• Title/Summary/Keyword: two antigens

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Detection of Circulating Melanoma Cells by a Two-marker Polymerase Chain Reaction Assay in Relation to Therapy

  • Bitisik, Ozlem;Camlica, Hakan;Duranyildiz, Derya;Tas, Faruk;Kurul, Sidika;Dalay, Nejat
    • BMB Reports
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    • v.36 no.2
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    • pp.173-178
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    • 2003
  • Malignant melanoma is one of the most rapidly increasing cancer types, and patients with metastatic disease have a very poor prognosis. Detection of metastatic melanoma cells in circulation may aid the clinician in assessing tumor progression, metastatic potential, and response to therapy. Tyrosinase is a key enzyme in melanine biosynthesis. The gene is actively expressed in melanocytes and melanoma cells. Melan A is a differentiation antigen that is expressed in melanocytes. The presence of these molecules in blood is considered a marker for circulating melanoma cells. In this study, we analyzed the usefulness of this marker combination I evaluating the response to therapy in the blood of 30 patients with malignant melanoma. Circulating cells were detected by a reverse-transcriptase-polymerase-chain reaction. The tyrosinase expression was observed in 9 (30%) patients and Melan A in 19 (63.3%) patients before therapy. Following treatment, the tyrosinase mRNA was detected in only one patient, while Melan A transcripts were still present in 14 patients. We suggest that this molecular assay can identify circulating melanoma cells that express melanoma-associated antigens and may provide an early indication of therapy effectiveness.

Characteristics of Thirty-Six Bacillus thuringiensis Isolates and a New Serovar of B. thuringiensis subsp. kim (Serotype H52)

  • Kim, Soo-Young;Kang, Min-Ho;Choi, Hee-Baeg;Lee, Jee-Un;Charles, Jean Francois;Dumanoir, Veronique Cosmao;Lecadet, Marguerite M.;Lee, Hyung-Hoan
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.534-540
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    • 1999
  • Thirty-seven strains of Bacillus thuringiensis were isolated from Korean soil and examined for H-antigen serotyping, toxicity, and different spectra of biological activities. The isolate HL-175 bore a specific H-antigen, different from the 51 known serotypes, a spherical $\delta$-endotoxin crystal, and minor different biochemical characteristics. It was resistant to ampicillin, colistin, and penicillin G. Therefore, it was classified as a new serotype, H52, with the name kim. The other 36 isolates also produced endotoxin crystals and endospores. The crystal shape of eight strains was cuboidal while the others were bipyramidal. Biochemical characteristics of the isolates were only slightly different from the known serotypes of B. thuringiensis. The flagellar (H) antigens of the 36 isolates were identified as: one colmeri (H21), three galleriae (H5a,5b); two pakistani (H13); one toumanoffi (H11a, 11b); and twenty-nine kurstaki (H3a,3b). All 36 isolates were resistant to ampicillin, colistin, penicillin, cephalothin, and chloramphenicol.

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Multi-Objective Optimum Shape Design of Rotor-Bearing System with Dynamic Constraints Using Immune-Genetic Algorithm (면역.유전 알고리듬을 이용한 로터 베어링시스템의 다목적 형상최적설계)

  • Choe, Byeong-Geun;Yang, Bo-Seok
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.24 no.7 s.178
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    • pp.1661-1672
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    • 2000
  • An immune system has powerful abilities such as memory, recognition and learning how to respond to invading antigens, and has been applied to many engineering algorithms in recent year. In this pap er, the combined optimization algorithm (Immune- Genetic Algorithm: IGA) is proposed for multi-optimization problems by introducing the capability of the immune system that controls the proliferation of clones to the genetic algorithm. The optimizing ability of the proposed combined algorithm is identified by comparing the result of optimization with simple genetic algorithm for two dimensional multi-peak function which have many local optimums. Also the new combined algorithm is applied to minimize the total weight of the shaft and the transmitted forces at the bearings. The inner diameter oil the shaft and the bearing stiffness are chosen as the design variables. The dynamic characteristics are determined by applying the generalized FEM. The results show that the combined algorithm and reduce both the weight of the shaft and the transmitted forces at the bearing with dynamic conatriants.

Diagnosis of bovine virus diarrhea-mucosal disease in breeding Holstein cows with diarrhea and vesicles (설사 및 수포발생 육성유우에서 바이러스성 설사.점막병의 진단)

  • 이성효;한수철;이종오;윤여백;송희종;채효석
    • Korean Journal of Veterinary Service
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    • v.20 no.4
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    • pp.331-338
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    • 1997
  • The purpose of this study was to establish early diagnosis for bovine virus diarrhea-mucosal disease(BVD-MD) Two Holstein among 22 breeding cows were shown ulcer in the mouth and watery diarrhea. Diarrheal feces and ulcerous lesion of the mouth from 2 cows were sampled for detection of viral antigen. BVD virus was isolated by inoculation of the samples to MDBK cells, and the cytopathic effects were observed in cultured MDBK cells which inoculated with virus isolates from the feces. Viral antigens were detected in the feces and ulceruous lesion by immunogold staining. The serum neutralization titers were shown 1 : 64 or greater in 8 blood samples by using BVD virus (NADL strain). By the RT-PCR, using reverse primer 5'-ACTCCATGTGCCATGTACAG-3', forward primer 5'-ACTCCATGTGCCATGTACAG-3', 285 base pair band specific to BVD virus was detected. In conclusions, the results of above tests which executed using the diarrheal feces and ulcerous lesion of the mouth and the isolates were conformed as BVD virus.

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Studies on the Serological Classification for Korean aspergilli (한국산 Aspergilli의 혈청학적 분류방법)

  • 문희주;이배함
    • Korean Journal of Microbiology
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    • v.12 no.4
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    • pp.180-187
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    • 1974
  • Of the Asp. spp. isolated by the Institute of Applied Microbiology, Kon-Kuk University, 7 strains were selected for the study of the immunological differencences among them using gel precipitation test. The strains were the following types : 1 type of flavus and 2 types of oryzae were isolated from Meju ; 1 type of flavus from Nuruk ; and each one type of flavus, parasiticus and oryzae from Kokja.Asp.flavus from ATCC, Asp. parasiticus nad Asp. niger NRRL strains were also used in the study as a standard. From this study, several points can be raised ; 1) There was no common antigenic property between Asp. niger and Asp. flavus, because of no formation of reaction line. Therefore, all strains could be easily distinguished. 2) There was common antigenic property, that is, the formation of reaction line between Asp. flavus and Asp. parasticus. Accordingly two strains could not be easily distinguished by the gel precipitation test. 3) Each type of oryzae, parasiticus and flavus of Asp. flavus group had common antigen one another as well as specific antigens only in the difference of the reaction lines, so they could be easily identified in the gel precipitation test. 4) Each isolated strain from Meju and Nuruk appeared to be identical. 5) It was shown that the gel precipitation test of serological methods was very useful for the classification of Asp. spp.

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Identification of Soybean Glycinin Precursor In Vitro (대두 세포내에서 Glycinin 전구체의 존재 확인)

  • 김정호
    • Journal of Plant Biology
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    • v.32 no.1
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    • pp.51-65
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    • 1989
  • Glycinin is the major storage protein in soybean. It has been known that a molecule of glycinin is composed of 6 subunits, each of which consists of two different kinds of polypeptides, acidic (A) and basic (B) one (NW 39K and 19K, respectively). To study the molecular origin and the relationship of glycinin subunit polypeptides, antibodies against A-and B-polypeptide were obtained by immunizing rabbits with either of the antigens purified by gel filtration and preparative electrophoresis. Each antibody was not only specific for its own antigen polypeptide in soybeans but also recoginzed the precursor which was synthesized in vivo and in vitro. The polyadenylated mRNAs were isolated from immature seeds and leaves and were translated in vitro using wheat germ extract. One of the seed-specific translation products. MW 60K, was identified to be the precursor of glycinin subunit by immunoprecipitation with antibodies against glycinin A- and B-polypeptide. Mature A- and B-polypeptides were not detected in the translte in vitro. These results suggest that the precursor polypeptide is synthesized from the mRNA and is cleaved to yield A- and B-polypeptides which from a glycinin subunit in the cell. Glycinin genes were expressed with the maturation of soybean seeds in a tissue-specific and developmental stage-specific manner.

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Characterization of Ubiquitinated Lysosomal Membrane Proteins in Acanthamieba castellanii

  • Oh, Sekyung;Ahn, Tae-In
    • Animal cells and systems
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    • v.4 no.2
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    • pp.165-171
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    • 2000
  • Ubiquitinated proteins in lysosomes were characterized by using two monoclonal antibodies (mAbs): LYS8-1, a mAb to lysosomal proteins, and NYA124, a mAb to ubiquitin. LYS8-1 stained lysosome-like vesicles in immunofluorescence microscopy of Amoeba proteus and Acanthamoeba castellanii. In immunoblotting, LYS8-1's antigens (LYS proteins) were detected as 68-kDa and 77-kDa proteins in A. proteus, and as 30-kDa and 39-kDa proteins in A. castellanii. In immunoprecipitation of A. castellanii, at least four distinct LYS proteins, LVS35p, LyS39p, LyS42p, and LYS46p, were detected and accumulated upon inhibition of lysosome functions but not upon that of 26S proteasome functions. They were all found to be ubiquitinated, and were recovered in the lysosome fractions in subcellular fractionation experiments. In chemical fractionation analyses, LYS35p and LYS39p were demonstrated to be peripherally associated with lysosome membrane, while LYS42p and LYS46p tightly bound to the membrane. These results suggest that the LYS proteins become associated to lysosomal membrane upon ubiquitination.

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Origin of Milled Wood Lignin : An Immunocytochemical Approaches in Combination with Transmission Electron Microscopy (면역전현법(免疫電顯法)에 의한 마쇄(磨碎) 리그닌의 기원(起源)에 관한 연구(硏究))

  • Kim, Yoon-Soo;Koh, Hong-Bum
    • Journal of the Korean Wood Science and Technology
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    • v.24 no.1
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    • pp.68-74
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    • 1996
  • Present work was undertaken to investigate the origin of milled wood lignin(MWL) in the wood cell wall using immunocytochemical techniques, which can provide the information on the localization of specific antigens(MWL in the present study) to be examined. Spruce MWL dissolved in DMSO and emulsified with Freund adjuvant was injected directly into the mouse spleen. The animals were boostered at two-week intervals after the initial immunization. Blood samples were purified in standard procedures. The characteristics of antibodies against MWL were tested by indirect ELISA. Visualization of MWL was carried out using conventional indirect immunogold-labelling methods on the ultrathin sections of spruce wood. Immuno-TEM observations showed that the immunogold probes were selectively attached to secondary cell walls of spruce wood. The most intense labelling was frequently observed in the S2 layer. In contrast, gold labelling in the lignin-rich regions, such as middle lamella and cell corner was not found. The immuno-TEM provides an indication that spruce MWL originates from the S2 layer.

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Phagocytic Uptake of Surface modified PLGA Microspheres Using Dendritic Cell

  • Kim, Ji-Seon;Lee, Young-Sung;Lee, Jung-Gil;Park, Jeong-Sook;Lee, Jong-Kil;Chung, Youn-Bok;Han, Kun
    • Journal of Pharmaceutical Investigation
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    • v.41 no.3
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    • pp.185-190
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    • 2011
  • The purpose of this study was to evaluate the phagocytic uptake of surface modified PLGA microspheres containing ovalbumin (OVA) into dendritic cell. In order to find the most suitable formulation for targeted delivery to antigen presenting cells (APC), OVA was encapsulated by a double emulsion solvent evaporation method with three PLGA microspheres (PLGA 50:50, PLGA 75:25 and PLGA 85:15) and two surface modified microspheres by chitosan and sodium dodecyl sulfate (SDS). Physicochemical properties were evaluated in terms of size, zeta potential, encapsulation efficiency, different scanning calorimeter (DSC), x-ray diffraction, morphology, and OVA release test from microspheres. Phagocytic activity was estimated using dendritic cells and analyzed by fluorescence activated cell sorter (FACS). The result showed that zeta potential of PLGA particles was changed to positive by the chitosan modification. The release profile of chitosan modified PLGA microspheres exhibited sustained release after initial burst. The chitosan modified microspheres had higher phagocytic uptake than the other microspheres. Such physicochemical properties and phagocytic uptake studies lead us to conclude that chitosan modified microspheres is more suitable formulation for the targeted delivery of antigens to APC compared with the other microspheres.

Immunological Properties of Recombinant Hepatitis B Surface Antigen Expressed in Mammalian Cell(C127)

  • Lee, Young-Soo;Kim, Byong-Kak;Choi, Eung-Chil
    • Archives of Pharmacal Research
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    • v.21 no.5
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    • pp.543-548
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    • 1998
  • We examined the immunological properties of the recombinant hepatitis B surface antigen (r-HBsAg) which was expressed in mammalian cell (C127). The cross-immunity of r-HBsAg and plasma-derived hepatitis B surface antigen (p-HBsAg) were tested using Western blotting and ELISA with guinea pig polyclonal antibody and naturally infected human-derived antibody and the both antigens show the same results in their response pattern and intensity, which indicate they have a good cross-immunity. from the measurement of $ED_{50}$ after formalin- or heat-inactivation, both r-HBsAg and p-HBsAg and p-HBsAg showed $ED_{50}$ of 0.2-0.3 in formalin-inactivaton, while r-HBsAg was 0.05-0.09 and p-HBsAg was 0.03-0.07 in heat-inactivation, which means heat-inactivation method is 3-4 times superior in immunogenicity. In the immunopersistency test performed in guinea pig for the period of 3 months with two different adjuvants, antibody titer was 34.2 with muramyl dipeptide adjuvant, which was 1.8 times greater than the antibody titer of 18.9 with $AIPO_{4}$ adjuvant. the mutagenicity of r-HBsAg has the same cross-immunity with p-HBsAg, and heat-inactivation method and muramyl dipeptide adjuvant allow development of r-HBsAg vaccine with excellent immunogenicity.

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