytochrome P450 (P450, CYP) are the superfamily of heme-containing monooxygenase enzymes, found throughout all nature including mammals, plants, and microorganisms. Mammalian P450 enzymes are involved in oxidative metabolism of a wide range of endo- and exogenous chemicals. Especially P450s involved in drug metabolisms are important for drug efficacy and polymorphisms of P450s in individuals reflect differences of drug responses between people. To study the functional differences of CYP2A6, CYP2D6, and CYP4A11 variants, we cloned the four CYP2A6, three CYP2D6, and three CYP4A11 variants, which were found in Korean populations, in mammalian expression vector pcDNA by PCR and examined their expressions in COS-7 mammalian cells using immunoblots using P450 specific polyclonal antibodies. Three of four CYP2A6, two of three CYP4A11, and two of three CYP2D6 variants showed expressions in COS-7 cells but the relative levels of expressions are remarkably different in those of each variants. Our findings may help to study and explain the differences between functions of CYP variants and drug responses in Korean populations.

High Levels of Antibodies to Plasmodium falciparum Liver Stage Antigen-1 in Naturally Infected Individuals in Myanmar

  • Lee, Hyeong-Woo;Moon, Sung-Ung;Kim, Yeon-Joo;Cho, Shin-Hyeong;Lin, Khin;Na, Byoung-Kuk;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.46 no.3
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    • pp.195-198
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    • 2008
  • Plasmodium falciparum liver stage antigen-1 (PfLSA-1) is one of the few antigens expressed exclusively in liver stage parasites. In this study, we evaluated the antibody responses against recombinant PfLSA-1 in naturally infected individuals in Myanmar. High levels of antibody responses (70.7%) were detected in 82 serum samples from 116 infected individuals, and IgG responses to PfLSA-1 principally composed of responses of IgG1 and IgG3 subclasses. These results show that PfLSA-1 elicits effective antibody responses in individuals infected with P. falciparum, and thus it could be not only an attractive candidate protein for vaccine development, but also a useful antigen for serodiagnosis of the infection.