• Title/Summary/Keyword: alpha-fetoprotein

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Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • Biomedical Science Letters
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    • v.7 no.3
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    • pp.103-110
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    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

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Five-Year Survival of Alpha-Fetoprotein-Producing Gastric Cancer with Synchronous Liver Metastasis: A Case Report

  • Koneri, Kenji;Hirono, Yasuo;Fujimoto, Daisuke;Sawai, Katsuji;Morikawa, Mitsuhiro;Murakami, Makoto;Goi, Takanori;Iida, Atsushi;Katayama, Kanji;Yamaguchi, Akio
    • Journal of Gastric Cancer
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    • v.13 no.1
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    • pp.58-64
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    • 2013
  • Alpha-fetoprotein-producing gastric cancer is associated with poor prognosis because of frequent liver and lymph node metastasis. We present a case with synchronous liver metastasis who survived for 5 years. A 69-year-old man with upper abdominal pain was referred to our hospital. Gastrointestinal endoscopy revealed a Borrmann II-like tumor in the lower part of the stomach. Computed tomography revealed a tumor in the left lobe of the liver. Serum alpha-fetoprotein levels were markedly increased. We performed distal gastrectomy after administering oral tegafur/gimeracil/oteracil potassium and administered hepatic intra-arterial cisplatin injection. Liver metastasis showed partial response on computed tomography. Despite left hepatic lobectomy, further metastases to the liver and mediastinal lymph nodes became difficult to control. After sorafenib tosylate administration, stabilization of the disease was observed for 4 months. We conclude that hepatic intra-arterial chemotherapy and oral administration of sorafenib tosylate may potentially improve the prognosis in such cases.

Altered Cell to Cell Communication, Autophagy and Mitochondrial Dysfunction in a Model of Hepatocellular Carcinoma: Potential Protective Effects of Curcumin and Stem Cell Therapy

  • Tork, Ola M;Khaleel, Eman F;Abdelmaqsoud, Omnia M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8271-8279
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    • 2016
  • Background: Hepato-carcinogenesis is multifaceted in its molecular aspects. Among the interplaying agents are altered gap junctions, the proteasome/autophagy system, and mitochondria. The present experimental study was designed to outline the roles of these players and to investigate the tumor suppressive effects of curcumin with or without mesenchymal stem cells (MSCs) in hepatocellular carcinoma (HCC). Materials and Methods: Adult female albino rats were divided into normal controls and animals with HCC induced by diethyl-nitrosamine (DENA) and $CCl_4$. Additional groups treated after HCC induction were: Cur/HCC which received curcumin; MSCs/HCC which received MSCs; and Cur+MSCs/HCC which received both curcumin and MSCs. For all groups there were histopathological examination and assessment of gene expression of connexin43 (Cx43), ubiquitin ligase-E3 (UCP-3), the autophagy marker LC3 and coenzyme-Q10 (Mito.Q10) mRNA by real time, reverse transcription-polymerase chain reaction, along with measurement of LC3II/LC3I ratio for estimation of autophagosome formation in the rat liver tissue. In addition, the serum levels of ALT, AST and alpha fetoprotein (AFP), together with the proinflammatory cytokines $TNF{\alpha}$ and IL-6, were determined in all groups. Results: Histopathological examination of liver tissue from animals which received DENA-$CCl_4$ only revealed the presence of anaplastic carcinoma cells and macro-regenerative nodules. Administration of curcumin, MSCs; each alone or combined into rats after induction of HCC improved the histopathological picture. This was accompanied by significant reduction in ${\alpha}$-fetoprotein together with proinflammatory cytokines and significant decrease of various liver enzymes, in addition to upregulation of Cx43, UCP-3, LC3 and Mito.Q10 mRNA. Conclusions: Improvement of Cx43 expression, nonapoptotic cell death and mitochondrial function can repress tumor growth in HCC. Administration of curcumin and/or MSCs have tumor suppressive effects as they can target these mechanisms. However, further research is still needed to verify their effectiveness.

Production of a Monoclonal Antibody to Human $\alpha$-Fetopotein and Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for Human $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein에 대한 모노클로날 항체의 제조 및 모노클로날 항체를 이용한 효소면역분석법의 개발)

  • Michung Yoon;Hyun-Hee Lee;Youngwon Lee
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.1-10
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    • 1999
  • This study was attempted to generate a monoclonal antibody against human $\alpha$-fetoprotein (AFP) and to produce an immunoassay, recognizing AFP in plasma and amniotic fluid. AFP was purified from human amniotic fluid and used to immunize mice. Spleens were taken from the mice and the cells were fused with mouse myeloma cells (Sp2/0-Ag-14) for the production of monoclonal antibodies by employing the hybridoma technology. As a result, a hybridoma cell line producing anti-AFP monoclonal antibody was cloned out and designated as MabF22. From isotyping analysis, it was found that monoclonal antibody MabF22 was IgG type with IgG1 heavy chain and k light chain. The binding specificity of MabF22 was analyzed by immunoblotting as well as by ELISA. MabF22 was highly specific, reacting with only AFP-containing samples. The binding affinity was determined by ELISA (free-capture mode) and Scatchard analysis. As a result, the value of Kd was 0.8$\times$10$^{-10}$M. The validity of the MabF22 for AFP assay was examined by two kinds of ELISAs, i.e., non-competitive and competitive ELISA. Both assays revealed that MabF22 reacted well with AFP in sample in a concentration-dependent manner. Standard curve and antibody titration curve were obtained by using purified AFP and MabF22. These results indicate that the monoclonal antibody produced in this study would be useful not only for research purposes but also for further development of immune-diagnostic kit for the measurement of AEP concentration.

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Effects of Hwangryeun Extract on the Recovery of Liver Function in $CCl_4$-exposed Rats (황련추출물이 $CCl_4$중독 Rat의 간기능 회복에 미치는 영향)

  • Kim, Byoung-Woo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.6
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    • pp.1020-1025
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    • 2011
  • Effects of Hwangryeun(Copitidis rhizoma) extract on recovery of liver function were researched in $CCl_4$ intoxicated rats. Concentration ${\alpha}$-fetoprotein, total protein in plasma showed a tendency to decrease in Hwangryeun(Copitidis rhizoma) extract applied groups while that of albumin showed a increase. Activities of plasma aspartate aminotransferase and alanine aminotransferase in Hwangryeun(Copitidis rhizoma) extract applied groups showed a lower value than those of controlled groups. Alkaline phosphatase, lactate dehydrogenase and ${\gamma}$-glutamyl transpeptidase activities showed a tendency to decrease in Hwangryeun(Copitidis rhizoma) extract applied groups. Concentration of plasma triglyceride and total cholesterol showed a lower values than that of control group.

Development of a One-step Two-site Enzyme Immunoassay for Measuring Human Alpha-fetoprotein by Eliminating Hook-effect

  • Kim, Se-Ho
    • BMB Reports
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    • v.34 no.1
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    • pp.47-50
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    • 2001
  • A one-step, two-site enzyme immunoassay was developed for measuring human alpha-fetoprotein (AFP) in serum and amniotic fluid using monoclonal antibodies (McAb) by eliminating the high-dose hook effect. Three McAbs that recognize different epitopes were selected among 16 different clones on the basis of epitope mapping, two for immobilization and one for horseradish peroxidase conjugation. This one-step immunoassay system is more convenient and rapid compared to a conventional two-step sandwich immunoassay system. It did not exhibit the hook effect to around 2.7 mg/ml of AFP, which is probably one of the highest concentrations of AFP in the serum. The dose-response curve of the system was linear to 500 mg/ml of AFP and the system could differentiate as low as 1 mg/ml of AFP The intra- and inter-assay variations were in an acceptable range; 95~104% and 97~105% respectively Its correlation with other commercial systems was around 95%.

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Effects of Administration of 2,2'-Azobis(2-amidinopropane) Dihydrochloride(AAPH) on Liver Function in Rats 1. Clinical Signs and Blood Chemical Values (2,2'-Azobis (2- amidinopropano) Dihydrochloride(AAPH)의 투여가 쥐의 간기능에 미치는 영향 1. 임상증상 및 혈액 화학치 소견)

  • 강정부;손호상;김철호
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.75-78
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    • 1998
  • This study was performed to estimate the clinical signs and changes of biochemical parameters in rats with hepatic injury induced by the administration of 2, 2'-azobis(2- amidinopropane)dihydrochloride (AAPH) . Minor behavioral change, brittleness of skin hair and decreased volume of water and feed intake were observed in ra% 2 hours after AAPH administration compared to control group. Concentration of serum albumin showed lower than that of control group. However, concentration of total bilirubin has shown higher than that of control group. As time goes on, . serum LDH activity increased significantly compared to control group, but senun CPK activity did not change compared to control group. Passive hemanglutination of $\alpha $-fetoprotein was negative in all the treaDent groups and control group.

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