• 제목/요약/키워드: Molecular Surface

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Intracellular Signaling Pathways for Type II IgE Receptor (CD23) Induction by Interleukin - 4 and Anti - CD40 Antibody

  • Kim, Hyun-Il;Park, Hee-Jeoung;Lee, Choong-Eun
    • BMB Reports
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    • 제30권6호
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    • pp.431-437
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    • 1997
  • Since the role of CD40 on the interleukin-4(IL-4) -induced B cell activation has been strongly implicated in the agumentation of IgE production and response, we have investigated the intracelluar signaling pathways utilized by IL-4 and CD40 for type II IgE receptor (CD23) expression. IL-4 and anti-CD40 antibody treatment of human B cells, independently caused a rapid induction of CD23 gene activation within 2 h. There was a noticeable synergism between the action of the two agents inducing CD23 expression: the addition of anti-CD40 to the IL-4-treated culture significantly agumented the IL-4-induced CD23 on both mRNA and surface protein levels, and the inclusion of IL-4 in the anti-CD40-treated cells caused a further increase of CD23 expression far above the maximal level induced by anti-CD40. Protein tyrosine kinase (PTK) inhibitors effectively suppressed the both IL-4- and anti -CD40-induced CD23 expression. whereas protein kinase C (PKC) inhibitors had no effects. Electrophoretic mobility shift assays (EMSA) have shown that IL-4 and anti-CD40 induce the activation of NF-IL-4 and $NF-_{K}B$, respectively, binding to the CD23 promoter, both in a PKC-independent and PTK-dependent manner. These data suggest that the synergistic activation of CD23 gene expression by IL-4 and anti-CD40 is mediated by co-operative action of distinct nuclear factors. each of which is rapidly activated via PKC-independent and PTK-dependent process.

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Unraveling Haplotype Diversity of the Apical Membrane Antigen-1 Gene in Plasmodium falciparum Populations in Thailand

  • Lumkul, Lalita;Sawaswong, Vorthon;Simpalipan, Phumin;Kaewthamasorn, Morakot;Harnyuttanakorn, Pongchai;Pattaradilokrat, Sittiporn
    • Parasites, Hosts and Diseases
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    • 제56권2호
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    • pp.153-165
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    • 2018
  • Development of an effective vaccine is critically needed for the prevention of malaria. One of the key antigens for malaria vaccines is the apical membrane antigen 1 (AMA-1) of the human malaria parasite Plasmodium falciparum, the surface protein for erythrocyte invasion of the parasite. The gene encoding AMA-1 has been sequenced from populations of P. falciparum worldwide, but the haplotype diversity of the gene in P. falciparum populations in the Greater Mekong Subregion (GMS), including Thailand, remains to be characterized. In the present study, the AMA-1 gene was PCR amplified and sequenced from the genomic DNA of 65 P. falciparum isolates from 5 endemic areas in Thailand. The nearly full-length 1,848 nucleotide sequence of AMA-1 was subjected to molecular analyses, including nucleotide sequence diversity, haplotype diversity and deduced amino acid sequence diversity and neutrality tests. Phylogenetic analysis and pair-wise population differentiation ($F_{st}$ indices) were performed to infer the population structure. The analyses identified 60 single nucleotide polymorphic loci, predominately located in domain I of AMA-1. A total of 31 unique AMA-1 haplotypes were identified, which included 11 novel ones. The phylogenetic tree of the AMA-1 haplotypes revealed multiple clades of AMA-1, each of which contained parasites of multiple geographical origins, consistent with the $F_{st}$ indices indicating genetic homogeneity or gene flow among geographically distinct populations of P. falciparum in Thailand's borders with Myanmar, Laos and Cambodia. In summary, the study revealed novel haplotypes and population structure needed for the further advancement of AMA-1-based malaria vaccines in the GMS.

PCR-RFLP for Rapid Subtyping of Plasmodium vivax Korean Isolates

  • Kang, Jung-Mi;Lee, Jinyoung;Kim, Tae Im;Koh, Eun-Ha;Kim, Tong-Soo;Sohn, Woon-Mok;Na, Byoung-Kuk
    • Parasites, Hosts and Diseases
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    • 제55권2호
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    • pp.159-165
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    • 2017
  • Vivax malaria reemerged in Korea in 1993 and the outbreak has been continued with fluctuating numbers of annual indigenous cases. Understanding the nature of the genetic population of Plasmodium vivax circulating in Korea is beneficial for the knowledge of the nationwide parasite heterogeneity and in the implementation of malaria control programs in the country. Previously, we analyzed polymorphic nature of merozoite surface protein-1 (MSP-1) and MSP-$3{\alpha}$ in Korean P. vivax population and identified the Korean P. vivax population has been diversifying rapidly, with the appearance of parasites with new genetic subtypes, despite the recent reduction of the disease incidence. In the present study, we developed simple PCR-RFLP methods for rapid subtyping of MSP-1 and MSP-$3{\alpha}$ of Korean P. vivax isolates. These PCR-RFLP methods were able to easily distinguish each subtype of Korean P. vivax MSP-1 and MSP-$3{\alpha}$ with high accuracy. The PCR-RFLP subtyping methods developed here would be easily applied to massive epidemiological studies for molecular surveillance to understand genetic population of P. vivax and to supervise the genetic variation of the parasite circulating in Korea.

Detecting the Honeycomb Sandwich Composite Material's Moisture Impregnating Defects by Using Infrared Thermography Technique

  • Kwon, Koo-Ahn;Park, Hee-Sang;Choi, Man-Yong;Park, Jeong-Hak;Choi, Won-Jae
    • 비파괴검사학회지
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    • 제37권2호
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    • pp.99-105
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    • 2017
  • Many composite materials are used in the aerospace industry because of their excellent mechanical properties. However, the nature of aviation exposes these materials to high temperature and high moisture conditions depending on climate, location, and altitude. Therefore, the molecular arrangement chemical properties, and mechanical properties of composite materials can be changed under these conditions. As a result, surface disruptions and cracks can be created. Consequently, moisture-impregnating defects can be induced due to the crack and delamination of composite materials as they are repeatedly exposed to moisture absorption moisture release, fatigue environment, temperature changes, and fluid pressure changes. This study evaluates the possibility of detecting the moisture-impregnating defects of CFRP and GFRP honeycomb structure sandwich composite materials, which are the composite materials in the aircraft structure, by using an active infrared thermography technology among non-destructive testing methods. In all experiments, it was possible to distinguish the area and a number of CFRP composite materials more clearly than those of GFRP composite material. The highest detection rate was observed in the heating duration of 50 mHz and the low detection rate was at the heating duration of over 500 mHz. The reflection method showed a higher detection rate than the transmission method.

직접모사법을 이용한 지구 저궤도 파라볼릭 안테나 탑재 위성의 항력 예측 (Prediction of Parabolic Antenna Satellite Drag Force in Low Earth Orbit using Direct Simulation Monte Carlo Method)

  • 신소민;나경수;이주영;조기대
    • 한국항공우주학회지
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    • 제42권7호
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    • pp.616-621
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    • 2014
  • 저궤도에서 운용되는 위성은 대기 저항에 의한 연료소모가 크며, 연료소모는 임무수명 및 발사무게에 영향을 미치게 되어 위성 형상에 따른 항력의 예측이 중요하다. 본 논문에서는 직접모사법을 이용하여 파라볼릭 안테나를 탑재한 저궤도 위성의 임무고도의 변화와 받음각에 따른 항력 및 항력 계수의 변화를 살펴보았다. 저궤도의 희박 기체의 거동을 모사하는 직접모사법의 적용성을 검증하기 위해 스타샤인(Starshine) 위성의 비행데이터를 이용하여 고도, 대기와 표면의 상호작용에 따른 항력 계수를 비교하였다. 결론적으로 계산결과로부터 저궤도 위성의 정밀한 궤도수명 계산에 적합한 항력 계수를 도출하였다.

Lack of O-Polysaccharide Renders Bradyrhizobium japonicum More Resistant to Organic Acid Stress

  • OH , EUN-TAEX;JU, YOUNG-JUN;KOH, SUNG-CHEOL;KIM, YONG-HWI;KIM, JONG-SUL;SO, JAE-SEONG
    • Journal of Microbiology and Biotechnology
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    • 제14권6호
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    • pp.1324-1326
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    • 2004
  • In previous studies, we isolated an isogenic LPS mutant of Bradyrhizobium japonicum 61A101C, which was completely devoid of O-polysaccharide and had altered cell surface characteristics. Subsequently, the mutated gene was identified, cloned, and used to complement the LPS mutant strain JS314 to restore the phenotype. Since it has been reported that in Escherichia coli LPS O-polysaccharide is involved in resistance to an organic acid such as acetic acid under low pH (Barna et al., Molecular Microbiology 43: 629-640, 2002), we compared the organic acid resistance of the three B. japonicum strains; wild-type 61A101C, the LPS mutant JS314, and the complemented strain to determine whether the role of O-polysaccharide in the resistance to organic acid could be generalized. Growth of all three strains was inhibited by the presence of 3 mM acetic acid under acidic condition (pH 5.5). To our surprise, however, in the presence of 2 mM acetic acid, wild-type and the complemented strains did not grow while the $LPS^-$ mutant showed a significant growth. Therefore, unlike in E. coli, the lack of O­polysaccharide of LPS appears to render B. japonicum more resistant to organic acid.

Cold-Seep Sediment Harbors Phylogenetically Diverse Uncultured Bacteria

  • Cho, Jae-Chang;Lee, Sang-Hoon;Oh, Hae-Ryun;Lee, Jung-Hyun;Kim, Sang-Jin
    • Journal of Microbiology and Biotechnology
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    • 제14권5호
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    • pp.906-913
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    • 2004
  • A culture-independent molecular phylogenetic survey was carried out on the bacterial community in cold-seep sediment at Edison Seamount, south of Lihir Island, Papua New Guinea. Small-subunit rRNA genes were amplified directly from the sediment DNA by PCR and cloned. The majority of the cloned 16S rRNA gene sequences were most closely related to as-yet-uncultivated microorganisms found in deep-sea sediments, and were primarily affiliated with one of four groups: the $\gamma$-, $\delta$-, and $\epsilon$-subdivisions of Proteobacteria, and Cytophaga-Flavobacterium-Bacteroides. We did not recover any sequences related to cyanobacteria, prochlorophytes, and $\alpha$-Proteobacteria, which are known to occur in great abundance within the surface mixed layer of the Atlantic and Pacific Oceans. The majority of the cloned $\gamma$-and $\epsilon$-Proteobacterial sequences were closely related to chemoautotrophic sulfur-oxidizing symbionts of marine benthic fauna, and the $\delta$-Proteobacterial sequences to sulfate- and sulfur-reducing bacteria, indicating that they might play an important role in chemoautotrophic primary production and the sulfur cycle in the cold-seep area. There results demonstrate the high diversity of the bacterial community in the cold-seep sediment, and substantially expand knowledge of the extent of bacterial diversity in this formidable and unique habitat.

Caco-2 세포 단층막 투과 실험시 교반이 약물의 투과계수에 미치는 염향 (Effect of Agitation on the in vitro Permeability of Xenobiotics across Caco-2 Cell Monolayers)

  • 홍순선;유호정;이홍;정석재;김대덕;심창구
    • Journal of Pharmaceutical Investigation
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    • 제35권2호
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    • pp.111-116
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    • 2005
  • The unstirred water layer (UWL), which has been known to exist in the boundary of the intestinal lumen and intestinal wall, often behaves as an absorption barrier especially for lipophilic drugs. The intestinal absorption of drugs is often characterized using Caco-2 cell monolayers grown on Transwell polycarbonate membranes. The permeability $(P_{app})$ of drugs across the cell monolayer might be influenced by the agitation of the donor compartment, since the width of UWL on the surface of the cell monolayer would be reduced by the agitation. In this study, the effect of agitation of the donor compartment with 60 rpm on the permeability was measured for 12 drugs with a wide range of lipophilicity and permeability. The $P_{app}$ of mannitol, tributylmethyl ammonium, cimetidine, ranitidine, hydrocortisone, benzylpenicillin and loxoprofen was not influenced by the agitation, while the $P_{app}$ of theophylline, propranolol, YH439, phenylpropanolamine and testosterone was increased by the agitation. There was a significant correlation between the increase of $P_{app}$ by agitation and the lipophilicity for the compounds having $P_{app}>2{\times}10^{-5}$ cm/sec. No correlation was observed for the difference in $P_{app}$ by agitation and the molecular weight, or lipophilicity of the drugs. Therefore, the agitation rate of the donor compartment in the Caco-2 cell monolayer study should be carefully controlled in order to estimate $P_{app}$ reproducibly especially for lipophilic drugs.

Interaction of Bovine Growth Hormone with Buffalo Adipose Tissue and Identification of Signaling Molecules in Its Action

  • Sodhi, R.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권7호
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    • pp.1030-1038
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    • 2007
  • Results on localization of growth hormone receptor (GHR), interaction of growth hormone (GH) with receptor in buffalo adipose tissue and identification of activated signaling molecules in the action of GH are presented. Bovine GH (bGH) was labeled with fluorescein or biotin. Fluorescein-labelled bGH was used for localization of GHRs in buffalo adipocytes. The receptors were present on the cell surface. The affinity of binding of GH to its receptor was determined by designing an experiment in which buffalo adipose tissue explants, biotinylated GH and streptavidin-peroxidase conjugate were employed. The affinity constant was calculated to be $2{\times}10^8M^{-1}$. The receptor density on adipose tissue was found to be 1 femto mole per mg of tissue. Signalling molecules generated in the action of GH were tentatively identified by employing Western blot and enhanced chemiluminescence techniques using anti-phosphotyrosine antibody. Based on molecular weights of proteins reactive to anti-phosphotyrosine antibody, three signaling molecules viz. insulin receptor substrate, Janus activated kinase (Jak) and mitogen activated protein were tentatively identified. These signaling molecules appeared in a time (incubation time of explants with growth hormone) dependent way. The activation of Jak2 was confirmed by employing anti-Jak2 antibody in a Western blot. The activation of Jak2 occurred during 5 min incubation of buffalo adipose tissue explants with GH and incubation for an additional period, viz. 30 min. or 60 min., resulted in a drastic reduction in activation. The results suggest that Jak2 activation is an early event in the action of GH in buffalo adipose tissue.

Bacteriocinogenic Potential of Newly Isolated Strains of Enterococcus faecium and Enterococcus faecalis from Dairy Products of Pakistan

  • Javed, Imran;Ahmed, Safia;Ali, Muhammad Ishtiaq;Ahmad, Bashir;Ghumro, Pir Bux;Hameed, Abdul;Chaudry, Ghulam Jilani
    • Journal of Microbiology and Biotechnology
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    • 제20권1호
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    • pp.153-160
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    • 2010
  • The present study was carried out for the isolation of bacteriocin-producing enterococci from indigenous sources. Gram-positive enterococci are known for having the ability to produce enterocins with good antimicrobial potential. A total of 34 strains were isolated from processed dairy products of Pakistan and seven out of them were found to be member of genus Enterococcus on selective enumeration. Biochemical and molecular characterization revealed that four of these isolates (IJ-03, IJ-07, IJ-11, and IJ-12) were Enterococcus faecalis and three (IJ-06, IJ-21, and IJ-31) were Enterococcus faecium. Local processed cheese was the source of all enterococcal isolates, except E. faecium IJ-21 and IJ-31, which were isolated from indigenous yoghurt and butter samples, respectively. Bacterial isolates were sensitive to commonly used antibiotics except methicillin and kanamycin. They also lacked critical virulence determinants, mainly cytolysin (cyl), gelatinase (gel), enterococcal surface protein (esp), and vancomycin resistance (vanA and vanB). Polymerase chain reaction amplification identified that enterocin A and P genes were present in the genome of E. faecium IJ-06 and IJ-21, whereas the E. faecium IJ-31 genome showed only enterocin P genes. No amplification was observed for genes that corresponded with the enterocins 31, AS-48, L50A, and L50B, and ent 1071A and 1071B. There were no signals of amplification found for E. faecalis IJ-11, indicating that the antimicrobial activity was because of an enterocin different from those checked by PCR. Hence, the indigenous bacterial isolates have great potential for bacteriocin production and they had antibacterial activity against a variety of closely related species.