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Sample records for alpha fetoprotein radioimmunoassay

  1. Radioimmunoassay of ovine alpha-fetoprotein

    International Nuclear Information System (INIS)

    Lai, P.C.W.

    1978-01-01

    Highly purified ovine alpha-fetoprotein (AFP) was used both for radioisotope labelling and as the reference standard in the double antibody radioimmunoassay of ovine AFP. The sensitivity of the assay is 2 ng/ml which is about 8000 times more sensitive than radioimmunodiffusion assay. The assay is of sufficient sensitivity to quantitate AFP in normal adult sheep serum, pregnancy serum, amniotic fluid and fetal lamb serum. (Auth.)

  2. Radioimmunoassay of alpha-fetoprotein, with special reference to iodination and purification techniques

    International Nuclear Information System (INIS)

    Schiller, H.S.; Kulchinski, L.; Luthy, D.A.

    1978-01-01

    We report a relatively simple and convenient method for iodinating human alpha-fetoprotein and for purifying the 125 I-labeled material. The label is incorporated into human alpha-fetoprotein enzymatically by use of lactoperoxidase. Before each assay the labeled material is purified over two successive short columns: Sephacryl S-200 Superfine and cellulose. This procedure removes both free iodine and damaged fetoprotein. With the purified material we developed a sensitive and reliable radioimmunoassay for alpha-fetoprotein in serum and amniotic fluid

  3. The radioimmunoassay of serum alpha-fetoprotein levels

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Y H; Choi, K A; Ahn, K S; Suh, W H; Lee, M J [Korea University College of Medicine, Seoul (Korea, Republic of)

    1982-06-15

    Alpha-fetoprotein (AFP) was first described in the human fetus in 1956 and became a marker protein of primary liver cancer in adults. Serum AFP levels were measured by radioimmunoassay in 212 patients with a variety of malignant and nonmalignant diseases to determine the incidence of leveis elevated above 40 ng/ml. The results obtained are as follows: In 44 cases of total 212 patients, abnormal AFP levels above 40 ng/ml in serum were measured; 24 of 31 patients with primary hepatocellular carcinoma and primary hepatocellular carcinoma with liver cirrhosis (77.4%), 7 of 51 patients with only liver cirrhosis (13.7%), 4 of 10 patients with metastatic liver cancer (40.0%), 4 of 15 patients with chronic hepatitis (26.7%), 2 of 23 patients with acute hepatitis (8.7%), and each one patient with 6 pancreatic carcinoma and 9 cholangiocarcinoma had elevated serum AFP levels. One pregnant woman with gestation 35 weeks had elevated level, but within normal limit during pregnancy.

  4. The radioimmunoassay of serum alpha-fetoprotein levels

    International Nuclear Information System (INIS)

    Kim, Y. H.; Choi, K. A.; Ahn, K. S.; Suh, W. H.; Lee, M. J.

    1982-01-01

    Alpha-fetoprotein (AFP) was first described in the human fetus in 1956 and became a marker protein of primary liver cancer in adults. Serum AFP levels were measured by radioimmunoassay in 212 patients with a variety of malignant and nonmalignant diseases to determine the incidence of leveis elevated above 40 ng/ml. The results obtained are as follows: In 44 cases of total 212 patients, abnormal AFP levels above 40 ng/ml in serum were measured; 24 of 31 patients with primary hepatocellular carcinoma and primary hepatocellular carcinoma with liver cirrhosis (77.4%), 7 of 51 patients with only liver cirrhosis (13.7%), 4 of 10 patients with metastatic liver cancer (40.0%), 4 of 15 patients with chronic hepatitis (26.7%), 2 of 23 patients with acute hepatitis (8.7%), and each one patient with 6 pancreatic carcinoma and 9 cholangiocarcinoma had elevated serum AFP levels. One pregnant woman with gestation 35 weeks had elevated level, but within normal limit during pregnancy

  5. Dosage of alpha-fetoprotein for radioimmunoassay in maternal blood

    International Nuclear Information System (INIS)

    Pandolfo, J.; Jaumandreu, C.A.; Aguirre de Garcia, B.; Robles, A.M.; Touya, E.

    1981-01-01

    Leigthon, Gordon and col. showed that the determination of serum levels of alpha-fetoprotein was useful as a method for the detection of fetal malformations of the neural tube. Due to its important clinical significance, the study of this simple test in women in different stages of pregnancy has been started. A total of 34 patients from the obstetrical external service were studied, with pregnancy stages between 10 and 40 weeks. Having established the incidence and implications of false positive and principally of false negative results, it has been concluded that this technique should be imposed as routine in the evolutive control of pregnancy. (M.B.) [es

  6. A new procedure for coupling antibody to paper discs for radioimmunoassay: application to the determination of alpha-fetoprotein

    International Nuclear Information System (INIS)

    Sasaki, T.; Tsukada, Y.; Hirai, H.

    1983-01-01

    Horse anti-alpha-fetoprotein was coupled to CM-cellulose discs by a modified carbodiimide reaction. The resulting coupled CM-discs were used in solid-phase radioimmunoassay of human alpha-fetoprotein. The sensitivity of these discs and conventional BrCN activated filter paper discs coupled anti-alpha-fetoprotein was approximately the same. A fair correlation between the alpha-fetoprotein levels determined by both methods was observed. The coupling procedure with carbodiimide is simple and the use of hazardous BrCN is eliminated. (Auth.)

  7. Measurement of alpha-fetoprotein in maternal serum: three commercial radioimmunoassay kits and two non-commercial radioimmunoassays compared

    International Nuclear Information System (INIS)

    Forest, J.C.; Verreault, F.; Pouliot, M.

    1982-01-01

    We evaluated three commercial radioimmunoassay kits (Amersham, Dainabot, Clinical Assays) and two non-commercial methods for determining alpha-fetoprotein in maternal serum during pregnancy. All five procedures were found to be acceptable with respect to practicability, sensitivity, linearity, and precision. Similar results were obtained with Dainabot, Clinical Assays, and the two non-commercial methods, but the Amersham method revealed a proportional error, results being about 20% lower than those by the other methods. Use of the international unit system is suggested for reporting results for AFP, to facilitate comparison between methods and laboratories

  8. The experience with setting-up radioimmunoassay for alpha-1 fetoprotein

    International Nuclear Information System (INIS)

    Fingerova, H.

    1981-01-01

    The decisive factor in the preparation of radioimmunological alpha-1-fetoprotein determination, provided sufficient commercial or own antisera and standards are available for calibration, is the quality of the preparation for labelling. Alpha-1-fetoprotein was separated by affinity chromatography using Sepharose with alpha-1-fetoprotein-bound antibodies. The isolates thus obtained were labelled with 125 I using enzyme and chloramine T and Iodogen techniques. The labelled alpha-1-fetoprotein can be used for RIA. In view of reduced immunoreactivity of the preparation, however, the performance of the radioimmunological determination has so far not matched the quality of imported kits. The technique is currently being optimized. (author)

  9. Immunoradiometric versus radioimmunoassay: a comparison using alpha-fetoprotein as the model analyte

    International Nuclear Information System (INIS)

    Hunter, W.M.; Budd, P.S.

    1981-01-01

    With alpha-fetoprotein as a model a formal comparison has been made between inhibition type radioimmunoassay with radioiodinated antigen, liquid-phase incubation and double antibody separation (RIA) and variants of the sandwich immunoradiometric assay (IRMA). 125 I-antibody was prepared (expensively) by labelling the IgG fraction and subsequently undertaking immunopurification to yield a reproducibly-high quality reagent, 70-75% being reactive with antigen. The same antiserum was coupled to Sepharose 4B for use in the IRMA and separation was by the sucrose layer procedure (Hunter, 1980) which obviates the need for centrifugation. The principal basis used for the comparison was computer-generated precision profiles (Ekins, 1976), each of which was derived from 10 assays of each kind. (Auth.)

  10. Significance of radioimmunoassay of human chorionic gonadotropin and alpha fetoprotein in nonseminomatous germ cell tumors of the testis

    International Nuclear Information System (INIS)

    Kausitz, J.; Hupka, S.; Cerny, V.; Bohunicky, L.; Korec, S.

    1980-01-01

    Radioimmunoassays human chorionic gonadotropin (HCG) and alpha fetoprotein (AFP) made in 49 patients with nonseminomatous testicular tumors showed that these investigations make the diagnosis more precise, permit to follow up the dynamics of the course of the disease and the effectiveness of treatment and may help to reveal the presence of otherwise undetectable tumorous metastases. The significance of these assays is enhanced if the two tumorous proteins are investigated in parallel. The results proved positive in 43 (87.8%) and false negative in 6 (12.2%) of the patients. The absence of HCG and AFP production in some patients with active disorder has not as yet been elucidated. (author)

  11. Significance of radioimmunoassay of human chorionic gonadotropin and alpha fetoprotein in nonseminomatous germ cell tumors of the testis

    Energy Technology Data Exchange (ETDEWEB)

    Kausitz, J.; Hupka, S. (Institute for Postgradual Training of Physicians and Pharmaceutists, Bratislava (Czechoslovakia)); Cerny, V.; Bohunicky, L.; Korec, S. (Ustav Klinickej Onkologie, Bratislava (Czechoslovakia))

    1980-01-01

    Radioimmunoassays human chorionic gonadotropin (HCG) and alpha fetoprotein (AFP) made in 49 patients with nonseminomatous testicular tumors showed that these investigations make the diagnosis more precise, permit to follow up the dynamics of the course of the disease and the effectiveness of treatment and may help to reveal the presence of otherwise undetectable tumorous metastases. The significance of these assays is enhanced if the two tumorous proteins are investigated in parallel. The results proved positive in 43 (87.8%) and false negative in 6 (12.2%) of the patients. The absence of HCG and AFP production in some patients with active disorder has not as yet been elucidated.

  12. Three-site sandwich radioimmunoassay with monoclonal antibodies for a sensitive determination of human alpha-fetoprotein

    International Nuclear Information System (INIS)

    Nomura, M.; Imai, M.; Takahashi, K.; Kumakura, T.; Tachibana, K.; Aoyagi, S.; Usuda, S.; Nakamura, T.; Miyakawa, Y.; Mayumi, M.

    1983-01-01

    Utilizing monoclonal antibodies against human alpha-fetoprotein, 3 distinct antigenic determinants were identified. These antigenic determinants, provisionally designated a, b and c, were arranged in such a manner that the binding of one determinant with the corresponding antibody did not inhibit, or only barely inhibited the binding of antibodies directed to the other 2 determinants. Monoclonal antibodies with 3 different specificities were, therefore, applied to develop a sandwich-type solid-phase radioimmunoassay of the antigen in which wells were coated with anti-a, and radiolabeled anti-b together with radiolabeled anti-c was employed to detect the bound antigen. The 3-site sandwich radioimmunoassay involving 3 different determinants gave a higher sensitivity than 2-site assays in which only anti-b or anti-c was employed as a radiolabeled reagent, because the radioactivity of the 2 labeled antibodies was added on the antigen bound to immobilized anti-a. (Auth.)

  13. Alpha-fetoprotein: methodology and clinical experiences with a new radioimmunoassay

    International Nuclear Information System (INIS)

    Lamerz, R.; Rjosk, H.; Schmalhorst, U.; Fateh-Moghadam, A.; Muenchen Univ.

    1976-01-01

    The clinical-diagnostical value of alpha-fetoproteins (AFP) compared with the use of common less sensitive methods was tested. The results obtained clearly showed that in the clinical-internal-surgical sphere the main importance of radioimmunological AFP-determinations lies in the improved supervision of the development of hepatomas in patients suffering from hepatic cirrhosis. In the pre-operative differential diagnosis of testicle tumors, these determinations help to optimize the therapeutic measures and are also useful for the case control of hepatoma and teratoma patients. (GSE) [de

  14. Preparation of Double Antibody Radioimmunoassay for Determination of Alpha fetoprotein as a Tumor Marker using BALB/C Mice as Host Animals

    International Nuclear Information System (INIS)

    Abdel-Ghany, I.Y.; EI- Mouhty, N.R.A.; Shafil, H.M.; Mehany, N.L.

    2009-01-01

    The aim of the present work was to prepare liquid phase radioimmunoassay system (RIA) reagents. Development as well as optimization and validation of this RIA system for the measurement of alpha fetoprotein (AFP) in human serum are described. The production of polyclonal antibodies was carried out by immunizing four BALB/C mice subcutaneously. The preparation of 125 I-AFP tracer was performed using chloramine-T oxidation method. The preparation of AFP standards was done by diluting cord sera using assay buffer. The results obtained provide a highly sensitive,precise and accurate RIA system of AFP based on liquid phase separation. In conclusion, this assay could be used for the diagnosis and management of patients with certain malignant diseases and in prenatal diagnosis of neural tube defects

  15. Radioimmunoassay for carcinoembryonic antigen and alpha1-fetoprotein in the qualitative evaluation of focal hepatic lesions in Japan

    International Nuclear Information System (INIS)

    Aburano, T.; Tonami, N.; Tada, A.; Hisada, K.

    1980-01-01

    The combined tests of serum alpha 1 -fetoprotein (AFP) und carcinoembryonic antigen (CEA) were routinely performed in 210 patients with focal hepatic lesions on a 99 sup(m)Tc-colloid liver scan in order to determine whether these could provide more useful information that AFP test alone in the qualitative evaluation of focal hepatic lesions. The predictive value of hepatoma with positive AFP alone remained 80%. However, when the negative CEA was combined with positive AFP, the predictive value of hepatoma (91%) with both was greatly increased. On the other hand, the predictive value of metastatic liver cancer with positive CEA showed 92%. The combined Test of AFP and CEA may be useful for preserving high predictive values of hepatoma and metastatic liver disease. (orig.) [de

  16. Alpha-fetoprotein in the routine clinical laboratory: evaluation of a simple radioimmunoassay and review of current concepts in its clinical application

    International Nuclear Information System (INIS)

    Brummund, W.; Mennuti, M.T.; Arvan, D.A.; Starkovsky, N.A.

    1980-01-01

    The authors have assessed the clinical utility of a radioimmunoassay for alpha-fetoprotein (AFP). The method, which relies on ammonium sulfate precipitation for the separation of 'bound' and 'free' radiolabeled antigen, can be completed in one working day. The assay is specific for AFP, has a sensitivity of <10 ng/ml, and has intra- and inter-assay precision of 5-8% and 9-11%, respectively. They have conducted a three-year study of 472 pregnancies in which physicians wished to detect neural tube defects, and of 400 non-pregnant patients to assess the value of serum AFP as a marker for certain benign and malignant diseases. Six of 6 fetal open neural-tube defects (NTD'S) and 3 of 3 intrauterine fetal deaths were correctly identified by their association with marked AFP elevations in both maternal serum and amniotic fluid. Thirty non-pregnant patients were found to have AFP elevations greater than 20 ng/ml. Malignancies associated with these elevations were hepatoma, germ cell tumors, Wilms' tumor, and carcinoma of unknown origin. Carcinoma metastatic to the liver was not associated with AFP elevations. In AFP-associated tumors they found serial measurements of serum AFP to be of value in assessing therapeutic response. (Auth.)

  17. Studies on local preparation of double antibody radioimmunoassay for determination of alpha fetoprotein as a tumor marker

    International Nuclear Information System (INIS)

    Elkolaly, M.T.; Mehany, N.L.; Ragab, M.T.; El-Mohty, A.A.

    2001-01-01

    The preparation of primary reagents of AFP- RIA technique with low cost is considered to be one of the main objective of labelled compounds Dept., Radioisotope production Division, Hot Labs Centre, Atomic Energy Authority, Egypt. The development of a radioimmunoassay system based on local production of basic reagents for the measurement of AFP in human serum is described. The assay followed by 100 μ1 of locally prepared 125 I-AFP to 100 μ1 standards and unknown samples. After overnight incubation at 4 degree, separating reagents were added in following sequence 100 μ1 non-immune rabbit serum (1:200) followed by addition of 100μ1 of donkey anti-rabbit serum, (1:20), then 500μ1 of 8 percentage polyethylene glycol (PEG) were added into all assay tubes. The tubes were allowed to stand for 30 minutes at room temperature and then centrifuged for 15 minutes. the tubes were decanted and counted in a gamma counter. The results of the present study showed that the assay is highly sensitive where the detection limit or minimal detectable dose of AFP assay approximately 1.5 I U/ml. The reliability of the present procedure was assessed by examining its reproducibility on pooled human serum samples selected to represent different levels of AFP. The data of intra-assay and intra-assay precision revealed the consistency of the results obtained by the method of the present study. The present technique for determination of serum AFP agreed well with commercially available kits (DPC, double antibody-diagnostic product corporation, los angeles, USA). The technical simplicity of this specific, accurate and reasonably precise method may suggest that the technique should be suited for routine laboratory use

  18. Alpha-fetoprotein determination and liver scintigraphy, in patients with hematologic diseases

    International Nuclear Information System (INIS)

    Silva, W.M.

    1977-12-01

    The serum alpha-fetoprotein is quantified in 30 patients by means of the radioimmunoassay method. They are divided into 2 newborn bobies, 5 control subjects, 1 pregnant woman and 22 with hematologic deseases. Liver scanning is also performed in all of them except the newborn bobies and the control subjects. In these last patients, the alpha - fetoprotein levels vary from below 6.25 to 14.0 mg/ml. High values are only found in the newborn babies, in the pregnant woman and in the patient with hysticcitic lymphoma. In these patients the alpha - fetoprotein levels are over 14,0 mg/ml [pt

  19. A case of alpha-fetoprotein-producing esophageal adenocarcinoma.

    Science.gov (United States)

    Chen, Yi-Yu; Hsu, Wen-Hung; Hu, Huang-Ming; Wu, Deng-Chyang; Lin, Wen-Yi

    2013-02-01

    Alpha-fetoprotein is a well-known tumor marker in the screening and follow-up of hepatocellular carcinoma. In Taiwanese society, a high prevalence of hepatitis and hepatoma and elevation of alpha-fetoprotein associated with liver function impairment usually suggested clinics undertake further examination for liver or genital tumor. We report the case of 45-year-old man who was found to have an alpha-fetoprotein-producing esophageal adenocarcinoma with an initial presentation of liver function impairment and rapid elevation of alpha-fetoprotein. Esophageal cancer was diagnosed via endoscope and a biopsy proved the presence of adenocarcinoma. A small endoscopic biopsy specimen failed to identify the alpha-fetoprotein positive tumor cell. Esophagectomy was performed and histopathological study of surgical specimen revealed grade II adenocarcinoma with regional metastatic lymphadenopathy. Immunohistochemical study was focal positive for alpha-fetoprotein. Serum alpha-fetoprotein declined transiently after esophagectomy and fluctuation of alpha-fetoprotein level was noted during the treatment with adjuvant chemotherapy. Finally, 19 months after the operation, the patient died due to multiple organ metastases with multiple organ failure. Thus, a small specimen for upper endoscopy may not be sufficient in the presence of alpha-fetoprotein-producing adenocarcinoma. Monitoring of serum alpha-fetoprotein may be useful in the evaluation and follow-up of esophageal alpha-fetoprotein-producing adenocarcinoma. Copyright © 2012. Published by Elsevier B.V.

  20. The evaluation of domestic immunoradiometric assay kit for alpha-fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Won, Kyoung Sook; Ryu, Jin Sook; Moon, Dae Hyuk; Lee, Hee Kyung [Ulsan Univ. College of Medicine, Seoul (Korea, Republic of)

    2000-08-01

    Although {alpha}-fetoprotein is one of the most commonly used tumor markers in Korea, most of the radioimmunoassay kits for {alpha}-fetoprotein have been imported from foreign countries. The purpose of this study was to evaluate the performance of a recently developed domestic immunoradiometric kit for {alpha}-fetoprotein (Riakey AFP IRMA CT{sup R}, Sin-Jin Medics, Seoul, Korea). We evaluated intra-and inter-assay precision, recovery rate, parallelism, and sensitivity of serum {alpha}-fetoprotein measurement using Riakey AFP IRMA CT{sup R} kit. The values of {alpha}-fetoprotein measured by Riakey AFP IRMA CT{sup R} kit were compared with those measured by two foreign commercial kits ({alpha}-fetoproteina of Radim and {alpha}-feto.riabead of Abbott). Intra-assay coefficients of variation on three different levels were 5.3% for 18.9 ng/ml, 3.4% for 133 ng/ml and 1.6% for 330 ng/ml. Inter-assay coefficients of variation were 9.7% for 20.9 ng/ml, 3.2% for 137 ng/ml and 4.1% for 330 ng/ml respectively. Recovery rate tests on all three different levels showed within 100{+-}10%. Parallelism was also good and the sensitivity was 0.63 ng/ml. There was strong correlation between the measurement of {alpha}-fetoprotein by Riakey AFP IRMA CT{sup R} and that by two foreign commercial kits(r=3D0.98). The first Korean domestic immunoradiometric kit for {alpha}-fetoprotein, Riakey AFP IRMA CT{sup R}, performed well for clinical use.

  1. User acceptability of an alpha-fetoprotein screening programme

    DEFF Research Database (Denmark)

    Jørgensen, Finn Stener

    1995-01-01

    The objective of the study was to determine user acceptability among women who were classified as false positives or test negatives in an alpha-fetoprotein screening programme. The study was performed as a questionnaire study over a one-year period from October 1, 1988 to September 30, 1989...... at Hvidovre University Hospital, Copenhagen, and the county hospitals of Sønderjylland, Denmark. The participating subjects were 4104 pregnant women who had had an alpha-fetoprotein test and had completed 30 weeks of gestation, when the questionnaire was delivered. Main outcome measures were degree...... and duration of anxiety, influence on daily life and whether the woman wanted the alpha-fetoprotein test again in a new pregnancy. Three thousand, three hundred and thirty-one questionnaires were analyzed. The participation rate was 81.2%. For 219 women (6.6%), the first alpha-fetoprotein test was abnormal...

  2. Diagnostic value of glypican-3 in alpha fetoprotein negative ...

    African Journals Online (AJOL)

    EB

    2013-09-03

    Sep 3, 2013 ... School of Basic Medical Sciences, Capital Medical University, 100069, China. Abstract ... alpha fetoprotein(AFP) negative HCC and cirrhotic nodules is still difficult. Objectives: ..... Asia-Pacific Working Party on Prevention of.

  3. Serum concentration of alpha-1-fetoprotein suggestive of, or pathognomonic for hepatoma

    International Nuclear Information System (INIS)

    Polterauer, P.; Horak, W.; Legenstein, E.; Mueller, M.

    1979-01-01

    A short review of alpha-1-fetoprotein (AFP), is followed by a presentation of the serum AFP concentrations obtained in healthy subjects and in patients with hepatoma, cirrhosis of the liver or metastatic liver cancer, measured by radioimmunoassay (RIA). A calculation is made from these results of the upper limit of normal (9 ng/ml), a limit which is suggestive of hepatome (215 ng/ml) and a limit which is pathognomonic for hepatoma (7500 ng/ml). It is concluded that the quantitative determination of AFP by RIA represents a sensitive method which provides valuable clinical information for the early diagnosis of hepatoma. (author)

  4. comparative study between different oxidizing agents to prepare radioiodinated alpha fetoprotein (AFP)

    International Nuclear Information System (INIS)

    El-Kolaly, M.T.; Ragab, M.T.; El-Mohty, A.A.; Sallam, K.M.; Arief, M.H.

    2004-01-01

    the aim of the present study was designed to prepare four different preparation of radioiodinated alpha fetoprotein ( 125 I-Afp) using different oxidizing agents. the oxidizing agents were chloramine-T (Ch-T), lodogen (1,3,4 , 6-tetrachloro 3 α ,6α diphenyl glycoluril ), N-bromosuccinimide (NBS) and lactoperoxidase (LPS). the product was purified by gel filtration using sephadex G-25. then the tracers obtained were tested by radioimmunoassay (RIA) technique. different affecting factors were extensively studied including reaction time, reaction volume, oxidizing agent content and Ph of reaction. it was found that the Ch-T method is the best one

  5. Alpha-fetoprotein (AFP) Test: MedlinePlus Lab Test Information

    Science.gov (United States)

    ... this page: https://medlineplus.gov/labtests/alphafetoproteinafptest.html Alpha-fetoprotein (AFP) Test To use the sharing features on this page, please enable JavaScript. What is an Alpha-fetoprotein (AFP) Test? Alpha-fetoprotein (AFP) is a protein ...

  6. Diagnostic value of glypican-3 in alpha fetoprotein negative ...

    African Journals Online (AJOL)

    Background: The prognosis of patients with hepatocellular carcinoma(HCC) is generally very poor with a 5-year survival rate of less than 15% since most of them are diagnosed clinically at their late stage.However,the differential diagnosis between alpha fetoprotein(AFP) negative HCC and cirrhotic nodules is still difficult.

  7. Alpha-fetoprotein detection by using a localized surface plasmon coupled fluorescence fiber-optic biosensor

    Science.gov (United States)

    Chang, Ying-Feng; Chen, Ran-Chou; Li, Ying-Chang; Yu, Chih-Jen; Hsieh, Bao-Yu; Chou, Chien

    2007-11-01

    Alpha-fetoprotein (AFP) detection by using a localized surface plasmon coupled fluorescence (LSPCF) fiber-optic biosensor is setup and experimentally demonstrated. It is based on gold nanoparticle (GNP) and coupled with localized surface plasmon wave on the surface of GNP. In this experiment, the fluorophores are labeled on anti-AFP which are bound to protein A conjugated GNP. Thus, LSPCF is excited with high efficiency in the near field of localized surface plasmon wave. Therefore, not only the sensitivity of LSPCF biosensor is enhanced but also the specific selectivity of AFP is improved. Experimentally, the ability of real time measurement in the range of AFP concentration from 0.1ng/ml to 100ng/ml was detected. To compare with conventional methods such as enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA), the LSPCF fiber-optic biosensor performs higher or comparable detection sensitivity, respectively.

  8. Negative interference by rheumatoid factor in alpha-fetoprotein chemiluminescent microparticle immunoassay.

    Science.gov (United States)

    Wang, Hui; Bi, Xiaohui; Xu, Lei; Li, Yirong

    2017-01-01

    Background Rheumatoid factor causes positive interference in multiple immunoassays. Recently, negative interference has also been found in immunoassays in the presence of rheumatoid factor. The chemiluminescent microparticle immunoassay is widely used to determine serum alpha-fetoprotein. However, it is not clear whether the presence of rheumatoid factor in the serum causes interference in the chemiluminescent microparticle immunoassay of alpha-fetoprotein. Methods Serum alpha-fetoprotein was determined using the ARCHITECT alpha-fetoprotein assay. The estimation of alpha-fetoprotein recovery was carried out in samples prepared by diluting high-concentration alpha-fetoprotein serum with rheumatoid factor-positive or rheumatoid factor-negative serum. Paramagnetic microparticles coated with hepatitis B surface antigen-anti-HBs complexes were used to remove rheumatoid factor from the serum. Results The average recovery of alpha-fetoprotein was 88.4% and 93.8% in the rheumatoid factor-positive and rheumatoid factor-negative serum samples, respectively. The recovery of alpha-fetoprotein was significantly lower in the rheumatoid factor-positive serum samples than in the rheumatoid factor-negative serum samples. In two of five rheumatoid factor-positive samples, a large difference was found (9.8%) between the average alpha-fetoprotein recoveries in the serially diluted and initial recoveries. Fourteen rheumatoid factor-positive serum samples were pretreated with hepatitis B surface antigen-anti-HBs complex-coated paramagnetic microparticles. The alpha-fetoprotein concentrations measured in the pretreated samples increased significantly. Conclusions It was concluded that the alpha-fetoprotein chemiluminescent microparticle immunoassay is susceptible to interference by rheumatoid factor, leading to significantly lower results. Eliminating the incidence of negative interference from rheumatoid factor should be an important goal for immunoassay providers. In the meantime

  9. Intrahepatic hematoma: hepatic lesion in a newborn with high {alpha}-fetoprotein level

    Energy Technology Data Exchange (ETDEWEB)

    Lam, Chiu Ying Flora; Chan, Kui Fai; Fan, Tsz Wo; Kwok, Chong Hei Philip; Chan, Chi Hum Susan; Tsang, Tsz Kan [Queen Elizabeth Hospital, Department of Radiology and Imaging, Hong Kong (China)

    2005-11-01

    Hepatic hematomas are relatively common in fetuses and neonates; most are subcapsular in location. Sometimes their imaging features can be non-specific, so differentiation from other aggressive lesions like hepatoblastoma can be difficult, especially if there is a concurrent high {alpha}-fetoprotein level. We report a case of intrahepatic hematoma with a rising {alpha}-fetoprotein level. (orig.)

  10. Decreased alpha-fetoprotein in amniotic fluid and maternal serum in diabetic pregnancy

    DEFF Research Database (Denmark)

    Henriques, C U; Damm, P; Tabor, A

    1993-01-01

    OBJECTIVE: To determine a reference level for alpha-fetoprotein (AFP) in the amniotic fluid (AF) in pregnant women with insulin-dependent diabetes mellitus in order to suggest an explanation for the observed decrease in maternal serum AFP. METHODS: Alpha-fetoprotein was measured in AF, maternal...

  11. Diagnostic value of alpha-fetoprotein in liver cancer

    International Nuclear Information System (INIS)

    Pervez, T.; Anwar, S.M.

    2001-01-01

    Objective: To determine diagnostic value of alpha-fetoproteins (alpha-FP) in liver cancer. Design: Prospective study. Place and duration of study: Department of clinical oncology services Hospital Lahore, during the period from February 1998 to February 2001. Subjects and Methods: Among 200 persons studied, 100 presented with liver mass, jaundice and other symptoms directing toward liver pathology, later confirmed histopathologically, as suffering from hepatocellular carcinoma (HCC) while the other 100 healthy subject came to the department for blood donation and were HBs Ag pasitive on blood screening. All these subjects under went blood test for alpha-FP. This tumor marker was analyzed by using enzyme immunoassay-based kit. Results: The alpha-FP positivity was statistically evaluated. In HCC this test was statistically significant with p value of <0.001. In this study sensitivity of alpha-FP was 72% specificity 89%, positive predictive value 86.7% and negative predictive value of 76.1%. Conclusion: This study showed that alpha-FP was a useful diagnostic tool in the diagnosis of HCC. (author)

  12. Characterizations of Anti-Alpha-Fetoprotein-Conjugated Magnetic Nanoparticles Associated with Alpha-Fetoprotein for Biomedical Applications.

    Science.gov (United States)

    Liao, Shu-Hsien; Huang, Han-Sheng; Chieh, Jen-Jie; Su, Yu-Kai; Tong, Yuan-Fu; Huang, Kai-Wen

    2017-09-03

    In this work, we report characterizations of biofunctionalized magnetic nanoparticles (BMNPs) associated with alpha-fetoprotein (AFP) for biomedical applications. The example BMNP in this study is anti-alpha-fetoprotein (anti-AFP) conjugated onto dextran-coated Fe₃O₄ labeled as Fe₃O₄-anti-AFP, and the target is AFP. We characterize magnetic properties, such as increments of magnetization ΔM H and effective relaxation time Δτ eff in the reaction process. It is found that both ΔM H and Δτ eff are enhanced when the concentration of AFP, Ф AFP , increases. The enhancements are due to magnetic interactions among BMNPs in magnetic clusters, which contribute extra M H after the association with M H and in turn enhance τ eff . The screening of patients carrying hepatocellular carcinoma (HCC) is verified via ΔM H /M H . The proposed method can be applied to detect a wide variety of analytes. The scaling characteristics of ΔM H /M H show the potential to develop a vibrating sample magnetometer system with low field strength for clinic applications.

  13. Intrinsic labeling of alpha-fetoprotein with 65Zn

    International Nuclear Information System (INIS)

    Preiss, I.L.

    1994-01-01

    Many enzymes, proteins, and large molecules bind a variety of inorganic ions. The replacement of a specific metal ion with another element can disrupt the biodistribution and biochemistry of the host molecule. Alpha-fetoprotein (AFP) is a known oncofetal antigen. The authors have demonstrated that the substitution of Cu or Pb on AFP inhibits the transplacental migration of AFP and may lead to fetal demise. It is evident that in labeling with a radiotracer in order to observe the true distribution of a molecule, one must select a radioisotope of an element which is naturally bound to the host molecule. It has been demonstrated that AFP binds Zn in vivo. Thus, substituting 65 Zn for natural Zn produces an intrinsic label for AFP. The authors discuss the methodology for stripping and labeling of AFP with 65 Zn. The biodistribution of labeled AFP is compared with that of 65 Zn 2+ in healthy and diseased (hepatoma) animals

  14. Clinical evaluation of anti-alpha-fetoprotein radioimmunodetection

    International Nuclear Information System (INIS)

    Chapman, C.E.; Keeling, A.A.; Bradwell, A.R.; Chandler, S.T.; Dykes, P.W.

    1986-01-01

    Anti-alpha-fetoprotein (AFP) radioimmunodetection was performed in response to clinical requests in 16 patients. In two patients, assessment of a known tumour was required; the anti-AFP scans were accurate and provided useful clinical information in both cases. In the remaining 14 patients the request was for localisation of suspected recurrent tumour. Accurate information was provided in four of these patients. In this latter group, various conventional methods of investigation had failed to disclose the site of recurrence. However, of a total of 21 sites reported as positive in these 14 patients, eight proved to be false positives. Two false negative results also occurred in this group and nine could not be evaluated. Although occasionally patients were usefully scanned, improvements are necessary before consistently reliable information can be obtained using this technique. (author)

  15. Prenatal radiographic diagnosis of alpha-fetoprotein positive malformations in early pregnancy

    International Nuclear Information System (INIS)

    Probst, F.P.; Sigurd, J.

    1980-01-01

    Early diagnosis of two malformed fetuses with positive alpha-fetoprotein test of the amniotic fluid with the aid of amniography is reported. Radiographic, diagnostic and prognostic aspects are discussed. (Auth.)

  16. Management strategy in pregnancies with elevated second-trimester maternal serum alpha-fetoprotein based on a second assay.

    Science.gov (United States)

    Spaggiari, Emmanuel; Ruas, Marie; Dreux, Sophie; Valat, Anne-Sylvie; Czerkiewicz, Isabelle; Guimiot, Fabien; Schmitz, Thomas; Delezoide, Anne-Lise; Muller, Françoise

    2013-04-01

    To assess maternal-fetal outcomes in pregnancies associated with persistently elevated second-trimester maternal serum alpha-fetoprotein. A retrospective cohort study in 658 patients with maternal serum alpha-fetoprotein ≥2.5 multiple of median, performed at routine Down syndrome screening. Maternal serum alpha-fetoprotein was assayed a second time in 341 of them. Outcomes were recorded in all cases. The group with unexplained maternal serum alpha-fetoprotein persistently ≥2.5 multiple of median was associated with more pregnancy complications 37 of 92 (40.2%) as fetal death, preeclampsia, intrauterine growth restriction, and congenital nephrotic syndrome, compared with the group with maternal serum alpha-fetoprotein that returned to a normal level 37 of 226 (16.4%) (P alpha-fetoprotein returns to a normal level on a second assay, the risk of adverse outcome significantly decreases, but these pregnancies are still at risk of complications and therefore need close surveillance. Repeat maternal serum alpha-fetoprotein assay allows identification of patients who should be offered amniocentesis to evaluate the risk of nephrotic syndrome and epidermolysis bullosa. Alpha-fetoprotein should be monitored in pregnancies associated with unexplained high maternal serum alpha-fetoprotein. A management strategy based on ultrasound examination, second maternal serum alpha-fetoprotein assay and amniocentesis is proposed to improve prenatal counseling and management of such pregnancies. However, a prospective study remains necessary to evaluate it. Copyright © 2013 Mosby, Inc. All rights reserved.

  17. Alpha?fetoprotein elevation in NUT midline carcinoma: a case report

    OpenAIRE

    D?Ambrosio, Lorenzo; Palesandro, Erica; Moretti, Marina; Pelosi, Giuseppe; Fabbri, Alessandra; Carnevale Schianca, Fabrizio; Aglietta, Massimo; Grignani, Giovanni

    2017-01-01

    Background Nuclear protein in testis (NUT) midline carcinoma is a rarely diagnosed and potentially under-recognized type of squamous carcinoma that is considered one of the most aggressive human solid tumors. Alpha-fetoprotein elevation has been associated with chronic liver diseases and a limited number of cancers. In particular, in presence of a mediastinal mass in a young man, alpha-fetoprotein elevation is considered nearly pathognomonic of a non-seminoma germ-cell tumor. Case presentatio...

  18. Rapid and label-free bioanalytical method of alpha fetoprotein detection using LSPR chip

    Science.gov (United States)

    Kim, Dongjoo; Kim, Jinwoon; Kwak, Cheol Hwan; Heo, Nam Su; Oh, Seo Yeong; Lee, Hoomin; Lee, Go-Woon; Vilian, A. T. Ezhil; Han, Young-Kyu; Kim, Woo-Sik; Kim, Gi-bum; Kwon, Soonjo; Huh, Yun Suk

    2017-07-01

    Alpha fetoprotein (AFP) is a cancer marker, particularly for hepatocellular carcinoma. Normal levels of AFP are less than 20 ng/mL; however, its levels can reach more than 400 ng/mL in patients with HCC. Enzyme linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) have been employed for clinical diagnosis of AFP; however, these methods are time consuming and labor intensive. In this study, we developed a localized surface plasmon resonance (LSPR) based biosensor for simple and rapid detection of AFP. This biosensor consists of a UV-Vis spectrometer, a cuvette cell, and a biosensor chip nanopatterned with gold nanoparticles (AuNPs). In our LSPR biosensor, binding of AFP to the surface of the sensor chip led to an increasing magnitude of the LSPR signals, which was measured by an ultraviolet-visible (UV-Vis) spectrometer. Our LSPR biosensor showed sufficient detectability of AFP at concentrations of 1 ng/mL to 1 μg/mL. Moreover, the overall procedure for detection of AFP was completed within 20 min. This biosensor could also be utilized for a point of care test (POCT) by employing a portable UV-Vis spectrometer. Owing to the simplicity and rapidity of the detection process, our LSPR biosensor is expected to replace traditional diagnostic methods for the early detection of diseases.

  19. Direct analysis of the lectin reactivity of alpha-fetoprotein in maternal serum by crossed affinity radio-immunoelectrophoresis

    International Nuclear Information System (INIS)

    Kerckaert, J.P.; Bayard, B.; Biserte, G.; Puech, F.; Codaccioni, X.

    1980-01-01

    Affinity experiments with the lentil (Lens culinaris) lectin have revealed the existence of two distinct molecular populations of alpha-fetoprotein: lectin reactive and lectin non-reactive. Using a combination of crossed lectin immunoelectrophoresis and radio-immunoelectrophoresis, it has been possible to obtain directly the lentil lectin affinity patterns of alpha-fetoprotein present in maternal sera. The lentil lectin reactivity of maternal alpha-fetoprotein decreases almost linearly with the gestational age from week 15 to 35. (Auth.)

  20. Quantitative Alpha Fetoprotein Detection with a Piezoelectric Microcantilever Mass Sensor

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sang Kyu; Cho, Jong Yun; Jeon, Sang Min; Cha, Hyung Joon; Moon, Won Kyu [Pohang University of Science and Technology, Pohang (Korea, Republic of); Lee, Yeol Ho [Samsung Advanced Institute of Technology, Yongin (Korea, Republic of)

    2011-10-15

    Alpha fetoprotein(AFP), which is serological marker for hepatocellular carcinoma, was quantitatively measured by its normal concentration, 10 ng/ml, with a label-free piezoelectric microcantilever mass sensor. The principle of detection is based on changes in the resonant frequency of the piezoelectric microcantilever before and after target molecules are attached to it, and its resonant frequency is measured electrically using a conductance spectrum. The resonant frequency of the developed sensor is approximately 1.34 MHz and the mass sensitivity is approximately 175 Hz/pg. The sensor has high reliability as mass sensor by reducing the effect of surface stress on resonant frequency due to attached proteins. 'Dip and dry' technique was used to react the sensor with reagents for immobilizing AFP antibody on the sensor and detecting AFP antigen. The measured mass of the detected AFP antigen was 6.02 pg at the concentration of 10 ng/ml, and 10.67 pg at 50 ng/ml when the immunoreaction time was 10 min.

  1. Alpha-fetoprotein, a fascinating protein and biomarker in neurology.

    Science.gov (United States)

    Schieving, J H; de Vries, M; van Vugt, J M G; Weemaes, C; van Deuren, M; Nicolai, J; Wevers, R A; Willemsen, M A

    2014-05-01

    Alpha-fetoprotein (AFP) is present in fetal serum in concentrations up to 5,000,000 μg/l. After birth, AFP gene expression is turned down with a subsequent fall of the serum concentrations of this albumin-like protein to 'adult values' of circa 0.5-15 μg/l from the age of 2 years onwards. Irrespective of its assumed important functions, individuals with AFP deficiency appear fully healthy. The other way around, the presence of AFP in the circulation after the first years of life doesn't seem to harm, since individuals with 'hereditary persistence of AFP' are also without clinical abnormalities. During pregnancy, AFP (in maternal serum) has long been recognized as a marker for congenital anomalies of the fetus. Equally well known is AFP as biomarker for hepatocellular carcinoma and some other malignancies. There are at least four neurodegenerative disorders, all inherited as autosomal recessive traits and characterized by the presence of cerebellar ataxia, abnormal ocular movements, and neuropathy, for which an elevated concentration of serum AFP is an important diagnostic biomarker. The availability of a reliable biomarker is not only important during screening or diagnostic processes, but is also relevant for objective follow-up during (future) therapeutic interventions. Copyright © 2013. Published by Elsevier Ltd.

  2. Quantitative Alpha Fetoprotein Detection with a Piezoelectric Microcantilever Mass Sensor

    International Nuclear Information System (INIS)

    Lee, Sang Kyu; Cho, Jong Yun; Jeon, Sang Min; Cha, Hyung Joon; Moon, Won Kyu; Lee, Yeol Ho

    2011-01-01

    Alpha fetoprotein(AFP), which is serological marker for hepatocellular carcinoma, was quantitatively measured by its normal concentration, 10 ng/ml, with a label-free piezoelectric microcantilever mass sensor. The principle of detection is based on changes in the resonant frequency of the piezoelectric microcantilever before and after target molecules are attached to it, and its resonant frequency is measured electrically using a conductance spectrum. The resonant frequency of the developed sensor is approximately 1.34 MHz and the mass sensitivity is approximately 175 Hz/pg. The sensor has high reliability as mass sensor by reducing the effect of surface stress on resonant frequency due to attached proteins. 'Dip and dry' technique was used to react the sensor with reagents for immobilizing AFP antibody on the sensor and detecting AFP antigen. The measured mass of the detected AFP antigen was 6.02 pg at the concentration of 10 ng/ml, and 10.67 pg at 50 ng/ml when the immunoreaction time was 10 min

  3. Intermethod discordance for alpha-fetoprotein measurements in Fanconi anemia.

    Science.gov (United States)

    Cassinat, B; Darsin, D; Guardiola, P; Toubert, M E; Rain, J D; Gluckman, E; Schlageter, M H

    2001-08-01

    The significantly higher serum alpha-fetoprotein (AFP) in patients with Fanconi anemia (FA) than in non-FA aplastic patients has potential diagnostic utility, but the increase is method-dependent. The aim of this study was to compare five AFP assays on FA and non-FA samples and to investigate possible explanations for FA-specific discrepancies. Two methods available in our laboratory (Kryptor and IMx) were compared on 59 FA and 27 non-FA patient samples. Kryptor, Immulite, Elecsys, Immuno-I, and Elsa-2 methods were then compared on 14 FA and 14 non-FA patient samples. The AFP glycosylation profile was analyzed by electrophoretic separation in a lectin-containing gel. With all six methods, AFP values were significantly higher in FA than in non-FA patients, but the diagnostic precision and optimal cutoff values varied. Indeed, two methods reached 100% sensitivity and specificity, but in other methods, one or both of these parameters were significantly <100%. Neither heterophilic antibodies nor a specific glycosylation profile was detected in FA samples. AFP results are method-dependent in FA. New methods must be evaluated before use in differential diagnosis of aplastic patients.

  4. The evaluation of domestic immunoradiometric assay kit for alpha-fetoprotein

    International Nuclear Information System (INIS)

    Won, Kyoung Sook; Ryu, Jin Sook; Moon, Dae Hyuk; Lee, Hee Kyung

    2000-01-01

    Although α-fetoprotein is one of the most commonly used tumor markers in Korea, most of the radioimmunoassay kits for α-fetoprotein have been imported from foreign countries. The purpose of this study was to evaluate the performance of a recently developed domestic immunoradiometric kit for α-fetoprotein (Riakey AFP IRMA CT R , Sin-Jin Medics, Seoul, Korea). We evaluated intra-and inter-assay precision, recovery rate, parallelism, and sensitivity of serum α-fetoprotein measurement using Riakey AFP IRMA CT R kit. The values of α-fetoprotein measured by Riakey AFP IRMA CT R kit were compared with those measured by two foreign commercial kits (α-fetoproteina of Radim and α-feto.riabead of Abbott). Intra-assay coefficients of variation on three different levels were 5.3% for 18.9 ng/ml, 3.4% for 133 ng/ml and 1.6% for 330 ng/ml. Inter-assay coefficients of variation were 9.7% for 20.9 ng/ml, 3.2% for 137 ng/ml and 4.1% for 330 ng/ml respectively. Recovery rate tests on all three different levels showed within 100±10%. Parallelism was also good and the sensitivity was 0.63 ng/ml. There was strong correlation between the measurement of α-fetoprotein by Riakey AFP IRMA CT R and that by two foreign commercial kits(r=3D0.98). The first Korean domestic immunoradiometric kit for α-fetoprotein, Riakey AFP IRMA CT R , performed well for clinical use.=20

  5. Association between alpha-fetoprotein and metabolic syndrome in a Chinese asymptomatic population: a cross-sectional study.

    Science.gov (United States)

    Chen, Yimin; Zhao, Ying; Feng, Linmin; Zhang, Jie; Zhang, Juanwen; Feng, Guofang

    2016-04-27

    Metabolic syndrome is closely associated with an increased risk for fatty liver disease morbidity and mortality. Recently, studies have reported that participants with fatty liver disease have higher serum alpha-fetoprotein levels than those without. We investigated the association between alpha-fetoprotein levels and the prevalence of metabolic syndrome in a Chinese asymptomatic population. A cross-sectional study was performed with 7,755 participants who underwent individual health examinations. Clinical and anthropometric parameters were collected and serum alpha-fetoprotein levels and other clinical and laboratory parameters were measured. Logistic regression analysis was used to examine associations between alpha-fetoprotein and metabolic syndrome. Participants with metabolic syndrome had significantly higher (p alpha-fetoprotein levels than those without, though all alpha-fetoprotein levels were within the reference interval. The association between the components of metabolic syndrome (central obesity, elevated blood pressure, elevated triglycerides, reduced high-density lipoprotein cholesterol, and elevated fasting plasma glucose) and alpha-fetoprotein levels was evaluated. Alpha-fetoprotein levels in the elevated triglycerides, reduced high-density lipoprotein cholesterol, and elevated fasting plasma glucose groups were significantly different (p=0.002, p alpha-fetoprotein in the normal triglycerides, high-density lipoprotein cholesterol, and fasting plasma glucose groups. Logistic regression analyses showed an association between alpha-fetoprotein levels and increased risk for metabolic syndrome, the presence of reduced high-density lipoprotein cholesterol, and elevated fasting plasma glucose, but not with obesity, elevated blood pressure, or triglycerides. These results suggest a significant association between alpha-fetoprotein and metabolic syndrome.

  6. Diagnosis of hepatocellular carcinoma by serial determination of serum alpha-fetoprotein concentration

    Energy Technology Data Exchange (ETDEWEB)

    Yumoto, Y; Namba, T; Tanaka, Y; Taketa, K; Ohta, Y [Okayama Univ. (Japan). School of Medicine

    1976-06-01

    This communication describes the clinical significance of serum ..cap alpha..-fetoprotein (S-AFP) determined by radioimmunoassay (RIA) in the diagnosis of hepatocellular carcinoma (HCC). The sensitivity of the AFP assay with the ..cap alpha..-Feto-125 kit produced by the Dainabot RI laboratories was over 2.5 ng/ml. Reproducibility of the kit was satisfactory; coefficient of variation was 6-10% within assays and 13% between assays. The mean and standard deviation of S-AFP in the sera of 30 healthy controls was 5.6+-2.6 ng/ml. When serum with a high level of S-AFP (2.9x10/sup 5/ ng/ml) was diluted with normal horse serum, the actual concentrations of AFP in the diluted serum were consistent with those determined. Thus, normal horse serum was employed to dilute the sera with high levels of AFP. Levels of S-AFP were over 2000 ng/ml in 75.5% of 49 cases with HCC, in 0.64% of 157 patients with hepatitis and cirrhosis of the liver, and in 3.8% of 52 cases of metastatic gastric cancer to the liver. In 20 patients with HCC, levels of S-AFP ranged widely from 7.5 to 9.5x10/sup 5/ ng/ml. In serial determinations of S-AFP in the clinical courses of patients such as those with S-AFP over 2000 ng/ml, the continuous increase of AFP was strongly suggestive of the presence of HCC. In 4 of 20 cases of HCC, the S-AFP levels remained under 300 ng/ml. Histological examination of these 4 cases of HCC showed Classes I and II, but no III in Edmondson's classification. Other cases of HCC with S-AFP levels over 300 ng/ml showed Edmondson's Classes II and III.

  7. Extremely elevated alpha-fetoprotein due to acute exacerbation of chronic hepatitis B without malignancy: a case report.

    Science.gov (United States)

    Yoon, Young-Min; Kang, Da-Yeong; Kim, Da-Young; Seo, Jun-Won; Lim, Hyun-Jong; Lee, Hee-Jeong; Park, Sang-Gon

    2016-06-01

    Alpha-fetoprotein is produced by a variety of tumors such as hepatocellular carcinoma, hepatoblastoma, and germ cell tumors of the ovary and testes. However, we present a case of significantly elevated serum alpha-fetoprotein without evidence of malignant disease in a patient who is a carrier of chronic hepatitis B. A 60-year-old Korean man presented with markedly increased alpha-fetoprotein (2350 ng/mL; normal 7 × 105 IU/mL). Our patient was diagnosed with acute exacerbation of chronic hepatitis B, and we presumed that this condition might be related to extremely elevated alpha-fetoprotein. When our patient was treated with entecavir, the serum alpha-fetoprotein level immediately decreased, in parallel with the hepatitis B virus deoxyribonucleic acid copy number. We report a rare case of extremely elevated alpha-fetoprotein due to acute exacerbation of chronic hepatitis B without any malignancy, and a decrease in this tumor marker simultaneous with a decrease in hepatitis B virus deoxyribonucleic acid copy number on entecavir treatment. This case report is important due to the rarity of the case; furthermore, it provides details of a diagnostic process for a variety of benign diseases and malignant tumors that should be considered in patients with elevated alpha-fetoprotein. Thus, we present a case report, along with a review, that will be helpful for diagnosis and treatment of patients with elevated alpha-fetoprotein.

  8. Replacing Alpha-Fetoprotein With Alpha-Fetoprotein-L3 Increases the Sensitivity of Prenatal Screening for Trisomy 21.

    Science.gov (United States)

    Huai, Lei; Leng, Jianhang; Ma, Shenglin; Huang, Fang; Shen, Junya; Ding, Yu

    This study aimed to investigate the serum concentration of alpha-fetoprotein (AFP)-L3 in midterm pregnancies and its potential application in prenatal trisomy screening. The serum samples from 27 women with trisomy 21 fetuses and 800 women with normal fetuses were examined to measure the concentrations of AFP, AFP-L3, human chorionic gonadotropin (hCG), unconjugated estriol (uE3), and inhibin-A. The screening results of various tests consisting of these markers were analyzed. In normal pregnancies within 15-20 weeks of gestation, the medians of serum AFP-L3 were 4.63, 5.70, 5.78, 6.58, 7.03, and 7.25 pg/mL. The median of AFP-L3 MoM in the trisomy 21 group was 0.46, which was significantly lower than the value of 1 in the normal group (P < 0.05). When using a cutoff value of 1/270, the sensitivity of the triple marker test (AFP, hCG, uE3) was improved from 74% to 81% by replacing AFP with AFP-L3, with the false-positive rate slightly increased from 5.4% to 6.8%. Similarly, the sensitivity of the quad marker test (AFP, hCG, uE3, inhibin-A) was improved from 81% to 89% by replacing AFP with AFP-L3, with the false-positive rate slightly increased from 4.6% to 5.6%. Serum AFP-L3 concentration increases along with more weeks of gestation in the midterm pregnancies. Trisomy 21 screening tests with AFP replaced by AFP-L3 have higher sensitivities at the expense of slightly increased false-positive rates. This improvement in screening may help to better prepare the parents and caregivers for the special needs of newborns with trisomy 21.

  9. Preoperative Alpha-Fetoprotein Slope is Predictive of Hepatocellular Carcinoma Recurrence after Liver Transplantation

    Directory of Open Access Journals (Sweden)

    Kathy Han

    2007-01-01

    Full Text Available BACKGROUND: Liver transplantation (LT offers a possible cure for patients with hepatocellular carcinoma (HCC and cirrhosis. However, tumour progression while on the waiting list and tumour recurrence after LT are common. The prognostic significance of various pre- and postoperative variables were investigated in regard to tumour recurrence, with an emphasis on the slope of preoperative serum alpha-fetoprotein (AFP levels.

  10. ORIGIN OF RAISED MATERNAL SERUM ALPHA-FETOPROTEIN LEVELS IN 2ND-TRIMESTER OLIGOHYDRAMNIOS

    NARCIS (Netherlands)

    LOS, FJ; BEEKHUIS, [No Value; MARRINK, J; HAGENAARS, AM; REUSS, A; SACHS, ES; JAHODA, MGJ; WLADIMIROFF, JW

    Concanavalin A (Con A) subtyping of alpha-fetoprotein (AFP) revealed higher concentrations of AFP non-reactive with Con A in sera of 12 pregnant women with second-trimester oligohydramnios and raised total serum AFP levels than in sera of 42 pregnant women with raised total serum AFP levels and a

  11. Interferometer immunosensor based on porous silicon for determining alpha-fetoprotein

    Science.gov (United States)

    Lv, Xiaoyi; Jiang, Jing; Lv, Guodong; Mo, Jiaqing; Jia, Zhenhong

    2016-10-01

    An increased level of alpha-fetoprotein ( AFP) in the blood may be a sign of liver cancer. Porous silicon based optical microcavities structure is prepared as a label-free immunosensor platform for detecting AFP. After the antigen-antibody reaction, it is monitored that the red shift of the reflection spectrum of the immunosensor increases

  12. 99MTC Alpha-Fetoprotein: A Novel, Specific Agent for the Detection of Human Breast Cancer

    National Research Council Canada - National Science Library

    Line, Bruce

    1998-01-01

    .... We have demonstrated that technetium-99m radiolabeled human alpha-fetoprotein (99mTc AFP) localizes in human breast cancer cells in-vivo, most likely concentrating in breast cancer cells due to a specific receptor not found in normal adult breast tissue...

  13. ALPHA-FETOPROTEIN IN FETAL SERUM, AMNIOTIC-FLUID, AND MATERNAL SERUM

    NARCIS (Netherlands)

    VANLITH, JMM; BEEKHUIS, [No Value; VANLOON, AJ; MANTINGH, A; DEWOLF, BTHM; BREED, ASPM

    In order to gain more insight into the association between alpha-fetoprotein (AFP) and fetal chromosomal disorders, especially Down's syndrome, we measured AFP in fetal serum, amniotic fluid, and maternal serum at cordocentesis. We compared the concentration and gradient of AFP in these three

  14. FIRST-TRIMESTER MATERNAL SERUM ALPHA-FETOPROTEIN AS A MARKER FOR FETAL CHROMOSOMAL DISORDERS

    NARCIS (Netherlands)

    VANLITH, JMM

    1994-01-01

    We evaluated first-trimester maternal serum alpha-fetoprotein (MS-AFP) as a marker for fetal chromosomal disorders. The multicentre study was performed under the auspices of the Dutch Working Party on Prenatal Diagnosis. MS-AFP was measured in 2404 normal pregnancies and 72 chromosomally abnormal

  15. DETECTION OF FETOMATERNAL HEMORRHAGE ASSOCIATED WITH CORDOCENTESIS USING SERUM ALPHA-FETOPROTEIN AND THE KLEIHAUER TECHNIQUE

    NARCIS (Netherlands)

    VANSELM, M; KANHAI, HHH; VANLOON, AJ

    Fetomaternal haemorrhage (FMH) was studied after 46 cordocenteses. alpha-Fetoprotein (AFP) concentration and Kleihauer staining of maternal blood, taken both before and after the procedure, revealed increases in AFP values of more than 40 per cent in 30 per cent of the patients examined; fetal

  16. Clinical studies on the radioimmunodetection of tumors containing alpha-fetoprotein

    International Nuclear Information System (INIS)

    Goldenberg, D.M.; Kim, E.E.; Deland, F.; Spremulli, E.; Nelson, M.O.; Gockerman, J.P.; Primus, F.J.; Corgan, R.L.; Alpert, E.

    1980-01-01

    This study reports the use of radiolabeled antibodies to alpha-fetoprotein for the detection and localization of hepatocellular and germ cell carcinomas. Twelve patients with histories of histologically-confirmed neoplasia received a total dose between 1.0 and 4.4 mCi of 131 I-labeled goat IgG prepared against human alpha-fetoprotein. Total-body photoscans were taken with a gamma scintillation camera at various intervals after injection of the radioactive antibody. Computer subtraction of radioactive technetium background images from the antibody 131 I scans permitted the visualization of all tumor sites known to be present in 4 patients with either primary hepatocellular cancer or metastatic germ cell carcinoma of the testis. Among 8 patients with diverse neoplasms not believed to contain alpha-fetoprotein, 5 of 19 tumor sites showed radioactive antibody accretion, although significantly less than in the patients with liver or testicular cancer. This investigation indicates that alpha-fetoprotein-containing tumors can be detected and localized in vivo by the method of radioimmunodetection

  17. 99MTC Alpha-Fetoprotein: A Novel, Specific Agent for the Detection of Human Breast Cancer

    National Research Council Canada - National Science Library

    Line, Bruce

    1999-01-01

    .... We have demonstrated that technetium-99m radiolabeled human alpha-fetoprotein (99mTc AFP) localizes in human breast cancer cells in-vivo, most likely concentrating in breast cancer cells due to a specific receptor not found in normal adult breast tissue...

  18. Improvement and multicenter evaluation of the analytical performance of an automated chemiluminescent immunoassay for alpha fetoprotein

    NARCIS (Netherlands)

    Morota, Kaori; Komori, Makoto; Fujinami, Ryo; Yamada, Koji; Kuribayashi, Kageaki; Watanabe, Naoki; Sokoll, Lori J.; Elliott, Debra; Chan, Daniel W.; Martens, Frans; Heijboer, Annemieke C.; Blankenstein, Marinus A.; Hershberger, Stefan J.; Pfeiffer, Zachary A.; Vaidya, Shyam V.; Dowell, Barry L.

    2012-01-01

    A new ARCHITECT® alpha fetoprotein (AFP) assay was developed to improve the linearity at the upper end of the calibration curve and to enhance other performance characteristics. In addition, this reformulation eliminated the possibility of falsely depressed samples at high AFP concentrations. The

  19. Alpha-fetoprotein is a predictor of outcome in acetaminophen-induced liver injury

    DEFF Research Database (Denmark)

    Schmidt, Lars E; Dalhoff, Kim

    2005-01-01

    An increase in alpha-fetoprotein (AFP) following hepatic necrosis is considered indicative of hepatic regeneration. This study evaluated the prognostic value of serial AFP measurements in patients with severe acetaminophen-induced liver injury. Prospectively, serial measurements of AFP were...

  20. Silencing alpha-fetoprotein inhibits VEGF and MMP-2/9 production in human hepatocellular carcinoma cell.

    Science.gov (United States)

    Meng, Wenbo; Li, Xun; Bai, Zhongtian; Li, Yan; Yuan, Jinqiu; Liu, Tao; Yan, Jun; Zhou, Wence; Zhu, Kexiang; Zhang, Hui; Li, Yumin

    2014-01-01

    Alpha-fetoprotein not only serves as a diagnostic marker for liver cancer, but also posses a variety of biological functions. However, the role of Alpha-fetoprotein on tumor angiogenesis and cell invasion remains incompletely understood. In this study, we aimed to evaluate if Alpha-fetoprotein can regulate the major angiogenic factors and matrix metalloproteinases in human liver cancer cells. Alpha-fetoprotein silencing was achieved by Stealth RNAi. Expression of Alpha-fetoprotein was examined by a full-automatic electrochemistry luminescence immunity analyzer. Expression of VEGF, VEGFR-2, MMP-9, and MMP-2 was examined by Western blot and immunocytochemistry. Apoptosis was detected by TUNEL assay. Angiogenesis was detected by in vitro angiogenesis assay kit. Silencing of Alpha-fetoprotein led to an increased apoptosis, which was associated with a decreased expression of vascular endothelial growth factor, vascular endothelial growth factor receptor 2, matrix metalloproteinases-2/9. These results suggest that Alpha-fetoprotein may play a regulatory role on angiogenesis and cell invasion during liver cancer development.

  1. Equine alpha-fetoprotein levels in Lipizzaner mares with normal pregnancies and with pregnancy loss.

    Science.gov (United States)

    Vincze, Boglárka; Gáspárdy, András; Kulcsár, Margit; Baska, Ferenc; Bálint, Ádám; Hegedűs, György Tamás; Szenci, Ottó

    2015-12-01

    Alpha-fetoprotein has proved to be a good indicator of fetal well-being in human medicine for decades. Although this molecule is present in most of the mammalian species including horses, reference values in healthy and high-risk pregnant mares have not yet been published. The aim of the present study was to determine whether equine alpha-fetoprotein (eqAFP) is a good indicator of complicated pregnancies in Lipizzaner mares. A total of 111 serum samples from 30 mares have been analyzed for eqAFP levels throughout gestation (Days 60-325). After the pregnancy was confirmed, 23 mares had normal pregnancies with viable foals, six had late embryonic loss, and one of the mares aborted in the ninth gestational month. Equine alpha-fetoprotein concentrations significantly differed in the normal group (72.93 ± 49.25 pg/mL; mean ± standard deviation) and in the complicated pregnancy loss group (152 ± 36.48 pg/mL; mean ± standard deviation). The mares' age, gestational age, and the conception rate significantly affected the alpha-fetoprotein concentrations in the normal group. Furthermore, notable individual differences occurred in eqAFP concentrations between mares. Equine alpha-fetoprotein seems to be an important indicator of fetal well-being in horses, but there are still some unanswered questions (levels in foals of different age, ponies, and draft horses) regarding this serum protein. Large-scale studies are needed to assess the specificity, sensitivity, and reliability of this test as a possible future diagnostic tool for fetal well-being in horses. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Immunohistochemical and radioimmunological demonstration of alpha/sub 1/-fetoprotein in nonmalignant changes of human gastric mucosa

    Energy Technology Data Exchange (ETDEWEB)

    Falser, N; Lederer, B; Reissigl, H [Innsbruck Univ. (Austria). Pathologisch-Anatomisches Lab.; Innsbruck Univ. (Austria). Gastroenterologisches Lab.)

    1977-07-01

    The occurence of ..cap alpha../sub 1/ fetoprotein in nonmalignant changes of the gastric mucosa was investigated by means of immunohistochemistry and radioimmunonoassay. The investigations were performed in tissue sections, cytological imprint preparations as well as in homogenized tissue samples (obtained by gastroscopy). ..cap alpha../sub 1/ fetoprotein could be demonstrated by immuno-histochemistry in about 90% of the samples originating from the surroundings of gastric ulcer and the region of gastrojejunostomy after B II-resection. The RIA was positive in about 75% of the tissue samples, whereas from gastric juice only 40% of positive results could be obtained. No ..cap alpha../sub 1/ fetoprotein-activity could be demonstrated in serum samples. These investigations indicate that ..cap alpha../sub 1/ fetoprotein is not exclusively synthesized by embryonic or neoplastic tissues and also can be synthesized also by regenerating cell-systems. It may be supposed that this synthesis represents an unspecific answer to growth-stimulation.

  3. Localisation of malignant germ-cell tumours by external scanning after injection of radiolabelled anti-alpha-fetoprotein

    International Nuclear Information System (INIS)

    Halsall, A.K.; Fairweather, D.S.; Bradwell, A.R.; Blackburn, J.C.; Howell, A.; Dykes, P.W.; Reeder, A.; Hine, K.R.

    1981-01-01

    Sheep IgG antibody to alpha-fetoprotein was labelled with 131 I and used to identify human germ-cell tumours by emission scanning. Eleven patients were studied after resection of their primary tumours. Ten had malignant teratoma and one an endodermal sinus tumour. All eight patients with raised serum alpha-fetoprotein concentrations had metastases apparent in the antibody scans. Of the remaining three patients with normal serum alpha-fetoprotein concentrations, two had positive scans. Three of the patients with positive results were scanned twice; the second scans were negative after treatment, when the alpha-fetoprotein concentrations had returned to normal. These results suggest that antibody scans are useful in the clinical management of patients with germ-cell tumours. (author)

  4. Fifty years of discovery of alpha-fetoprotein as the first tumor marker

    Directory of Open Access Journals (Sweden)

    Nikulina Dina

    2015-01-01

    Full Text Available Alpha-fetoprotein represents the most prominent oncobiomarker, widely used in the diagnosis of hepatocellular carcinoma for monitoring of tumor progression, presence of metastasis, assessment of cancer prognosis and successful antitumor therapeutic measures. Yuri Semenovich Tatarinov is a Russian scientist who first published antigen specific for human hepatocellular carcinoma in 1963. To commemorate the 50th anniversary of the discovery of alpha-fetoprotein, 9th International Scientific- Practical Conference entitled “Achievements of fundamental science and translational medicine capabilities in solving actual problems of practical public health”, was held from May 6-8th, 2013 in Astrakhan, Russia. The conference was held in memory of historical scientific work of Yuri Semenovich Tatarinov. [Projekat Ministarstva nauke Republike Srbije, br. 175056

  5. Alpha-fetoprotein-producing Gastric Cancer with Metastasis to the Scrotum: Case Report

    International Nuclear Information System (INIS)

    Cho, Young Joon; Chung, Jae Joon; Yu, Jeong Sik; Kim, Joo Hee; Kim, Dae Jung; Ahn, Jhii Hyun; Cho, Eun Suk

    2010-01-01

    An alpha-fetoprotein-producing gastric cancer is rare, making up only about 3% of all gastric cancers. Further, gastric cancer with metastasis to the scrotum via a transperitoneal route is extremely rare. We report a case of metastatic scrotal mass in a 68-year-old man who had undergone a subtotal gastrectomy and gastroduodenostomy due to signet ring cell type gastric cancer with a description focusing on the radiologic findings

  6. Alpha-fetoprotein-producing Gastric Cancer with Metastasis to the Scrotum: Case Report

    Energy Technology Data Exchange (ETDEWEB)

    Cho, Young Joon [Yonsei University College of Medicine, Seoul (Korea, Republic of); Chung, Jae Joon; Yu, Jeong Sik; Kim, Joo Hee; Kim, Dae Jung; Ahn, Jhii Hyun; Cho, Eun Suk [Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul (Korea, Republic of)

    2010-11-15

    An alpha-fetoprotein-producing gastric cancer is rare, making up only about 3% of all gastric cancers. Further, gastric cancer with metastasis to the scrotum via a transperitoneal route is extremely rare. We report a case of metastatic scrotal mass in a 68-year-old man who had undergone a subtotal gastrectomy and gastroduodenostomy due to signet ring cell type gastric cancer with a description focusing on the radiologic findings

  7. Alpha-fetoprotein as a tool to distinguish amniotic fluid from urine, vaginal discharge, and semen.

    Science.gov (United States)

    Mor, Amir; Tal, Reshef; Haberman, Shoshana; McCalla, Sandra; Irani, Mohamad; Perlman, Jaqueline; Seifer, David B; Minkoff, Howard

    2015-02-01

    To estimate whether alpha-fetoprotein (AFP) can be used to distinguish amniotic fluid absorbed in sanitary pads from other similarly absorbed substances (semen, urine, and normal vaginal discharge). A prospective cohort study. Urine and amniotic fluid specimens were collected from 52 pregnant women admitted for labor. Semen specimens were collected from 17 men undergoing infertility evaluation. Alpha-fetoprotein concentrations were measured directly from urine, amniotic fluid, and semen and from pads instilled with samples from these specimens. Alpha-fetoprotein concentrations were also measured from pads absorbed with normal vaginal discharge collected from 27 pregnant women. Alpha-fetoprotein levels in amniotic fluid (245.38 ± 21.03 ng/mL, n = 52) were significantly higher than those measured in maternal urine (0.84 ± 0.17 ng/mL, n = 52, P < .001), or semen (1.52 ± 0.35 ng/mL, n = 17, P < .001). The same trend was seen when AFP was extracted from pads: amniotic fluid levels (19.44 ± 1.98 ng/mL, n=52) were significantly higher than those of urine (undetectable, n=52), semen (undetectable, n = 17), or normal vaginal discharge (0.53 ± 0.16 ng/mL, n = 27, P < .001). Receiver operator characteristic curve analysis demonstrated 96.2% sensitivity and 100% specificity for distinguishing the presence of amniotic fluid from normal vaginal discharge on sanitary pads (cutoff 3.88 ng/mL, area under the curve 0.99). When the diagnosis of rupture of membranes is in doubt, AFP levels can assist in differentiating amniotic fluid from other bodily fluids. A method that utilizes sanitary pads and an assay for AFP quantification may be an accurate and convenient way to confirm the diagnosis of rupture of membranes.

  8. RIA of alpha-fetoprotein in serum and amniotic fluid

    International Nuclear Information System (INIS)

    Fingerova, H.; Talas, M.; Stroufova, A.

    1979-01-01

    An own modification of the double antibody radioimmunoassay for AFP using 125 I-labelled AFP as a tracer, rabbit anti-AFP obtained from SEVAC, Prague and precipitating antibodies prepared by the authors is described. The AFP levels measured in the serum and the amniotic fluid using the method were in agreement with those obtained by the means of the AFPK RIA kit by SORIN in the Institute for the Care of Mother and Child in Prague. The AFP concentrations found in the cord serum and the amniotic fluid were confirmed also by the rocket electroimmunoassay according to Laurell. The described AFP RIA seems suitable for the clinical application in prenatal screening for congenital malformations, in difficult pregnancies, in hepatology and the diagnosis and the evaluation of therapy of some human malignancies. (author)

  9. RIA of alpha-fetoprotein in serum and amniotic fluid

    Energy Technology Data Exchange (ETDEWEB)

    Fingerova, H; Talas, M; Stroufova, A [Palackeho Univ., Olomouc (Czechoslovakia). Lekarska Fakulta; Santavy, J; Krikal, Z [Ustav pro Peci o Matku a Dite, Prague (Czechoslovakia)

    1979-01-01

    An own modification of the double antibody radioimmunoassay for AFP using /sup 125/I-labelled AFP as a tracer, rabbit anti-AFP obtained from SEVAC, Prague and precipitating antibodies prepared by the authors is described. The AFP levels measured in the serum and the amniotic fluid using the method were in agreement with those obtained by the means of the AFPK RIA kit by SORIN in the Institute for the Care of Mother and Child in Prague. The AFP concentrations found in the cord serum and the amniotic fluid were confirmed also by the rocket electroimmunoassay according to Laurell. The described AFP RIA seems suitable for the clinical application in prenatal screening for congenital malformations, in difficult pregnancies, in hepatology and the diagnosis and the evaluation of therapy of some human malignancies.

  10. The progressive elevation of alpha fetoprotein for the diagnosis of hepatocellular carcinoma in patients with liver cirrhosis

    International Nuclear Information System (INIS)

    Arrieta, Oscar; Cacho, Bernardo; Morales-Espinosa, Daniela; Ruelas-Villavicencio, Ana; Flores-Estrada, Diana; Hernández-Pedro, Norma

    2007-01-01

    Hepatocellular carcinoma is the most common cause of primary liver neoplasms and is one of the main causes of death in patients with liver cirrhosis. High Alpha fetoprotein serum levels have been found in 60–70% of patients with Hepatocellular carcinoma; nevertheless, there are other causes that increase this protein. Alpha fetoprotein levels ≥200 and 400 ng/mL in patients with an identifiable liver mass by imaging techniques are diagnostic of hepatocellular carcinoma with high specificity. We analysed the sensitivity and specificity of the progressive increase of the levels of alpha fetoprotein for the detection of hepatocellular carcinoma in patients with liver cirrhosis. Seventy-four patients with cirrhosis without hepatocellular carcinoma and 193 with hepatic lesions diagnosed by biopsy and shown by image scans were included. Sensitivity and specificity of transversal determination of alpha fetoprotein ≥ 200 and 400 ng/mL and monthly progressive elevation of alpha fetoprotein were analysed. Areas under the ROC curves were compared. Positive and negative predictive values adjusted to a 5 and 10% prevalence were calculated. For an elevation of alpha fetoprotein ≥ 200 and 400 ng/mL the specificity is of 100% in both cases, with a sensitivity of 36.3 and 20.2%, respectively. For an alpha fetoprotein elevation rate ≥7 ng/mL/month, sensitivity was of 71.4% and specificity of 100%. The area under the ROC curve of the progressive elevation was significantly greater than that of the transversal determination of alpha fetoprotein. The positive and negative predictive values modified to a 10% prevalence are of: 98.8% and 96.92%, respectively; while for a prevalence of 5% they were of 97.4% and 98.52%, respectively. The progressive elevation of alpha fetoprotein ≥7 ng/mL/month in patients with liver cirrhosis is useful for the diagnosis of hepatocellular carcinoma in patients that do not reach αFP levels ≥200 ng/mL. Prospective studies are required to

  11. [The alpha-fetoprotein in prognosis of survival of and functional rehabilitation of patients with ischemic stroke].

    Science.gov (United States)

    Arkhipkin, A A; Liang, O V; Kochetov, A G

    2014-10-01

    The study was carried out to determine the prognostic value of alpha-fetoprotein in development of lethal outcome and degree of functional rehabilitation of patients with ischemic stroke. The sampling included 216 patients in acute period of ischemic stroke. At the first day of development of disease they were measured the level of human alpha-fetoprotein. At the second day of disease patients were evaluated the degree of functional rehabilitation and the rate of lethal outcomes was calculated. Previously, the reference interval for alpha-fetoprotein was calculated according the guidelines of the International federation of clinical chemistry and national standard. The reference interval amounted to 0.59-3.78 mE/l. The study results demonstrated that low level of alpha-fetoprotein is related to higher risk of lethal outcome (SE=1.7, p=0.012). The increasing of level of alpha-fetoprotein over mentioned threshold value statistically significant increases probability of survival of patients. The further increasing more than 2.28 mE/l is related to subsequent good functional rehabilitation according the modifies Rankine scale (SE=1.4, p=0.001) and Barthel index (SE=1.49, p<0.001).

  12. Performance of Alpha Fetoprotein in Combination with Alpha-1-acid Glycoprotein for Diagnosis of Hepatocellular Carcinoma Among Liver Cirrhosis Patients

    Directory of Open Access Journals (Sweden)

    Rino A Gani

    2016-05-01

    Full Text Available Aim: to evaluate the use of alpha-1-acid glycoprotein (AAG for diagnosing hepatocellular carcinoma (HCC, and to combine with alpha fetoprotein (AFP as part of routine examination in liver cirrhosis patients. Methods: this is a diagnostic study using cross-sectional design. A hundred and six patients were included in this study. Baseline data such as age, gender, AFP, AAG, peripheral blood count, AST and ALT were consecutively collected from liver cirrhosis patients with or without HCC. Serum AAG were measured quantitatively using immunoturboditimetric assay and AFP with enzyme immune assay (EIA. Statistical analysis were done using SPSS 13.0. Data comparisons between group were done using Mann-Whitney test. Diagnostic performance for each marker alone was compared to the surrogate use of both markers (combined parallel approach in HCC cases. Results: receiver operating characteristic (ROC analysis showed that area under the curve for AFP AAG combination was 88.1% and higher than AFP only (86.2% or AAG only (76.5% with sensitivity of 83%, 73% and 44%, respectively, at specificity of >80%. Conclusion: our study showed that combination of AFP and AAG is superior than either marker alone in diagnosing HCC in liver cirrhosis patients. Combination of AFP and AAG may be used to prompt early diagnosis screening of HCC. Key words: alpha fetoprotein, alpha-1-acid glycoprotein, biomarker, liver cancer

  13. Autism Spectrum Disorders and Maternal Serum alpha-Fetoprotein Levels During Pregnancy

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Grove, Jakob; Hougaard, David M

    2011-01-01

    Objective: Numerous studies have been trying to disentangle the complex pathophysiology of autism spectrum disorders (ASD). In our study, we explored the potential role of maternal serum (MS) alpha-fetoprotein (AFP) in the prediction and the pathophysiology of ASD. Methods: A total of 112 patients...... role in the pathophysiology of ASD makes AFP a good candidate for further larger-scale studies to confirm such an association and to determine whether this pattern is unique to ASD or related to other psychiatric disorders as well....

  14. Subtle Hepatocellular Carcinoma: A Persisting Role for Alpha-Fetoprotein Monitoring in High-Risk Patients with Cirrhosis

    Directory of Open Access Journals (Sweden)

    Barry Schlansky

    2011-10-01

    Full Text Available Hepatocellular carcinoma (HCC is a common, aggressive malignancy that usually develops in a background of liver cirrhosis. Practice guidelines recommend screening of cirrhotic patients with ultrasound and more detailed imaging (computed tomography or magnetic resonance imaging if abnormalities are detected. The utility of alpha-fetoprotein levels in HCC surveillance is controversial. Although HCC risk differs by etiology of cirrhosis, screening and surveillance guidelines are uniform after cirrhosis is established. We report a case of rapidly progressive HCC occurring in a cirrhotic patient with multiple unique risk factors for neoplasia, detected by a rising alpha-fetoprotein level without imaging features of liver cancer.

  15. Removal Of Labeled ALPHA-Fetoprotein (AFP) Using Rice Husk-Based Activated Carbon

    International Nuclear Information System (INIS)

    ABDEL-MOUHTY, N.R.

    2009-01-01

    Biomass agricultural waste materials, rice husk (RH) or saw dust (SD), were used for the preparation of activated carbons. RH was activated by chemical activation using phosphoric acid or potassium hydroxide. The prepared activated carbons were characterized and used for the adsorption of labeled alpha-fetoprotein ( 125 I-AFP) from the lab waste of iodine labeled alpha-fetoprotein tracer. The effects of various factors, e.g. carbon type, carbon dosage, temperature, particle size of carbon, effect of different waste volumes on the adsorption capacity, were quantitatively determined. Desorption of activated carbon was also investigated. From the experimental results, it was found that SDK had the lowest ability for adsorption of 125I-AFP and the highest uptake was 83% by carbon RHH. The amount of adsorption accomplished per unit weight of a solid adsorbent was greater, the more finely divided and the more porous the solid. 0.5 g for RHH carbon was found to be optimum dose of adsorbent for the removal of 125I-AFP. The optimum volume of waste with 0.5 g dose of RHH was 15 ml. The increased adsorption with temperature may be due to the increase of the intra-particle diffusion rate of sorbate ions into the pores at higher temperature as diffusion is an endothermic process.

  16. Regression of hepatocarcinoma cells using RNA aptamer specific to alpha-fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Young Ju [Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Yongin 448-701 (Korea, Republic of); Lee, Seong-Wook, E-mail: SWL0208@dankook.ac.kr [Department of Molecular Biology, Institute of Nanosensor and Biotechnology, Dankook University, Yongin 448-701 (Korea, Republic of)

    2012-01-06

    Highlights: Black-Right-Pointing-Pointer Identification of RNA aptamer specific to AFP with high affinity. Black-Right-Pointing-Pointer Specific induction of HCC proliferation by AFP. Black-Right-Pointing-Pointer Efficient increase in oncogene expression by AFP. Black-Right-Pointing-Pointer Efficient inhibition of AFP-mediated HCC proliferation by the aptamer. Black-Right-Pointing-Pointer Efficient suppression of AFP-induced oncogene expression of by the aptamer. -- Abstract: Alpha-fetoprotein (AFP) is a cancer-associated fetal protein and has long been utilized as a serum fetal defect/tumor marker to monitor distress/disease progression. In addition, AFP is closely associated with the proliferation of hepatocellular carcinoma. Thus, direct targeting of AFP has been recommended for a therapeutic strategy against hepatocellular carcinoma. In this study, we developed and characterized an RNA aptamer that specifically bound to the alpha-fetoprotein using SELEX technology. The aptamer interacted with the AFP with a K{sub D} of {approx}33 nM. Importantly, the identified aptamer specifically and efficiently inhibited the AFP-mediated proliferation of hepatocarcinoma cells in a dose dependent manner. Moreover, the aptamer efficiently down-regulated AFP-induced expression of oncogenes in the cells. These results indicate that an AFP-specific RNA aptamer could be a useful therapeutic and diagnostic agent against AFP-related hepatocellular carcinoma.

  17. Fabrication of protein microarrays for alpha fetoprotein detection by using a rapid photo-immobilization process

    Directory of Open Access Journals (Sweden)

    Sirasa Yodmongkol

    2016-03-01

    Full Text Available In this study, protein microarrays based on sandwich immunoassays are generated to quantify the amount of alpha fetoprotein (AFP in blood serum. For chip generation a mixture of capture antibody and a photoactive copolymer consisting of N,N-dimethylacrylamide (DMAA, methacryloyloxy benzophenone (MaBP, and Na-4-styrenesulfonate (SSNa was spotted onto unmodified polymethyl methacrylate (PMMA substrates. Subsequently to printing of the microarray, the polymer and protein were photochemically cross-linked and the forming, biofunctionalized hydrogels simultaneously bound to the chip surface by short UV- irradiation. The obtained biochip was incubated with AFP antigen, followed by biotinylated AFP antibody and streptavidin-Cy5 and the fluorescence signal read-out. The developed microarray biochip covers the range of AFP in serum samples such as maternal serum in the range of 5 and 100 ng/ml. The chip production process is based on a fast and simple immobilization process, which can be applied to conventional plastic surfaces. Therefore, this protein microarray production process is a promising method to fabricate biochips for AFP screening processes. Keywords: Photo-immobilization, Protein microarray, Alpha fetoprotein, Hydrogel, 3D surface, Down syndrome

  18. Regression of hepatocarcinoma cells using RNA aptamer specific to alpha-fetoprotein

    International Nuclear Information System (INIS)

    Lee, Young Ju; Lee, Seong-Wook

    2012-01-01

    Highlights: ► Identification of RNA aptamer specific to AFP with high affinity. ► Specific induction of HCC proliferation by AFP. ► Efficient increase in oncogene expression by AFP. ► Efficient inhibition of AFP-mediated HCC proliferation by the aptamer. ► Efficient suppression of AFP-induced oncogene expression of by the aptamer. -- Abstract: Alpha-fetoprotein (AFP) is a cancer-associated fetal protein and has long been utilized as a serum fetal defect/tumor marker to monitor distress/disease progression. In addition, AFP is closely associated with the proliferation of hepatocellular carcinoma. Thus, direct targeting of AFP has been recommended for a therapeutic strategy against hepatocellular carcinoma. In this study, we developed and characterized an RNA aptamer that specifically bound to the alpha-fetoprotein using SELEX technology. The aptamer interacted with the AFP with a K D of ∼33 nM. Importantly, the identified aptamer specifically and efficiently inhibited the AFP-mediated proliferation of hepatocarcinoma cells in a dose dependent manner. Moreover, the aptamer efficiently down-regulated AFP-induced expression of oncogenes in the cells. These results indicate that an AFP-specific RNA aptamer could be a useful therapeutic and diagnostic agent against AFP-related hepatocellular carcinoma.

  19. Serum alpha fetoprotein surge after the initiation of chemotherapy for non-seminomatous testicular cancer has an adverse prognostic significance

    NARCIS (Netherlands)

    de Wit, R; Collette, L; Sylvester, R; de Mulder, PHM; Sleijfer, DT; Huinink, WWT; Kaye, SB; van Oosterom, AT; Boven, E; Stoter, G

    It has been recognized that the tumour markers alpha-fetoprotein (AFP) and human chorionic gonadotrophin (HCG) may show a transient elevation after the initiation of chemotherapy in non-seminomatous testicular cancer. We investigated the prognostic importance of these so-called marker surges in a

  20. CONCANAVALIN-A VARIANTS OF ALPHA-FETOPROTEIN IN FIRST TRIMESTER FETUSES WITH TRISOMY-21 AND WITH NORMAL KARYOTYPES

    NARCIS (Netherlands)

    LOS, FJ; JANSE, HC; BRANDENBURG, H; DEVRIJ, RW; DEBRUIJN, HWA

    1995-01-01

    Total alpha-fetoprotein (AFP) concentrations and proportions of AFP non-reactive with the lectin concanavalin A (Con A) were studied in extracellular fluid of 22 first-trimester fetuses. Total AFP concentrations were significantly lower in fetuses with Down's syndrome than in those with

  1. MATERNAL SERUM ALPHA-FETOPROTEIN LEVELS AND FETAL-OUTCOME IN EARLY 2ND-TRIMESTER OLIGOHYDRAMNIOS

    NARCIS (Netherlands)

    LOS, FJ; HAGENAARS, AM; MARRINK, J; COHENOVERBEEK, TE; GAILLARD, JLJ; BRANDENBURG, H

    Early second-trimester oligohydramnios was associated with normal maternal serum alpha-fetoprotein (MSAFP) levels in nine out of 26 cases (35 per cent). Congenital malformations of the fetal urinary tract resulting in fetal anuria were present in nine cases; in seven of them, normal MSAFP levels

  2. Female Mice Deficient in Alpha-Fetoprotein Show Female-Typical Neural Responses to Conspecific-Derived Pheromones

    NARCIS (Netherlands)

    Brock, O.; Keller, M.; Douhard, Q.; Bakker, J.

    2012-01-01

    The neural mechanisms controlling sexual behavior are sexually differentiated by the perinatal actions of sex steroid hormones. We recently observed using female mice deficient in alpha-fetoprotein (AFP-KO) and which lack the protective actions of AFP against maternal estradiol, that exposure to

  3. Transcriptional switch from albumin to alpha-fetoprotein and changes in transcription of other genes during carbon tetrachloride induced liver regeneration

    International Nuclear Information System (INIS)

    Panduro, A.; Shalaby, F.; Weiner, F.R.; Biempica, L.; Zern, M.A.; Shafritz, D.A.

    1986-01-01

    During liver regeneration induced by CCl 4 administration to rats, changes in the relative transcription rates of albumin and alpha-fetoprotein genes have been measured in conjunction with other liver-specific and general cellular function genes. Within 24 h following CCl 4 administration, albumin gene transcription decreases by 85%, whereas alpha-fetoprotein transcription increases from undetectable levels to 50% of that observed for albumin. These changes precede maximal [ 3 H]thymidine incorporation into DNA which peaks at 48 h. Other genes related to liver-specific functions, such as ligandin, alpha 1-antitrypsin, and cytochrome P-450's, as well as general cellular genes pro alpha 1- and pro alpha 2-collagen, beta-actin, and alpha-tubulin, respond in kinetic patterns often distinct from each other and from albumin and alpha-fetoprotein. Changes in the steady-state levels of albumin and alpha-fetoprotein mRNA correlate with changes in transcription, but there is a lag in alpha-fetoprotein mRNA accumulation, which peaks at 72 h following CCl 4 administration. These studies indicate that reciprocal changes in albumin and alpha-fetoprotein gene transcription occur during CCl 4 -induced liver regeneration, leading to changes in the level of these specific mRNAs. These changes precede DNA synthesis and would appear to represent an alteration in differentiated function of hepatocytes in conjunction with the liver regenerative process

  4. Nonpalpable testicular pure seminoma with elevated serum alpha-fetoprotein presenting with retroperitoneal metastasis: a case report.

    Science.gov (United States)

    Iwatsuki, Shoichiro; Naiki, Taku; Kawai, Noriyasu; Etani, Toshiki; Iida, Keitaro; Ando, Ryosuke; Nagai, Takashi; Okada, Atsushi; Tozawa, Keiichi; Sugiyama, Yosuke; Yasui, Takahiro

    2016-05-05

    Patients with a primary pure seminoma in the testis who have elevated serum alpha-fetoprotein are rare and should be treated as patients with nonseminomatous germ cell tumors. However, nonpalpable testicular tumors in this condition have never been reported. We describe a case of nonpalpable pure testicular seminoma with elevated serum alpha-fetoprotein presenting retroperitoneal metastasis. A 29-year-old Asian man was referred to our hospital with right flank pain. Computed tomography showed a mass located between his aorta and inferior vena cava, but a testicular tumor was not detected. His serum levels of lactate dehydrogenase, alpha-fetoprotein, and DUPAN-2 were high. Although no tumor or nodule was palpable in his testis, ultrasonography revealed multiple low echoic lesions in his right testicular parenchyma. He was diagnosed with right testicular cancer with retroperitoneal lymph node metastasis and underwent right high orchiectomy. A pathological examination revealed pure seminoma and no nonseminomatous components were found in the specimen. Three courses of induction systemic chemotherapy (cisplatin, etoposide, and bleomycin) normalized his serum alpha-fetoprotein and DUPAN-2 levels. Three additional courses of chemotherapy (etoposide and bleomycin) were performed, and treatment was completed with laparoscopic retroperitoneal lymph node dissection. Pathology of the dissected specimen showed fibrous and necrotic tissue with no viable cells. He is alive without recurrence 54 months after orchiectomy. We report a case of pure testicular seminoma with elevated serum alpha-fetoprotein and DUPAN-2 presenting retroperitoneal metastasis. We recommend an ultrasound examination of bilateral testes when large retroperitoneal tumors are detected in young men, even if a mass is not palpable in the scrotum.

  5. Validation of the alpha-fetoprotein model for hepatocellular carcinoma recurrence after transplantation in an Asian population.

    Science.gov (United States)

    Rhu, Jinsoo; Kim, Jong Man; Choi, Gyu Seong; Kwon, Choon Hyuck David; Joh, Jae-Won

    2018-02-20

    This study was designed to validate the alpha-fetoprotein model for predicting recurrence after liver transplantation in Korean hepatocellular carcinoma patients. Patients who underwent liver transplantation for hepatocellular carcinoma at Samsung Medical Center between 2007 and 2015 were included. Recurrence, overall survival, and disease-specific survival of patients divided by both the Milan criteria and the alpha-fetoprotein model were compared using Kaplan-Meier log-rank test. The predictability of the alpha-fetoprotein model compared to the Milan criteria was tested by means of net reclassification improvement analysis applied to patients with a follow-up of at least 2 years. A total of 400 patients were included in the study. Patients within Milan criteria had 5-year recurrence, and overall survival rates of 20.9% and 76.3% respectively, compared to corresponding rates of 50.3% and 55.7%, respectively, for patients who were beyond Milan criteria. Alpha-fetoprotein model low risk patients had 5-year recurrence and overall survival rates of 21.1% and 76.2%, respectively, compared to corresponding rates of 57.7% and 52.2%, respectively, in high risk patients (P<0.001, all). Although overall net reclassification improvements were statistically nonsignificant for recurrence (NRI=1.7%, Z=0.30, p=0.7624), and overall survival (NRI=9.0%, Z=1.60, p=0.1098), they were significantly better for predicting no recurrence (NRI=6.6%, Z=3.16, p=0.0016) and no death. (NRI=7.7%, Z=3.65, p=0.0003) CONCLUSIONS: The alpha-fetoprotein model seems to be a promising tool for liver transplantation candidacy, but further investigation is needed.

  6. Alpha-Fetoprotein: From a Diagnostic Biomarker to a Key Role in Female Fertility

    Directory of Open Access Journals (Sweden)

    Christelle De Mees

    2006-01-01

    Full Text Available Alpha-fetoprotein (AFP is a well-known diagnostic biomarker used in medicine to detect fetal developmental anomalies such as neural tube defects or Down's syndrome, or to follow up the development of tumors such as hepatocellular carcinomas. However, and despite the fact that the protein was discovered almost half a century ago, little was known about its physiological function. The study of Afp knock-out mice uncovered a surprising function of AFP: it is essential for female fertility and for expression of normal female behaviors, and this action is mediated through its estrogen binding capacity. AFP sequestrates estrogens and by so doing protects the female developing brain from deleterious (defeminizing/ masculinizing effects of these hormones

  7. Purification and characterization of a bioactive alpha-fetoprotein produced by HEK-293 cells.

    Science.gov (United States)

    Lin, Bo; Peng, Guoqing; Feng, Haipeng; Li, Wei; Dong, Xu; Chen, Yi; Lu, Yan; Wang, Qiaoyun; Xie, Xieju; Zhu, Mingyue; Li, Mengsen

    2017-08-01

    Alpha-fetoprotein (AFP) is a biomarker that is used to diagnose hepatocellular carcinoma (HCC) and can promote malignancy in HCC. AFP is an important target in the treatment of liver cancer. To obtain enough AFP to screen for AFP inhibitors, we expressed and purified AFP in HEK-293 cells. In the present study, we produced AFP in the cells and harvested highly pure rAFP (or recombinant expression AFP in HEK-293 cells). We also analysed the bioactivity of rAFP and found that rAFP promoted growth of the human HCC cells, antagonize paclitaxel inhibition of HCC cell proliferation, suppress expression of active caspase-3, and promote expression of Ras and survivin. This study provides a method to produce significant amounts of AFP for use in biochemical assays and functional studies and to screen AFP inhibitors for use in HCC therapy. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Late solitary pelvic metastasis of hepatocellular carcinoma mimicking alpha-fetoprotein-producing gynaecologic tumour

    Directory of Open Access Journals (Sweden)

    Ji He Kim

    2018-01-01

    Full Text Available Extrahepatic spread of hepatocellular carcinoma (HCC is uncommon; and, pelvic metastasis, in particular, is extremely rare. A 71-year-old woman was admitted for evaluation of pelvic solitary solid mass. She had undergone a left lobectomy 28 years previously. Magnetic resonance imaging of the abdomen and pelvis demonstrated a heterogeneous mass in the right pelvic cavity, whereas no space-occupying lesions or ascites were detected in the liver. CA 125 levels were within normal limits; however, serum alpha-fetoprotein levels were markedly elevated. She underwent laparoscopic pelvic mass excision, total hysterectomy, and bilateral salpingo-oophorectomy. Histopathologic findings and immunochemical staining results indicated metastatic HCC. Herein, we report an unusual case of a patient with solitary recurrence in the pelvic cavity 28 years after initial diagnosis and treatment.

  9. Elevated alpha-fetoprotein: differential diagnosis - hepatocellular carcinoma and other disorders.

    Science.gov (United States)

    Wong, Robert J; Ahmed, Aijaz; Gish, Robert G

    2015-05-01

    The incidence of cirrhosis-related hepatocellular carcinoma (HCC) is rising. Curative surgical options are available; outcomes are acceptable with early diagnosis. Lens culinaris agglutinin-reactive fraction of alpha-fetoprotein (AFP-L3) and des-gamma-carboxy prothrombin (DCP) are HCC risk markers. A high or increasing serum biomarker level can be predictive of the eventual development of HCC, large tumor size, advanced stage, extrahepatic metastases, portal vein thrombosis, and postoperative HCC recurrence. Based on FDA guidelines for HCC risk assessment, clinicians can consider using either the combination of AFP-L3 with DCP, or the combination of AFP-L3 with AFP and DCP. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Human alpha-fetoprotein and prostaglandins suppress human lymphocyte transformation by different mechanisms

    International Nuclear Information System (INIS)

    Yachnin, S.; Lester, E.P.

    1979-01-01

    The capacity of human alpha-fetoprotein (HAFP) to suppress human lymphocyte transformation is well established, although some investigators have reported negative results in their efforts to demonstrate this phenomenon. This discrepancy may reside in the fact that not all isolates of HAFP are potent inhibitors of lymphocyte transformation and that the immunosuppressive potency of various HAFP isolates may be correlated with the proportion of certain negatively charged HAFP isomers which they contain. The possibility was considered that noncovalent binding of low-molecular-weight, negatively charged molecules might be partially responsible. Since fatty acids, including certain prostaglandins (PG), are capable of binding to a partly related serum protein, namely, human serum albumin, and since certain prostaglandins are known to be potent suppressors of human lymphocyte transformation, a study was undertaken of the role which prostaglandins might play in HAFP-induced suppression of human lymphocyte transformation

  11. Validity of Alpha Fetoprotein for Diagnosis of Hepatocellular Carcinoma in Cirrhosis

    International Nuclear Information System (INIS)

    Sarwar, S.; Tarique, S.; Khan, A. A.

    2014-01-01

    Objective: To determine the accuracy of serum alpha fetoprotein (AFP) for diagnosis of hepatocellular carcinoma (HCC) in patients with cirrhosis. Study Design: Observational study. Place and Duration of Study: Department of Medicine, The King Edward Medical University, Lahore, from November 2007 to August 2011. Methodology: Consecutive patients, diagnosed with HCC by contrast enhanced CT, MRI or biopsy were included as cases. Patients of cirrhosis with no evidence of HCC were enrolled as controls. Demographic, laboratory and radiological data were recorded. Serum AFP was determined in all patients at outset and was analyzed using ROC curve for its accuracy in diagnosing HCC. Results: A total of 275 patients were included; of them 173 had HCC and 102 had cirrhosis. One hundred and thirty nine cases (80.3%) with HCC and 86 (84.3%) without HCC had cirrhosis due to HCV. Stage of liver disease, as determined by Child Turcotte Pugh (CTP) score, was comparable; mean CTP value 7.97 A +- 2.1A +- 2.21 in control group (p 0.41). Area under curve (AUC) for AFP was 0.85 (95%, CI: 0.80 - 0.90) with optimum cut off value of 20.85 ng/ml which showed 72.2% sensitivity, 86.2% specificity, 89.9% positive predictive value, 64.7% negative predictive value and 77.4% overall accuracy in diagnosing patients with HCC. Conclusion: Despite sub-optimal sensitivity, alpha fetoprotein is still a valid screening test for diagnosis of hepatocellular carcinoma till other more sensitive markers are developed. Till then, it should be used in conjunction with ultrasound. (author)

  12. Concentration of carcinoembryonic antigen alpha-fetoprotein and beta-subunit of human chorionic gonadotropin in the serum of coke oven workers

    Energy Technology Data Exchange (ETDEWEB)

    Snit, M. [Silesian Medical Academy, Zabrze (Poland)

    1993-01-01

    Increased levels of carcinoembryonic antigen and {alpha}-fetoprotein were found in blood serum of coke oven workers, and also to some extent in smokers and in residents of industrial cities. The {beta} subunit of chorionic gonadotropin was barely detectable.

  13. Evaluation on a radioimmunoassay of. alpha. /sub 1/ microglobulin (. alpha. /sub 1/-m) with simplified procedures

    Energy Technology Data Exchange (ETDEWEB)

    Matsui, Kazuyo; Moriuma, Hatsuko; Mishima, Chiho; Honda, Minoru; Tomonobu, Masahiro; Kanao, Keisuke; Fushimi, Hisako (Sumitomo Hospital, Osaka (Japan))

    1989-06-01

    A newly established double antibody radioimmunoassay (RIA) was fundamentally and clinically evaluated. Original procedures were partially modified as follows: Sample volume for serum and urine was changed to 25{mu}l, and thus 200 mg/l of {alpha}/sub 1/-m standard was prepared using 50 {mu}l of original standard solution (100 mg/l). The results were satisfactory in sensitivity (0.3 mg/l obtained from -2SD method), intraassay precision with its coefficient variation (CV) ranging from 3.0 to 7.4%, interassay precision with its CV ranging from 3.0 to 10.7%, and recovery with the mean value of 102.4% in serum and 108.2% in urine respectively. There were no changes about {alpha}/sub 1/-m value between diluted (2 times) and undiluted with high concentration samples. Normal levels of {alpha}/sub 1/-m were less than 25 mg/l is serum and less than 10 mg/l in urine. The present results indicate that the determination of {alpha}/sub 1/-m could be very simple and useful for the most sensitive screening test for the evaluation of renal function. (author).

  14. [Engineering of a System for the Production of Mutant Human Alpha-Fetoprotein in the Methylotrophic Yeast Pichia pastoris].

    Science.gov (United States)

    Morozkina, E V; Vavilova, E A; Zatsepin, S S; Klyachko, E V; Yagudin, T A; Chulkin, A M; Dudich, I V; Semenkova, L N; Churilova, I V; Benevolenskii, S V

    2016-01-01

    A system for the production of mutant recombinant human alpha-fetoprotein (rhAFPO) lacking the glycosylation site has been engineered in the yeast Pichia pastoris. A strain of the methylotrophic yeast Pichia pastoris GS 115/pPICZ?A/rhAFP0, which produces unglycosylated rhAFPO and secretes it to the culture medium, has been constructed. Optimization and scale-up of the fermentation technology have resulted in an increase in the rhAFP0 yield to 20 mg/L. A scheme of isolation and purification of biologically active rhAFP0 has been developed. The synthesized protein has the antitumor activity, which is analogous to the activity of natural human embryonic alpha-fetoprotein.

  15. Alpha-fetoprotein-L3 and Golgi protein 73 may serve as candidate biomarkers for diagnosing alpha-fetoprotein-negative hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    Zhang ZG

    2015-12-01

    Full Text Available Zhiguo Zhang,1 Yanying Zhang,2 Yeying Wang,1 Lingling Xu,3 Wanju Xu3 1Department of Clinical Laboratory, Zhangqiu Maternity and Child Care Hospital, Zhangqiu, 2Department of Clinical Laboratory, Zaozhuang City Wangkai Infection Hospital, Zaozhuang, 3Department of Clinical Laboratory, Qianfoshan Hospital, Jinan, People’s Republic of China Abstract: Currently, there is no reliable biomarker for use in diagnosing alpha-fetoprotein (AFP-negative hepatocellular carcinoma (HCC. Such a biomarker would aid in making an early diagnosis of AFP-negative HCC, ensuring the timely initiation of treatment. This study examined AFP-L3 and Golgi protein 73 (GP73 as candidate biomarkers for AFP-negative HCC. The affinity adsorption method and enzyme-linked immunoassays were separately used to determine serum levels of AFP-L3 and GP73 in 50 patients with AFP-negative HCC, 30 non-HCC patients, and 50 healthy subjects. Fifty percent of patients with AFP-negative HCC tested positive for AFP-L3, while 3.33% of non-HCC patients and 2.00% of healthy subjects were AFP-L3 positive. Patients with AFP-negative HCC had significantly higher serum levels of AFP-L3 compared to non-HCC patients and healthy individuals; however, there was no significant difference in the AFP-L3 levels of non-HCC patients and healthy subjects. Sixty-six percent of patients with AFP-negative HCC tested positive for GP73, while 10% of non-HCC patients and 0% of healthy subjects were GP73-positive. Patients with AFP-negative HCC had significantly higher serum levels of GP73 compared to non-HCC patients and healthy subjects, but there was no significant difference between the GP73 levels of non-HCC patients and healthy individuals. Moreover, 20 patients with AFP-negative HCC were both AFP-L3- and GP73-positive, while no non-HCC patients or healthy subjects tested positive for both markers. Either AFP-L3 or GP73 may be used as a biomarker for diagnosing AFP-negative HCC, while their combined use

  16. Hepatocellular carcinoma (HCC) and diagnostic significance of alpha-fetoprotein (AFP)

    International Nuclear Information System (INIS)

    Baig, J.A.; Alam, J.M.; Baig, M.; Mahmood, S.R.; Shaheen, R.; Waheed, A.

    2009-01-01

    Alpha-fetoprotein (alpha-fetoprotein, AFP) is a Glycoprotein, belonging to the intriguing class of onco-development protein. Generally designated as tumour marker, AFP is recognized as an important blood component, having specific diagnostic utilities Elevation of its level up to pathological range in adults correlate with the appearance of several malignant and chronic conditions, such as hepatocellular carcinoma (HCC) and chronic liver disease, respectively. To evaluate the diagnostic significance of AFP in HCC, a study was carried out for a period of two years (Jan. 2004 to Dec. 2005) A brief history of Patients was taken with clinical symptoms and signs and initial diagnosis. Patients admitted in wards or visiting OPDs with diagnosis or suspicions of HCC and additional conditions of Chronic Liver disease (CLDs), hepatitis C (HCV) and hepatitis B viral (HBV) infections, were selected and classified according to gender. When confirmed, their HCC status was evaluated and classified according to clinical condition. In 1012 adults including, males 762 (75.3%) and females 250 (24.7%) patients suspected of or diagnosed with HCC and presence of HBV and HCV infections. Out of 480 males, who depicted elevated AFP levels, 39 (8.13%) were diagnosed with HCC. Similarly, 7 (5.34%) females out of 131 with elevated levels of AFP were diagnosed with HCC. Mean elevated AFP levels in all HCC patients were, 421+-59 mu g/ml (range 157-4019 mu g/ml) in males and 163+-32 mu g/ml (range 101-2341 mu g/ml) in females. In males, the overall estimated mean AFP elevated values were analyzed to be 514 mu g/ml (range 67-4019+-59 mu g/ml), whereas in females it was 396+-42 mu g/ml (range 21-2341 mu g/ml). It was also noted that 43 (8.96%) males and 7 (5.34%) female patients, exhibited elevated levels of AFP, however, found negative for HCV and HBV infections. It is concluded that AFP is a significant markers for Hepatocellular carcinoma, helpful in assessing problems in management of HCC and

  17. Specific Genetic Immunotherapy Induced by Recombinant Vaccine Alpha-Fetoprotein-Heat Shock Protein 70 Complex

    Science.gov (United States)

    Wang, Xiaoping; Lin, Huanping; Wang, Qiaoxia

    Purposes: To construct a recombinant vaccine alpha-fetoprotein (AFP)-heat shock protein (HSP70) complex, and study its ability to induce specific CTL response and its protective effect against AFP-producing tumor. Material/Methods: A recombinant vaccine was constructed by conjugating mouse alpha-fetoprotein to heat shock protein 70. By way of intracutaneous injection, mice were primed and boosted with recombinant vaccine mAFP/HSP70, whereas single mAFP or HSP70 injection as controls. The ELISPOT and ELISA were used to measure the frequency of cells producing the cytokine IFN-γ in splenocytes and the level of anti-AFP antibody of serum from immunized mice respectively. In vivo tumor challenge were carried out to assess the immune effect of the recombinant vaccine. Results: By recombinant mAFP/HSP70 vaccine immunization, the results of ELISPOT and ELISA showed that the number of splenic cells producing IFN-γ and the level of anti-AFP antibody of serum were significantly higher in mAFP/HSP70 group than those in mAFP and HSP70 groups (108.50±11.70 IFN-γ spots/106 cells vs 41.60±10.40 IFN-γ spots/106 cells, 7.32±3.14 IFN-γ spots/106 cells, P<0.01; 156.32±10.42 μg/mL vs 66.52±7.35 μg/mL, 5.73±2.89 μg/mL, P<0.01). The tumor volume in mAFP/HSP70 group was significantly smaller than that in mAFP and HSP70 groups (42.44±7.14 mm3 vs 392.23±12.46 mm3, 838.63±13.84 mm3, P<0.01). Conclusions: The study further confirmed the function of heat shock protein 70's immune adjuvant. Sequential immunization with recombinant mAFP/HSP70 vaccine could generate effective antitumor immunity on AFP-producing tumor. The recombined mAFP/HSP70 vaccine may be suitable for serving as an immunotherapy for hepatocellular carcinoma.

  18. Localization of radioiodinated antibody to alpha-fetoprotein in rats with transplanted hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Koji, T; Ishii, N; Munehisa, T; Kusumoto, Y; Nakamura, S; Tamenishi, A [Nagasaki Univ. (Japan). School of Medicine; Kobayashi, K; Hara, A; Tsukada, Y; Nishi, S

    1980-01-01

    Total body scintigraphy, organ and subcellular distribution of radioactivity and autoradiography of tissue sections has been assessed in an animal model using radioiodinated horse antibody to rat alpha-fetoprotein (AFP). Rats bearing subcutaneous transplants of AH-7974 ascites hepatoma were injected with /sup 125/I-labeled anti-AFP and scintigraphed. Localization of radioactivity in the tumors was observed 48-168 h after injection. Scintigraphy using /sup 125/I-labeled F(ab')/sub 2/ fragment of the antibody gave approximately the same results as that with the intact anti-AFP antibody. /sup 125/I-labeled normal horse IgG was used as control. The tumor/blood radioactivity ratio after a week after injection was approximately four times higher in the antibody group than that in the control group. This ratio suggested an active accumulation of radioactive antibody in the tumor tissue. In its subcellular distribution, about 30 to 60% of the total radioactivity administered was found in a fraction of the cell membrane plus nucleus. The specific activity of this fraction increased in the antibody group with time over 10 days. In autoradiograms of the fixed tissue sections specific localization of the antibody was observed on the tumor cell surface. The specific uptake of radiolabeled antibody to AFP into AFP producing tumor cells was confirmed.

  19. Clinicopathologic and prognostic characteristics of alpha-fetoprotein-producing gastric cancer.

    Science.gov (United States)

    He, Ruji; Yang, Qinyi; Dong, Xuqiang; Wang, Yao; Zhang, Weiming; Shen, Lizong; Zhang, Zhihong

    2017-04-04

    Alpha-fetoprotein-producing gastric cancer (AFPGC) accounts for 1.5%-7.1% of all gastric cancer cases. Compared with other types of gastric cancer, AFPGC is more aggressive and prone to liver and lymph node (LN) metastasis, with extremely poor prognosis. To improve understanding of AFPGC we reviewed a consecutive series of 82 AFPGC patients and investigated the prognostic factors. The incidence of AFPGC among our gastric cancer patients was 1.95%, and 29.27% of AFPGCs were diagnosed with metastasis at the time of presentation, mainly liver metastasis. The serum AFP level of patients with AFPGC was significantly associated with tumor differentiation. Histologically, these AFPGC patients were composed of 34.55% hapatiod type, 58.18% fetal gastrointestinal type, 9.09% yolk sac tumor-like type, and 14.55% mixed type. Patient gender, tumor differentiation, Lauren classification, and number of metastatic lymph nodes showed significant differences among these four subtypes. The overall survival time was 42.02 months and the 3-year cumulative survival rate was 53.13%. Age, American Joint Committee on Cancer (AJCC) TNM staging classification (TNM stage), serum AFP level, and surgery were prognostic factors for overall survival; however, TNM stage was the only independent risk factor for prognosis of AFPGC. In short, AFPGC is a rare, unique, and heterogeneous entity, and its proper identification and treatment remain a challenge. More attention should be paid to AFPGC to improve patient care and the dismal prognosis.

  20. The prognostic utility of baseline alpha-fetoprotein for hepatocellular carcinoma patients.

    Science.gov (United States)

    Silva, Jack P; Gorman, Richard A; Berger, Nicholas G; Tsai, Susan; Christians, Kathleen K; Clarke, Callisia N; Mogal, Harveshp; Gamblin, T Clark

    2017-12-01

    Alpha-fetoprotein (AFP) has a valuable role in postoperative surveillance for hepatocellular carcinoma (HCC) recurrence. The utility of pretreatment or baseline AFP remains controversial. The present study hypothesized that elevated baseline AFP levels are associated with worse overall survival in HCC patients. Adult HCC patients were identified using the National Cancer Database (2004-2013). Patients were stratified according to baseline AFP measurements into the following groups: Negative (2000). The primary outcome was overall survival (OS), which was analyzed by log-rank test and graphed using Kaplan-Meier method. Multivariate regression modeling was used to determine hazard ratios (HR) for OS. Of 41 107 patients identified, 15 809 (33.6%) were Negative. Median overall survival was highest in the Negative group, followed by Borderline, Elevated, and Highly Elevated (28.7 vs 18.9 vs 8.8 vs 3.2 months; P < 0.001). On multivariate analysis, overall survival hazard ratios for the Borderline, Elevated, and Highly Elevated groups were 1.18 (P = 0.267), 1.94 (P < 0.001), and 1.77 (P = 0.007), respectively (reference Negative). Baseline AFP independently predicted overall survival in HCC patients regardless of treatment plan. A baseline AFP value is a simple and effective method to assist in expected survival for HCC patients. © 2017 Wiley Periodicals, Inc.

  1. p55PIK regulates alpha-fetoprotein expression through the NF-κB signaling pathway.

    Science.gov (United States)

    Ye, Guoguo; Sun, Ge; Cheng, Zhikui; Zhang, Lei; Hu, Kanghong; Xia, Xianmin; Zhou, Yin

    2017-12-15

    Alpha-fetoprotein (AFP) is regarded as a diagnostic and prognostic biomarker and a potential therapeutic target for hepatocellular carcinoma (HCC). However, the regulation of AFP expression in HCC remains poorly understood. This study aimed to investigate the mechanism by which AFP expression is regulated by p55PIK, an isoform of PI3K. Human HCC cell lines (HepG2 and Huh-7) were treated with p55PIK specific competitive inhibitor or shRNA, or p55PIK overexpression vector, in the absence or presence of NF-κB inhibitor PDTC. AFP expression was detected by quantitative real-time PCR and Western blotting. NF-κB responsive elements in AFP enhancer region were characterized by luciferase reporter assay. p55PIK significantly stimulated the expression of AFP by activating NF-κB signaling pathway in HCC cells. Furthermore, two NF-κB binding sites in AFP enhancer region were identified to be primarily responsible for p55PIK mediated upregulation of AFP expression. p55PIK/NF-κB signaling plays an important role in the upregulation of AFP expression in HCC. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Antibody modified gold nano-mushroom arrays for rapid detection of alpha-fetoprotein.

    Science.gov (United States)

    Li, Wanbo; Jiang, Xueqin; Xue, Jiancai; Zhou, Zhangkai; Zhou, Jianhua

    2015-06-15

    Localized surface plasmon resonance (LSPR) combined with immunoassay shows greatly potential in fast detection of tumor markers. In this paper, a highly sensitive LSPR substrate has been fabricated and modified for direct detection of alpha-fetoprotein (AFP). The biosensor was prepared by interference lithography, and modified by covalently immobilizing anti-AFP on the surface of gold nano-mushroom arrays (GNMA). The modification process was investigated by Vis-NIR reflectance spectra and cyclic voltammogram measurements. We revealed the optical properties of the modified GNMA by measuring the Vis-NIR reflectance spectra and simulating its electric intensity field distribution under light illumination. The GNMA substrate was highly sensitive, with a refractive index sensitivity of ~465 nm/RIU. The substrate can be applied to label-free detection of AFP, with the linear range and the limit of detection determined to be 20-200 ng/mL and 24 ng/mL (S/N=3), respectively. We also demonstrated its clinical application by directly detecting AFP in human serum samples. It is expected that our biosensor could be integrated on microfluidic chips for high-throughput detection in portable early diagnosis, post-operative and point-of-care (POC) in clinical applications. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Phenylboronic acid immunoaffinity reactor coupled with flow injection chemiluminescence for determination of {alpha}-fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Wu Yafeng [Jiangsu Provincial Key Lab of Biomaterials and Biodevices, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 210096 (China); Zhuang Yafeng [Department of Chemistry, Changzhou Institute of Technology, Changzhou 213022 (China); Liu Songqin [Jiangsu Provincial Key Lab of Biomaterials and Biodevices, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 210096 (China)], E-mail: liusq@seu.edu.cn; He Lin [Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204 (United States)

    2008-12-23

    A reusable and sensitive immunoassay based on phenylboronic acid immunoaffinity reactor in combination with flow injection chemiluminescence (CL) for determination of glycoprotein was described. The reactor was fabricated by immobilizing 3-aminophenylboronic acid (APBA) on glass microbeads with {gamma}-glycidoxypropyltrimethoxysilane (GPMS) as linkage. The {alpha}-fetoprotein (AFP) could be easily immobilized on the APBA coated beads through sugar-boronic interaction. After an off-line incubation, the mixture of the analyte AFP with horseradish peroxidase-labeled AFP antibody (HRP-anti-AFP) was injected into the reactor. This led the trapping of free HRP-anti-AFP by the surface coated AFP on glass beads. The trapped HRP-anti-AFP was detected by chemiluminescence due to its sensitizing effect on the reaction of luminol and hydrogen peroxide. Under optimal conditions, the chemiluminescent signal was proportional to AFP concentration in the range of 10-100 ng mL{sup -1}. The whole assay process including regeneration of the reactor could be completed within 31 min. The proposed system showed acceptable detection and fabrication reproducibility, and the results obtained with the present method were in acceptable agreement with those from parallel single-analyte test of practical clinical sera. The described method enabled a low-cost, time saving and was potential to detect the serum AFP level in clinical diagnosis.

  4. Label-free immunosensor based on Pd nanoplates for amperometric immunoassay of alpha-fetoprotein.

    Science.gov (United States)

    Wang, Huan; Li, He; Zhang, Yihe; Wei, Qin; Ma, Hongmin; Wu, Dan; Li, Yan; Zhang, Yong; Du, Bin

    2014-03-15

    In this paper, Pd nanoplates were used as a kind of electrode materials for fabrication of an electrochemical immunosensor, which was applied for detection of cancer biomarker alpha-fetoprotein (AFP). Thanks to the unique structure and properties of Pd nanoplates, the antibody of AFP (Ab) was effectively immobilized onto the surface of the Pd nanoplates modified glassy carbon electrode (GCE). Moreover, the good electrochemical properties of Pd nanoplates greatly improved the electronic transmission rate and enhanced the electrochemical signal, which led to an increase of the detection sensitivity. Based on the specific antibody-antigen interaction, a label-free immunosensor based on Pd nanoplates was developed for sensing of AFP. The current method allows us to detect AFP over a wide concentration range from 0.01 to 75.0 ng/mL with a detection limit of 4 pg/mL. The proposed immunosensor has been used to determine AFP in human serum with satisfactory results. © 2013 Elsevier B.V. All rights reserved.

  5. Alpha-fetoprotein as a prognostic marker in acute liver failure: a pilot study.

    Science.gov (United States)

    Varshney, Anshul; Gupta, Rohit; Verma, Sanjiv K; Ahmad, Sohaib

    2017-07-01

    Prognostic markers of acute liver failure (ALF) are based on clinical, laboratory or radiological parameters. Most of the biochemical markers are based on hepatic degeneration. We studied the impact of serial serum alpha-fetoprotein (AFP) levels, a marker of liver regeneration, on the outcome of the patients with ALF. AFP levels were estimated on days 1 and 3 of hospitalisation of 32 patients with ALF and the ratio (AFP day3/day1) was calculated. All subjects were categorised as group A (expired) or group B (survived). The AFP ratio was 0.84  +  0.15 in group A (n = 20) versus 1.55  +  0.70 in group B (n = 10); P < 0.001. However, the absolute initial AFP values were not associated with the outcome, favourable or unfavourable. We conclude that AFP levels change dynamically during ALF and have the potential to be used as a predictor of outcome in isolation or in combination with well-established prognostic markers.

  6. The utility of alpha-fetoprotein screening in Beckwith-Wiedemann syndrome.

    Science.gov (United States)

    Duffy, Kelly A; Deardorff, Matthew A; Kalish, Jennifer M

    2017-03-01

    Beckwith-Wiedemann syndrome (BWS) is one of the most common cancer predisposition disorders. As a result, BWS patients receive tumor screening as part of their clinical management. Until recently, this screening has been employed uniformly across all genetic and epigenetic causes of BWS, including the utilization of ultrasonography to detect abdominal tumors and alpha-fetoprotein (AFP) to detect hepatoblastoma. The advancements in our understanding of the genetics and epigenetics leading to BWS has evolved over time, and has led to the development of genotype/phenotype correlations. As tumor risk appears to correlate with genetic and epigenetic causes of BWS, several groups have proposed alterations to tumor screening protocols based on the etiology of BWS, with the elimination of AFP as a screening measure and the elimination of all screening measures in BWS patients with loss of methylation at the KCNQ1OT1:TSS-DMR 2 (IC2). There are many challenges to this suggestion, as IC2 patients may have additional factors that contribute to risk of hepatoblastoma including fetal growth patterns, relationship with assisted reproductive technologies, and the regulation of the IC2 locus. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  7. Structural basis for alpha fetoprotein-mediated inhibition of caspase-3 activity in hepatocellular carcinoma cells.

    Science.gov (United States)

    Lin, Bo; Zhu, Mingyue; Wang, Wenting; Li, Wei; Dong, Xu; Chen, Yi; Lu, Yan; Guo, Junli; Li, Mengsen

    2017-10-01

    Alpha-fetoprotein (AFP) is an early serum growth factor in the foetal liver development and hepatic carcinogenesis; However, the precise biological role of cytoplasmic AFP remains elusive. Although we recently demonstrated that cytoplasmic AFP might interact with caspase-3 and inhibit the signal transduction of apoptosis in human hepatocellular carcinoma (HCC) cells, the details of this interaction are not clear. To reveal the molecular relationship between AFP and caspase-3, we performed molecular docking, co-immunoprecipitation (Co-IP), laser confocal microscopy, site-directed mutagenesis and functional experiments to analyse the key amino acid residues in the binding site of caspase-3. The results of Co-IP, laser confocal microscopy and functional analyses were consistent with the computational model. We also used the model to explain why AFP cannot bind to caspase-8. These results provide the molecular basis for the AFP-mediated inhibition of caspase-3 activity in HCC cells. Altogether, we found that AFP interacts with caspase-3 through precise amino acids, namely loop-4 residues Glu-248, Asp-253 and His-257. The results further demonstrated that AFP plays a critical role in the inhibition of the apoptotic signal transduction that mediated by caspase-3. Thus, AFP might represent a novel biotarget for the therapy of HCC patients. © 2017 UICC.

  8. Alpha-fetoprotein-targeted reporter gene expression imaging in hepatocellular carcinoma.

    Science.gov (United States)

    Kim, Kwang Il; Chung, Hye Kyung; Park, Ju Hui; Lee, Yong Jin; Kang, Joo Hyun

    2016-07-21

    Hepatocellular carcinoma (HCC) is one of the most common cancers in Eastern Asia, and its incidence is increasing globally. Numerous experimental models have been developed to better our understanding of the pathogenic mechanism of HCC and to evaluate novel therapeutic approaches. Molecular imaging is a convenient and up-to-date biomedical tool that enables the visualization, characterization and quantification of biologic processes in a living subject. Molecular imaging based on reporter gene expression, in particular, can elucidate tumor-specific events or processes by acquiring images of a reporter gene's expression driven by tumor-specific enhancers/promoters. In this review, we discuss the advantages and disadvantages of various experimental HCC mouse models and we present in vivo images of tumor-specific reporter gene expression driven by an alpha-fetoprotein (AFP) enhancer/promoter system in a mouse model of HCC. The current mouse models of HCC development are established by xenograft, carcinogen induction and genetic engineering, representing the spectrum of tumor-inducing factors and tumor locations. The imaging analysis approach of reporter genes driven by AFP enhancer/promoter is presented for these different HCC mouse models. Such molecular imaging can provide longitudinal information about carcinogenesis and tumor progression. We expect that clinical application of AFP-targeted reporter gene expression imaging systems will be useful for the detection of AFP-expressing HCC tumors and screening of increased/decreased AFP levels due to disease or drug treatment.

  9. Detection of alpha-fetoprotein in magnetic immunoassay of thin channels using biofunctional nanoparticles

    Science.gov (United States)

    Tsai, H. Y.; Gao, B. Z.; Yang, S. F.; Li, C. S.; Fuh, C. Bor

    2014-01-01

    This paper presents the use of fluorescent biofunctional nanoparticles (10-30 nm) to detect alpha-fetoprotein (AFP) in a thin-channel magnetic immunoassay. We used an AFP model biomarker and s-shaped deposition zones to test the proposed detection method. The results show that the detection using fluorescent biofunctional nanoparticle has a higher throughput than that of functional microparticle used in previous experiments on affinity reactions. The proposed method takes about 3 min (versus 150 min of previous method) to detect 100 samples. The proposed method is useful for screening biomarkers in clinical applications, and can reduce the run time for sandwich immunoassays to less than 20 min. The detection limits (0.06 pg/ml) and linear ranges (0.068 pg/ml-0.68 ng/ml) of AFP using fluorescent biofunctional nanoparticles are the same as those of using functional microparticles within experimental errors. This detection limit is substantially lower and the linear range is considerably wider than those of enzyme-linked immunosorbent assay (ELISA) and other methods in sandwich immunoassay methods. The differences between this method and an ELISA in AFP measurements of serum samples were less than 12 %. The proposed method provides simple, fast, and sensitive detection with a high throughput for biomarkers.

  10. Association between serum alpha-fetoprotein levels and fatty liver disease: A cross-sectional study

    Science.gov (United States)

    Xu, Ping; Xu, Cheng-Fu; Wan, Xing-Yong; Yu, Chao-Hui; Shen, Chao; Chen, Peng; Xu, Gen-Yun; Li, You-Ming

    2014-01-01

    AIM: To investigate the association between serum alpha-fetoprotein (AFP) levels and fatty liver disease (FLD) in a Chinese population. METHODS: A cross-sectional study was performed among subjects who presented for a health examination at the First Affiliated Hospital, College of Medicine, Zhejiang University in 2013. FLD was diagnosed based on an ultrasonography examination. Serum AFP levels were measured with a chemiluminescence immunoassay. RESULTS: Of the 9800 subjects enrolled, 2601 were diagnosed with FLD. Subjects with FLD had higher serum AFP levels than those without the disease. Subjects with high serum AFP levels had a higher prevalence of FLD, metabolic syndrome, and its components. Univariate logistic analysis showed that elevated serum AFP levels were associated with an increased risk of FLD (OR = 1.057, 95%CI: 1.031-1.084). However, after adjusting for covariates, AFP no longer remained significantly associated with the risk factors for FLD. CONCLUSION: Our results suggest that serum AFP levels are significantly associated with FLD and that AFP acts as a cofactor, but not as an independent factor, for FLD. PMID:25206293

  11. Nanobody medicated immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein.

    Science.gov (United States)

    Chen, Jing; He, Qing-hua; Xu, Yang; Fu, Jin-heng; Li, Yan-ping; Tu, Zhui; Wang, Dan; Shu, Mei; Qiu, Yu-lou; Yang, Hong-wei; Liu, Yuan-yuan

    2016-01-15

    Immunoassay for cancer biomarkers plays an important role in cancer prevention and early diagnosis. To the development of immunoassay, the quality and stability of applied antibody is one of the key points to obtain reliability and high sensitivity for immunoassay. The main purpose of this study was to develop a novel immunoassay for ultrasensitive detection of cancer biomarker alpha-fetoprotein (AFP) based on nanobody against AFP. Two nanobodies which bind to AFP were selected from a phage display nanobody library by biopanning strategy. The prepared nanobodies are clonable, thermally stable and applied in both sandwich enzyme linked immunoassay (ELISA) and immuno-PCR assay for ultrasensitive detection of AFP. The limit detection of sandwich ELISA setup with optimized nanobodies was 0.48ng mL(-1), and the half of saturation concentration (SC50) value was 6.68±0.56ng mL(-1). These nanobodies were also used to develop an immuno-PCR assay for ultrasensitive detection of AFP, its limit detection values was 0.005ng mL(-1), and the linear range was 0.01-10,000ng mL(-1). These established immunoassays based on nanobodies were highly specific to AFP and with negligible cross reactivity with other tested caner biomarkers. Furthermore, this novel concept of nanobodies mediated immunoassay may provide potential applications in a general method for the ultrasensitive detection of various cancer biomarkers. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Alpha-fetoprotein-producing esophageal adenocarcinoma: a mimicker of hepatocellular carcinoma.

    Science.gov (United States)

    Wang, Jeremy; Liu, Wendy; Parikh, Keyur; Post, Anthony Benjamin

    2017-02-01

    Alpha-fetoprotein (AFP)-producing esophageal adenocarcinoma (EAC) is a rare occurrence. Elevation of serum AFP is commonly associated with hepatocellular carcinoma and yolk sac tumors, but rarely with esophageal carcinoma. Here, we report a rare case of AFP-producing EAC. A 51-year-old man presented with two weeks of acid reflux and a 35-lb weight loss. Laboratory data were notable for transaminitis and AFP was 2524 ng/mL. Computed tomography of the abdomen revealed abnormal thickening of the esophagus and multiple metastatic masses throughout the liver. Biopsy of one of the masses revealed adenocarcinoma of gastrointestinal origin. Subsequent upper endoscopy revealed an esophageal mass with biopsy notable for ulcerated dysplastic glandular mucosa with likely underlying malignancy. The patient underwent palliative esophageal stent placement but died two months later. Elevated AFP levels are an unusual occurrence in EAC. Prognosis is poor given its advanced presenting stage and high metastatic potential. Most cases are unsuccessfully treated with surgery and chemotherapy. Serial measurement of serum AFP may be useful for monitoring clinical status and treatment response. Clinicians should consider AFP-producing EAC in their differential diagnosis in the work-up of a liver mass in the setting of elevated AFP or liver function impairment, especially in the absence of chronic liver disease.

  13. Capillary electrochromatography immunoassay for alpha-fetoprotein based on poly(guanidinium ionic liquid) monolithic material.

    Science.gov (United States)

    Liu, Cuicui; Deng, Qiliang; Fang, Guozhen; Dang, Meng; Wang, Shuo

    2017-08-01

    Alpha-fetoprotein (AFP) is widely used as a tumor marker for the serum diagnosis of primary hepatoma. Sensitive detection of AFP level plays an important role in the early diagnosis of disease and highly reliable prediction. In this study, a novel non-competitive immunoassay (IA) based on poly(guanidinium ionic liquid) monolithic material was developed for detecting ultra trace levels of AFP in capillary electrochromatography (CEC) mode. The AFP was mixed with an excess amount of fluorescently labeled antibody. After incubation, the immunocomplex was separated from the free labeled antibody and detected by CEC coupled with laser-induced fluorescence detector. Under the optimized conditions, the developed CEC-IA performed a low detection limit of 0.05 μg L -1  (S/N = 3) and a wide linearity ranging from 0.1 to 1000 μg L -1 for AFP, which can be largely attributed to the high separation and enrichment efficiency of poly(guanidinium ionic liquid) monolithic material for the targets. The application of this method was demonstrated by determining AFP in human serum. Copyright © 2017. Published by Elsevier Inc.

  14. Reference Intervals of Alpha-Fetoprotein and Carcinoembryonic Antigen in the Apparently Healthy Population.

    Science.gov (United States)

    Zhang, Gao-Ming; Guo, Xu-Xiao; Ma, Xiao-Bo; Zhang, Guo-Ming

    2016-12-12

    BACKGROUND The aim of this study was to calculate 95% reference intervals and double-sided limits of serum alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) according to the CLSI EP28-A3 guideline. MATERIAL AND METHODS Serum AFP and CEA values were measured in samples from 26 000 healthy subjects in the Shuyang area receiving general health checkups. The 95% reference intervals and upper limits were calculated by using MedCalc. RESULTS We provided continuous reference intervals from 20 years old to 90 years old for AFP and CEA. The reference intervals were: AFP, 1.31-7.89 ng/ml (males) and 1.01-7.10 ng/ml (females); CEA, 0.51-4.86 ng/ml (males) and 0.35-3.45ng/ml (females). AFP and CEA were significantly positively correlated with age in both males (r=0.196 and r=0.198) and females (r=0.121 and r=0.197). CONCLUSIONS Different races or populations and different detection systems may result in different reference intervals for AFP and CEA. Continuous reference intervals of age changes are more accurate than age groups.

  15. Serial Changes in Alpha-Fetoprotein Levels During Therapy for Chronic Hepatitis C

    Directory of Open Access Journals (Sweden)

    Altug Senol

    2014-12-01

    Full Text Available Aim: Alpha-fetoprotein (AFP has been widely used as a diagnostic marker for hepatocellular carcinoma. Some patients with hepatitis C show high AFP values, but no evidence of hepatocellular carcinoma. The aim of this study is to assess the influence of antiviral treatment on the serum AFP in patients with chronic hepatitis C without hepatocellular carcinoma. Material and Method: Thirty seven chronic hepatitis C patients (20 females and 17 males were included in the study. All patients were given a combined treatment of pegylated or conventional interferon (IFN and ribavirin. Serum AFP was measured at baseline and on months 3-6-12 of the therapy. Results: Compared to the pretreatment levels of ALT (88,59 ± 57,22 IU, those at 3, 6 and 12 months were statistically lower (p0,05, to 4,34 ± 4,64 (p>0,05 and to 2,63 ± 2,17 (p10 ng/ml. In these patients, mean serum AFP levels were decreased from pretreatment level of 15,09 ± 5,92 ng/ml to 11,39±3,30, to 6,97±2,53 (p

  16. Clinical value of imaging using antibody to alpha fetoprotein in germ cell tumours

    International Nuclear Information System (INIS)

    Hitchins, R.N.; Begent, R.H.J.; Green, A.J.; Searle, F.; Bagshawe, K.D.; Charing Cross Group of Hospitals, London; Van Heyningen, V.

    1989-01-01

    Germ cell tumours (GCT) producing alpha fetoprotein (aFP) can be imaged by external scintigraphy after intraveneous administration of radiolabelled antibody directed against aFP. Antibody imaging (AI) by this method was used in an attempt to guide surgical resection of deposits of drug-resistant or recurrent GCT. 30 patients with GCT and raised aFP in whom site of tumour was not known were investigated by AI and conventional imaging methods. All but one were heavily pretreated. Where tumour appeared localised, resection was attempted. Tumour was found in all sites positive by both AI and conventional imaging. AI produced false-positive results in one of 30 patients and false-negative results in 9 patients. Computerised tomography was false-positive in one case and false-negative in three. In these patients, AI gave true-negative and true-positive results, respectively. Of 11 patients with positive AI in whom resection was attempted, 6 achieved sustained complete response with up to 5 years follow-up. We conclude AI and conventional imaging methods to be complementary in selection for surgery of patients with drug-resistant or recurrent GCT. (orig.) [de

  17. Alpha-fetoprotein and Fanconi Anemia: Relevance to DNA Repair and Breast Cancer Susceptibility.

    Science.gov (United States)

    Lakhi, Nisha A; Mizejewski, Gerald J

    2017-02-01

    Elevations of serum alpha-fetoprotein (sAFP) have been reported in fetal and infant states of anemia. Fanconi anemia (FA) belongs to a family of genetic instability disorders which lack the capability to repair DNA breaks. The lesion occurs at a checkpoint regulatory step of the G2 to mitotic transition, allowing FA cells to override cell-cycle arrest. FA DNA repair pathways contain complementation groups known as FANC proteins. FANC proteins form multi-protein complexes with BRCA proteins and are involved in homologous DNA repair. An impaired cascade in these events imparts an increased breast cancer susceptibility to female FA patients. Elevations of sAFP have availed this fetal protein to serve as a biomarker for FA disease. However, the origin of the synthesis of sAFA has not been determined in FA patients. We hypothesize that hematopoietic multipotent progenitor stem cells in the bone marrow are the source of sAFP production in FA patients.

  18. Immunohistochemical and radioimmunological demonstration of alpha1-fetoprotein in nonmalignant changes of human gastric mucosa

    International Nuclear Information System (INIS)

    Falser, N.; Lederer, B.; Reissigl, H.; Innsbruck Univ.

    1977-01-01

    The occurence of α 1 fetoprotein in nonmalignant changes of the gastric mucosa was investigated by means of immunohistochemistry and radioimmunonoassay. The investigations were performed in tissue sections, cytological imprint preparations as well as in homogenized tissue samples (obtained by gastroscopy). α 1 fetoprotein could be demonstrated by immuno-histochemistry in about 90% of the samples originating from the surroundings of gastric ulcer and the region of gastrojejunostomy after B II-resection. The RIA was positive in about 75% of the tissue samples, whereas from gastric juice only 40% of positive results could be obtained. No α 1 fetoprotein-activity could be demonstrated in serum samples. These investigations indicate that α 1 fetoprotein is not exclusively synthesized by embryonic or neoplastic tissues and also can be synthesized also by regenerating cell-systems. It may be supposed that this synthesis represents an unspecific answer to growth-stimulation. (orig.) [de

  19. Radioimmunodetection of hepatocellular carcinoma with a radiolabeled antibody to alpha fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Ishii, N; Nakata, K; Muro, T [Nagasaki Univ. (Japan). School of Medicine

    1982-03-01

    The present study was undertaken to investigate whether or not radiolabeled antibody to Alpha-fetoprotein (AFP) intravenously administered accumulates in AFP-producing tumors. Rats with subcutaneously transplanted hepatoma or with heptoma produced in liver by 3'-Me-DAB, and 12 patients with hepatocellular carcinoma (HCC) were studied. In animal experiments, the tumor localization was clearly demonstrated by total body scintigraphy 48 to 168 hours after the injection of the radioiodinated antibody. The rats were sacrificed and radioactivity in tissues was counted. Tumor tissues showed the highest counts. In patients with HCC, the imaging was made with a gamma camera usually 24 and 48 hours after injection of I-131-labeled antibody. In order to make the contrast of tumor image clear, nonspecific background images obtained by administration of Tc-99m-labeled serum albumin were subtracted from the images obtained by I-131 labeled antibody with a computer. In 6 of 12 patients with HCC, the tumor location could successfully demonstrated. However, tumor with sizes below 2 cm in diameter which were able to be detected by the celiac angiography were not detectable. Interestingly the images were more clear in patients with relatively higher serum AFP levels. In the blood of patient given injection of AFP radioantibody, both immune complex and free antibody were able to be detected at least for 120 hours so that the presence of free antigen (AFP) did not appear to prevent tumor radioimmunodetection. The information obtained concerning the biochemical factors which influence antibody localization in the tumor would provide better method for radioimmunodetection with antitumor antibodies.

  20. Clinicopathological features of alpha-fetoprotein producing early gastric cancer with enteroblastic differentiation.

    Science.gov (United States)

    Matsumoto, Kohei; Ueyama, Hiroya; Matsumoto, Kenshi; Akazawa, Yoichi; Komori, Hiroyuki; Takeda, Tsutomu; Murakami, Takashi; Asaoka, Daisuke; Hojo, Mariko; Tomita, Natsumi; Nagahara, Akihito; Kajiyama, Yoshiaki; Yao, Takashi; Watanabe, Sumio

    2016-09-28

    To investigate clinicopathological features of early stage gastric cancer with enteroblastic differentiation (GCED). We retrospectively investigated data on 6 cases of early stage GCED and 186 cases of early stage conventional gastric cancer (CGC: well or moderately differentiated adenocarcinoma) who underwent endoscopic submucosal dissection or endoscopic mucosal resection from September 2011 to February 2015 in our hospital. GCED was defined as a tumor having a primitive intestine-like structure composed of cuboidal or columnar cells with clear cytoplasm and immunohistochemical positivity for either alpha-fetoprotein, Glypican 3 or SALL4. The following were compared between GCED and CGC: age, gender, location and size of tumor, macroscopic type, ulceration, depth of invasion, lymphatic and venous invasion, positive horizontal and vertical margin, curative resection rate. Six cases (5 males, 1 female; mean age 75.7 years; 6 lesions) of early gastric cancer with a GCED component and 186 cases (139 males, 47 females; mean age 72.7 years; 209 lesions) of early stage CGC were investigated. Mean tumor diameters were similar but rates of submucosal invasion, lymphatic invasion, venous invasion, and non-curative resection were higher in GCED than CGC (66.6% vs 11.4%, 33.3% vs 2.3%, 66.6% vs 0.4%, 83.3% vs 11% respectively, P < 0.01). Deep submucosal invasion was not revealed endoscopically or by preoperative biopsy. Histologically, in GCED the superficial mucosal layer was covered with a CGC component. The GCED component tended to exist in the deeper part of the mucosa to the submucosa by lymphatic and/or venous invasion, without severe stromal reaction. In addition, Glypican 3 was the most sensitive marker for GCED (positivity, 83.3%), immunohistochemically. Even in the early stage GCED has high malignant potential, and preoperative diagnosis is considered difficult. Endoscopists and pathologists should know the clinicopathological features of this highly malignant type

  1. Serum alpha-fetoprotein response can predict prognosis in hepatocellular carcinoma patients undergoing radiofrequency ablation therapy

    Energy Technology Data Exchange (ETDEWEB)

    Kao, W.-Y. [Division of Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taiwan (China); Chiou, Y.-Y., E-mail: yychiou@vghtpe.gov.tw [Department of Radiology, Taipei Veterans General Hospital, Taiwan (China); Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Hung, H.-H. [Division of Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taiwan (China); Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Su, C.-W., E-mail: cwsu2@vghtpe.gov.tw [Division of Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taiwan (China); Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Institute of Clinical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Chou, Y.-H. [Department of Radiology, Taipei Veterans General Hospital, Taiwan (China); Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Wu, J.-C. [Institute of Clinical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Department of Medical Research and Education, Taipei Veterans General Hospital, Taiwan (China); Huo, T.-I. [Division of Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taiwan (China); Institute of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Huang, Y.-H. [Division of Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taiwan (China); Institute of Clinical Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan (China); Wu, W.-C. [Division of Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taiwan (China)

    2012-05-15

    Aims: To evaluate the clinical inference of serum alpha-fetoprotein (AFP) response in hepatocellular carcinoma (HCC) patients undergoing percutaneous radiofrequency ablation (RFA). Materials and methods: Three hundred and thirteen previously untreated HCC patients were enrolled in the study. The optimal AFP response was defined as >20% decrease from baseline after 1 month of RFA for those with a baseline AFP level of {>=}100 ng/ml. The impact of AFP response on prognosis was analysed and prognostic factors were assessed. Results: After a median follow-up of 26.7 {+-} 19.1 months, 49 patients died and 264 patients were alive. The cumulative 5 year survival rates were 75.3 and 57.4% in patients with an initial AFP of <100 ng/ml and {>=}100 ng/ml, respectively (p = 0.003). In the 58 patients with a baseline AFP of {>=}100 ng/ml and initial completed tumour necrosis after RFA, the cumulative 5 year survival rates were 62.4 and 25.7% in optimal and non-optimal AFP responders, respectively (p = 0.001). By multivariate analysis, the prothrombin time international normalized ratio >1.1 (p = 0.009), non-optimal AFP response (p = 0.023), and creatinine >1.5 mg/dl (p = 0.021) were independent risk factors predictive of poor overall survival. Besides, the cumulative 5 year recurrence rates were 83.4 and 100% in optimal and non-optimal AFP responders, respectively (p < 0.001). Multivariate analysis demonstrated platelet count {<=}10{sup 5}/mm{sup 3} (p = 0.048), tumour size >2 cm (p = 0.027), and non-optimal AFP response (p < 0.001) were independent risk factors associated with tumour recurrence after RFA. Conclusions: Serum AFP response may be a useful marker for predicting prognosis in HCC patients undergoing RFA.

  2. Alpha-fetoprotein and hepatitis B virus infection in chronic liver disease

    International Nuclear Information System (INIS)

    Solina, A.

    1987-01-01

    Among 177 patients with chronic non-malignant liver disease, the prevalence of raised levels of alpha-fetoprotein (AFP) was 11.2 percent. A close association between the severity of the liver disease and AFP concentrations was observed. The most frequent pattern of hepatitis B virus (HBV) infection in these patients was the presence of antibodies to hepatitis B surface antigen (anti-HBs) and hepatitis B core antigen (anti-HBc), whilst only 5 cases were positive for hepatitis B surface antigen (HBsAg). Sixty seven patients, 18 of whom with high AFP, were longitudinally studied over a period ranging from 1 to 6 years. The evaluation of AFP was transient in 6 cases and persistent in 12 cases. The patients in the former group had high activity of liver disease and none of them developed primary liver cell cancer during the follow-up. In contrast, only two patients in the latter group had high activity, and 3 developed PLC. Active HBV replication was significantly more frequent among those with transient elevation of AFP, whereas anti-HBs and anti-HBc positive cases were significantly more frequent among those with persistent elevation of AFP. It is concluded that the severity of the liver disease is the major factor of elevation of AFP. Transiently raised levels of AFP are not related with PLC, whereas persistently high levels represent a premalignant condition. At variance with data obtained in areas with high prevalence of PLC, HBsAg seems not to be related with persistent elevation of AFP. The high rate positive for anti-HBs and anti-HBc in this group remains to be clarified

  3. Radioimmunodetection of hepatocellular carcinoma with a radiolabeled antibody to alpha fetoprotein

    International Nuclear Information System (INIS)

    Ishii, Nobuko; Nakata, Keisuke; Muro, Toyokichi

    1982-01-01

    The present study was undertaken to investigate whether or not radiolabeled antibody to Alpha-fetoprotein (AFP) intravenously administered is accumulate in AFP-producing tumors. Rats with subcutaneously transplanted hepatoma or with heptoma produced in liver by 3'-Me-DAB, and 12 patients with hepatocellular carcinoma (HCC) were studied. In animal experiments, the tumor localization was clearly demonstrated by total body scintigraphy 48 to 168 hours after the injection of the radioiodinated antibody. The rats were sacrificed and radioactivity in tissues was counted. Tumor tissues showed the highest counts. In patients with HCC, the imaging was made with a gamma camera usually 24 and 48 hours after injection of I-131-labeled antibody. In order to make the contrast of tumor image clear, nonspecific background images obtained by administration of Tc-99m-labeled serum albumin were subtracted from the images obtained by I-131 labeled antibody with a computer. In 6 of 12 patients with HCC, the tumor location could successfully demonstrated. However, tumor with sizes below 2 cm in diameter which were able to be detected by the celiac angiography were not detectable. Interes tingly the images were more clear in patients with relatively higher serum AFP levels. In the blood of patient given injection of AFP radioantibody, both immune complex and free antibody were able to be detected at least for 120 hours so that the presence of free antigen (AFP) did not appear to prevent tumor radioimmunodetection. The information obtained concerning the biochemical factors which influence antibody localization in the tumor would provide better method for radioimmunodetection with antitumor antibodies. (author)

  4. Immunohistochemical expression profiles of solute carrier transporters in alpha-fetoprotein-producing gastric cancer.

    Science.gov (United States)

    Shimakata, Takaaki; Kamoshida, Shingo; Kawamura, Jumpei; Ogane, Naoki; Kameda, Yoichi; Yanagita, Emmy; Itoh, Tomoo; Takeda, Risa; Naka, Ayano; Sakamaki, Kuniko; Hayashi, Yurie; Kuwao, Sadahito

    2016-11-01

    Alpha-fetoprotein (AFP)-producing gastric cancer (GC) is an aggressive tumour with high rates of liver metastasis and poor prognosis, and for which a validated chemotherapy regimen has not been established. Drug uptake by solute carrier (SLC) transporters is proposed as one of the mechanisms involved in sensitivity to chemotherapy. In this study, we aimed to develop important insights into effective chemotherapeutic regimens for AFP-producing GC. We evaluated immunohistochemically the expression levels of a panel of SLC transporters in 20 AFP-producing GCs and 130 conventional GCs. SLC transporters examined were human equilibrative nucleoside transporter 1 (hENT1), organic anion transporter 2 (OAT2), organic cation transporter (OCT) 2, OCT6 and organic anion-transporting polypeptide 1B3 (OATP1B3). The rates of high expression levels of hENT1 (hENT1 high ) and OAT2 (OAT2 high ) were statistically higher in AFP-producing GC, compared with conventional GC. When analysing hENT1 and OAT2 in combination, hENT1 high /OAT2 high was the most particular expression profile for AFP-producing GC, with a greater significance than hENT1 or OAT2 alone. However, no significant differences in OCT2, OCT6 or OATP1B3 levels were detected between AFP-producing and conventional GCs. However, immunoreactivity for hENT1, OAT2 and OCT6 tended to be increased in GC tissues compared with non-neoplastic epithelia. Because hENT1 and OAT2 are crucial for the uptake of gemcitabine and 5-fluorouracil, respectively, our results suggest that patients with AFP-producing GC could potentially benefit from gemcitabine/fluoropyrimidine combination chemotherapy. Increased expression of hENT1, OAT2 and OCT6 may also be associated with the progression of GC. © 2016 John Wiley & Sons Ltd.

  5. Development of graphite carbon nitride based fluorescent immune sensor for detection of alpha fetoprotein

    Science.gov (United States)

    Li, Yike; Dong, Lingyu; Wang, Xiangfeng; Liu, Yuan; Liu, Hailing; Xie, Mengxia

    2018-05-01

    A novel fluorescent immunosensor for determination of alpha fetoprotein (AFP) in serum samples has been developed based on the nano graphite carbon nitride (g-C3N4) as fluorophore and immunomagnetic beads (MBs) as separation material. The bulk g-C3N4 was obtained by thermal polymerization of melamine, and then carboxylated and exfoliated to acquire the carboxylated nano g-C3N4 (c-n-g-C3N4), which has been characterized and the results showed that it had excellent fluorescent properties. The antibodies of AFP (Ab1, Ab2) were conjugated to the MBs and the c-n-g-C3N4, respectively. In assay of AFP detection, the magnetic part of the immunosensor, MBs-Ab1, would form the sandwich type complex with the signal part of the sensor, c-n-g-C3N4-Ab2. The developed immunosensor could simplify the process of separation due to the MBs. The results illustrated that proposed approach held a good linearity between the fluorescence intensity of the sensor and the AFP concentration ranging from 5-600 ng/mL with the limit of detection as low as 0.43 ng/mL, and its spiking recoveries ranged from 98.2% to 105.9% with RSD from 2.1% to 3.5%. The fabricated fluorescent immunosensor possesses the merits of good sensitivity, excellent selectivity, high biocompatibility and low cost, and the results provide a novel clue to develop immunosensor for determination of the biomarkers in complex matrices.

  6. Appraisal of radioimmunotherapy with 131I anti-alpha fetoprotein monoclonal antibody in patients with primary liver cancer

    International Nuclear Information System (INIS)

    Huang Yaozhang

    1992-01-01

    Mixed anti-alpha-fetoprotein monoclonal antibodies (AFPMcAb) labeled with 131 I were used in the treatment of 23 patients of moderate to advanced primary liver cancer. In 16 cases treated with 24 doses, the survival periods were 18-605 days with a mean of 135 days. Two patients with moderately advanced liver cancer had a mean survival period of 465 days. According to our experience, the larger dose of 131 I and of anti-AFPMcAb, the longer the survival period and the better the therapeutic results were observed. The relationship between the ratio of cancer/liver radioactivity and the survival period remains to be elucidated

  7. Diagnostic utility of alpha-fetoprotein and des-gamma-carboxyprothrombin in nigerians with hepatocellular carcinoma.

    Science.gov (United States)

    Ette, Akpakip Ikpong; Ndububa, D A; Adekanle, O; Ekrikpo, U

    2017-10-01

    Alpha-fetoprotein (AFP) and Des-gamma-carboxyprothrombin (DCP) have been extensively studied as biomarkers for the diagnosis of and prognostication in hepatocellular carcinoma (HCC). However there are only few reports on the clinical characteristics of hepatocellular carcinoma in relation to the combination of the two tumor markers in hepatitis B virus-related HCC. The aim of this study was to investigate the clinical characteristics of HBV-related HCC in relation to different sets of AFP and DCP values. Sixty-two patients with untreated HCC were studied. The positive value of AFP was set at 20 1U/L while DCP positive value was set at 150 mAU/ml. Patients were divided into three groups: Group 1(n=36) with AFP ≥ 20 IU/L and DCP ≥ 150 mAU/ml. Group 2(n=24) with AFP <20 1U/L and DCP ≥ 150 mAU/ml. Group 3 (n=2) with AFP < 20 1U/L and DCP < 150 mAU/ml. There were no patients in group 4 meant for those with AFP ≥ 20 1U/L and DCP < 150 mAU/ml. Clinical and laboratory variables were compared among the groups. Clinical and laboratory variables were comparable among the groups with the exception of gender and values of serum alanine aminotransferase (ALT). Males were significantly more than females among the groups (p<0.03). ALT values were significantly different among the groups (p<0.006). Paired comparisons between the groups showed the mean values of serum ALT were significantly higher in group 2 than in group 1(p<0.003). The mean serum ALT values were also higher in group 2 than in group 3 (p <0.014). There was no significant difference between group 1 and group 3 (P = 0.124). HCC patients who are sero-positive for DCP and sero-negative for AFP have significantly higher levels of serum ALT; serum ALT levels may be of diagnostic importance in AFP-negative, HBV-related HCC patients.

  8. Measurement of Glycosylated Alpha-Fetoprotein Improves Diagnostic Power over the Native Form in Hepatocellular Carcinoma

    Science.gov (United States)

    Jin, Jonghwa; Park, Jiyoung; Yu, Su Jong; Yoon, Jung-Hwan; Kim, Youngsoo

    2014-01-01

    Serum alpha-fetoprotein (AFP) has long been used as a diagnostic marker for hepatocellular carcinoma (HCC), albeit controversially. Although it remains widely used in clinics, the value of AFP in HCC diagnosis has recently been challenged due to its significant rates of false positive and false negative findings. To improve the efficacy of AFP as HCC diagnostic marker, we developed a method of measuring total and glycosylated AFP by multiple reaction monitoring (MRM)-MS. In this study, we verified the total amount of AFP (nonglycopeptide levels) and the degree of glycosylated AFP (deglycopeptide levels) in 60 normal (41 men and 19 women; mean age 53 years; range 32–74 years), 35 LC (23 men and 12 women; mean age 56 years; range 43–78 years; HBV-related), and 60 HCC subjects (42 men and 18 women; mean age 58 years; range 38–76 years; HBV-related; 30 stage I, 15 stage II, and 10 stage III). By MRM-MS analysis, the nonglycopeptide had 56.7% sensitivity, 68.3% specificity, and an AUC of 0.687 [cutoff value: ≥0.02 (light/heavy ratio)], comparing the normal and HCC group, whereas the deglycopeptide had 93.3% sensitivity, 68.3% specificity, and an AUC of 0.859 [cutoff value: ≥0.02 (light/heavy ratio)]. In comparing the stage I HCC subgroup with the LC group, the nonglycopeptide had a sensitivity of 66.7%, specificity of 80.0%, and an AUC of 0.712 [cutoff value: ≥0.02 (light/heavy ratio)], whereas the deglycopeptide had a sensitivity of 96.7%, specificity of 80.0%, and an AUC of 0.918 [cutoff value: ≥0.02 (light/heavy ratio)]. These data demonstrate that the discriminatory power of the deglycopeptide is greater than that of the nonglycopeptide. We conclude that deglycopeptide can distinguish cancer status between normal subjects and HCC patients better than nonglycopeptide. PMID:25310463

  9. Quantum-dot-based homogeneous time-resolved fluoroimmunoassay of alpha-fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Chen Meijun; Wu Yingsong; Lin Guanfeng; Hou Jingyuan; Li Ming [Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, 510515 (China); Liu Tiancai, E-mail: liutc@smu.edu.cn [Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, 510515 (China)

    2012-09-05

    Highlights: Black-Right-Pointing-Pointer QDs-based homogeneous time-resolved fluoroimmunoassay was developed to detect AFP. Black-Right-Pointing-Pointer The conjugates were prepared with QDs-doped microspheres and anti-AFP McAb. Black-Right-Pointing-Pointer The conjugates were prepared with LTCs and another anti-AFP McAb. Black-Right-Pointing-Pointer Excess amounts of conjugates were used for detecting AFP without rinsing. Black-Right-Pointing-Pointer The wedding of QPs and LTCs was suitable for HTRFIA to detect AFP. - Abstract: Quantum dots (QDs) with novel photoproperties are not widely used in clinic diagnosis, and homogeneous time-resolved fluorescence assays possess many advantages over current methods for alpha-fetoprotein (AFP) detection. A novel QD-based homogeneous time-resolved fluorescence assay was developed and used for detection of AFP, a primary marker for many cancers and diseases. QD-doped carboxyl-modified polystyrene microparticles (QPs) were prepared by doping oil-soluble QDs possessing a 605 nm emission peak. The antibody conjugates (QPs-E014) were prepared from QPs and an anti-AFP monoclonal antibody, and luminescent terbium chelates (LTCs) were prepared and conjugated to a second anti-AFP monoclonal antibody (LTCs-E010). In a double-antibodies sandwich structure, QPs-E014 and LTCs-E010 were used for detection of AFP, serving as energy acceptor and donor, respectively, with an AFP bridge. The results demonstrated that the luminescence lifetime of these QPs was sufficiently long for use in a time-resolved fluoroassay, with the efficiency of time-resolved Foerster resonance transfer (TR-FRET) at 67.3% and the spatial distance of the donor to acceptor calculated to be 66.1 Angstrom-Sign . Signals from TR-FRET were found to be proportional to AFP concentrations. The resulting standard curve was log Y = 3.65786 + 0.43863{center_dot}log X (R = 0.996) with Y the QPs fluorescence intensity and X the AFP concentration; the calculated sensitivity was 0

  10. A novel label-free voltammetric immunosensor for the detection of {alpha}-fetoprotein using functional titanium dioxide nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Liang Wenbin [Chongqing Key Laboratory of Analytical Chemistry, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); Yuan Ruo [Chongqing Key Laboratory of Analytical Chemistry, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)], E-mail: yuanruo@swu.edu.cn; Chai Yaqin; Li Yan; Zhuo Ying [Chongqing Key Laboratory of Analytical Chemistry, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

    2008-01-01

    A highly sensitive label-free voltammetric immunosensor was developed based on the functional titanium dioxide nanoparticles (PV-NTiP), which was prepared by capping 1,1'-bis-(2-phosphonoethyl)-4,4'-bipyridinium dibromide (PV) on the surface of the titanium dioxide nanoparticles (NTiP) with covalent attachment. The PV-NTiP has prominent biocompatibility, good electron transfer ability, primarily excellent adsorption, large specific surface area and positively charged environment. As a result, the negatively charged gold nanoparticles (NGP) could be adsorbed on the PV-NTiP modified electrode surface by electrostatic adsorption, and then to immobilize {alpha}-1-fetoprotein antibody (anti-AFP) for the assay of {alpha}-1-fetoprotein (AFP). The fabricated procedures and electrochemical behaviors of the immunosensor were characterized by electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM) and cyclic voltammetry (CV). The anti-AFP/NGP/PV-NTiP modified electrode was sensitive to AFP in linear relation between 1.25 and 200 ng/mL with the correlation coefficient of 0.9982, and the detection limit (S/N = 3) is 0.6 ng/mL under the optimal conditions. In addition, the proposed immunosensor exhibits good sensitivity, selectivity, stability and long-term maintenance of bioactivity and it may be used to immobilize other biomoleculars to develop biosensor for the detection of other antigens or biocompounds.

  11. Gold nanoparticles and polyethylene glycols functionalized conducting polyaniline nanowires for ultrasensitive and low fouling immunosensing of alpha-fetoprotein.

    Science.gov (United States)

    Hui, Ni; Sun, Xiaotian; Song, Zhiling; Niu, Shuyan; Luo, Xiliang

    2016-12-15

    An ultrasensitive biosensor for alpha-fetoprotein was developed based on electrochemically synthesized polyaniline (PANI) nanowires, which were functionalized with gold nanoparticles (AuNPs) and polyethylene glycols (PEG). The prepared PEG/AuNPs/PANI composite, combining the electrical conductivity of the AuNPs/PANI with the robust antifouling ability of PEG, offered an ideal substrate for the development of low fouling electrochemical biosensors. Alpha-fetoprotein (AFP), a well-known hepatocellular carcinoma biomarker, was used as a model analyte, and its antibody was immobilized on the PEG/AuNPs/PANI for the construction of the AFP immunosensor. Using the redox current of PANI as the sensing signal, in addition to the good biocompatibility of PEG/AuNPs and the anti-biofouling property of PEG, the developed immunosensor showed improved biosensing performances, such as wide linear range and ultralow detection limit (0.007pgmL(-1)). More importantly, it is label-free, reagentless and low fouling, making it capable of assaying AFP in real serum samples without suffering from significant interference or biofouling. Copyright © 2016 Elsevier B.V. All rights reserved.

  12. Sensitive radioimmunoassay for detection of antibodies to recombinant human interferon-alpha A

    International Nuclear Information System (INIS)

    Palleroni, A.V.; Trown, P.W.

    1986-01-01

    A radioimmunoassay (RIA) for the detection of antibodies to recombinant human leukocyte interferon A (rHuIFN-alpha A) in human serum has been developed and validated against the standard antiviral neutralization bioassay (ANB). The assay measures the binding of 125 I-labeled rHuIFN-alpha A to immunoglobulins in serum. Aliquots of patients' sera are incubated with 125 I-rHuIFN-alpha A and the complexes formed between antibodies in the sera and the 125 I-rHuIFN-alpha A are precipitated with goat anti-human IgG serum. The radioactivity in the immune precipitate is a measure of the quantity of antibody (if present) in the serum. The sensitivity of this RIA is 5 ng of IgG/ml of serum

  13. Acute Exacerbation of Hepatitis in Liver Cirrhosis with Very High Levels of alpha-Fetoprotein But No Occurrence of Hepatocellular Carcinoma

    Science.gov (United States)

    Park, Sang Jong; Park, Kwang Bo; Paik, So Ya; Ryu, Jin Kyung; Choi, Chang Kyu; Hwang, Tae Joon

    2005-01-01

    Aminotransferase levels do not always increase during acute hepatitis or during an acute flare-up of chronic hepatitis. Persistently increased levels of serum alpha-Fetoprotein in an adult with liver disease suggest not only the presence or progression of hepatocellular carcinoma or its recurrence after hepatic resection or after other therapeutic approaches such as chemotherapy or chemoembolization, but also it suggests that there is an acute exacerbation of hepatitis or liver cirrhosis. We report here on two unusual cases of HBV- & HCV-related liver cirrhosis with acute exacerbation of hepatitis in which there was an insignificant elevation of the aminotransferase levels, but there were markedly increased alpha-Fetoprotein levels observed. The levels of alpha-Fetoprotein decreased gradually in both cases since the beginning of antiviral therapy, which implies that the increased levels were due to aggravation of the accompanying hepatitis. These cases also emphasize that using only the measurement of alpha-Fetoprotein is not sufficient for the diagnosis of hepatocellular carcinoma, and that this diagnosis also requires a more specific measurement such as AFP L3 along with the standard imaging studies. PMID:15906959

  14. INCREASED MATERNAL SERUM ALPHA-FETOPROTEIN AND HUMAN CHORIONIC-GONADOTROPIN IN COMPROMISED PREGNANCIES OTHER THAN FOR NEURAL-TUBE DEFECTS OR DOWN-SYNDROME

    NARCIS (Netherlands)

    BEEKHUIS, [No Value; VANLITH, JMM; DEWOLF, BTHM; MANTINGH, A

    Intrauterine fetal death occurred in four women who were 'screen-positive' in a screening programme for neural tube defects (NTDs) and Down syndrome (DS). These women had very high levels of maternal serum alpha-fetoprotein (MSAFP) and maternal serum human chorionic gonadotropin (MShCG). Therefore,

  15. Ovarian Sertoli-Leydig cell tumor with heterologous elements of gastrointestinal type associated with elevated serum alpha-fetoprotein level: an unusual case and literature review.

    Science.gov (United States)

    Horta, Mariana; Cunha, Teresa Margarida; Marques, Rita Canas; Félix, Ana

    2014-11-01

    Here we describe the case of a 19-year-old woman with a poorly differentiated ovarian Sertoli-Leydig cell tumor and an elevated serum alpha-fetoprotein level. The patient presented with diffuse abdominal pain and bloating. Physical examination, ultrasound, and magnetic resonance imaging revealed a right ovarian tumor that was histopathologically diagnosed as a poorly differentiated Sertoli-Leydig cell tumor with heterologous elements. Her alpha-fetoprotein serum level was undetectable after tumor resection. Sertoli-Leydig cell tumors are rare sex cord-stromal tumors that account for 0.5% of all ovarian neoplasms. Sertoli-Leydig cell tumors tend to be unilateral and occur in women under 30 years of age. Although they are the most common virilizing tumor of the ovary, about 60% are endocrine-inactive tumors. Elevated serum levels of alpha-fetoprotein are rarely associated with Sertoli-Leydig cell tumors, with only approximately 30 such cases previously reported in the literature. The differential diagnosis should include common alpha-fetoprotein-producing ovarian entities such as germ cell tumors, as well as other non-germ cell tumors that have been rarely reported to produce this tumor marker.

  16. ABNORMAL LEVELS OF MATERNAL SERUM HUMAN CHORIONIC-GONADOTROPIN AND ALPHA-FETOPROTEIN IN THE 2ND-TRIMESTER - RELATION TO FETAL WEIGHT AND PRETERM DELIVERY

    NARCIS (Netherlands)

    MORSSINK, LP; KORNMAN, LH; BEEKHUIS, [No Value; DEWOLF, BTHM; MANTINGH, A

    The aim of this prospective descriptive cross-sectional study was to examine the clinical significance of abnormal maternal serum human chorionic gonadotropin (MShCG) and alpha-fetoprotein (MSAFP) in the second trimester of pregnancy. The study group comprised 8892 women with a singleton pregnancy,

  17. Correlation between preoperative serum alpha-fetoprotein levels and survival with respect to the surgical treatment of hepatocellular carcinoma at a tertiary care hospital in Veracruz, Mexico

    Directory of Open Access Journals (Sweden)

    G. Martínez-Mier

    2017-10-01

    Full Text Available Introduction: Preoperative serum alpha-fetoprotein levels can have predictive value for hepatocellular carcinoma survival. Aim: Our aim was to analyze the correlation between preoperative serum alpha-fetoprotein levels and survival, following the surgical treatment of hepatocellular carcinoma. Methods: Nineteen patients were prospectively followed (07/2005-01/2016. An ROC curve was created to determine the sensitivity and specificity of alpha-fetoprotein in relation to survival (Kaplan-Meier. Results: Of the 19 patients evaluated, 57.9% were men. The mean patient age was 68.1 ± 8.5 years and survival at 1, 3, and 5 years was 89.4, 55.9, and 55.9%. The alpha-fetoprotein cutoff point was 15.1 ng/ml (sensitivity 100%, specificity 99.23%. Preoperative alpha-fetoprotein levels below 15.1, 200, 400, and 463 ng/ml correlated with better 1 and 5-year survival rates than levels above 15.1, 200, 400, and 463 ng/ml (P<.05. Conclusions: Elevated preoperative serum alpha-fetoprotein levels have predictive value for hepatocellular carcinoma survival. Resumen: Introducción: Los niveles séricos de alfafetoproteína (AFP preoperatoria pueden tener valor predictivo para la sobrevida del hepatocarcinoma (HCC. Objetivo: Analizar la correlación entre los niveles séricos de AFP preoperatoria y la sobrevida posterior al tratamiento quirúrgico del HCC. Métodos: Diecinueve pacientes fueron seguidos prospectivamente (julio del 2005-enero del 2016. Se realizó una curva ROC para determinar la sensibilidad y la especificidad de la AFP con relación con la sobrevida (Kaplan-Meier. Resultados: Se evaluó a 19 pacientes, 57.9% hombres, edad media 68.1 ± 8.5 años con sobrevida a 1, 3 y 5 años del 89.4, el 55.9 y el 55.9%. El punto de corte de AFP fue 15.1 ng/ml (sensibilidad 100%, especificidad 99.23%. Los niveles preoperatorios de AFP menores de 15.1, 200, 400 y 463 ng/ml correlacionaron con mejor sobrevida a 1 y 5 años que niveles mayores de AFP (p < 0

  18. Ascites and alpha-fetoprotein improve prognostic performance of Barcelona Clinic Liver Cancer staging.

    Science.gov (United States)

    Gomaa, Asmaa I; Al-Khatib, Alzhraa; Abdel-Razek, Wael; Hashim, Mohammed Saad; Waked, Imam

    2015-05-14

    To assess how ascites and alpha-fetoprotein (AFP) added to the Barcelona Clinic Liver Cancer (BCLC) staging predict hepatocellular carcinoma survival. The presence of underlying cirrhosis, ascites and encephalopathy, Child-Turcotte-Pugh (CTP) score, the number of nodules, and the maximum diameter of the largest nodule were determined at diagnosis for 1060 patients with hepatocellular carcinoma at a tertiary referral center for liver disease in Egypt. Demographic information, etiology of liver disease, and biochemical data (including serum bilirubin, albumin, international normalized ratio, alanine and aspartate aminotransferases, and AFP) were evaluated. Staging of the tumor was determined at the time of diagnosis using the BCLC staging system; 496 patients were stage A and 564 patients were stage B. Patients with mild ascites on initial ultrasound, computed tomography, or clinical examination, and who had a CTP score ≤ 9 were included in this analysis. All patients received therapy according to the recommended treatment based on the BCLC stage, and were monitored from the time of diagnosis to the date of death or date of data collection. The effect of the presence of ascites and AFP level on survival was analyzed. At the time the data were censored, 123/496 (24.8%) and 218/564 (38.6%) patients with BCLC stages A and B, respectively, had died. Overall mean survival of the BCLC A and B patients during a three-year follow-up period was 31 mo [95% confidence interval (95%CI): 29.7-32.3] and 22.7 mo (95%CI: 20.7-24.8), respectively. The presence of ascites, multiple focal lesions, large tumor size, AFP level and CTP score were independent predictors of survival for the included patients on multivariate analysis (P < 0.001). Among stage A patients, 18% had ascites, 33% had AFP ≥ 200 ng/mL, and 8% had both. Their median survival in the presence of ascites was shorter if AFP was ≥ 200 ng/mL (19 mo vs 24 mo), and in the absence of ascites, patients with AFP ≥ 200

  19. Comparative studies of Iodo-bead and chloramine-T methods for the radioiodination of human alpha-fetoprotein

    International Nuclear Information System (INIS)

    Lee, D.S.C.; Griffiths, B.W.

    1984-01-01

    Human alpha-fetoprotein (h-AFP) of iodination grade was radiolabeled with Na 125 I by chloramine-T (conventional method) and by Iodo-bead (solid-phase method) under standard and varied conditions, such as quantity of h-AFP and volume of the reaction mixture. The authors have modified the chloramine-T method and developed a protocol for the Iodo-bead method of iodination. The results show that while the chloramine-T method yields radiolabeled h-AFP of apparently 40% higher specific radioactivity (SR) than those of the Iodo-bead method, the latter appears to be a simpler, more controllable and milder method, giving rise to a more stable population of [ 125 I]h-AFP. This is supported by the results of radioactive counting of [ 125 I]h-AFP collected from a Sephadex G-50 column after radioiodination, storage stability and immunoreactivity with anti-h-AFP. (Auth.)

  20. Increasing of sensitivity of fluorescent immunoassay analysis of alpha-fetoprotein by means of plasmonical silver nanoparticles

    International Nuclear Information System (INIS)

    Vashchenko, S.V.; Min'ko, A.A.; Romanenko, A.A.; Gaponenko, S.V.; Kulakovich, O.S.

    2014-01-01

    A test system is proposed based on metal enhanced fluorescence to analyze low concentrations of alpha-fetoprotein (AFP), a tumor marker. Antigen-antibody reaction was performed on polystyrene plates coated with silver nanoparticles to increase sensitivity of fluorescent immunoassay and signal-to-noise ratio as compared to silver-free system. As compared to widely used ELISA technique and other immunoassay techniques the proposed approach is characterized by smaller probe volume, fast analysis and simplicity. The proposed test system uses layer-by-layer assembly approach, LED excitation and nanowatt photodetection set-up. The proposed test system offers AFP detection at concentrations used in clinical practice. Fluorescence enhancement for labeled AFP antibodies on a silver substrate was found to depend on antibodies concentration and was up to 6 times. (authors)

  1. Binding of alpha-fetoprotein by immobilized monoclonal antibodies during episodes of zero-gravity obtained by parabolic flight

    Science.gov (United States)

    Spooner, Brian S.; Guikema, James A.; Barnes, Grady

    1990-01-01

    Alpha-fetoprotein (AFP), a single-chain polypeptide which is synthesized by the liver and yolk sac of the human fetus, provided a model ligand for assessing the effects of microgravity on ligand binding to surface-immobilized model receptor molecules. Monoclonal antibodies, used as receptors for AFP, were immobilized by covalent attachment to latex microparticles. Zero gravity environment was obtained by parabolic flight aboard NASA 930, a modified KC-135 aircraft. Buring the onset of an episode of zero gravity, ligand and receptor were mixed. Timed incubation (20 s) was terminated by centrifugation, the supernatant removed, and microparticies were assessed for bound AFP by immunochemical methods. The extent of binding was not influenced by microgravity, when compared with 1-G controls, which suggests that aberrant cellular activities observed in microgravity are not the simple expression of altered macromolecular interactions.

  2. Fluorescence Quenching of Alpha-Fetoprotein by Gold Nanoparticles: Effect of Dielectric Shell on Non-Radiative Decay

    Science.gov (United States)

    Zhu, Jian; Li, Jian-Jun; Wang, A.-Qing; Chen, Yu; Zhao, Jun-Wu

    2010-09-01

    Fluorescence quenching spectrometry was applied to study the interactions between gold colloidal nanoparticles and alpha-fetoprotein (AFP). Experimental results show that the gold nanoparticles can quench the fluorescence emission of adsorbed AFP effectively. Furthermore, the intensity of fluorescence emission peak decreases monotonously with the increasing gold nanoparticles content. A mechanism based on surface plasmon resonance-induced non-radiative decay was investigated to illuminate the effect of a dielectric shell on the fluorescence quenching ability of gold nanoparticles. The calculation results show that the increasing dielectric shell thickness may improve the monochromaticity of fluorescence quenching. However, high energy transfer efficiency can be obtained within a wide wavelength band by coating a thinner dielectric shell.

  3. Gastric Composite Tumor of Alpha Fetoprotein-Producing Carcinoma/Hepatoid Adenocarcinoma and Endocrine Carcinoma with Reference to Cellular Phenotypes

    Directory of Open Access Journals (Sweden)

    Akira Suzuki

    2012-01-01

    Full Text Available Alpha-fetoprotein-producing carcinoma (AFPC/hepatoid adenocarcinoma (HAC and neuroendocrine carcinoma (NEC are uncommon in the stomach. Composite tumors consisting of these carcinomas and their histologic phenotypes are not well known. Between 2002 and 2007, to estimate the prevalence of composite tumors consisting of tubular adenocarcinoma, AFPC/HAC and NEC, we reviewed specimens obtained from 294 consecutive patients treated surgically for gastric cancer. We examined histological phenotype of tumors of AFPC or NEC containing the composite tumor by evaluating immunohistochemical expressions of MUC2, MUC5AC, MUC6, CDX2, and SOX2. Immunohistochemically, AFPC/HAC dominantly showed the intestinal or mixed phenotype, and NEC frequently showed the gastric phenotype. In the composite tumor, the tubular and hepatoid components showed the gastric phenotype, and the neuroendocrine component showed the mixed type. The unique composite tumor predominantly showed the gastric phenotype, and the hepatoid and neuroendocrine components were considered to be differentiated from the tubular component.

  4. Radioimmunological assay of alpha-fetoprotein concentrations in blood serum samples from women in the first half of gestation

    International Nuclear Information System (INIS)

    Chomanski, M.; Grzes, A.

    1977-01-01

    In 66 unpregnant women and in 199 pregnant subjects (in the first half of pregnancy) the alpha-fetoprotein (α-FP) concentrations were determined in peripheric blood-serum samples. The α-FP was determined using radioimmunoo-assay technique where the addition of I 125 labelled protein was preceded by 3 hours lasting preincubation period. The free protein was separated from the bound by means of precipitation with a mixture of 96% ethanol and 34.5% ammonium acetate in a ratio 77.5 : 22.5. The patients were grouped into 5 subgroups according to the gestational age. The mean values of α-FP concentration demonstrated a steady increase along with the advancement of gestation. It is suggested that the concentration of this protein in the blood-serum samples may serve as a valuable parameter for the study of gestation development and the state of the fetus. (author)

  5. Clinical value of combined measurement of serum alpha-fetoprotein, alpha-L-fucosidase and ferritin levels in the diagnosis of primary liver cancer

    International Nuclear Information System (INIS)

    Zhang Aimin; Chai Xiaohong; Jin Ying; Dong Xuemei

    2005-01-01

    Objective: To investigate the clinical value of combined measurement of serum alpha-fetoprotein (AFP), alpha-L-fucosidase (AFU) and ferritin (SF) levels in the diagnosis of primary liver cancer. Methods: Serum AFP, AFU (with RIA) and SF (with biochemical method) were determined in 52 patients with primary liver cancer and 40 controls. Results: The positive rates of AFP, AFU and SF in patient with liver cancer were 82.7%, 86.6% and 76.9%, respectively. Positive rates with combined measurement of AFP plus AFU, AFP plus SF, and AFP plus AFU, SF were 94.2%, 90.4% and 98.1% respectively. Conclusion: Combined measurement of AFP, AFU and SF can significantly increase the positive rate in the diagnosis of primary liver cancer. (authors)

  6. Diagnostic performance of alpha-fetoprotein, lens culinaris agglutinin-reactive alpha-fetoprotein, des-gamma carboxyprothrombin, and glypican-3 for the detection of hepatocellular carcinoma: a systematic review and meta-analysis protocol

    Science.gov (United States)

    2013-01-01

    Background Diagnosis of early-stage hepatocellular carcinoma (HCC) followed by curative resection or liver transplantation offers the best chance for long-term patient survival. Clinically, ultrasonography has suboptimal sensitivity for detecting early-stage HCC. Several serological tests including alpha-fetoprotein (AFP), the ratio of lens culinaris agglutinin-reactive alpha-fetoprotein to total AFP (AFP-L3/AFP), des-gamma carboxyprothrombin (DCP), and glypican-3 (GPC-3) have been widely investigated as diagnostic biomarkers for early-stage HCC in at-risk populations. However, these tests are not recommended for routine HCC screening. Our objective is to determine the diagnostic performance of AFP, AFP-L3/AFP, DCP, and GPC-3 for the detection of HCC, particularly early-stage tumors meeting the Milan criteria. Methods/design We will include cross-sectional studies that consecutively or randomly recruit target populations. We will search the Cochrane Library, Medline, Embase, Science Citation Index, and the Chinese National Knowledge Infrastructure. We will also search the MEDION and ARIF databases to identify diagnostic systematic reviews that include primary studies. Reference lists of relevant reviews will be searched for additional trials. Language restrictions will not be applied. Two reviewers will independently screen study eligibility and extract data. Methodological quality will be assessed according to the revised tool for the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2). Two authors will apply the QUADAS-2 assessment to all the included studies, and any discrepancies will be resolved by the third author. The following test characteristics will be extracted into 2 × 2 tables for all included studies: true positives, false positives, true negatives, and false negatives. Study-specific estimates of sensitivity and specificity with 95% confidence intervals will be displayed in forest plots. When possible, we will use the bivariate random

  7. Studies of an alpha-fetoprotein assay using dry blood-spot samples to be used for the detection of fetal neural tube defects.

    Science.gov (United States)

    Wong, P Y; Mee, A V; Doran, T A

    1982-06-01

    We modified the Pharmacia serum alpha-fetoprotein (AFP) kit to enable its use with dry blood-spots on filter paper. Reference values were established for blood from 253 women in the 16th to 18th weeks of gestation. The result by the present technique in a woman with a confirmed anencephalic fetus was elevated, and in agreement with the results of AFP assays in serum and amniotic fluid. Blood AFP was stable on dried filter paper sent by mail.

  8. Frequency of deaths in hepatitis C virus infected hepatocellular carcinoma patients and its relationship with raised serum alpha-fetoprotein levels.

    Science.gov (United States)

    Shaikh, Fida Hussain; Zeb, Shaista; Chandio, Sultan Ahmed; Munaf, Alvina; Ghori, Muhamad Aamir; Memon, Mohammad Sadik; Burney, Asif Ali

    2016-01-01

    To determine the frequency of deaths in hepatitis C virus infected hepatocellular carcinoma patients, and its relationship with raised serum alpha-fetoprotein levels. The cross-sectional study was conducted at Isra University Hospital, Hyderabad, Pakistan, between March 2013 and April 2014, and comprised all patients diagnosed with hepatitis C virus and hepatocellular carcinoma over 30 years ofage. Blood sample was drawn for the measurement of serum Alfa fetoprotein levels. Data was analysed using SPSS 16. The mean age of the 165 patients was55.49±11.67 years. The mean tumour size was 5.63 ± 2.14cm. Of the total, 31(18.8%) patients had tumour size 5cm. The mean serum Alfa fetoprotein level was 7641.0±3665.32 IU/ml. Overall mortality rate was 70(41.9%). Tumour size >5cm was significantly associated with mortality (p=0.016). Serum Alfa fetoprotein levels were a useful tool for the detection of hepatocellular carcinoma in hepatitis C virus patients.

  9. Radioimmunoassay of alpha-foeto-protein in hepatology. Observation of 151 cases

    International Nuclear Information System (INIS)

    Dao, Mouhamadou.

    1976-03-01

    Alpha-foeto-protein, although discovered several years ago, was considered as only able to exist in primary adult liver cancer. With progress in science and in research it was observed that AFP can be found in normal subjects, without any pathological significance. APF has been found moreover in illnesses other than primary liver cancer. This range of diseases, restricted at first, has continued to widen, a development due to the discovery of new AFP detection methods and radioimmunoassay. Our work, covering 151 cases discussed in detail in the 'Material and Methods' chapter, is interesting from two viewpoints: in hepatology it is useful to know whether the very sensitive radio-immunological method reveals abnormal AFP content variations in different liver diseases; given the large number of hepatomas in our series it is also interesting to have the value of this determination confirmed in primary liver cancer diagnosis. Radioimmunoassay now replaces other quantitative methods (MANCINI's radial immunodiffusion and LAURELL's elec. diffusion) and is the most accurate [fr

  10. Elevated serum alpha-fetoprotein in poorly differentiated adenocarcinoma with neuroendocrine differentiation of the ascending colon: a case report.

    Science.gov (United States)

    Lin, Hung-Hsin; Chang, Chia-Chu; Yang, Shung-Haur; Chang, Shih-Ching; Chen, Wei-Shone; Liang, Wen-Yih; Lin, Jen-Kou; Jiang, Jeng-Kai

    2016-03-15

    Colorectal cancer (CRC) is the most common form of cancer and the third leading cause of death in Taiwan. Serum alpha-fetoprotein (AFP) has been extensively used as a biomarker for hepatocellular carcinoma (HCC) and yolk sac tumors. This case report presents a 90-year-old woman with right abdominal pain and poor appetite for 1 week. The computed tomography (CT) showed wall thickening in the proximal ascending colon with ruptured appendicitis. Preoperative serum AFP was high. There was no definite liver metastasis or other abnormal findings in the hepatobiliary systems. After initial empirical antibiotic treatment, we performed laparoscopic right hemicolectomy. The pathological assessment was poorly differentiated adenocarcinoma with neuroendocrine differentiation in the ascending colon. The tumor cells did not produce AFP. Amazingly, the follow-up serum AFP level 1 month after the surgery declined to normal range. The patient had an uneventful course after the surgery and was free of recurrence or metastasis within 5 months of follow-up. AFP may be a useful tumor marker in poorly differentiated colorectal cancer with neuroendocrine component patients and a prediction of early treatment response.

  11. The alpha-fetoprotein third domain receptor binding fragment: in search of scavenger and associated receptor targets.

    Science.gov (United States)

    Mizejewski, G J

    2015-01-01

    Recent studies have demonstrated that the carboxyterminal third domain of alpha-fetoprotein (AFP-CD) binds with various ligands and receptors. Reports within the last decade have established that AFP-CD contains a large fragment of amino acids that interact with several different receptor types. Using computer software specifically designed to identify protein-to-protein interaction at amino acid sequence docking sites, the computer searches identified several types of scavenger-associated receptors and their amino acid sequence locations on the AFP-CD polypeptide chain. The scavenger receptors (SRs) identified were CD36, CD163, Stabilin, SSC5D, SRB1 and SREC; the SR-associated receptors included the mannose, low-density lipoprotein receptors, the asialoglycoprotein receptor, and the receptor for advanced glycation endproducts (RAGE). Interestingly, some SR interaction sites were localized on the AFP-derived Growth Inhibitory Peptide (GIP) segment at amino acids #480-500. Following the detection studies, a structural subdomain analysis of both the receptor and the AFP-CD revealed the presence of epidermal growth factor (EGF) repeats, extracellular matrix-like protein regions, amino acid-rich motifs and dimerization subdomains. For the first time, it was reported that EGF-like sequence repeats were identified on each of the three domains of AFP. Thereafter, the localization of receptors on specific cell types were reviewed and their functions were discussed.

  12. Radiolabelled monoclonal antibodies against alpha-fetoprotein for in vivo localization of human hepatocellular carcinoma by immunotomoscintigraphy

    International Nuclear Information System (INIS)

    Bergmann, J.F.; Lumbroso, J.D.; Manil, L.; Saccavini, J.C.; Rougier, P.; Assicot, M.; Mathieu, A.; Bellet, D.; Bohuon, C.

    1987-01-01

    Two high affinity monoclonal antibodies, designated AF01 and AF04, directed against distinct epitopes of human alpha-fetoprotein (AFP) and the Fab fragments of one of them, were labelled with 131 I and injected into 18 patients with AFP producing hepatocellular carcinoma (HCC) in order to carry out imaging studies by tomoscintigraphy. Twelve patients were injected with whole antibody, only three of seven patients injected with AF01 and two of five patients injected with AF04 had a positive scan. In contrast, five out of six patients injected with labelled Fab fragments of AF04 had positive imaging. These results confirm that tumour imaging of HCC using 131 I labelled monoclonal antibody against AFP is feasible. Moreover, utilization of tomoscintigraphy in place of linear scintigraphy and Fab fragments instead of whole immunoglobulin may improve the sensitivity of radioimmunolocalization. This technique provides useful information on the in vivo distribution of monoclonal antibodies directed against AFP and on the practicability of the eventual therapeutic use of anti-AFP antibodies in HCC. (orig.)

  13. Horseradish peroxidase functionalized gold nanorods as a label for sensitive electrochemical detection of alpha-fetoprotein antigen.

    Science.gov (United States)

    Guo, Jinjin; Han, Xiaowei; Wang, Junchun; Zhao, Junqing; Guo, Zilin; Zhang, Yuzhong

    2015-12-15

    In this study, a novel tracer, horseradish peroxidase (HRP) functionalized gold nanorods (Au NRs) nanocomposites (HRP-Au NRs), was designed to label the signal antibodies for sensitive electrochemical measurement of alpha-fetoprotein (AFP). The preparation of HRP-Au NRs nanocomposites and the labeling of secondary antibody (Ab2) were performed by one-pot assembly of HRP and Ab2 on the surface of Au NRs. The immunosensor was fabricated by assembling carbon nanotubes (CNTs), Au NRs, and capture antibodies (Ab1) on the glassy carbon electrode. In the presence of AFP antigen, the labels were captured on the surface of the Au NRs/CNTs via specific recognition of antigen-antibody, resulting in the signal intensity being clearly increased. Differential pulse voltammetry (DPV) was employed to record the response signal of the immunosensor in phosphate-buffered saline (PBS) containing hydrogen peroxide (H2O2) and 3,3',5,5'-tetramethylbenzidine (TMB). Under optimal conditions, the signal intensity was linearly related to the concentration of AFP in the range of 0.1-100 ng ml(-1), and the limit of detection was 30 pg ml(-1) (at signal/noise [S/N] = 3). Furthermore, the immunoassay method was evaluated using human serum samples, and the recovery obtained was within 99.0 and 102.7%, indicating that the immunosensor has potential clinical applications. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Development and application of a fluorescence protein microarray for detecting serum alpha-fetoprotein in patients with hepatocellular carcinoma.

    Science.gov (United States)

    Zhang, Aiying; Yin, Chengzeng; Wang, Zhenshun; Zhang, Yonghong; Zhao, Yuanshun; Li, Ang; Sun, Huanqin; Lin, Dongdong; Li, Ning

    2016-12-01

    Objective To develop a simple, effective, time-saving and low-cost fluorescence protein microarray method for detecting serum alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma (HCC). Method Non-contact piezoelectric print techniques were applied to fluorescence protein microarray to reduce the cost of prey antibody. Serum samples from patients with HCC and healthy control subjects were collected and evaluated for the presence of AFP using a novel fluorescence protein microarray. To validate the fluorescence protein microarray, serum samples were tested for AFP using an enzyme-linked immunosorbent assay (ELISA). Results A total of 110 serum samples from patients with HCC ( n = 65) and healthy control subjects ( n = 45) were analysed. When the AFP cut-off value was set at 20 ng/ml, the fluorescence protein microarray had a sensitivity of 91.67% and a specificity of 93.24% for detecting serum AFP. Serum AFP quantified via fluorescence protein microarray had a similar diagnostic performance compared with ELISA in distinguishing patients with HCC from healthy control subjects (area under receiver operating characteristic curve: 0.906 for fluorescence protein microarray; 0.880 for ELISA). Conclusion A fluorescence protein microarray method was developed for detecting serum AFP in patients with HCC.

  15. Hepatoid adenocarcinoma of the stomach: an unusual case of elevated alpha-fetoprotein with prior treatment for hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    Joon Seong Ahn

    2013-06-01

    Full Text Available Hepatoid adenocarcinoma (HAC is a rare type of extrahepatic carcinoma whose morphology is similar to that of hepatocellular carcinoma (HCC. Metachronous HCC and HAC in the same patient is extremely rare. The case of a 68-year-old man with chronic hepatitis B infection who had both HCC and HAC of the stomach is reported herein. Nine years previously this patient had been diagnosed with HCC and received a right lobectomy. HCC that recurred at the caudate lobe at 6 months after the operation was successfully treated with transarterial chemoembolization. The patient was followed up regularly thereafter without evidence of tumor recurrence for 9 years. In July 2010 his serum alpha-fetoprotein (AFP level elevated from 6.5 ng/mL to 625.4 ng/mL, and he developed a probable single metastatic lymph node around the hepatic artery without intrahepatic lesions. Subsequent evaluation with upper endoscopy revealed a 4-cm ulcerative lesion on the antrum of the stomach. Subtotal gastrectomy was performed with lymph-node dissection. Histologic examination revealed a special type of extrahepatic AFP-producing adenocarcinoma-HAC with lymph-node metastasis-which indicates that HAC can be a cause of elevated AFP even in patients with HCC. HAC should be considered if a patient with stable HCC exhibits unusual elevation of AFP.

  16. Golgi protein 73 versus alpha-fetoprotein as a biomarker for hepatocellular carcinoma: a diagnostic meta- analysis

    International Nuclear Information System (INIS)

    Zhou, Ying; Yin, Xin; Ying, Jun; Zhang, BoHeng

    2012-01-01

    There have been conflicting reports about serum golgi protein 73 (GP73) as one of the most promising serum markers for the diagnosis of hepatocellular carcinoma (HCC). This study was to make a systematic review about the diagnostic accuracy of serum GP73 versus alpha-fetoprotein (AFP) for HCC. After a systematic review of related studies, sensitivity, specificity and other measures about the accuracy of serum GP73 and AFP in the diagnosis of HCC were pooled using random-effects models. Summary receiver operating characteristic curve analysis was used to summarize the overall test performance. Eight studies were included in our meta-analysis. The summary estimates for serum GP73 and AFP in diagnosing HCC in the studies included were as follows: sensitivity, 76% (95% confidence interval (CI) 51-91%) vs. 70% (47-86%); specificity, 86% (95%CI 65-95%) vs. 89% (69-96%); diagnostic odds ratio (DOR), 18.59 (95%CI 5.33-64.91) vs. 18.00(9.41-34.46); and area under sROC, 0.88 (95%CI 0.77-0.99) vs. 0.86 (95%CI 0.84-0.87). The current evidence indicates that serum GP73 has a comparable accuracy to AFP for the diagnosis of HCC, while the value of serum GP73 in combination with AFP for HCC detection deserves further investigation

  17. Alpha-fetoprotein is a biomarker of unfolded protein response and altered proteostasis in hepatocellular carcinoma cells exposed to sorafenib.

    Science.gov (United States)

    Houessinon, Aline; Gicquel, Albane; Bochereau, Flora; Louandre, Christophe; Nyga, Rémy; Godin, Corinne; Degonville, James; Fournier, Emma; Saidak, Zuzana; Drullion, Claire; Barbare, Jean-Claude; Chauffert, Bruno; François, Catherine; Pluquet, Olivier; Galmiche, Antoine

    2016-01-28

    Sorafenib is the treatment of reference for advanced hepatocellular carcinoma (HCC). A decrease in the serum levels of Alpha-fetoprotein (AFP) is reported to be the biological parameter that is best associated with disease control by sorafenib. In order to provide a biological rationale for the variations of AFP, we analyzed the various steps of AFP production in human HCC cell lines exposed to sorafenib. Sorafenib dramatically reduced the levels of AFP produced by HCC cells independently of its effect on cell viability. The mRNA levels of AFP decreased upon sorafenib treatment, while the AFP protein remained localized in the Golgi apparatus. Sorafenib activated the Regulated Inositol-Requiring Enzyme-1α (IRE-1α) and the PKR-like ER Kinase (PERK)-dependent arms of the Unfolded Protein Response (UPR). The inhibition of IRE-1α partially restored the mRNA levels of AFP upon treatment with sorafenib. The inhibition of both pathways partially prevented the drop in the production of AFP induced by sorafenib. The findings provide new insights on the regulation of AFP, and identify it as a biomarker suitable for the exploration of HCC cell proteostasis in the context of therapeutic targeting. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  18. Intrahepatic bile duct adenoma in a patient with chronic hepatitis B accompanied by elevation of alpha-fetoprotein.

    Science.gov (United States)

    Ahn, Jem Ma; Paik, Yong-Han; Lee, Jun Hee; Cho, Ju Yeon; Sohn, Won; Gwak, Geum-Youn; Choi, Moon Seok; Lee, Joon Hyeok; Koh, Kwang Cheol; Paik, Seung Woon; Yoo, Byung Chul

    2015-12-01

    A 51-year-old male patient with chronic hepatitis B was referred to our hospital due to a 1-cm liver nodule on ultrasonography. Alpha-fetoprotein (AFP) was slightly elevated. The nodule showed prolonged enhancement on dynamic liver magnetic resonance imaging and appeared as a hyperintensity on both diffusion-weighted and T2-weighted imaging. The nodule was followed up because it was small and typical findings of hepatocellular carcinoma (HCC) were not observed in the dynamic imaging investigations. However, liver contrast-enhanced ultrasonography performed 1 month later showed enhancement during the arterial phase and definite washout during the delayed phase. Also, AFP had increased to over 200 ng/mL even though AST and ALT were decreased after administering an antiviral agent. He was presumptively diagnosed as HCC and underwent liver segmentectomy. Microscopy findings of the specimen indicated bile duct adenoma. After resection, the follow-up AFP had decreased to within the normal range. This patient represents a case of bile duct adenoma with AFP elevation mimicking HCC on contrast-enhanced ultrasonography.

  19. Morphologic alterations and immunohistochemical analysis of alpha-fetoprotein and CD34 in chorionic villi of anembryonic pregnancy

    International Nuclear Information System (INIS)

    Aydin, S.; Yenilmez, E.; Yulug, E.; Arvas, H.; Ozeren, M.; Cobanoglu, U.

    2006-01-01

    To investigate the morphology of chorionic villi using light and electron microscopy, especially the expression of alpha-fetoprotein (AFP) in trophoblastic cells and the process of maturation and margination vasculogenesis proper using CD34 immunohistochemistry in tissues from the first trimester of pregnancy loss due to anembryonic pregnancy in comparison with embryonic pregnancy. The study consisted of 2 groups: 9 patients with anembryonic pregnancies and 9 patients with embryonic pregnancies between 6 and 10 weeks of gestational age registered at the Department of Gynecology and Obstetrics, University Hospital of Karadeniz Technical University, Turkey, from March 2003 to December 2004. We examined the chorionic villi using light and electron microscopy. For immunohistochemical staining, we used AFP and CD 34. Microscopically, pathologic changes were shown in syncytiotrophoblast cells of anembryonic pregnancies and AFP was strongly expressed by villous trophoblastic cells compared to embryonic pregnancies. We determined the CD34 positivity in both groups. In anembryonic pregnancies, vascular elements were much fewer in number compared with embryonic pregnancies (p<0.001) and were located in the formed of hemangioblastic cords. Placental vasculogenesis is a basic feature in all types of pregnancy and a relationship exists between trophoblast cells and vessels in the chorionic villi with the potential to influence each other's functions. Defective chorionic villus vascularization is associated with embryonic death. This study may support the hypothesis, as suggested by previous studies, that anembryonic pregnancy results from early embryonic death and subsequent reabsorption rather than from the nondevelopment of an embryo. (author)

  20. A sensitive fluorescent sensor for quantification of alpha-fetoprotein based on immunosorbent assay and click chemistry.

    Science.gov (United States)

    Xie, Qunfang; Weng, Xiuhua; Lu, Lijun; Lin, Zhenyu; Xu, Xiongwei; Fu, Caili

    2016-03-15

    A novel fluoresencent immunosensor for determination of cancer biomarkers such as alpha-fetoprotein (AFP) was designed by utilizing both the high specificity of antigen-antibody sandwich structure and the high sensitivity of the click chemistry based fluorescence detection. Instead of an enzyme or fluorophore, the CuO nanoparticles are labeled on the detection antibody, which was not susceptible to the change of the external environments. The CuO nanoparticles which were modified on the sandwich structure can be dissolved to produce Cu(2+) ions with the help of HCl and then the Cu(2+) ions were reduced by sodium ascorbate to produce Cu(+) ions which triggered the Cu(+) catalyzed alkyne-azide cycloaddition (CuAAC) reaction between the weak fluorescent compound (3-azido-7-hydroxycoumarin) and propargyl alcohol to form a strong fluorescent compound. A good linear relationship was observed between the fluorescence increase factor of the system and the concentration of AFP in the range of 0.025-5.0 ng/mL with a detection limit of 12 pg/mL (S/N=3). The proposed fluorescent sensor had been applied to detect AFP in the human serum samples and gave satisfactory results. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Recombinant heat shock protein 70 functional peptide and alpha-fetoprotein epitope peptide vaccine elicits specific anti-tumor immunity.

    Science.gov (United States)

    Wang, Xiao-Ping; Wang, Qiao-Xia; Lin, Huan-Ping; Xu, Bing; Zhao, Qian; Chen, Kun

    2016-11-01

    Alpha-fetoprotein (AFP) is a marker of hepatocellular carcinoma (HCC) and serves as a target for immunotherapy. However, current treatments targeting AFP are not reproducible and do not provide complete protection against cancer. This issue may be solved by developing novel therapeutic vaccines with enhanced immunogenicity that could effectively target AFP-expressing tumors. In this study, we report construction of a therapeutic peptide vaccine by linking heat shock protein 70 (HSP70) functional peptide to the AFP epitope to obtain HSP70-P/AFP-P. This novel peptide was administered into BALB/c mice to observe the effects. Quantification of AFP-specific CD8 + T cells that secrete IFN-γ in these mice via ELISPOT revealed the synergistic effects of HSP70-P/AFP-P with increased numbers of AFP-specific CD8 + T cells. Similarly, ELISA analysis showed increased granzyme B and perforin released by natural killer cells. Moreover, in vitro cytotoxic T-lymphocyte assays and in vivo tumor preventive experiments clearly showed the higher antitumor effects of HSP70-P/AFP-P against AFP-expressing tumors. These results show that treatment of BALB/c mice with HSP70-P/AFP-P induced stronger T-cells responses and improved protective immunity. Our data suggest that HSP70-P/AFP-P may be used as a therapeutic approach in the treatment of AFP-expressing cancers.

  2. A peptide derived from alpha-fetoprotein inhibits the proliferation induced by estradiol in mammary tumor cells in culture.

    Science.gov (United States)

    Sierralta, Walter D; Epuñan, Maria J; Reyes, José M; Valladares, Luis E; Andersen, Thomas T; Bennett, James A; Jacobson, Herbert I; Pino, Ana M

    2008-01-01

    This study was aimed to obtain additional information on the activity of a cyclized 9-amino acid peptide (cP) containing the active site of alpha fetoprotein, which inhibits the estrogen-stimulated proliferation of tumor cells in culture and of xenografts in immunodeficient mice. Breast cancer cells cultured in the presence of 2 nM estradiol were exposed to cP for different periods and their proliferation, estradiol binding parameters, clustering tendency and expression of E-cadherin and p21Cip1 were analyzed by biochemical and cell biology methods. The proliferation of MCF7 cells was significantly decreased by the addition of 2 microg/ml cP to the medium. cP did not increase cell death rate nor alter the number of binding sites for estradiol nor the endogenous aromatase activity of MCF7 cells. cP also decreased the proliferation of estrogen-dependent ZR75-1 cells but had no effect on estrogen-independent MDA-MB-231 cells. An increased nuclear p21Cip1 expression detected after cP treatment suggests that cP slows MCF7 cell proliferation via this regulator. We propose that cP could represent a novel breast cancer therapeutic agent whose mechanism of action is different from that of tamoxifen or of inhibitors of aromatase.

  3. Serum Adiponectin, Vitamin D, and Alpha-Fetoprotein in Children with Chronic Hepatitis C: Can They Predict Treatment Response?

    Directory of Open Access Journals (Sweden)

    Mohamed Ahmed Khedr

    2015-01-01

    Full Text Available Background & Aims. The currently available treatment for chronic hepatitis C (CHC in children is costly and with much toxicity. So, predicting the likelihood of response before starting therapy is important. Methods. Serum adiponectin, vitamin D, and alpha-fetoprotein (AFP were measured before starting pegylated-interferon/ribavirin therapy for 50 children with CHC. Another 21 healthy children were recruited as controls. Results. Serum adiponectin, vitamin D, and AFP were higher in the CHC group than healthy controls (p<0.0001, p=0.071, and p=0.87, resp.. In univariate analysis, serum adiponectin was significantly higher in responders than nonresponders (p<0.0001 and at a cutoff value ≥8.04 ng/mL it can predict treatment response by 77.8% sensitivity and 92.9% specificity, while both AFP and viremia were significantly lower in responders than nonresponders, p<0.0001 and p=0.0003, respectively, and at cutoff values ≤3.265 ng/mL and ≤235,384 IU/mL, respectively, they can predict treatment response with a sensitivity of 83.3% for both and specificity of 85.7% and 78.6%, respectively. In multivariate analysis, adiponectin was found to be the only independent predictor of treatment response (p=0.044. Conclusions. The pretreatment serum level of adiponectin can predict the likelihood of treatment response, thus avoiding toxicities for those unlikely to respond to therapy.

  4. Serum Adiponectin, Vitamin D, and Alpha-Fetoprotein in Children with Chronic Hepatitis C: Can They Predict Treatment Response?

    Science.gov (United States)

    Khedr, Mohamed Ahmed; Sira, Ahmad Mohamed; Saber, Magdy Anwar; Raia, Gamal Yousef

    2015-01-01

    Background & Aims. The currently available treatment for chronic hepatitis C (CHC) in children is costly and with much toxicity. So, predicting the likelihood of response before starting therapy is important. Methods. Serum adiponectin, vitamin D, and alpha-fetoprotein (AFP) were measured before starting pegylated-interferon/ribavirin therapy for 50 children with CHC. Another 21 healthy children were recruited as controls. Results. Serum adiponectin, vitamin D, and AFP were higher in the CHC group than healthy controls (p < 0.0001, p = 0.071, and p = 0.87, resp.). In univariate analysis, serum adiponectin was significantly higher in responders than nonresponders (p < 0.0001) and at a cutoff value ≥8.04 ng/mL it can predict treatment response by 77.8% sensitivity and 92.9% specificity, while both AFP and viremia were significantly lower in responders than nonresponders, p < 0.0001 and p = 0.0003, respectively, and at cutoff values ≤3.265 ng/mL and ≤235,384 IU/mL, respectively, they can predict treatment response with a sensitivity of 83.3% for both and specificity of 85.7% and 78.6%, respectively. In multivariate analysis, adiponectin was found to be the only independent predictor of treatment response (p = 0.044). Conclusions. The pretreatment serum level of adiponectin can predict the likelihood of treatment response, thus avoiding toxicities for those unlikely to respond to therapy.

  5. Screening and Identifying a Novel ssDNA Aptamer against Alpha-fetoprotein Using CE-SELEX

    Science.gov (United States)

    Dong, Lili; Tan, Qiwen; Ye, Wei; Liu, Dongli; Chen, Haifeng; Hu, Hongwei; Wen, Duo; Liu, Yang; Cao, Ya; Kang, Jingwu; Fan, Jia; Guo, Wei; Wu, Weizhong

    2015-01-01

    Alpha-fetoprotein (AFP) is a liver cancer associated protein and has long been utilized as a serum tumor biomarker of disease progression. AFP is usually detected in HCC patients by an antibody based system. Recently, however, aptamers generated from systematic evolution of ligands by exponential enrichment (SELEX) were reported to have an alternative potential in targeted imaging, diagnosis and therapy. In this study, AFP-bound ssDNA aptamers were screened and identified using capillary electrophoresis (CE) SELEX technology. After cloning, sequencing and motif analysis, we successfully confirmed an aptamer, named AP273, specifically targeting AFP. The aptamer could be used as a probe in AFP immunofluorescence imaging in HepG2, one AFP positive cancer cell line, but not in A549, an AFP negative cancer cell line. More interesting, the aptamer efficiently inhibited the migration and invasion of HCC cells after in vivo transfection. Motif analysis revealed that AP273 had several stable secondary motifs in its structure. Our results indicate that CE-SELEX technology is an efficient method to screen specific protein-bound ssDNA, and AP273 could be used as an agent in AFP-based staining, diagnosis and therapy, although more works are still needed. PMID:26497223

  6. Intrahepatic bile duct adenoma in a patient with chronic hepatitis B accompanied by elevation of alpha-fetoprotein

    Directory of Open Access Journals (Sweden)

    Jem Ma Ahn

    2015-12-01

    Full Text Available A 51-year-old male patient with chronic hepatitis B was referred to our hospital due to a 1-cm liver nodule on ultrasonography. Alpha-fetoprotein (AFP was slightly elevated. The nodule showed prolonged enhancement on dynamic liver magnetic resonance imaging and appeared as a hyperintensity on both diffusion-weighted and T2-weighted imaging. The nodule was followed up because it was small and typical findings of hepatocellular carcinoma (HCC were not observed in the dynamic imaging investigations. However, liver contrast-enhanced ultrasonography performed 1 month later showed enhancement during the arterial phase and definite washout during the delayed phase. Also, AFP had increased to over 200 ng/mL even though AST and ALT were decreased after administering an antiviral agent. He was presumptively diagnosed as HCC and underwent liver segmentectomy. Microscopy findings of the specimen indicated bile duct adenoma. After resection, the follow-up AFP had decreased to within the normal range. This patient represents a case of bile duct adenoma with AFP elevation mimicking HCC on contrast-enhanced ultrasonography.

  7. Expression of β-catenin protein in hepatocellular carcinoma and its relationship with alpha-fetoprotein.

    Science.gov (United States)

    Ren, Ya-Jun; Huang, Tao; Yu, Hong-Lu; Zhang, Li; He, Qian-Jin; Xiong, Zhi-Fan; Peng, Hua

    2016-12-01

    This study aimed to investigate the expression of β-catenin in hepatocellular carcinoma (HCC) tissues and its relationship with α-fetoprotein (AFP) in HCC. Immunohistochemistry was used to determine the expression of β-catenin in normal liver tissues (n=10), liver cirrhosis tissues (n=20), and primary HCC tissues (n=60). The relationship between β-catenin expression and clinical parameters of HCC was investigated. Real-time PCR and Western blotting were used to detect the mRNA and protein expression levels of β-catenin in the liver cancer cell line SMMC-7721 transfected with a plasmid encoding AFP, and also the mRNA and protein expression levels of β-catenin were measured in the liver cancer cell line Huh7 before and after the transfection with AFP shRNA plasmids. The results showed that β-catenin was only expressed on the cell membrane in normal liver tissues. Its localization to the cytoplasm and nucleus of cells was observed in a small proportion of cirrhotic tissues or adjacent HCC tissues, and such ectopic expression of β-catenin was predominant in HCC tissues. The abnormal expression of β-catenin was correlated with serum AFP levels, cancer cell differentiation and vascular invasion (P<0.05). Additionally, the increased expression of AFP resulted in the upregulation of β-catenin mRNA and protein levels, while knockdown of AFP with AFP shRNA led to significantly decreased β-catenin mRNA and protein levels (P<0.05). It was suggested that the abnormal expression of β-catenin is implicated in hepatic carcinogenesis and development. AFP can lead to increased expression of β-catenin, which may account for the poor prognosis of AFP-associated HCC patients.

  8. Radioimmunoimaging using F(ab')2 fragment of monoclonal antibodies against human alpha-fetoprotein

    International Nuclear Information System (INIS)

    Sakahara, Harumi; Endo, Keigo; Nakashima, Tetsuo; Koizumi, Mitsuru; Ohta, Hitoya; Torizuka, Kanji; Okada, Kenichiro; Yoshida, Osamu; Nishi, Shinzo.

    1985-01-01

    Using monoclonal antibodies against human α-fetoprotein (AFP), radioiodinated F(ab') 2 fragments were compared with whole IgG as a radiotracer for radioimmunoimaging of cancer. F(ab') 2 fragments were obtained by pepsin digestion of whole IgG (IgGl). IgG and F(ab') 2 were labeled with 125 I or 131 I by the chloramine-T method with almost full retention of antibody activity. F(ab') 2 fragments were cleared more rapidly from the circulation in normal mice with a half life of 6.3 hours than whole IgG with a half life of 5.5 days. Radioactivity of F(ab') 2 in various organs also decreased faster than IgG. In nude mice transplanted with AFP-producing human testicular tumor, F(ab') 2 fragments demonstrated superior scintigrams to whole IgG at 2 days after the injection, because of the fast disappearance of background radioactivity. Although absolute accumulation of 131 I labeled F(ab') 2 in the tumor was less than that of 131 I labeled IgG, tumor to other organ ratios were much higher with F(ab') 2 than those of IgG. The tumor to blood ratio of 131 I labeled F(ab') 2 was 1.04 at day 2, whereas tumor to blood ratio of 131 I labeled IgG was 0.55 at day 2 and 0.92 at day 4, respectively. These results indicated that for the radiolabeling of monoclonal antibodies, F(ab') 2 fragments would be superior to whole IgG in the radioimmunoimaging of cancer. (author)

  9. Basic studies on the tumor imaging using antibodies to human alpha-fetoprotein

    International Nuclear Information System (INIS)

    Sakahara, Harumi; Endo, Keigo; Nakashima, Tetsuo; Ohta, Hitoya; Torizuka, Kanji

    1984-01-01

    Using polyclonal antibodies to human α-fetoprotein (AFP), the effect of iodination on the antibody activity and tumor accumulation of radioiodinated antibodies in tumor bearing nude mice were examined. Antibodies, obtained from horse antiserum and purified by affinity chromatography, were iodinated by the chloramine-T method and their antibody activity was evaluated using RIA and Scatchard plot analysis. When high concentrations of chloramine-T were used or more than 2.6 iodine atoms were incorporated per antibody molecule, the antigen binding capacity rather than the affinity constant was affected by the iodination. The antibody activity was completely destroyed at an iodine to antibody molar ratio of 15.4. Antibodies, however, which were iodinated under low concentrations of chloramine-T and contained less than 0.8 iodines per antibody molecule, showed almost full retention of their antibody activity. Nude mice transplanted with AFP producing human testicular tumor or AFP non-producing human urinary bladder tumor were administered intravenously with 131 I-labeled antibodies to human AFP. Scintigrams were taken at 1, 2, 4 and 7 days after the injection of labeled antibodies. At day 7, nude mice were sacrificed and organs and tumor were removed, weighed and counted. In nude mice bearing testicular tumor, tumor image became gradually clear with decreasing background activity and tumor to blood ratio, obtained, was 0.82 for testicular tumor compared to 0.42 for bladder tumor. These results indicated a specific in vivo localization of 131 I-labeled antihuman AFP antibodies in AFP producing tumor. (author)

  10. Assessment of the correlation between serum prolidase and alpha-fetoprotein levels in patients with hepatocellular carcinoma.

    Science.gov (United States)

    Ilikhan, Sevil Uygun; Bilici, Muammer; Sahin, Hatice; Akca, Ayşe Semra Demir; Can, Murat; Oz, Ibrahim Ilker; Guven, Berrak; Buyukuysal, M Cagatay; Ustundag, Yucel

    2015-06-14

    To determine the predictive value of increased prolidase activity that reflects increased collagen turnover in patients with hepatocellular carcinoma (HCC). Sixty-eight patients with HCC (mean age of 69.1 ± 10.1), 31 cirrhosis patients (mean age of 59.3 ± 6.3) and 33 healthy volunteers (mean age of 51.4 ± 12.6) were enrolled in this study. Univariate and multivariate analysis were used to evaluate the association of serum α-fetoprotein (AFP) values with HCC clinicopathological features, such as tumor size, number and presence of vascular and macrovascular invasion. The patients with HCC were divided into groups according to tumor size, number and presence of vascular invasion (diameters; ≤ 3 cm, 3-5 cm and ≥ 5 cm, number; 1, 2 and ≥ 3, macrovascular invasion; yes/no). Barcelona-clinic liver cancer (BCLC) criteria were used to stage HCC patients. Serum samples for measurement of prolidase and alpha-fetoprotein levels were kept at -80 °C until use. Prolidase levels were measured spectrophotometrically and AFP concentrations were determined by a chemiluminescence immunometric commercial diagnostic assay. In patients with HCC, prolidase and AFP values were evaluated according to tumor size, number, presence of macrovascular invasion and BCLC staging classification. Prolidase values were significantly higher in patients with HCC compared with controls (P < 0.001). Prolidase levels were significantly associated with tumor size and number (P < 0.001, P = 0.002, respectively). Prolidase levels also differed in patients in terms of BCLC staging classification (P < 0.001). Furthermore the prolidase levels in HCC patients showed a significant difference compared with patients with cirrhosis (P < 0.001). In HCC patients grouped according to tumor size, number and BCLC staging classification, AFP values differed separately (P = 0.032, P = 0.038, P = 0.015, respectively). In patients with HCC, there was a significant correlation (r = 0.616; P < 0.001) between

  11. A hypoxia- and {alpha}-fetoprotein-dependent oncolytic adenovirus exhibits specific killing of hepatocellular carcinomas.

    Science.gov (United States)

    Kwon, Oh-Joon; Kim, Pyung-Hwan; Huyn, Steven; Wu, Lily; Kim, Minjung; Yun, Chae-Ok

    2010-12-15

    Oncolytic adenoviruses (Ad) constitute a new promising modality of cancer gene therapy that displays improved efficacy over nonreplicating Ads. We have previously shown that an E1B 19-kDa-deleted oncolytic Ad exhibits a strong cell-killing effect but lacks tumor selectivity. To achieve hepatoma-restricted cytotoxicity and enhance replication of Ad within the context of tumor microenvironment, we used a modified human α-fetoprotein (hAFP) promoter to control the replication of Ad with a hypoxia response element (HRE). We constructed Ad-HRE(6)/hAFPΔ19 and Ad-HRE(12)/hAFPΔ19 that incorporated either 6 or 12 copies of HRE upstream of promoter. The promoter activity and specificity to hepatoma were examined by luciferase assay and fluorescence-activated cell sorting analysis. In addition, the AFP expression- and hypoxia-dependent in vitro cytotoxicity of Ad-HRE(6)/hAFPΔ19 and Ad-HRE(12)/hAFPΔ19 was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cytopathic effect assay. In vivo tumoricidal activity on subcutaneous and liver orthotopic model was monitored by noninvasive molecular imaging. Ad-HRE(12)/hAFPΔ19 exhibited enhanced tumor selectivity and cell-killing activity when compared with Ad-hAFPΔ19. The tumoricidal activity of Ad-HRE(12)/hAFPΔ19 resulted in significant inhibition of tumor growth in both subcutaneous and orthotopic models. Histologic examination of the primary tumor after treatment confirmed accumulation of viral particles near hypoxic areas. Furthermore, Ad-HRE(12)/hAFPΔ19 did not cause severe inflammatory immune response and toxicity after systemic injection. The results presented here show the advantages of incorporating HREs into a hAFP promoter-driven oncolytic virus. This system is unique in that it acts in both a tissue-specific and tumor environment-selective manner. The greatly enhanced selectivity and tumoricidal activity of Ad-HRE(12)/hAFPΔ19 make it a promising therapeutic agent in the treatment

  12. Clinical characteristics of hepatocellular carcinoma patients with normal serum alpha-fetoprotein level: A study of 112 consecutive cases.

    Science.gov (United States)

    Li, Li; Chen, Jinglong; Xu, Weiran; Ding, Xiaosheng; Wang, Xiangyi; Liang, Jun

    2017-10-26

    Serum alpha-fetoprotein (AFP) level is normal in 30-40% of hepatocellular carcinoma (HCC) patients, and knowledge on its characteristics and clinical outcome is limited. The purpose of this observational study was to determine the clinical presentation, biological behavior and outcome of HCC patients with normal AFP level. Data of 112 consecutive HCC patients with normal AFP level were analyzed retrospectively. Statistical analysis including survival and factors associated with serum AFP level were performed by Kaplan-Meier method and t-test, respectively. Hepatitis B virus infection exited in 83.0% of all 112 HCC patients with normal AFP level. During a mean 52 ± 20 months (range 5-85 months) follow-up, the 1-, 2-, 3-year overall survival (OS) rate was 97.2%, 85.3% and 81.7%, respectively. The OS rates at 3 years stratified by stages at diagnosis were 100%, 96.2%, 85.7%, 11.1% and 0%, respectively for Barcelona Clinic Liver Cancer (BCLC) stage 0-D diseases. Significant difference in OS was observed among patients with BCLC stage 0-D diseases, P level elevated beyond normal figure during follow-up (AFP conversion) in 16 patients, which related with deterioration of liver function, quantitative changes of T helper cell subsets, rapid tumor progression and shorter survival. Patients with sustained normal AFP level had better survival than patients with AFP conversion, P level elevation and the time of AFP elevation to death, P level was relatively optimal. Serum AFP level elevation during follow-up was significantly associated with clinical outcome in terms of OS. © 2017 John Wiley & Sons Australia, Ltd.

  13. Immunoprecipitation assay of alpha-fetoprotein synthesis by cultured mouse hepatoma cells treated with estrogens and glucocorticoids.

    Science.gov (United States)

    Rosebrock, J A; Parker, C L; Kute, T E

    1981-01-01

    This investigation was to study the biosynthesis of 3H-labeled alpha-fetoprotein (AFP) by cultured mouse hepatoma (HEPA-2) cells. Both the function and regulation of this oncodevelopmental gene are unknown. However, evidence indicates that mechanisms controlling the expression of AFP involve aspects of both normal embryonic development and neoplastic transformation. the secretion of AFP was analyzed during different phases of the growth cycle to provide information on AFP production using standard culture conditions. The highest rate of secretion occurred during the stationary phase, followed by the late logarithmic and early logarithmic phases of growth, respectively. The production of AFP was then determined following the addition of glucocorticoids and estrogens in an attempt to understand hormonal factors that may be involved. Studies utilizing estradiol-17 beta indicated that the secretion of AFP did not appear to be sensitive to this steroid even though sucrose density gradient analysis of HEPA-2 cytosol, for estrogenic receptors, revealed competitive binding moieties on the 8S and 4S regions of the gradient. In contrast, the secretion of the total complement of proteins, including AFP, was significantly stimulated by the glucocorticoids, dexamethasone and corticosterone. Analysis of HEPA-2 cytosol for glucocorticoid receptors revealed binding components in the 7S and 3-4S regions of the gradient. The 3H-dexamethasone binding appeared to be stereospecific since nonlabeled dexamethasone, but not nonlabeled estradiol-17 beta, effectively displaced the bound radioactivity. The glucocorticoid-binding component in HEPA-2 therefore displayed characteristics reported for glucocorticoid receptors in normal liver and other hepatomas.

  14. Time of hepatocellular carcinoma recurrence after liver resection and alpha-fetoprotein are important prognostic factors for salvage liver transplantation.

    Science.gov (United States)

    Lee, Sanghoon; Hyuck David Kwon, Choon; Man Kim, Jong; Joh, Jae-Won; Woon Paik, Seung; Kim, Bong-Wan; Wang, Hee-Jung; Lee, Kwang-Woong; Suh, Kyung-Suk; Lee, Suk-Koo

    2014-09-01

    Salvage liver transplantation (LT) is considered a feasible option for the treatment of recurrent hepatocellular carcinoma (HCC). We performed this multicenter study to assess the risk factors associated with the recurrence of HCC and patient survival after salvage LT. Between January 2000 and December 2011, 101 patients who had previously undergone liver resection (LR) for HCC underwent LT at 3 transplant centers in Korea. Sixty-nine patients' data were retrospectively reviewed for the analysis. The recurrence of HCC was diagnosed at a median of 10.6 months after the initial LR, and patients underwent salvage LT. Recurrences were within the Milan criteria in 48 cases and were outside the Milan criteria in 21 cases. After salvage LT, 31 patients had HCC recurrence during a median follow-up period of 24.5 months. There were 24 deaths, and 20 were due to HCC recurrence. The 5-year overall survival rate was approximately 54.6%, and the 5-year recurrence-free survival rate was 49.3%. HCC recurrence within the 8 months after LR [hazard ratio (HR) = 3.124, P = 0.009], an alpha-fetoprotein level higher than 200 ng/mL (HR = 2.609, P = 0.02), and HCC outside the Milan criteria at salvage LT (HR = 2.219, P = 0.03) were independent risk factors for poor recurrence-free survival after salvage LT. In conclusion, the timing and extent of HCC recurrence after primary LR both play significant roles in the outcome of salvage LT. © 2014 American Association for the Study of Liver Diseases.

  15. Prognostic significance of an early decline in serum alpha-fetoprotein during chemotherapy for ovarian yolk sac tumors.

    Science.gov (United States)

    de la Motte Rouge, Thibault; Pautier, Patricia; Genestie, Catherine; Rey, Annie; Gouy, Sébastien; Leary, Alexandra; Haie-Meder, Christine; Kerbrat, Pierre; Culine, Stéphane; Fizazi, Karim; Lhommé, Catherine

    2016-09-01

    The ovarian yolk sac tumor (OYST) is a very rare malignancy arising in young women. Our objective was to determine whether an early decline in serum alpha-fetoprotein (AFP) during chemotherapy has a prognostic impact. This retrospective study is based on prospectively recorded OYST cases at Gustave Roussy (Cancer Treatment Center). Survival curves were estimated using the Kaplan-Meier method. The serum AFP decline was calculated with the formula previously developed and validated in male patients with poor prognosis non-seminomatous germ cell tumors. Univariate and multivariate analyses were performed using the log-rank test and logistic regression, respectively. Data on AFP were available to calculate an early AFP decline in 57 patients. All patients had undergone surgery followed by chemotherapy. The 5-year overall survival (OS) and event-free survival (EFS) rates were 86% (95% CI: 74%-93%) and 84% (95% CI: 73%-91%), respectively. The disease stage, presence of ascites at presentation, use of the BEP regimen, serum AFP half-life and an early AFP decline were significantly predictive factors for OS and EFS in the univariate analysis. The OS rate was 100% and 49% (95% CI: 26%-72%) in patients with a favorable AFP decline and in those with an unfavorable decline, respectively (p<0.001). In the multivariate analysis, only the presence of ascites at diagnosis (RR=7.3, p=0.03) and an unfavorable early AFP decline (RR=16.9, p<0.01) were significant negative predictive factors for OS. An early AFP decline during chemotherapy is an independent prognostic factor in patients with OYSTs. No conflict of interest. Copyright © 2016. Published by Elsevier Inc.

  16. Targeting Alpha-Fetoprotein (AFP)-MHC Complex with CAR T-Cell Therapy for Liver Cancer.

    Science.gov (United States)

    Liu, Hong; Xu, Yiyang; Xiang, Jingyi; Long, Li; Green, Shon; Yang, Zhiyuan; Zimdahl, Bryan; Lu, Jingwei; Cheng, Neal; Horan, Lucas H; Liu, Bin; Yan, Su; Wang, Pei; Diaz, Juan; Jin, Lu; Nakano, Yoko; Morales, Javier F; Zhang, Pengbo; Liu, Lian-Xing; Staley, Binnaz K; Priceman, Saul J; Brown, Christine E; Forman, Stephen J; Chan, Vivien W; Liu, Cheng

    2017-01-15

    The majority of tumor-specific antigens are intracellular and/or secreted and therefore inaccessible by conventional chimeric antigen receptor (CAR) T-cell therapy. Given that all intracellular/secreted proteins are processed into peptides and presented by class I MHC on the surface of tumor cells, we used alpha-fetoprotein (AFP), a specific liver cancer marker, as an example to determine whether peptide-MHC complexes can be targets for CAR T-cell therapy against solid tumors. We generated a fully human chimeric antigen receptor, ET1402L1-CAR (AFP-CAR), with exquisite selectivity and specificity for the AFP 158-166 peptide complexed with human leukocyte antigen (HLA)-A*02:01. We report that T cells expressing AFP-CAR selectively degranulated, released cytokines, and lysed liver cancer cells that were HLA-A*02:01 + /AFP + while sparing cells from multiple tissue types that were negative for either expressed proteins. In vivo, intratumoral injection of AFP-CAR T cells significantly regressed both Hep G2 and AFP 158 -expressing SK-HEP-1 tumors in SCID-Beige mice (n = 8 for each). Moreover, intravenous administration of AFP-CAR T cells in Hep G2 tumor-bearing NSG mice lead to rapid and profound tumor growth inhibition (n = 6). Finally, in an established intraperitoneal liver cancer xenograft model, AFP-CAR T cells showed robust antitumor activity (n = 6). This study demonstrates that CAR T-cell immunotherapy targeting intracellular/secreted solid tumor antigens can elicit a potent antitumor response. Our approach expands the spectrum of antigens available for redirected T-cell therapy against solid malignancies and offers a promising new avenue for liver cancer immunotherapy. Clin Cancer Res; 23(2); 478-88. ©2016 AACR. ©2016 American Association for Cancer Research.

  17. Hepatocellular carcinoma in children and young patients with chronic HBV infection and the usefulness of alpha-fetoprotein assessment.

    Science.gov (United States)

    Tajiri, Hitoshi; Takano, Tomoko; Tanaka, Hideo; Ushijima, Kosuke; Inui, Ayano; Miyoshi, Yoko; Ozono, Keiichi; Abukawa, Daiki; Endo, Takeshi; Brooks, Stephen; Tanaka, Yasuhito

    2016-11-01

    The aims of the study were to elucidate the clinical characteristics of patients who developed hepatocellular carcinoma (HCC) related to persistent HBV infection since childhood and to investigate usefulness of assessing alpha-fetoprotein (AFP) in this population. A nationwide multicenter survey of children with chronic HBV infection was performed. Among 548 patients, 15 patients developed HCC at the median age of 15 years (range 9-36), including 13 males and 2 females. A case-control comparison showed that HBeAg seroconversion and liver cirrhosis were associated with the occurrence of HCC. Of the 15 HCC patients, 5 were treated with interferon and none of them responded to interferon therapy as compared with 12 of the 17 responders in the control group. Of the 15 patients, 10 died and 9 of the 10 who died never visited any medical facilities until diagnosis of HCC, while the remaining 5 surviving patients never stopped their clinic visits. The usefulness of AFP assessment was shown by the findings that AFP levels were elevated in all HCC cases, that elevations in AFP levels were detected prior to the diagnosis in the surviving patients, and that sensitivity of AFP as a diagnostic test for HCC was very high among 40 patients including our 14 and an additional 26 collected from the literature. HBeAg seroconversion and liver cirrhosis are associated with the occurrence of HCC. Regular measurement of AFP might be helpful to watch for the occurrence of HCC when following children and young patients with chronic HBV infection since childhood. © 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  18. Nonsecreted cytoplasmic alpha-fetoprotein: a newly discovered role in intracellular signaling and regulation. An update and commentary.

    Science.gov (United States)

    Mizejewski, G J

    2015-12-01

    The concept of a non-secreted cytoplasmic-bound form of alpha-fetoprotein is not a new notion in AFP biological activities. Cytoplasmic AFP (CyAFP) is a long known but forgotten protein in search of a function other than a histochemical biomarker. In this report, CyAFP is presented as an "old" protein with a newly described intracellular function. In 1976, CyAFP was shown to be a product of hepatoma cells utilizing 14Cleucine incorporation and demonstrated by autoradiographic procedures. The synthesis of CyAFP without secretion was demonstrated to occur in both malignant and non-malignant cells encompassing hepatomas, ascite fluid cells, immature rodent uterus, MCF-7 breast cancers, and cytosols from human breast cancer patients. Using computer protein matching and alignments in AFP versus members of the nuclear receptor superfamily, a consecutive series of leucine zipper (heptad) repeats in AFP was previously reported, suggesting the possibility for protein-to-protein interactions. The potential for heptad heterodimerization between protein-binding partners provided the rationale for proposing that CyAFP might have the capability to form molecular hetero-complexes with cytoplasmic based transcription factors. More recent investigations have now provided experimental evidence that CyAFP is capable of colocalizing and interacting with transcription-associated factors. Such proteins can modulate intracellular signaling leading to regulation of transcription factors and initiation of growth in human cancer cells. Although circulating serum AFP is known as a growth-enhancing factor during development, cytoplasmic AFP has a lethal role in the oncogenesis, growth, and metastasis of adult liver cancer.

  19. Liver transplantation for hepatocellular carcinoma: evaluation of the alpha-fetoprotein model in a multicenter cohort from Latin America.

    Science.gov (United States)

    Piñero, Federico; Tisi Baña, Matías; de Ataide, Elaine Cristina; Hoyos Duque, Sergio; Marciano, Sebastian; Varón, Adriana; Anders, Margarita; Zerega, Alina; Menéndez, Josemaría; Zapata, Rodrigo; Muñoz, Linda; Padilla Machaca, Martín; Soza, Alejandro; McCormack, Lucas; Poniachik, Jaime; Podestá, Luis G; Gadano, Adrian; Boin, Ilka S F Fatima; Duvoux, Christophe; Silva, Marcelo

    2016-11-01

    The French alpha-fetoprotein (AFP) model has recently shown superior results compared to Milan criteria (MC) for prediction of hepatocellular carcinoma (HCC) recurrence after liver transplantation (LT) in European populations. The aim of this study was to explore the predictive capacity of the AFP model for HCC recurrence in a Latin-American cohort. Three hundred twenty-seven patients with HCC were included from a total of 2018 patients transplanted at 15 centres. Serum AFP and imaging data were both recorded at listing. Predictability was assessed by the Net Reclassification Improvement (NRI) method. Overall, 82 and 79% of the patients were within MC and the AFP model respectively. NRI showed a superior predictability of the AFP model against MC. Patients with an AFP score >2 points had higher risk of recurrence at 5 years Hazard Ratio (HR) of 3.15 (P = 0.0001) and lower patient survival (HR = 1.51; P = 0.03). Among patients exceeding MC, a score ≤2 points identified a subgroup of patients with lower recurrence (5% vs 42%; P = 0.013) and higher survival rates (84% vs 45%; P = 0.038). In cases treated with bridging procedures, following restaging, a score >2 points identified a higher recurrence (HR 2.2, P = 0.12) and lower survival rate (HR 2.25, P = 0.03). A comparative analysis between HBV and non-HBV patients showed that the AFP model performed better in non-HBV patients. The AFP model could be useful in Latin-American countries to better select patients for LT in subgroups presenting with extended criteria. However, particular attention should be focused on patients with HBV. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  20. The alpha-fetoprotein (AFP) third domain: a search for AFP interaction sites of cell cycle proteins.

    Science.gov (United States)

    Mizejewski, G J

    2016-09-01

    The carboxy-terminal third domain of alpha-fetoprotein (AFP-3D) is known to harbor binding and/or interaction sites for hydrophobic ligands, receptors, and binding proteins. Such reports have established that AFP-3D consists of amino acid (AA) sequence stretches on the AFP polypeptide that engages in protein-to-protein interactions with various ligands and receptors. Using a computer software program specifically designed for such interactions, the present report identified AA sequence fragments on AFP-3D that could potentially interact with a variety of cell cycle proteins. The cell cycle proteins identified were (1) cyclins, (2) cyclin-dependent kinases, (3) cell cycle-associated proteins (inhibitors, checkpoints, initiators), and (4) ubiquitin ligases. Following detection of the AFP-3D to cell cycle protein interaction sites, the computer-derived AFP localization AA sequences were compared and aligned with previously reported hydrophobic ligand and receptor interaction sites on AFP-3D. A literature survey of the association of cell cycle proteins with AFP showed both positive relationships and correlations. Previous reports of experimental AFP-derived peptides effects on various cell cycle proteins served to confirm and verify the present computer cell cycle protein identifications. Cell cycle protein interactions with AFP-CD peptides have been reported in cultured MCF-7 breast cancer cells subjected to mRNA microarray analysis. After 7 days in culture with MCF-7 cells, the AFP-derived peptides were shown to downregulate cyclin E, SKP2, checkpoint suppressors, cyclin-dependent kinases, and ubiquitin ligases that modulate cyclin E/CdK2 transition from the G1 to the S-phase of the cell cycle. Thus, the experimental data on AFP-CD interaction with cell cycle proteins were consistent with the "in silico" findings.

  1. Diagnostic value of PIVKA-II and alpha-fetoprotein in hepatitis B virus-associated hepatocellular carcinoma

    Science.gov (United States)

    Seo, Seung In; Kim, Hyoung Su; Kim, Won Jin; Shin, Woon Geon; Kim, Doo Jin; Kim, Kyung Ho; Jang, Myoung Kuk; Lee, Jin Heon; Kim, Joo Seop; Kim, Hak Yang; Kim, Dong Joon; Lee, Myung Seok; Park, Choong Kee

    2015-01-01

    AIM: To determine the cutoff values and to compare the diagnostic role of alpha-fetoprotein (AFP) and prothrombin induced by vitamin K absence-II (PIVKA-II) in chronic hepatitis B (CHB). METHODS: A total of 1255 patients with CHB, including 157 patients with hepatocellular carcinoma (HCC), 879 with non-cirrhotic CHB and 219 with cirrhosis without HCC, were retrospectively enrolled. The areas under the receiver operating characteristic (AUROC) curves of PIVKA-II, AFP and their combination were calculated and compared. RESULTS: The optimal cutoff values for PIVKA-II and AFP were 40 mAU/mL and 10 ng/mL, respectively, for the differentiation of HCC from nonmalignant CHB. The sensitivity and specificity were 73.9% and 89.7%, respectively, for PIVKA-II and 67.5% and 90.3% for AFP, respectively. The AUROC curves of both PIVKA-II and AFP were not significantly different (0.854 vs 0.853, P = 0.965) for the differentiation of HCC from nonmalignant CHB, whereas the AUROC of PIVKA-II was significantly better than that of AFP in patients with cirrhosis (0.870 vs 0.812, P = 0.042). When PIVKA-II and AFP were combined, the diagnostic power improved significantly compared to either AFP or PIVKA-II alone for the differentiation of HCC from nonmalignant CHB (P < 0.05), especially when cirrhosis was present (P < 0.05). CONCLUSION: Serum PIVKA-II might be a better tumor marker than AFP, and its combination with AFP may enhance the early detection of HCC in patients with CHB. PMID:25852278

  2. Maternal serum alpha-fetoprotein at 12, 22 and 32 weeks' gestation in screening for pre-eclampsia.

    Science.gov (United States)

    Bredaki, F E; Matalliotakis, M; Wright, A; Wright, D; Nicolaides, K H

    2016-04-01

    To examine the distribution of maternal serum alpha-fetoprotein (AFP) at 12, 22 and 32 weeks' gestation in singleton pregnancies which develop pre-eclampsia (PE) and examine the performance of this biomarker in screening for PE. Serum AFP was measured in 17 071 cases at 11-13 weeks, in 8583 cases at 19-24 weeks and 8609 cases at 30-34 weeks' gestation. Bayes' theorem was used to combine the a-priori risk from maternal characteristics and medical history with AFP. The performance of screening for PE requiring delivery < 32, at 32 + 0 to 36 + 6, < 37 and ≥ 37 weeks' gestation was estimated. In pregnancies that developed PE, serum AFP multiples of the median (MoM) was increased at 11-13 and 19-24 weeks' gestation, but not at 30-34 weeks, and the values were inversely related to gestational age at delivery. Combined screening with maternal factors and serum AFP improved the prediction provided by maternal factors alone for PE delivering < 37 weeks, but not for PE delivering ≥ 37 weeks. The performance of screening for preterm PE was better at 19-24 weeks than at 11-13 weeks and the detection rate (DR) for a given false-positive rate (FPR) was higher for PE delivering < 32 weeks than for PE delivering at 32 + 0 to 36 + 6 weeks. The DRs, at 10% FPR, of combined screening at 11-13 weeks for PE delivering < 32 and at 32 + 0 to 36 + 6 weeks were 54% and 45%, respectively, and these improved to 72% and 53% with screening at 19-24 weeks. Measurement of serum AFP at 11-13 and 19-24 weeks' gestation improves the prediction of preterm PE provided by maternal factors alone. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd.

  3. Serum alpha-fetoprotein in the three trimesters of pregnancy: effects of maternal characteristics and medical history.

    Science.gov (United States)

    Bredaki, F E; Sciorio, C; Wright, A; Wright, D; Nicolaides, K H

    2015-07-01

    To define the contribution of maternal variables which influence the measured level of maternal serum alpha-fetoprotein (AFP) in screening for pregnancy complications. Maternal characteristics and medical history were recorded and serum AFP was measured in women with a singleton pregnancy attending for three routine hospital visits at 11 + 0 to 13 + 6, 19 + 0 to 24 + 6 and 30 + 0 to 34 + 6 weeks' gestation. For pregnancies delivering phenotypically normal live births or stillbirths ≥ 24 weeks' gestation, variables from maternal demographic characteristics and medical history that are important in the prediction of AFP were determined from a linear mixed-effects multiple regression. Serum AFP was measured in 17 071 cases in the first trimester, 8583 in the second trimester and 8607 in the third trimester. Significant independent contributions to serum AFP were provided by gestational age, maternal weight, racial origin, gestational age at delivery and birth-weight Z-score of the neonate of the previous pregnancy and interpregnancy interval. Cigarette smoking was found to significantly affect serum AFP in the first trimester only. The machine used to measure serum AFP was also found to have a significant effect. Random-effects multiple regression analysis was used to define the contribution of maternal variables that influence the measured level of serum AFP and express the values as multiples of the median (MoMs). The model was shown to provide an adequate fit of MoM values for all covariates, both in pregnancies that developed pre-eclampsia and in those without this pregnancy complication. A model was fitted to express measured serum AFP across the three trimesters of pregnancy as MoMs, after adjusting for variables from maternal characteristics and medical history that affect this measurement. Copyright © 2015 ISUOG. Published by John Wiley & Sons Ltd.

  4. [The Clinical Significance of Serum Alpha-fetoprotein in Diagnosing Hepatocellular Carcinoma in a Health Screening Population].

    Science.gov (United States)

    Ko, Young Sun; Bae, Joo Hwan; Sinn, Dong Hyun; Gwak, Geum Youn; Kang, Wonseok; Paik, Yong Han; Choi, Moon Seok; Lee, Joon Hyeok; Koh, Kwang Cheol; Paik, Seung Woon

    2017-04-25

    Serum alpha-fetoprotein (AFP) measurement is commonly included in a health check-up program in Korea. However, its benefits remain uncertain. We analyzed whether AFP measurement should be included in a general health check-up program to screen for hepatocellular carcinoma (HCC). A total of 36,552 adults aged 18 years or older-who participated in a routine health examination including AFP determination between January 2009 and December 2009 at the Health Promotion Center, Samsung Medical Center, South Korea-were analyzed. High risk of HCC was defined as positivity for hepatitis B surface antigen, anti-hepatitis C virus antibody or having liver cirrhosis. AFP level >10 ng/mL was observed in 27 participants (0.1%) and primary liver cancer was diagnosed in 9 patients (6 HCC and 3 cholangiocarcinoma). Among 1,619 participants with high risk factors of HCC, AFP level >10 ng/mL was observed in 16 participants, of which, 4 diagnoses were made. Sensitivity, specificity, positive predictive value, and negative predictive value of AFP for HCC was 0.66, 0.99, 0.25 and 0.99, respectively, for high risk participants. Among 34,933 participants without risk factors for HCC, 11 patients (<0.1%) showed elevated AFP levels above 10 ng/mL, and no case was diagnosed with primary liver cancer during a median follow-up period of 36 months (range: 0-48 months). AFP elevation was rare in participants without risk factors for HCC, and was unable to screen for HCC in this population. We discourage routine AFP measurements for asymptomatic adults without risk factors of HCC.

  5. Changes in highly sensitive alpha-fetoprotein for the prediction of the outcome in patients with hepatocellular carcinoma after hepatectomy

    International Nuclear Information System (INIS)

    Toyoda, Hidenori; Kumada, Takashi; Tada, Toshifumi; Ito, Takanori; Maeda, Atsuyuki; Kaneoka, Yuji; Kagebayashi, Chiaki; Satomura, Shinji

    2014-01-01

    We investigated changes in highly sensitive lens culinaris agglutinin A-reactive fraction of alpha-fetoprotein (hsAFP-L3) measured using a novel method and its predictive ability for prognosis in patients with hepatocellular carcinoma (HCC) who underwent curative hepatectomy, comparing to other HCC tumor markers, that is, AFP, des-gamma-carboxy prothrombin (DCP), and AFP-L3 measured with conventional method (cAFP-L3). AFP, DCP, and AFP-L3 including both cAFP-L3 and hsAFP-L3 were measured before and after curative hepatectomy in 187 patients. The percentage of patients with elevated tumor marker levels pre- and postoperatively was compared, and recurrence-free and overall survival rates were analyzed based on changes in tumor markers. The percentages of patients with elevated AFP, DCP, and cAFP-L3 decreased postoperatively. In contrast, the percentage of patients with elevated hsAFP-L3 did not decrease postoperatively. Both recurrence-free and overall survival rates were significantly lower in patients whose tumor marker levels remained elevated postoperatively than patients without tumor marker elevation postoperatively. Recurrence-free and overall survival rates of patients in whom hsAFP-L3 became elevated postoperatively despite normal preoperative hsAFP-L3 levels were significantly lower than those of patients with normal hsAFP-L3 postoperatively, and were similar to those of patients with persistent elevation. Preoperative elevations of AFP, DCP, and cAFP normalized in many patients postoperatively, but not for hsAFP-L3. The elevation of hsAFP-L3 identifies patients with poor prognosis despite the normalization of AFP and DCP

  6. Alpha-fetoprotein-producing ovarian clear cell adenocarcinoma with fetal gut differentiation: a rare case report and literature review.

    Science.gov (United States)

    Chao, Wei-Ting; Liu, Chia-Hao; Lai, Chiung-Ru; Chen, Yi-Jen; Chuang, Chi-Mu; Wang, Peng-Hui

    2018-06-22

    Alpha-fetoprotein (AFP) is a useful tumor marker for ovarian germ cell tumors, particularly yolk sac tumor (YST). It is valuable for both diagnosis and further follow-up. Epithelial ovarian carcinoma (EOC) rarely secretes AFP, especially for clear cell type and in the postmenopausal women. Based on the limited knowledge about AFP-producing clear cell type EOC, a case and literature review on this topic is extensively reviewed. We report a 55-year-old postmenopausal woman experienced vaginal spotting for one month, and serum level of AFP was 60,721 ng/ml initially. Histological examination was clear cell type EOC. Tumor cells revealed strong immunoreactivity for glypican-3 (GPC3) and AFP and weak for hepatocyte nuclear factor-1 beta (HNF-1 beta), but negative for CD30, making the diagnosis of AFP-producing clear cell type EOC with fetal gut differentiation in focal areas, FIGO (International Federation of Gynecology and Obstetrics) IIIc. Although the patient underwent an intensive treatment, including optimal debulking surgery and multi-agent chemotherapy, the patient died of disease. To provide a better understanding of clinical and molecular characteristics of the AFP-producing clear cell type EOC, we conducted a systematic literature review. A total of three papers described the AFP-producing clear cell type EOC are available. The overall survival rate of these cases, including the current case is 50%. Although immunohistochemical examination is not always needed in routine for the diagnosis of clear cell type EOC, to distinguish from other tumors, especially germ cell tumors, or to provide the better way to monitor therapeutic response or to evaluate the disease status, immunostaining, including GPC3, HNF-1 beta, CD30, cytokeratin 7 or 20, and AFP is taken into account. Due to rarity, the appropriate chemotherapy regimen and the biological behavior of AFP-producing clear cell type EOC are still unclear.

  7. Female mice deficient in alpha-fetoprotein show female-typical neural responses to conspecific-derived pheromones.

    Directory of Open Access Journals (Sweden)

    Olivier Brock

    Full Text Available The neural mechanisms controlling sexual behavior are sexually differentiated by the perinatal actions of sex steroid hormones. We recently observed using female mice deficient in alpha-fetoprotein (AFP-KO and which lack the protective actions of AFP against maternal estradiol, that exposure to prenatal estradiol completely defeminized the potential to show lordosis behavior in adulthood. Furthermore, AFP-KO females failed to show any male-directed mate preferences following treatment with estradiol and progesterone, indicating a reduced sexual motivation to seek out the male. In the present study, we asked whether neural responses to male- and female-derived odors are also affected in AFP-KO female mice. Therefore, we compared patterns of Fos, the protein product of the immediate early gene, c-fos, commonly used as a marker of neuronal activation, between wild-type (WT and AFP-KO female mice following exposure to male or estrous female urine. We also tested WT males to confirm the previously observed sex differences in neural responses to male urinary odors. Interestingly, AFP-KO females showed normal, female-like Fos responses, i.e. exposure to urinary odors from male but not estrous female mice induced equivalent levels of Fos protein in the accessory olfactory pathways (e.g. the medial part of the preoptic nucleus, the bed nucleus of the stria terminalis, the amygdala, and the lateral part of the ventromedial hypothalamic nucleus as well as in the main olfactory pathways (e.g. the piriform cortex and the anterior cortical amygdaloid nucleus, as WT females. By contrast, WT males did not show any significant induction of Fos protein in these brain areas upon exposure to either male or estrous female urinary odors. These results thus suggest that prenatal estradiol is not involved in the sexual differentiation of neural Fos responses to male-derived odors.

  8. Zhx2 and Zbtb20: Novel regulators of postnatal alpha-fetoprotein repression and their potential role in gene reactivation during liver cancer

    Science.gov (United States)

    Peterson, Martha L.; Ma, Chunhong; Spear, Brett T.

    2012-01-01

    The mouse alpha-fetoprotein (AFP) gene is abundantly expressed in the fetal liver, normally silent in the adult liver but is frequently reactivated in hepatocellular carcinoma. The basis for AFP expression in the fetal liver has been studied extensively. However, the basis for AFP reactivation during hepatocarcinogenesis is not well understood. Two novel factors that control postnatal AFP repression, Zhx2 and Zbtb20, were recently identified. Here, we review the transcription factors that regulate AFP in the fetal liver, as well as Zhx2 and Zbtb20, and raise the possibility that the loss of these postnatal repressors may be involved in AFP reactivation in liver cancer. PMID:21216289

  9. Radioimmunological analysis of the levels of the carcinoembryonic antigen and alpha-fetoprotein in the blood of patients with lung cancer

    International Nuclear Information System (INIS)

    Barbolin, V.I.; Abramov, V.F.; Tkacheva, G.A.

    1980-01-01

    The level of the carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) are determined in the blood of 41 patients with lung cancer and 70 healthy individuals (donors). It has been found that the CEA content in the blood of patients with lung cancer 2.6-4 times exceeds normal. No variations in the CEA and AFP levels depending upon the age and sex of the donors are revealed. A histological diagnosis of lung cancer has been made in 83.8% of the patients with CEA high concentration in the blood. Determination of the CEA level in the blood may be of diagnostic significance in the clinical picture of lung cancer

  10. Triple composite tumor of stomach: A rare combination of alpha fetoprotein positive hepatoid adenocarcinoma, tubular adenocarcinoma and large cell neuroendocrine carcinoma

    Directory of Open Access Journals (Sweden)

    Lipika Lipi

    2014-01-01

    Full Text Available A 50-year-old male patient presented with pain abdomen of 6 months duration. Computed tomography scan revealed a large mass in the stomach occluding the lumen. Histopathology revealed a triple composite tumor comprising of tubular adenocarcinoma arising on a background of high-grade dysplasia, hepatoid adenocarcinoma (positive for Hep Par-1 and alpha fetoprotein and large cell neuroendocrine carcinoma (positive for synaptophysin and chromogranin with nodal metastasis.Triple composite tumors are distinctly rare with few reports in literature.

  11. Serum albumin and globulin analysis for hepatocellular carcinoma detection avoiding false-negative results from alpha-fetoprotein test negative subjects

    Science.gov (United States)

    Wang, Jing; Feng, Shangyuan; Lin, Juqiang; Zeng, Yongyi; Li, Ling; Huang, Zufang; Li, Buhong; Zeng, Haishan; Chen, Rong

    2013-11-01

    Surface-enhanced Raman spectroscopy (SERS) of serum albumin and globulin were employed to detect hepatocellular carcinoma (HCC). Tentative assignments of SERS bands show specific biomolecular changes associated with cancer development. These changes include a decrease in relative amounts of tryptophan, glutamine, glycine, and serine, indicating excessive consumption of amino acids for protein duplication. Principal component analysis was also introduced to analyze the obtained spectra, resulting in both diagnostic sensitivity and specificity of 100%. More importantly, it reveals that this method can detect HCC patients with alpha-fetoprotein negative test results, suggesting its great potential as a new alternative to detect HCC.

  12. Experimental study on 131I-labelled anti-alpha-fetoprotein antibodies in the diagnosis of rat hepatoma

    International Nuclear Information System (INIS)

    Terashima, Hiromi

    1980-01-01

    The tumor-specificity of 131 I-labelled anti-α-fetoprotein antibodies was evaluated in rats using α-fetoprotein-producing AH66C4 rat hepatoma as a model. 1) Following the 12 hour incubation of 125 I-labelled anti-α-fetoprotein antibodies and tumor cells, microautoradiography revealed marked radioactivity in and around the tumor cells. This suggested that the labelled antibodies accumulated around the cells and were combined with the α-fetoprotein secreted from the cells. 2) The tumor was transplanted subcutaneously into the thighs of rats. There was marked accumulation of 131 I-antibodies in the tumor with cyst formation, but there was none in the tumor without cyst formation. The accumulation was enhanced by the administration of non-labelled antibodies to the rats before the administration of 131 I-antibodies. The α-fetoprotein level was higher in the cyst than in any other organ. 131 I-labelled horse-γ-globulins administered as a control, also accumulated in the tumor with cyst but the degree of accumulation did not exceed that of the 131 I-antibodies. The amount of 131 I-antibodies accumulated increased, while that of 131 I-horse-γ-globulins decreased with time. This indicated that the accumulation of the γ-globulins in the tumor was nonspecific and that it was related to the blood pool. These results strongly suggest that the accumulation of 131 I-antibodies in the tumor with cyst formation was a specific antigen-antibody reaction, and the present procedure reported is applicable in the specific diagnosis of such kinds of α-fetoprotein secreting tumor. (author)

  13. Preparation of radioiodinated insulin and thyroid stimulating hormone using 1,3,4,6-tetrachloro-3. cap alpha. , 6. cap alpha. -diphenylglycouril (iodogen) for radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Pillai, M R.A.; Gupte, J H; Jyotsna, T; Mani, R S

    1987-11-01

    Radioiodinated insulin and thyroid stimulating hormones were prepared using 1,3,4,6-tetrachloro-3..cap alpha.., 6..cap alpha..-diphenylglycouril (iodogen). Conditions of iodination like concentration of iodogen, reaction time, etc. were optimized to get maximum yield. Stability studies of iodogen coated tubes were carried out over a period of time. The dependence of iodination yield on varying amounts of activity and protein concentration were investigated. Iodination yield over a range of pH was also studied. The radiolabelled hormones prepared by this method were used in radioimmunoassay and compared with tracers prepared by the Chloramine-T method. (author) 16 refs.; 5 tabs.

  14. Direct-acting antiviral-based triple therapy on alpha-fetoprotein level in chronic hepatitis C patients.

    Science.gov (United States)

    Takayama, Koji; Furusyo, Norihiro; Ogawa, Eiichi; Ikezaki, Hiroaki; Shimizu, Motohiro; Murata, Masayuki; Hayashi, Jun

    2015-04-21

    To investigate the impact of telaprevir-based triple therapy on the serum alpha-fetoprotein (AFP) level of chronic hepatitis C patients. A total of 210 patients with chronic hepatitis C genotype 1 of high viral load (baseline serum hepatitis C virus RNA > 5.0 log10 IU/mL) were divided into two groups by type of treatment: triple therapy with telaprevir, pegylated-interferon-α (PEG-IFNα), and ribavirin (RBV) for 24 wk (n = 88), or dual therapy with PEG-IFNα and RBV for 48 wk (n = 122). The relationship between virological response and the change in the serum AFP level from baseline to 24 wk after the end of treatment was examined. No significant difference in mean baseline AFP level was found between the triple and dual therapy groups (8.8 ng/mL vs 7.8 ng/mL). Triple therapy produced significant declines in the AFP level in sustained virological response (SVR) and non-SVR patients (7.8 ng/mL at baseline to 3.5 ng/mL at 24 wk after the end of treatment, P < 0.001 and 14.3 ng/mL to 9.5 ng/mL, P = 0.004, respectively). In contrast, dual therapy resulted in a significant decline in AFP level only in SVR patients (4.7 ng/mL to 2.8 ng/mL, P < 0.001), but not in non-SVR patients (10.2 ng/mL to 10.1 ng/mL). Among patients with a high-baseline AFP level (≥ 10 ng/mL), the decline in the AFP level was significantly higher in the triple therapy than in the dual therapy group (15.9 ng/mL vs 1.6 ng/mL, P = 0.037). Regardless of virological response, telaprevir-based triple therapy reduced the serum AFP level.

  15. Radioimmunoassay of alpha-foetoprotein in the eluate of dried blood. A method for antenatal screening of neural tube defects

    International Nuclear Information System (INIS)

    Travert, G.; Herlicoviez, M.; Laroche, D.

    1979-01-01

    A radioimmunoassay for alpha-foetoprotein (AFP) in dried blood spots is reported. The main technical characteristics (reproducibility, sensitivity, recovery of exogenous AFP added and AFP stability in dried blood) are evaluated. They indicate that this method is feasible and well adapted to AFP measurement during pregnancy. AFP determination in maternal serum allows early detection of at least 80% of neural tube defects. The use of dried blood spots as samples for AFP assay makes our method a possible mass screening test for these malformations, which occur with an incidence of 12 for 10,000 [fr

  16. Alcoholic hepatitis with negligible sup(99m)Tc uptake and transient elevation of serum alpha-fetoprotein

    International Nuclear Information System (INIS)

    Hoshino, Hirosuke; Okumura, Makoto; Shimizu, Masanori; Eimoto, Tadaaki

    1981-01-01

    A 35 year old male with typical alcoholic hepatitis presented almost negligible uptake of sup(99m)Tc on the liver scan. Electron microscopic findings disclosing decreased number of Kupffer cells and impaired blood flow in the sinusoids may elucidate extremely diminshed uptake of isotope by the liver. Transient elevation of serum α-fetoprotein up to 3200 ng/ml observed during the active stage may indicate a regeneration process of hepatic necrosis occurred following the acute alcoholic hepatitis. (author)

  17. Development of an alpha-fetoprotein and Golgi protein 73 multiplex detection assay using xMAP technology.

    Science.gov (United States)

    Wu, Yun; Ma, Jie; Wang, Yipeng; Zhang, Yonghong; Hou, Yongjin; Zhang, Chunming; Sun, Huanqin; Sun, Jianping; Wang, Zikang; Li, Ning

    2018-04-06

    Development of a new method to simultaneously detect Alpha-fetoprotein (AFP) and Golgi protein 73 (GP73) from peripheral blood. Anti human AFP and GP73 monoclonal antibodies was used to develop a sandwich immunity reaction using xMAP technology for the simultaneous detection of plasma AFP and GP73. The assay evaluated the sensitivity, cross reactivity, range of detection, precision, recovery and linearity dilution effect. The assay utilized plasma samples and compared its performance with commercially available Enzyme Linked Immunosorbent Assay (ELISA) kits. The assay was successful in detecting AFP and GP73 simultaneously. Validation experiments demonstrated the limit of detection was AFP 0.006 μg/l and GP73 0.482 μg/l. The functional sensitivity was AFP 0.010 μg/l and GP73 0.640 μg/l. The range of detection was AFP 0.01-50 μg/l and GP73 0.64-100 μg/l. No cross reactivity was observed. The intra- and inter-assay variation for AFP was 0.19-3.46% and 3.1-5.8% and for GP73 was 1.5-3.2% and 1.1-7.6% respectively. The recovery for AFP was 96-106% and GP73 was 89-110%. 80 clinical plasma samples from healthy controls, and patients with liver cirrhosis and Hepatocellular Carcinoma (HCC) were evaluated. For healthy controls (n = 25), the AFP was 2.40 (1.55, 3.30) μg/l and GP73 was 42.60 (39.10, 57.40) μg/l. For patients with liver cirrhosis (n = 19), the AFP was 2.60 (1.70, 4.20) μg/l and GP73 was 136.10 (92.10, 261.70) μg/l, and for HCC patients (n = 36), the AFP was 13.65 (3.35, 158.88) μg/l and GP73 was 186.25 (96.73, 262.03) μg/l. The new assay demonstrated a good correlation with commercially available ELISA kits (correlation coefficients (r) were 0.997 (AFP, p < 0.001) and 0.959 (GP73, p < 0.001). The method demonstrated a sensitive, effective and accurate method for the simultaneous detection of AFP and GP73, and could be used clinically for routine detection and monitoring of patients with chronic hepatitis B

  18. Synthesis of CdTe/CdS/ZnS quantum dots and their application in imaging of hepatocellular carcinoma cells and immunoassay for alpha fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Tian Jianniao; Liu Rongjun; Zhao Yanchun; Peng Yan; Hong Xue; Zhao Shulin [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), College of Chemistry and Chemical Engineering of Guangxi Normal University, Guilin 541004 (China); Xu Qing, E-mail: tianjn58@yahoo.com.cn [Pharmacology Department of Guilin Medical College, Guilin 541004 (China)

    2010-07-30

    We report the imaging of hepatocellular carcinoma cells and the immunoassay for alpha fetoprotein (AFP) using CdTe/CdS/ZnS core-shell-shell QDs. Stable and high PLQY (20%-48%) CdTe/CdS/ZnS core-shell-shell QDs were synthesized by a stepwise process. Bioconjugation of the core-shell-shell QDs with streptavidin (SA) was successfully applied in immunofluorescent imaging of the human hepatocellular carcinoma (HCC) cell line HepG2.2.15. Furthermore, the thioglycolic acid (TGA)-capped CdTe/CdS/ZnS core-shell-shell QDs fluorescence lifetime is longer than fluorescein, so it was first engaged to conjugate with antigen for the determination of protein (AFP) by fluorescence polarization immunoassay.

  19. Gallium-67-citrate scanning in primary cancer of the liver: diagnostic value in the presence of cirrhosis and relation to alpha-fetoprotein

    International Nuclear Information System (INIS)

    Levin, J.; Kew, M.C.

    1975-01-01

    Gallium-67-citrate and /sup 99m/Tc-sulfur colloid scans were performed in 38 South African blacks with primary hepatocellular cancer. Selective uptake of the radionuclide by the tumor occurred in 27 patients (70 percent). In 12 out of 18 patients with associated cirrhosis, 67 Ga was concentrated in the defect or defects visible on the /sup 99m/Tc-sulfur colloid scan, but in the remaining 6 cases (33 percent), the 2 scans were identical and the defects may have been attributed wrongly to cirrhosis. Alpha-fetoprotein (AFP) was detected by immunodiffusion in the serum of 26 patients. Twenty-one of these showed selective uptake of 67 Ga by the tumor as compared with 6 out of 12 patients in whom this protein could not be detected. We were therefore unable to confirm a previous finding of a greater uptake of the radionuclide in AFP-negative primary liver cancer. (auth)

  20. Diagnosis and screening of small hepatocellular carcinomas. Comparison of radionuclide imaging, ultrasound, computed tomography, hepatic angiography, and alpha 1-fetoprotein assay

    International Nuclear Information System (INIS)

    Takashima, T.; Matsui, O.; Suzuki, M.; Ida, M.

    1982-01-01

    Twenty-nine small (less than 5 cm) hepatocellular carcinomas in 18 patients were examined by radionuclide imaging (RN), ultrasound (US), computed tomography (CT), hepatic angiography, and serum alpha 1-fetoprotein (AFP) assay. Sensitivity was 39% with RN, 50% with US, 56% with CT, and 94% with angiography, including infusion hepatic angiography (IHA). Lesions larger than 3 cm could be detected by all of these methods; those between 2 and 3 cm were generally shown by US and CT but not RN. IHA was essential for diagnosis of lesions less than 2 cm, which were otherwise difficult or impossible to detect except with angiography. As a screening method, AFP was best, followed by US and CT. The authors recommend using AFP and US to minimize expense and radiation exposure. In questionable cases, IHA should be performed

  1. Maternal serum alpha-fetoprotein levels are normal in Fanconi anemia: Can it be a lack of postnatal inhibition of AFP gene resulting in the elevation?

    Science.gov (United States)

    Aslan, Deniz; Karabacak, Recep Onur; Aslan, Oner Deniz

    2017-04-01

    We investigated the feasibility of using serum alpha-fetoprotein (AFP) levels as a screening test for prenatal diagnosis of Fanconi anemia (FA). Serial measurements in maternal serum were recorded. Parents, both heterozygous for FA, had declined prenatal molecular testing. The infant was born with no somatic abnormalities, and FA was confirmed by postnatal molecular analysis. Maternal serum AFP levels during each trimester of pregnancy were normal indicating that these levels cannot be used as a screening test in prenatal diagnosis. Three-year follow-up after birth showed constantly elevated serum levels in the patient from the start, suggesting a lack of postnatal inhibition on AFP gene. © 2016 Wiley Periodicals, Inc.

  2. Synthesis of CdTe/CdS/ZnS quantum dots and their application in imaging of hepatocellular carcinoma cells and immunoassay for alpha fetoprotein

    International Nuclear Information System (INIS)

    Tian Jianniao; Liu Rongjun; Zhao Yanchun; Peng Yan; Hong Xue; Zhao Shulin; Xu Qing

    2010-01-01

    We report the imaging of hepatocellular carcinoma cells and the immunoassay for alpha fetoprotein (AFP) using CdTe/CdS/ZnS core-shell-shell QDs. Stable and high PLQY (20%-48%) CdTe/CdS/ZnS core-shell-shell QDs were synthesized by a stepwise process. Bioconjugation of the core-shell-shell QDs with streptavidin (SA) was successfully applied in immunofluorescent imaging of the human hepatocellular carcinoma (HCC) cell line HepG2.2.15. Furthermore, the thioglycolic acid (TGA)-capped CdTe/CdS/ZnS core-shell-shell QDs fluorescence lifetime is longer than fluorescein, so it was first engaged to conjugate with antigen for the determination of protein (AFP) by fluorescence polarization immunoassay.

  3. A cyclic peptide derived from alpha-fetoprotein inhibits the proliferative effects of the epidermal growth factor and estradiol in MCF7 cells.

    Science.gov (United States)

    Torres, Cristian; Antileo, Elmer; Epuñán, Maráa José; Pino, Ana María; Valladares, Luis Emilio; Sierralta, Walter Daniel

    2008-06-01

    A cyclic peptide derived from the active domain of alpha-fetoprotein (AFP) significantly inhibited the proliferation of MCF7 cells stimulated with the epidermal growth factor (EGF) or estradiol (E2). The action of these three agents on cell growth was independent of the presence of calf serum in the culture medium. Our results demonstrated that the cyclic peptide interfered markedly with the regulation of MAPK by activated c-erbB2. The cyclic peptide showed no effect on the E2-stimulated release of matrix metalloproteinases 2 and 9 nor on the shedding of heparin-binding EGF into the culture medium. We propose that the AFP-derived cyclic peptide represents a valuable novel antiproliferative agent for treating breast cancer.

  4. Small and asymptomatic hepatocellular carcinoma detected by alpha-fetoprotein screening in black hepatitus B carriers. A report of 2 cases

    Energy Technology Data Exchange (ETDEWEB)

    Dusheiko, G M; Bowyer, S M; Epstein, B; Levien, L J; Beale, P; Kew, M C; Conradie, J C; Simjee, A

    1986-01-01

    Two patients in whom asymptomatic and small hepatocellular carcinomas (HCC) were detected by ..cap alpha..-fetoprotein (AFP) screening are reported. Both patients were hepatitis B surface antigen-positive. In the first patient, the tumour grew slowly and was resected more than 2 years after a significant elevation in serum AFP levels had been first detected and 13 months after hepatic angiography confirmed the presence of a vascular tumour. In the second patient, a small encapsulated HCC was diagnosed by AFP screening and hepatic imaging. Phosphorus 32 was used. The clinical course in these 2 patients illustrates that small, asymptomatic and encapsulated HCCs do occur in Southern African black hepatitis B carriers. Regular screening of patients at risk may be justified.

  5. Proteomic data show an increase in autoantibodies and alpha-fetoprotein and a decrease in apolipoprotein A-II with time in sera from senescence-accelerated mice

    Energy Technology Data Exchange (ETDEWEB)

    Guo, S.J. [Beijing Institute of Pharmacology and Toxicology, Beijing (China); Shanghai Key Laboratory of Hypertension, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai (China); Qi, C.H.; Zhou, W.X.; Zhang, Y.X. [Beijing Institute of Pharmacology and Toxicology, Beijing (China); Zhang, X.M.; Wang, J.; Wang, H.X. [National Center of Biomedical Analysis, Beijing (China)

    2013-04-12

    We evaluated changes in levels by comparing serum proteins in senescence-accelerated mouse-prone 8 (SAMP8) mice at 2, 6, 12, and 15 months of age (SAMP8-2 m, -6 m, -12 m, -15 m) to age-matched SAM-resistant 1 (SAMR1) mice. Mice were sacrificed, and blood was analyzed by 2-dimensional electrophoresis combined with mass spectrometry. Five protein spots were present in all SAMP8 serum samples, but only appeared in SAMR1 samples at 15 months of age except for spot 3, which also showed a slight expression in SAMR1-12 m sera. Two proteins decreased in the sera from SAMP8-2 m, -6 m, and -12 m mice, and divided into 2 spots each in SAMP8-15 m sera. Thus, the total number of altered spots in SAMP8 sera was 7; of these, 4 were identified as Ig kappa chain V region (M-T413), chain A of an activity suppressing Fab fragment to cytochrome P450 aromatase (32C2-A), alpha-fetoprotein, and apolipoprotein A-II. M-T413 is a monoclonal CD4 antibody, which inhibits T cell proliferation. We found that M-T413 RNA level was significantly enhanced in splenocytes from SAMP8-2 m mice. This agreed with serum M-T413 protein alterations and a strikingly lower blood CD4{sup +} T cell count in SAMP8 mice when compared to the age-matched SAMR1 mice, with the latter negatively correlating with serum M-T413 protein volume. Age-related changes in serum proteins favored an increase in autoantibodies and alpha-fetoprotein and a decrease of apolipoprotein A-II, which occurred in SAMP8 mice at 2 months of age and onwards. These proteins may serve as candidate biomarkers for early aging.

  6. Nanosilver-penetrated polyion graphene complex membrane for mediator-free amperometric immunoassay of alpha-fetoprotein using nanosilver-coated silica nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Tang Juan [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Tang Dianping, E-mail: dianping.tang@fzu.edu.c [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Su Biling; Li Qunfang; Qiu Bin [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China); Chen Guonan, E-mail: gnchen@fzu.edu.c [Ministry of Education Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108 (China)

    2011-04-15

    Research highlights: {yields} We fabricate a polyion graphene complex membrane-based immunosensing platform for sensitive electrochemical immunoassay of alpha-fetoprotein. {yields} Nanosilver-coated silica nanocomposites as bionanolabels. {yields} Graphene nanosheets, single-stranded DNA and silver nanoparticles as matrices. {yields} Direct electron transfer without electron mediator. {yields} Analysis of real samples and method comparison. - Abstract: A facile and sensitive mediator-free electrochemical immunosensor for detection of alpha-fetoprotein (AFP) was designed by using nanosilver-coated silica nanoparticles (Ag-SiO{sub 2}) as bionanolabels. To construct such an electrochemical immunosensor, silver ions/single-stranded DNA/graphene nanosheets were initially immobilized on a gold electrode in turn, then silver ions were in situ reduced to silver nanoparticles with the aid of NaBH{sub 4}, and anti-AFP antibodies conjugated to silver nanoparticles were used. In the presence of AFP analyte, the sandwiched immunocomplex was formed on the electrode surface by using horseradish peroxidase-anti-AFP conjugate-labeled Ag-SiO{sub 2} (HRP-anti-AFP-Ag-SiO{sub 2}) as secondary antibodies. Compared with pure silver nanoparticles, Ag-SiO{sub 2} nanocomposites could provide a large room for the immobilization of HRP-anti-AFP, and improve the electrochemical responses of the immunosensor. Meanwhile, the presence of highly conductive graphene nanosheets and silver nanoparticles provided a good pathway for electron transfer. Under optimal conditions, the immunosensor exhibited good electrochemical responses toward AFP ranging from 0.3 to 200 ng/mL with a detection limit (LOD) of 0.05 ng/mL (at 3{sigma}) in pH 6.0 PBS-H{sub 2}O{sub 2} system. Intra- and inter-assay displayed good precisions with coefficient of variation below 9.5%. In addition, the method was evaluated with 23 clinical serum samples, receiving good correlation with results from commercially available

  7. Nanosilver-penetrated polyion graphene complex membrane for mediator-free amperometric immunoassay of alpha-fetoprotein using nanosilver-coated silica nanoparticles

    International Nuclear Information System (INIS)

    Tang Juan; Tang Dianping; Su Biling; Li Qunfang; Qiu Bin; Chen Guonan

    2011-01-01

    Research highlights: → We fabricate a polyion graphene complex membrane-based immunosensing platform for sensitive electrochemical immunoassay of alpha-fetoprotein. → Nanosilver-coated silica nanocomposites as bionanolabels. → Graphene nanosheets, single-stranded DNA and silver nanoparticles as matrices. → Direct electron transfer without electron mediator. → Analysis of real samples and method comparison. - Abstract: A facile and sensitive mediator-free electrochemical immunosensor for detection of alpha-fetoprotein (AFP) was designed by using nanosilver-coated silica nanoparticles (Ag-SiO 2 ) as bionanolabels. To construct such an electrochemical immunosensor, silver ions/single-stranded DNA/graphene nanosheets were initially immobilized on a gold electrode in turn, then silver ions were in situ reduced to silver nanoparticles with the aid of NaBH 4 , and anti-AFP antibodies conjugated to silver nanoparticles were used. In the presence of AFP analyte, the sandwiched immunocomplex was formed on the electrode surface by using horseradish peroxidase-anti-AFP conjugate-labeled Ag-SiO 2 (HRP-anti-AFP-Ag-SiO 2 ) as secondary antibodies. Compared with pure silver nanoparticles, Ag-SiO 2 nanocomposites could provide a large room for the immobilization of HRP-anti-AFP, and improve the electrochemical responses of the immunosensor. Meanwhile, the presence of highly conductive graphene nanosheets and silver nanoparticles provided a good pathway for electron transfer. Under optimal conditions, the immunosensor exhibited good electrochemical responses toward AFP ranging from 0.3 to 200 ng/mL with a detection limit (LOD) of 0.05 ng/mL (at 3σ) in pH 6.0 PBS-H 2 O 2 system. Intra- and inter-assay displayed good precisions with coefficient of variation below 9.5%. In addition, the method was evaluated with 23 clinical serum samples, receiving good correlation with results from commercially available electrochemiluminescent analyzer.

  8. Proteomic data show an increase in autoantibodies and alpha-fetoprotein and a decrease in apolipoprotein A-II with time in sera from senescence-accelerated mice

    International Nuclear Information System (INIS)

    Guo, S.J.; Qi, C.H.; Zhou, W.X.; Zhang, Y.X.; Zhang, X.M.; Wang, J.; Wang, H.X.

    2013-01-01

    We evaluated changes in levels by comparing serum proteins in senescence-accelerated mouse-prone 8 (SAMP8) mice at 2, 6, 12, and 15 months of age (SAMP8-2 m, -6 m, -12 m, -15 m) to age-matched SAM-resistant 1 (SAMR1) mice. Mice were sacrificed, and blood was analyzed by 2-dimensional electrophoresis combined with mass spectrometry. Five protein spots were present in all SAMP8 serum samples, but only appeared in SAMR1 samples at 15 months of age except for spot 3, which also showed a slight expression in SAMR1-12 m sera. Two proteins decreased in the sera from SAMP8-2 m, -6 m, and -12 m mice, and divided into 2 spots each in SAMP8-15 m sera. Thus, the total number of altered spots in SAMP8 sera was 7; of these, 4 were identified as Ig kappa chain V region (M-T413), chain A of an activity suppressing Fab fragment to cytochrome P450 aromatase (32C2-A), alpha-fetoprotein, and apolipoprotein A-II. M-T413 is a monoclonal CD4 antibody, which inhibits T cell proliferation. We found that M-T413 RNA level was significantly enhanced in splenocytes from SAMP8-2 m mice. This agreed with serum M-T413 protein alterations and a strikingly lower blood CD4 + T cell count in SAMP8 mice when compared to the age-matched SAMR1 mice, with the latter negatively correlating with serum M-T413 protein volume. Age-related changes in serum proteins favored an increase in autoantibodies and alpha-fetoprotein and a decrease of apolipoprotein A-II, which occurred in SAMP8 mice at 2 months of age and onwards. These proteins may serve as candidate biomarkers for early aging

  9. Evaluation of a second trimester triple marker screening test for fetal status using alpha-fetoprotein (aFP), human chorionic gonadotropin (hCG) and unconjugated estriol (uE3)

    International Nuclear Information System (INIS)

    Mi, Seong Young; Kim, Jong Ho; Choi, Seung Hun

    1997-01-01

    Our purpose was to assess the utility of maternal serum triple-marker screening test using alpha-fetoprotein (aFP), human Chorionic Gonadotropin (hCG) and unconjugated Estriol (uE 3 ) for fetal chromosomal abnormalities. 1,767 venous blood samples (4ml) between 15 and 20 week's gestation for maternal serum screening from January to October 1996, were tested with Kodak Amerix-M triple marker radioimmunoassay kits. Risk analysis was achieved with interpretive software such as Alpha (LMS, Kodak Clinical Diagnostics). Marker levels are transformed into multiples of median (MOM), which represent an interpretation of (weight regressed) patient marker levels relative to regressed median levels for stated gestation. By multivariate anaysis, the three MOM values are combined to generate a liklihood ratio. Calculation of a patient, risk is the product of liklihood ratio and age-related risk. Risk assessment is weight for maternal age. The median values of aFP, hCG and uE 3 were well correlated with gestational age, respectively (r=0.94, p=0.003; r=-0.97, p=0.029; r=0.99, p 3 weren't (r=-0.17, p=0.22; r=0.36, p=0.09, respectively). The values of aFP, CG and uE 3 between pregnancy younger than 35 years-old (n=87) and older than that (n=1640) were 51.67±27.44, vs 54.65±126.36, 46.45±30.08 vs 51.33±38.50 and 8.01±11.01 vs 6.68±7.23, respectively but all of them failed to show significant differences. A second-trimester risk for trisomy 21 > or = 1:270 was considered screen positive. Patients were screen positive for trisomy 21 if aFP or 2.1 MOM and E 3 2.5 MOM. The initial screen-positive rate for both Down' syndrome and neural tube defect were 1.46% (26/1767); 0.73% (13/1767) with each other. Among screen positive 26 patients, three and nine were normal karyotype and normal phenotype, respectively and five patients had still births. Reamining 9 patients underwent terminations. In conclusion, compared with the other group's data even in Koreans (Whang et al, and Song et al

  10. A synthetic peptide derived from alpha-fetoprotein inhibits the estradiol-induced proliferation of mammary tumor cells in culture through the modulation of p21.

    Science.gov (United States)

    Sierralta, Walter D; Epuñan, María J; Reyes, José M; Valladares, Luis E; Pino, Ana M

    2008-01-01

    A stable cyclized 9-mer peptide (cP) containing the active site of alpha-alpha fetoprotein (alphaFP) has been shown to be effective for prevention of estrogen-stimulated tumor cell proliferation in culture or of xenographt growth in immunodeficient mice. cP does not block 17beta-estradiol (E2) binding to its receptors, but rather appears to interfere with intracellular processing of the signal that supports growth. To obtain insight on that mechanism we studied the effect of cP on the proliferation of MCF-7 cells in culture. Proliferation in the presence of 2 microM E2 is decreased up to 40% upon addition of 2 microg ml(-1) cP to the medium; the presence of cP did not increase cell death, cP reduced also the proliferation of estrogen-dependent ZR75-1 cells but had no effect on autonomous MDA-MB-231 cells, cP did not modify the number of binding sites for labeled E2 or affected cell death. We detected increased nuclear p21Cip1 immunoreactivity after cP treatment. Our results suggest that cP acts via p21Cip1 to slow the process of MCF-7 cells through the cycle.

  11. Prognostic value of determination of carcinoembryonic antigen and α-fetoprotein level in blood plasma in patients with cancer stomach

    International Nuclear Information System (INIS)

    Smyslova, V.N.; Vygonnyj, I.I.

    1986-01-01

    60 donors and 129 patients with cancer stomach were examined. Tumor antigens were determined in blood plasma by the method of radioimmunoassay. The upper boundary of the norm of alpha-fetoprotein (AFP) and carcino-embryonic antigen (CEA) is 12 ng/ml. Increased concentration of antigens studied is detected in most patients. It is established that the level of antigens increases depending on generalization of the process, cancer stage, tumor propagation in the stomach wall, patient's age. High volumes of AFP and CEA after operation give evidence about non-radicality of operation and bad prognosis

  12. Non-specific interference of certain components of tissue culture media with the radioimmunoassay of rat alpha-foetoprotein

    International Nuclear Information System (INIS)

    Dambuyant, C.; Sizaret, Ph.

    1975-01-01

    Interferences of 'Williams' tissue culture medium used for cultivating rat hepatocytes upon rat alpha-foetoprotein (AFP) radioimmunoassay have been investigated. They are not due to foetal calf serum proteins which are added as growth factor and can be abolished by dialysis which appears to be necessary for the distinction between AFP non-producer and low-producer cell lines. Of the three major groups of non-mineral components examined, amino acid solution played a major role. When individual amino acids were examined using the double antibody technique, arginine was found to interfere predominantly; its dose-response curve was parallel to that of rat AFP which confirmed that an immunological identity between two substances cannot be established on the basis of parallelism as the only criterion

  13. A sensitive label–free amperometric immunosensor for alpha-fetoprotein based on gold nanorods with different aspect ratio

    Science.gov (United States)

    Zhou, Chunyang; Liu, Dali; Xu, Lin; Li, Qingling; Song, Jian; Xu, Sai; Xing, Ruiqing; Song, Hongwei

    2015-01-01

    A simple and accurate label–free amperometric immunosensor for α–fetoprotein (AFP) detection is developed based on gold nanorods (GNRs) with different aspect ratio and compared with gold particles (GNPs). The positively charged GNRs and GNPs due to the surface immobilized cetyltrimethyl ammonium bromide (CTAB) can adsorb the negatively charged AFP antibody (Ab) directly. The presence of the GNRs not only enhanced the immobilized amount of biomolecules, but also improved the electrochemical properties of the immunosensor. With the aid of GNRs, the electrochemical signal was greatly enhanced in comparison with GNPs. Under optimal conditions, the proposed immunosensor could detect AFP in a linear range from 0.1 to 200 ng/mL with a detection limit of 0.04 ng/mL (signal–to–noise ratio = 3), and it also possessed good reproducibility and storage stability. Moreover, the detection of AFP in five human serum samples also showed satisfactory accuracy. The proposed methodology was potentially attractive for clinical immunoassay. PMID:25909588

  14. A novel signal-on photoelectrochemical immunosensor for detection of alpha-fetoprotein by in situ releasing electron donor.

    Science.gov (United States)

    Chen, Jiexia; Zhao, Guang-Chao

    2017-12-15

    A signal-on photoelectrochemical (PEC) immunosensor was constructed for detecting tumor marker in this work. α-fetoprotein (AFP) was chosen as a model analyte to investigate the prepared procedure and the analytical performance of the exploited sensor. In order to construct the sensor, CdSe QDs were used as photoactive material, biotin conjugated AFP antibody (Bio-anti-AFP) as detecting probe, streptavidin (SA) as signal capturing unit, biotin functionalized apoferritin encapsulated ascorbic acid (Bio-APOAA) as amplification unit, which were assembled onto the electrodes. The sensing strategy was based on in situ enzymatic hydrolysis of Bio-APOAA to release ascorbic acid (AA) as sacrificial electron donor to produce photocurrent. The photocurrent from the immunosensor was monitored as a result of AFP concentrations. The constructed sensing platform displayed high selectivity and good sensitivity for detecting AFP. Under optimal conditions, a wide linear range from 0.001 to 1000ng/mL and a low detection limit of 0.31pg/mL were obtained. The developed immunosensor is expected to be used to determine AFP and other tumor markers in human plasma in clinical laboratories either for pre-cancer screening or cancer monitoring. Moreover, this sensing platform further has the potential to use for the detection of trypsin activity and the corresponding inhibitor-screening. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Possible application of tumor marker radioimmunoassay in diagnosis of testicular and prostatic carcinomas

    Energy Technology Data Exchange (ETDEWEB)

    Kausitz, J

    1988-10-01

    Determinations of alpha fetoprotein and chorionic gonadotropin levels by radioimmunoassay in 340 patients with germ cell tumors of the testes have confirmed that tumor markers are suitable prognostic parameters, facilitate assessment of the clinical stage, are sensitive parameters of the efficacy of chemotherapy and enable early detection of relapses. In a group of 71 patients with prostate cancer, systematic determination of prostate specific antigen levels proved to be a reliable method of monitoring and a sensitive method of detecting remote metastases. (author). 5 figs., 13 tabs., 23 refs.

  16. Radioimmunoassay of alpha-foeto protein in the amniotic fluid in normal and pathological pregnancies

    International Nuclear Information System (INIS)

    Degueldre, M.; Golstein, J.; Rodesch, F.; L'Hermite, M.

    1975-01-01

    A radioimmunoassay of AFP was developed and the normal amniotic concentrations of this protein were measured as a function of the gestation age. The specific reagents used included a rabbit anti-AFP serum and a highly purified preparation of AFP labelled with 125 I, which also served as a standard. Amniotic fluid was drawn off by amniocentesis between the 11th and 40th week following the last menstrual period. Samples of amniotic fluid were also obtained in 2 cases of foetal death in utero and 3 cases of anencephaly. The normal amniotic AFP concentration limits were particularly well established between the 14th and 18th weeks, an ideal period to perform an amniocentesis both from a technical viewpoint and with regard to the bulk of information obtainable for the antenatal diagnosis of many congenital diseases. The concentrations observed in pathological cases were distinctly higher than the upper 95% confidence limit of normal values for the gestation age considered [fr

  17. Magnetic immunoassay coupled with inductively coupled plasma mass spectrometry for simultaneous quantification of alpha-fetoprotein and carcinoembryonic antigen in human serum

    Science.gov (United States)

    Zhang, Xing; Chen, Beibei; He, Man; Zhang, Yiwen; Xiao, Guangyang; Hu, Bin

    2015-04-01

    The absolute quantification of glycoproteins in complex biological samples is a challenge and of great significance. Herein, 4-mercaptophenylboronic acid functionalized magnetic beads were prepared to selectively capture glycoproteins, while antibody conjugated gold and silver nanoparticles were synthesized as element tags to label two different glycoproteins. Based on that, a new approach of magnetic immunoassay-inductively coupled plasma mass spectrometry (ICP-MS) was established for simultaneous quantitative analysis of glycoproteins. Taking biomarkers of alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) as two model glycoproteins, experimental parameters involved in the immunoassay procedure were carefully optimized and analytical performance of the proposed method was evaluated. The limits of detection (LODs) for AFP and CEA were 0.086 μg L- 1 and 0.054 μg L- 1 with the relative standard deviations (RSDs, n = 7, c = 5 μg L- 1) of 6.5% and 6.2% for AFP and CEA, respectively. Linear range for both AFP and CEA was 0.2-50 μg L- 1. To validate the applicability of the proposed method, human serum samples were analyzed, and the obtained results were in good agreement with that obtained by the clinical chemiluminescence immunoassay. The developed method exhibited good selectivity and sensitivity for the simultaneous determination of AFP and CEA, and extended the applicability of metal nanoparticle tags based on ICP-MS methodology in multiple glycoprotein quantifications.

  18. Facile fabrication of a silicon nanowire sensor by two size reduction steps for detection of alpha-fetoprotein biomarker of liver cancer

    International Nuclear Information System (INIS)

    Pham, Van Binh; Pham, Xuan ThanhTung; Phan, Thanh Nhat Khoa; Le, Thi Thanh Tuyen; Dang, Mau Chien

    2015-01-01

    We present a facile technique that only uses conventional micro-techniques and two size-reduction steps to fabricate wafer-scale silicon nanowire (SiNW) with widths of 200 nm. Initially, conventional lithography was used to pattern SiNW with 2 μm width. Then the nanowire width was decreased to 200 nm by two size-reduction steps with isotropic wet etching. The fabricated SiNW was further investigated when used with nanowire field-effect sensors. The electrical characteristics of the fabricated SiNW devices were characterized and pH sensitivity was investigated. Then a simple and effective surface modification process was carried out to modify SiNW for subsequent binding of a desired receptor. The complete SiNW-based biosensor was then used to detect alpha-fetoprotein (AFP), one of the medically approved biomarkers for liver cancer diagnosis. Electrical measurements showed that the developed SiNW biosensor could detect AFP with concentrations of about 100 ng mL"−"1. This concentration is lower than the necessary AFP concentration for liver cancer diagnosis. (paper)

  19. A novel electrochemical immunosensor using β-cyclodextrins functionalized silver supported adamantine-modified glucose oxidase as labels for ultrasensitive detection of alpha-fetoprotein.

    Science.gov (United States)

    Gao, Jian; Ma, Hongmin; Lv, Xiaohui; Yan, Tao; Li, Na; Cao, Wei; Wei, Qin

    2015-09-17

    In this work, a novel sandwich-type electrochemical immunosensor based on host-guest interaction was fabricated for the detection of alpha-fetoprotein (AFP). Due to the large specific surface area of multiwalled carbon nanotubes and the unique supramolecular recognition ability of β-cyclodextrins, ferrocenecarboxylic acid (Fc) was incorporated into this sensor platform by host-guest interaction to generate an electrochemical signal. And β-cyclodextrins functionalized silver supported adamantine-modified glucose oxidase (GOD-CD-Ag), was used as a label to improve the analytical performance of the immunosensor by the dual amplification strategy. The obtained GOD-CD-Ag conjugates could convert glucose into gluconic acid with the formation of hydrogen peroxide (H2O2). And then silver nanoparticles could in situ catalyze the reduction of the generated H2O2, dramatically improving the oxidation reaction of Fc. The developed immunosensor shows a wide linear calibration range from 0.001 to 5.0 ng/mL with a low detection limit (0.2 pg/mL) for the detection of AFP. The method, with ideal reproducibility and selectivity, has a wide application prospect in clinical research. Copyright © 2015 Elsevier B.V. All rights reserved.

  20. Serum reference interval of ARCHITECT alpha-fetoprotein in healthy Chinese Han adults: Sub-analysis of a prospective multi-center study.

    Science.gov (United States)

    Yan, Cunling; Yang, Jia; Wei, Lianhua; Hu, Jian; Song, Jiaqi; Wang, Xiaoqin; Han, Ruilin; Huang, Ying; Zhang, Wei; Soh, Andrew; Beshiri, Agim; Fan, Zhuping; Zheng, Yijie; Chen, Wei

    2018-02-01

    Alpha-fetoprotein (AFP) has been widely used in clinical practice for decades. However, large-scale survey of serum reference interval for ARCHITECT AFP is still absent in Chinese population. This study aimed to measure serum AFP levels in healthy Chinese Han subjects, which is a sub-analysis of an ongoing prospective, cross-sectional, multi-center study (ClinicalTrials.gov Identifier: NCT03047603). This analysis included a total of 530 participants (41.43±12.14years of age on average, 48.49% males), enrolled from 5 regional centers. Serum AFP level was measured by ARCHITECT immunoassay. Statistical analysis was performed using SAS 9.4 and R software. AFP distribution did not show significant correlation with age or sex. The overall median and interquartile range of AFP was 2.87 (2.09, 3.83) ng/mL. AFP level did not show a trend of increasing with age. The new reference interval was 2.0-7.07ng/mL (LOQ- 97.5th percentiles). The reference interval for ARCHITECT AFP is updated with the data of adequate number of healthy Han adults. This new reference interval is more practical and applicable in Chinese adults. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  1. Amperometric immunosensor for {alpha}-fetoprotein antigen in human serum based on co-immobilizing dinuclear copper complex and gold nanoparticle doped chitosan film

    Energy Technology Data Exchange (ETDEWEB)

    Gan Ning; Meng Linghua; Wang Feng [State Key Laboratory Base of Novel Functional Materials and Preparation science, Faculty of Material Science and Chemical Engineering of Ninbo University, Ningbo, 315211 (China)], E-mail: ganning@nbu.edu.cn

    2009-09-01

    A sensitive amperometric immunosensor for {alpha}-fetoprotein (AFP), a tumor marker for the diagnosis of hepatocellular carcinoma (HCC), was constructed, The immunosensor is prepared by co-immobilizing [Cu{sub 2}(phen){sub 2}Cl{sub 2}] ({mu}-Cl){sub 2} (CuL), nano-Au/Chitosan(Chit) composite, horseradish peroxidase (HRP) and AFP antibody(anti-AFP) on a glassy carbon electrode (GCE). Firstly, CuL was irreversibly absorb on GCE electrode through {pi}-{pi} stacking interaction; then nano-Au/Chit composite was immobilized onto the electrode because of its excellent membrane-forming ability, finally HRP and anti-AFP was adsorbed onto the surface of the gold nanoparticles to construct GCE | CuL/nanoAu-chit/HRP/anti-AFP immunosensor. The preparation procedure of the electrode was characterized by electrochemical and spectroscopy method. The results showed that this immunosensor exhibited an excellent electrocatalytic response to the reduction of hydrogen peroxide (H{sub 2}O{sub 2}) without the aid of an electron mediator, offers a high-sensitivity (1710 nA {center_dot} ng{sup -1} {center_dot} ml{sup -1}) for the detection of AFP and has good correlation for detection of AFP in the range of 0.2 to 120.0 ng/ml with a detection limit of 0.05 ng/ml. The biosensor showed high selectivity as well as good stability and reproductivity.

  2. Fullerol-fluorescein isothiocyanate-concanavalin agglutinin phosphorescent sensor for the detection of alpha-fetoprotein and forecast of human diseases

    Science.gov (United States)

    Liu, Jia-ming; Lin, Li-ping; Jiang, Shu-Lian; Cui, Ma Lin; Jiao, Li; Zhang, Xiao Yang; Zhang, Li-hong; Zheng, Zhi Yong; Lin, Xuan; Lin, Shao-qin

    2013-11-01

    Based on the reaction of the active -OH group in fullerol (F) with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form an F-FITC and the enhanced effect of N, N-dimethylaniline (DMA) on phosphorescence signal of F-FITC, a new phosphorescent labeling reagent (DMA-F-FITC) was developed. What's more, a phosphorescent sensor for the determination of alpha-fetoprotein variant (AFP-V) has been designed via the coupling technique of the high sensitivity for affinity adsorption-solid substrate-room temperature phosphorimetry (AA-SS-RTP) with the strong specificity reaction between DMA-F-FITC-Con A and AFP-V. The DMA-F-FITC increased the number of luminescent molecules in the biological target which improved the sensitivity of phosphorescent sensor. The proposed sensor was responsive, simple, selective and sensitive, and it has been applied to the determination of trace AFP-V in human serum and the forecast of human diseases using phosphorescence emission wavelength of F or FITC, with the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Meanwhile, the mechanisms for the labeling reaction and the sensing detection of AFP-V were discussed.

  3. Mechanism of Cancer Growth Suppression of Alpha-Fetoprotein Derived Growth Inhibitory Peptides (GIP): Comparison of GIP-34 versus GIP-8 (AFPep). Updates and Prospects

    Energy Technology Data Exchange (ETDEWEB)

    Mizejewski, Gerald J. [Division of Translational Medicine, Wadsworth Center, New York State Department of Health, Empire State Plaza, Albany, NY 12201 (United States)

    2011-06-20

    The Alpha-fetoprotein (AFP) derived Growth Inhibitory Peptide (GIP) is a 34-amino acid segment of the full-length human AFP molecule that inhibits tumor growth and metastasis. The GIP-34 and its carboxy-terminal 8-mer segment, termed GIP-8, were found to be effective as anti-cancer therapeutic peptides against nine different human cancer types. Following the uptake of GIP-34 and GIP-8 into the cell cytoplasm, each follows slightly different signal transduction cascades en route to inhibitory pathways of tumor cell growth and proliferation. The parallel mechanisms of action of GIP-34 versus GIP-8 are demonstrated to involve interference of signaling transduction cascades that ultimately result in: (1) cell cycle S-phase/G2-phase arrest; (2) prevention of cyclin inhibitor degradation; (3) protection of p53 from inactivation by phosphorylation; and (4) blockage of K{sup +} ion channels opened by estradiol and epidermal growth factor (EGF). The overall mechanisms of action of both peptides are discussed in light of their differing modes of cell attachment and uptake fortified by RNA microarray analysis and electrophysiologic measurements of cell membrane conductance and resistance. As a chemotherapeutic adjunct, the GIPs could potentially aid in alleviating the negative side effects of: (1) tamoxifen resistance, uterine hyperplasia/cancer, and blood clotting; (2) Herceptin antibody resistance and cardiac (arrest) arrhythmias; and (3) doxorubicin's bystander cell toxicity.

  4. Prussian blue-gold nanoparticles-ionic liquid functionalized reduced graphene oxide nanocomposite as label for ultrasensitive electrochemical immunoassay of alpha-fetoprotein.

    Science.gov (United States)

    Gao, Qi; Liu, Na; Ma, Zhanfang

    2014-06-04

    In this work, poly(diallyldimethylammonium chloride) (PDDA) protected Prussian blue/gold nanoparticles/ionic liquid functionalized reduced graphene oxide (IL-rGO-Au-PDDA-PB) nanocomposite was fabricated. The resulting nanocomposite exhibited high biocompatibility, conductivity and catalytic activity. To assess the performance of the nanocomposite, a sensitive sandwich-type immunosensor was constructed for detecting alpha-fetoprotein (AFP). Greatly enhanced sensitivity for this immunosensor was based on triple signal amplification strategies. Firstly, IL-rGO modified electrode was used as biosensor platform to capture a large amount of antibody due to its increased surface area, thus amplifying the detection response. Secondly, a large number of Au-PDDA-PB was conjugated on the surface of IL-rGO, which meant the enrichment of the signal and the more immobilization of label antibody. Finally, the catalytic reaction between H2O2 and the IL-rGO-Au-PDDA-PB nanocomposite further enhanced the signal response. The signals increased linearly with AFP concentrations in the range of 0.01-100 ng mL(-1). The detection limit for AFP was 4.6 pg mL(-1). The immunosensor showed high sensitivity, excellent selectivity and good stability. Moreover, the immunosensor was applied to the analysis of AFP in serum sample with satisfactory result. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Facile fabrication of a silicon nanowire sensor by two size reduction steps for detection of alpha-fetoprotein biomarker of liver cancer

    Science.gov (United States)

    Binh Pham, Van; ThanhTung Pham, Xuan; Nhat Khoa Phan, Thanh; Thanh Tuyen Le, Thi; Chien Dang, Mau

    2015-12-01

    We present a facile technique that only uses conventional micro-techniques and two size-reduction steps to fabricate wafer-scale silicon nanowire (SiNW) with widths of 200 nm. Initially, conventional lithography was used to pattern SiNW with 2 μm width. Then the nanowire width was decreased to 200 nm by two size-reduction steps with isotropic wet etching. The fabricated SiNW was further investigated when used with nanowire field-effect sensors. The electrical characteristics of the fabricated SiNW devices were characterized and pH sensitivity was investigated. Then a simple and effective surface modification process was carried out to modify SiNW for subsequent binding of a desired receptor. The complete SiNW-based biosensor was then used to detect alpha-fetoprotein (AFP), one of the medically approved biomarkers for liver cancer diagnosis. Electrical measurements showed that the developed SiNW biosensor could detect AFP with concentrations of about 100 ng mL-1. This concentration is lower than the necessary AFP concentration for liver cancer diagnosis.

  6. Shrink-induced graphene sensor for alpha-fetoprotein detection with low-cost self-assembly and label-free assay

    Science.gov (United States)

    Sando, Shota; Zhang, Bo; Cui, Tianhong

    2017-12-01

    Combination of shrink induced nano-composites technique and layer-by-layer (LbL) self-assembled graphene challenges controlling surface morphology. Adjusting shrink temperature achieves tunability on graphene surface morphology on shape memory polymers, and it promises to be an alternative in fields of high-surface-area conductors and molecular detection. In this study, self-assembled graphene on a shrink polymer substrate exhibits nanowrinkles after heating. Induced nanowrinkles on graphene with different shrink temperature shows distinct surface roughness and wettability. As a result, it becomes more hydrophilic with higher shrink temperatures. The tunable wettability promises to be utilized in, for example, microfluidic devices. The graphene on shrink polymer also exhibits capability of being used in sensing applications for pH and alpha-fetoprotein (AFP) detection with advantages of label free and low cost, due to self-assembly technique, easy functionalization, and antigen-antibody reaction on graphene surface. The detection limit of AFP detection is down to 1 pg/mL, and therefore the sensor also has a significant potential for biosensing as it relies on low-cost self-assembly and label-free assay.

  7. Forkhead box protein A2, a pioneer factor for hepatogenesis, is involved in the expression of hepatic phenotype of alpha-fetoprotein-producing adenocarcinoma.

    Science.gov (United States)

    Yamamura, Nobuhisa; Fugo, Kazunori; Kishimoto, Takashi

    2017-09-01

    Alpha-fetoprotein (AFP)-producing adenocarcinoma is a high-malignant variant of adenocarcinoma with a hepatic or fetal-intestinal phenotype. The number of cases of AFP-producing adenocarcinomas is increasing, but the molecular mechanism underlying the aberrant production of AFP is unclear. Here we sought to assess the role of Forkhead box A (FoxA)2, which is a pioneer transcription factor in the differentiation of hepatoblasts. FoxA2 expression was investigated in five cases of AFP-producing gastric adenocarcinomas by immunohistochemistry, and all cases showed FoxA2 expression. Chromatin immunoprecipitation revealed the DNA binding of FoxA2 on the regulatory element of AFP gene in AFP-producing adenocarcinoma cells. The inhibition of FoxA2 expression with siRNA reduced the mRNA expression of liver-specific proteins, including AFP, albumin, and transferrin. The inhibition of FoxA2 also reduced the expressions of liver-enriched nuclear factors, i.e., hepatocyte nuclear factor (HNF) 4α and HNF6, although the expressions of HNF1α and HNF1β were not affected. The same effect as FoxA2 knockdown in AFP producing adenocarcinoma cells was also observed in hepatocellular carcinoma cells. Our results suggest that FoxA2 plays a key role in the expression of hepatic phenotype of AFP-producing adenocarcinomas. Copyright © 2017 Elsevier GmbH. All rights reserved.

  8. Sandwich immunoassay for alpha-fetoprotein in human sera using gold nanoparticle and magnetic bead labels along with resonance Rayleigh scattering readout

    International Nuclear Information System (INIS)

    Lu, Yao; Huang, Xiangyi; Ren, Jicun

    2013-01-01

    We describe a sensitive sandwich immunoassay for alpha-fetoprotein (AFP). It is making use of gold nanoparticles (GNPs) and magnetic beads (MBs) as labels, and of resonance Rayleigh scattering for detection. Two antibodies were labeled with GNPs and MBs, respectively, and MB-antigen-GNP complexes were formed in the presence of antigens. The MB labels also serve as solid phase carriers that can be used to magnetically separate the immuno complex. The GNP labels are used as optical probes, and Rayleigh scattering was used to determine the concentration of free GNPs-antibody after separation of the MB-antigen-GNP complexes. The concentration of AFP is related to the intensity of light scattered by free GNPs in the 13.6 pM to 436 pM concentration range, and the limit of detection is 13.6 pM. The method was applied to the determination of AFP in sera of cancer patients, and the results agree well with those obtained by conventional ELISA. (author)

  9. Improvement of insertion loss and quality factor of flexural plate-wave-based alpha-fetoprotein biosensor using groove-type reflective grating structures

    Science.gov (United States)

    Lin, Chang-Yu; Huang, I.-Yu; Lan, Je-Wei

    2013-01-01

    Conventional flexural plate-wave (FPW) transducers have limited applications in biomedical sensing due to their disadvantages such as high insertion loss and low quality factor. To overcome these shortcomings, we propose a FPW transducer on a low phase velocity insulator membrane (5-μm-thick SiO2) with a novel groove-type reflective grating structure design. Additionally, a cystamine self-assembly monolayer and a glutaraldehyde cross-linking layer are implemented on the backside of the FPW device to immobilize alpha-fetoprotein (AFP) antibody. A FPW-based AFP biosensor with low detection limit (5 ng/mL) can be achieved and used to measure the extreme low concentration of AFP antigen in human serum for early detection of hepatocellular carcinoma. The proposed FPW-based AFP biosensor also demonstrates a very high quality factor (206), low insertion loss (-40.854 dB), low operating frequency (6.388 MHz), and high sensing linearity (90.7%).

  10. Upconversion nanoparticle as elemental tag for the determination of alpha-fetoprotein in human serum by inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Liu, Zhengru; Yang, Bin; Chen, Beibei; He, Man; Hu, Bin

    2016-12-19

    Upconversion nanoparticles (UCNPs) have received increasing attention due to their unique optical properties. Recognizing that UCNPs are lanthanide-doped nanoparticles, we incorporated UCNPs into an immunoassay with inductively coupled plasma mass spectrometry (ICP-MS) detection for the determination of specific proteins, e.g., alpha-fetoprotein (AFP). The sensitivity of the assay was enhanced because of the ICP-MS detection of UCNPs that contained large numbers of lanthanide elemental tags. Conjugates of UCNPs and antibodies were prepared and the morphology of the conjugates was characterized by transmission electron microscopy. After a sandwich immunoreaction, the AFP was determined by the ICP-MS analysis of UCNPs. Under the optimized conditions, a limit of detection (3σ) of 0.31 ng mL -1 based on 89 Y signal and 0.22 ng mL -1 based on 174 Yb signal was obtained for AFP, with a dynamic range of 0.5-35 ng mL -1 and a relative standard deviation of 4.8% (c = 5 ng mL -1 , n = 9). The developed method was applied to the determination of AFP in human serum and the recovery for the spiked sample was in the range of 98.6-123%. The proposed method is simple, rapid, selective and sensitive, and has a good tolerance for the complex biological matrix, indicating great potential for the application of UCNP in biological research as an elemental tag.

  11. Nanogram per milliliter-level immunologic detection of alpha-fetoprotein with integrated rotating-resonance microcantilevers for early-stage diagnosis of heptocellular carcinoma.

    Science.gov (United States)

    Liu, Yongjing; Li, Xinxin; Zhang, Zhixiang; Zuo, Guomin; Cheng, Zhenxing; Yu, Haitao

    2009-02-01

    Nanogram per milliliter-level ultra-low concentration detection of alpha-fetoprotein (AFP), which is an important marker for heptocellular carcinoma, is in favor of early-stage prognosis and disease diagnosis. On-the-spot rapid detection of such antigens as AFP highly requires innovative micro/nano techniques. To meet this requirement, an advanced resonant microcantilever is developed and used for screening the tumor marker at nanogram per milliliter level. The sensing principle of the resonant microcantilever is measuring frequency-shift versus specific-adsorbed mass. With both electromagnetic resonance-exciting and piezoresistive readout elements on-chip integrated, the microcantilever sensor is operated in a rotating resonance mode to improve sensitivity and resolution to specific mass adsorption. Prior to detection of AFP with previously immobilized anti-AFP antibody, the antigen-antibody specific-binding is confirmed with an enzyme linked immunosorbent assay experiment. By implementing the specific reaction in liquid and reading out the sensor signal in lab air environment, the micromechanical sensor has achieved the sensitive scale between 2 and 20 ng/ml. To effectively depress cross-talk signal and improve resolution, the insensitive regions of the cantilever surface are pre-modified with 2-[methoxy (polyethyleneoxy) propyl] trimethoxysilane for nonspecific bio-adsorption minimization. Finally, a better AFP detecting limit than 2 ng/mL is experimentally achieved. The label-free resonant microcantilever sensor is promising in low-cost or even disposable early-stage prognosis and diagnosis of tumors.

  12. Elevated second-trimester maternal serum β-human chorionic gonadotropin and amniotic fluid alpha-fetoprotein as indicators of adverse obstetric outcomes in fetal Turner syndrome.

    Science.gov (United States)

    Alvarez-Nava, Francisco; Soto, Marisol; Lanes, Roberto; Pons, Hector; Morales-Machin, Alisandra; Bracho, Ana

    2015-12-01

    The objective of this study was to determine the ability of biochemical analytes to identify adverse outcomes in pregnancies with Turner syndrome. Maternal serum and amniotic fluid (AF) marker concentrations were measured in 73 singleton pregnancies with Turner syndrome (10-22 weeks of gestation). Fetal Turner syndrome was definitively established by cytogenetic analysis. Two subgroups, fetuses with hydrops fetalis versus fetuses with cystic hygroma, were compared. Receiver operating characteristic curves and relative risk were established for a cut-off multiples of the median ≥3.5 for β-subunit of human chorionic gonadotropin (hCG) or AF alpha-fetoprotein (AFP). Forty-nine (67%) of 73 pregnant women had an abnormal maternal serum. While levels of pregnancy-associated plasma protein-A and free β-subunit (fβ)-hCG were not different to those of the control group, AFP, unconjugated estriol and β-hCG concentrations were significantly different in the study group (P Turner syndrome pregnancies with the highest risk of fetal death. © 2015 Japan Society of Obstetrics and Gynecology.

  13. Gold nanoparticle labeling with tyramide signal amplification for highly sensitive detection of alpha fetoprotein in human serum by ICP-MS.

    Science.gov (United States)

    Li, Xiaoting; Chen, Beibei; He, Man; Xiao, Guangyang; Hu, Bin

    2018-01-01

    In this work, we developed an immunoassay based on tyramide signal amplification (TSA) and gold nanoparticles (Au NPs) labeling for highly sensitive detection of alpha fetoprotein (AFP) by inductively coupled plasma mass spectrometry (ICP-MS). AFP was captured by anti-AFP1 coating on the 96-well plate and labeled by anti-AFP2-horseradish peroxidase (HRP), in which the HRP can catalyze the deposition of biotinylated tyramine on the nearby protein. Then the streptavidin (SA)-Au NPs was labeled on the deposited biotinylated tyramine as the intensive signal probe for ICP-MS measurement. Under the optimal experimental conditions, the limit of detection of the developed method for AFP was 1.85pg/mL and the linear range was 0.005-2ng/mL. The relative standard deviation for seven replicate detections of 0.01ng/mL AFP was 5.2%. The proposed method was successfully applied to the detection of AFP in human serum with good recoveries. This strategy is highly sensitive and easy to operate, and can be extended to the sensitive detection of other biomolecules in human serum. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. An Ultrasensitive Electrochemical Immunosensor for Alpha-Fetoprotein Using an Envision Complex-Antibody Copolymer as a Sensitive Label

    Science.gov (United States)

    Xiong, Ping; Gan, Ning; Cao, Yuting; Hu, Futao; Li, Tianhua; Zheng, Lei

    2012-01-01

    A novel strategy is presented for sensitive detection of alfa-fetoprotein (AFP), using a horseradish peroxidase (HRP)-functionalized Envision antibody complex (EVC) as the label. The Envision-AFP signal antibody copolymer (EVC-AFP Ab2) was composed of a dextran amine skeleton anchoring more than 100 molecules of HRP and 15 molecules of secondary antibody, and acted as a signal tag in the immunosensor. The sensor was constructed using the following steps: First, gold electrode (GE) was modified with nano-gold (AuNPs) by electro-deposition in HAuCl4 solution. The high affinity of the AuNPs surface facilitates direct formation of a self-assembled thiolated protein G layer. Next, the coated GE was incubated in a solution of AFP capture antibody (AFP Ab1); these antibodies attach to the thiolated protein G layer through their non-antigenic regions, leaving the antigen binding sites for binding of target analyte. Following a sandwich immunoreaction, an EVC-AFP Ab2-AFP-AFP Ab1 immunocomplex was formed on the electrode surface, allowing large amounts of HRP on the complex to produce an amplified electrocatalytic current of hydroquinone (HQ) in the presence of hydrogen peroxide (H2O2). Highly amplified detection was achieved, with a detection limit of 2 pg/mL and a linear range of 0.005–0.2 ng/mL for AFP in 10 μL undiluted serum; this is near or below the normal levels of most cancer biomarker proteins in human serum. Measurements of AFP in the serum of cancer patients correlated strongly with standard enzyme-linked immunosorbent assays. These easily fabricated EVC-modified immunosensors show excellent promise for future fabrication of bioelectronic arrays. By varying the target biomolecules, this technique may be easily extended for use with other immunoassays, and thus represents a versatile design route.

  15. Evaluation of a second trimester triple marker screening test for fetal status using alpha-fetoprotein (aFP), human chorionic gonadotropin (hCG) and unconjugated estriol (uE{sub 3})

    Energy Technology Data Exchange (ETDEWEB)

    Mi, Seong Young; Kim, Jong Ho; Choi, Seung Hun [Chung Ang Gil Hospital, Inchon (Korea, Republic of)

    1997-07-01

    Our purpose was to assess the utility of maternal serum triple-marker screening test using alpha-fetoprotein (aFP), human Chorionic Gonadotropin (hCG) and unconjugated Estriol (uE{sub 3}) for fetal chromosomal abnormalities. 1,767 venous blood samples (4ml) between 15 and 20 week's gestation for maternal serum screening from January to October 1996, were tested with Kodak Amerix-M triple marker radioimmunoassay kits. Risk analysis was achieved with interpretive software such as Alpha (LMS, Kodak Clinical Diagnostics). Marker levels are transformed into multiples of median (MOM), which represent an interpretation of (weight regressed) patient marker levels relative to regressed median levels for stated gestation. By multivariate anaysis, the three MOM values are combined to generate a liklihood ratio. Calculation of a patient, risk is the product of liklihood ratio and age-related risk. Risk assessment is weight for maternal age. The median values of aFP, hCG and uE{sub 3} were well correlated with gestational age, respectively (r=0.94, p=0.003; r=-0.97, p=0.029; r=0.99, p<0.001, respectively). The median value of hCG were correlated withmateral age (r=0.13, p=0.04) but those of aFP and eU{sub 3} weren't (r=-0.17, p=0.22; r=0.36, p=0.09, respectively). The values of aFP, CG and uE{sub 3} between pregnancy younger than 35 years-old (n=87) and older than that (n=1640) were 51.67{+-}27.44, vs 54.65{+-}126.36, 46.45{+-}30.08 vs 51.33{+-}38.50 and 8.01{+-}11.01 vs 6.68{+-}7.23, respectively but all of them failed to show significant differences. A second-trimester risk for trisomy 21 > or = 1:270 was considered screen positive. Patients were screen positive for trisomy 21 if aFP < or 0.7 multiples of median (MOM), hCG> or 2.1 MOM and E{sub 3} < or 0.7 MOM. Patients were screen positive for neural tube defect if aFP >2.5 MOM. The initial screen-positive rate for both Down' syndrome and neural tube defect were 1.46% (26/1767); 0.73% (13/1767) with each other

  16. In vitro radionuclide therapy and in vivo scintigraphic imaging of alpha fetoprotein producing hepatocellular carcinoma by targeted sodium iodide symporter gene expression

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Kwang Il; Lee, Yong Jin; Lee, Tae Sup; Song, Inho; Cheon, Gi Jeong; Lim, Sang Moo; Kang, Joo Hyun [Korea Institute of Radiological and Medical and Medical Sciences, Seoul (Korea, Republic of); Chung, June Key [Seoul National Univ. College of Medicine, Seoul (Korea, Republic of)

    2012-03-15

    This study aimed to develop a gene expression targeting method for specific imaging and therapy of alpha fetoprotein (AFP) producing hepatocellular carcinoma (HCC) cells, using an adenovirus vector containing the human sodium/iodide symporter (hNIS) gene driven by an AFP enhancer/promoter. The recombinant adenovirus vector, AdAFPhNIS (containing the hNIS gene driven by human AFP enhancer/promoter) was prepared. After in vitro infection by the adenovirus, hNIS gene expression in AFP producing cells and in AFP nonproducing cells was investigated using {sup 125}I uptake assay and semi quantitative reverse transcription polymerase chain reaction (RT-PCR). The killing effect of {sup 131}I vitro clonogenic assay. In addition, tumor bearing mice were intravenously injected with the adenovirus, and scintigraphic images were obtained. The expression of hNIS was efficiently demonstrated by {sup 125}I uptake assay in AFP producing cells, but not in AFP nonproducing cells. AFP producing HCC targeted gene expression was confirmed at the mRNA level. Furthermore, in vitro clonogenic assay showed that hNIS gene expression induced by AdAFPhNIS infection in AFP producing cells caused more sensitivity to {sup 131}I than that in AFP nonproducing cells. Injected intravenously in HuH-7 tumor xenografts mice by adenovirus, the functional hNIS gene expression was confirmed in tumor by in vivo scintigraphic imaging. An AFP producing HCC was targeted with an adenovirus vector containing the hNIS gene using the AFP enhancer/promoter in vitro and in vivo. These findings demonstrate that AFP producing HCC specific molecular imaging and radionuclide gene therapy are feasible using this recombinant adenovirus vector system.

  17. Characterization of Alpha-fetoprotein Levels in Three Dolphin Species: Development of Sensitive Immunoassays for Analysis of the Pregnancy-associated Variations

    Science.gov (United States)

    MORITA, Yuka; HIRAMATSU, Naoshi; FUJITA, Toshiaki; AMANO, Haruna; KATSUMATA, Etsuko; ARAI, Kazutoshi; IWASAKI, Toshihide; TODO, Takashi; HARA, Akihiko

    2013-01-01

    Abstract A single radial immunodiffusion (SRID) assay and a chemiluminescent immunoassay (CLIA) were initially developed for alpha-fetoprotein (AFP) of the striped dolphin. Utilizing these developed assays, we investigated pregnancy-associated changes in the levels of AFP in the sera of fetuses and pregnant females of three dolphin species; samples were either collected from captive individuals or obtained as fishery by-products. The concentrations of AFP in the fetal serum ranged from 419.0 to 2026.3 μg/ml in the striped dolphin, 12.6 to 1218.7 μg/ml (for an AFP equivalent; eqAFP) in the common bottlenose dolphin and 770.6 to 3129.1 μg eqAFP/ml in the Risso's dolphin. AFP levels decreased with increased fetal size in fetuses over 20 cm in length. The concentrations of AFP in sera of pregnant females ranged from 7.18 to 8068.7 ng/ml in the striped dolphin, 6.6 to 1241.1 ng eqAFP/ml in the common bottlenose dolphin and 3.4 to 2868.7 ng eqAFP/ml in the Risso's dolphin. The levels in most pregnant females were equal to or lower than those found in males and nonpregnant individuals, although a few pregnant females exhibited extremely high levels (in the range of hundreds to thousands of nanograms per milliliter). Such high levels of AFP were not observed during pseudopregnancy. To our knowledge, this is the first report on basal profiles for serum AFP levels in small odontocetes. The profiles indicated that AFP may play a significant role during embryonic development, although maternal levels do not appear to be a diagnostic biomarker for monitoring pregnancy. PMID:23656975

  18. Propensity Score Matching Analysis of Changes in Alpha-Fetoprotein Levels after Combined Radiotherapy and Transarterial Chemoembolization for Hepatocellular Carcinoma with Portal Vein Tumor Thrombus.

    Directory of Open Access Journals (Sweden)

    Yuri Jeong

    Full Text Available To investigate the value of changes in alpha-fetoprotein (AFP levels for the prediction of radiologic response and survival outcomes in hepatocellular carcinoma (HCC patients with portal vein tumor thrombus (PVTT who received combined treatment of 3-dimensional conformal radiotherapy (3D-CRT and transarterial chemoembolization (TACE.A database of 154 HCC patients with PVTT and elevated AFP levels (>20 ng/mL treated with 3D-CRT and TACE as an initial treatment between August 2002 and August 2008 was retrospectively reviewed. AFP levels were determined 1 month after radiotherapy, and AFP response was defined as an AFP level reduction of >20% from the initial level. Radiologic response, overall survival (OS, and progression-free survival (PFS rates were compared between AFP responders and non-responders. Propensity-score based matching analysis was performed to minimize the effect of potential confounding bias.The median follow-up period was 11.1 months (range, 3.1-82.7 months. In the propensity-score matching cohort (92 pairs, a best radiologic response of CR or PR occurred in more AFP responders than AFP non-responders (41.3% vs. 10.9%, p < 0.001. OS and PFS were also longer in AFP responders than in non-responders (median OS 13.2 months vs. 5.6 months, p < 0.001; median PFS 8.7 months vs. 3.5 months, p < 0.001.AFP response is a significant predictive factor for radiologic response. Furthermore, AFP response is significant for OS and PFS outcomes. AFP evaluation after combined radiotherapy and TACE appears to be a useful predictor of clinical outcomes in HCC patients with PVTT.

  19. Alpha-Fetoprotein, Identified as a Novel Marker for the Antioxidant Effect of Placental Extract, Exhibits Synergistic Antioxidant Activity in the Presence of Estradiol

    Science.gov (United States)

    Choi, Hye Yeon; Kim, Seung Woo; Kim, BongWoo; Lee, Hae Na; Kim, Su-Jeong; Song, Minjung; Kim, Sol; Kim, Jungho; Kim, Young Bong; Kim, Jin-Hoi; Cho, Ssang-Goo

    2014-01-01

    Placenta, as a reservoir of nutrients, has been widely used in medical and cosmetic materials. Here, we focused on the antioxidant properties of placental extract and attempted to isolate and identify the main antioxidant factors. Porcine placental extracts were prepared through homogenization or acid hydrolysis, and their antioxidant activity was investigated in the human keratinocyte HaCaT cell line. Treatment with homogenized placental extract (H-PE) increased the cell viability of H2O2-treated HaCaT cells more than two-fold. H-PE treatment suppressed H2O2-induced apoptotic and necrotic cell death and decreased intracellular ROS levels in H2O2-treated HaCaT cells. The antioxidant factors in H-PE were found to be thermo-unstable and were thus expected to include proteins. The candidate antioxidant proteins were fractionated with cation-exchange, anion-exchange, and size-exclusion chromatography, and the antioxidant properties of the chromatographic fractions were investigated. We obtained specific antioxidant fractions that suppressed ROS generation and ROS-induced DNA strand breaks. From silver staining and MALDI-TOF analyses, alpha-fetoprotein (AFP) precursor was identified as a main marker for the antioxidant effect of H-PE. Purified AFP or ectopically expressed AFP exhibited synergistic antioxidant activity in the presence of estradiol. Taken together, our data suggest that AFP, a serum glycoprotein produced at high levels during fetal development, is a novel marker protein for the antioxidant effect of the placenta that exhibits synergistic antioxidant activity in the presence of estradiol. PMID:24922551

  20. Alpha-fetoprotein (AFP) modulates the effect of serum albumin on brain development by restraining the neurotrophic effect of oleic acid.

    Science.gov (United States)

    García-García, Alejandro G; Polo-Hernández, Erica; Tabernero, Arantxa; Medina, José M

    2015-10-22

    We have previously shown that serum albumin controls perinatal rat brain development through the regulation of oleic acid synthesis by astrocytes. In fact, oleic acid synthesized and released by astrocytes promoted neurite growth, neuron migration and the arrangement of prospective synapses. In this work we show that alpha-fetoprotein (AFP) is also present in the brain during embryonic development, its concentrations peaking at E15.5 and at E19.5. However, after E19.5 AFP concentrations plummeted concurrently with a sharp increase in serum albumin concentrations. At E15.5, AFP is present in caudal regions of the brain, particularly in brain areas undergoing differentiation during this period, such as the thalamic reticular nucleus of the thalamus, the hypothalamus, the amygdala and the hippocampus. Albumin was not detected in the brain at E15.5 but stained brain cells substantially on day E19.5, showing a very similar distribution to that of AFP under the same circumstances. The concentrations of free oleic acid in the brain were inversely correlated with those of AFP, suggesting that the signals elicited by AFP and oleic acid can be inversely associated. GAP-43, a marker of axonal growth that is highly expressed by the presence of oleic acid, was not co-localized with AFP except in the marginal zone and areas delimiting the subplate. AFP prevented the increase in GAP-43 expression caused by the presence of oleic acid in neurons in primary culture in vitro and in organotypic cultures of embryonic rat brain ex vivo, suggesting that AFP may modulate the effect of serum albumin on brain development. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Alpha-fetoprotein is present in the fetal fluids and is increased in plasma of mares with experimentally induced ascending placentitis.

    Science.gov (United States)

    Canisso, Igor F; Ball, Barry A; Scoggin, Kirsten E; Squires, Edward L; Williams, Neil M; Troedsson, Mats H

    2015-03-01

    The objectives of this study were to: (i) determine alpha-fetoprotein (AFP) concentrations in fetal fluids (FF), and (ii) compare plasma concentrations of AFP in mares with placentitis (n=17) and gestationally age-matched control mares (n=17). Fetal fluid sampling (FFS, n=7/group) was performed at 0, 5 and 12 days post inoculation (DPI) or until abortion. Plasma was harvested daily for 12 days or until abortion. Placentitis was induced via intracervical inoculation of Streptococcus equi ssp. zooepidemicus. Proteins present in the FF were resolved by 1D-SDS-PAGE, and immunoblotting was used to detect the presence of AFP in fetal fluids. Concentrations of AFP in FF and plasma were determined with a chemiluminescence immunoassay. Mixed models for DPI, and for days from abortion (DFA) were used to analyze plasma concentrations of AFP. A protein band ∼68kDa consistent with the AFP size was present in all samples of fetal fluids examined. Immunoblotting for AFP revealed a single protein band (∼68kDa) in all samples. Concentrations of AFP in FF appeared higher than those in maternal plasma. There were effects of time (DPI p<0.0001; DFA p=0.0002) and time-by-group interactions (DPI*Group p<0.06; Group*DFA p<0.001). This study confirmed that AFP is present in the FF of mares during the third trimester of pregnancy. Experimentally induced placentitis was associated with an elevation in maternal plasma concentrations of AFP. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Glypican-3 level assessed by the enzyme-linked immunosorbent assay is inferior to alpha-fetoprotein level for hepatocellular carcinoma diagnosis.

    Science.gov (United States)

    Jeon, Yejoo; Jang, Eun Sun; Choi, Yun Suk; Kim, Jin-Wook; Jeong, Sook-Hyang

    2016-09-01

    Glypican-3 (GPC3) protein is highly expressed in hepatocellular carcinoma (HCC) tissue. It has been suggested as a diagnostic biomarker, but its inconsistent performance means that it requires further assessment. We therefore investigated the diagnostic value of the plasma GPC3 level compared to the alpha-fetoprotein (AFP) level as a diagnostic biomarker of HCC. We enrolled 157 consecutive patients with newly diagnosed HCC and 156 patients with liver cirrhosis (LC) as the control group. GPC3 plasma levels were measured using two commercially available enzyme-linked immunosorbent assays (ELISAs, named as Assay 1 and 2), and AFP levels were measured using an enzyme-linked chemiluminescent immunoassay. The diagnostic accuracy was analyzed using the receiver operating characteristics (ROC) curve. Plasma GPC3 levels in HCC patients were very low (0-3.09 ng/mL) in Assay 1, while only 3 of the 157 patients (1.9%) showed detectable GPC3 levels in Assay 2. The median GPC3 level was not significantly elevated in the HCC group (0.80 ng/mL) compared with the LC group (0.60 ng/mL). The area under the ROC curve (AUC) for GPC3 was 0.559 in Assay 1. In contrast, the median AFP level was significantly higher in HCC (27.72 ng/mL) than in LC (4.74 ng/mL), with an AUC of 0.729. The plasma level of GPC3 is a poor diagnostic marker for HCC, being far inferior to AFP. The development of a consistent detection system for the blood level of GPC3 is warranted.

  3. Magnetic immunoassay coupled with inductively coupled plasma mass spectrometry for simultaneous quantification of alpha-fetoprotein and carcinoembryonic antigen in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Xing; Chen, Beibei; He, Man; Zhang, Yiwen; Xiao, Guangyang; Hu, Bin, E-mail: binhu@whu.edu.cn

    2015-04-01

    The absolute quantification of glycoproteins in complex biological samples is a challenge and of great significance. Herein, 4-mercaptophenylboronic acid functionalized magnetic beads were prepared to selectively capture glycoproteins, while antibody conjugated gold and silver nanoparticles were synthesized as element tags to label two different glycoproteins. Based on that, a new approach of magnetic immunoassay-inductively coupled plasma mass spectrometry (ICP-MS) was established for simultaneous quantitative analysis of glycoproteins. Taking biomarkers of alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) as two model glycoproteins, experimental parameters involved in the immunoassay procedure were carefully optimized and analytical performance of the proposed method was evaluated. The limits of detection (LODs) for AFP and CEA were 0.086 μg L{sup −1} and 0.054 μg L{sup −1} with the relative standard deviations (RSDs, n = 7, c = 5 μg L{sup −1}) of 6.5% and 6.2% for AFP and CEA, respectively. Linear range for both AFP and CEA was 0.2–50 μg L{sup −1}. To validate the applicability of the proposed method, human serum samples were analyzed, and the obtained results were in good agreement with that obtained by the clinical chemiluminescence immunoassay. The developed method exhibited good selectivity and sensitivity for the simultaneous determination of AFP and CEA, and extended the applicability of metal nanoparticle tags based on ICP-MS methodology in multiple glycoprotein quantifications. - Highlights: • 4-Mercaptophenylboronic acid functionalized magnetic beads were prepared and characterized. • ICP-MS based magnetic immunoassay approach was developed for quantification of glycoproteins. • AFP and CEA were quantified simultaneously with Au and Ag NPs as element tags. • The developed method exhibited good selectivity and sensitivity for target glycoproteins.

  4. 8-Quinolineboronic acid as a potential phosphorescent molecular switch for the determination of alpha-fetoprotein variant for the prediction of primary hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Liu Jiaming, E-mail: zzsyliujiaming@163.com [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Xianzhiqian Street, 36 Zhangzhou, Fujian 363000 (China); Li Feiming [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Xianzhiqian Street, 36 Zhangzhou, Fujian 363000 (China); Liu Zhenbo [Third Hospital of Xiamen, Xiamen 316000 (China); Lin Changqing [Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou 363000 (China); Lin Shaoqin [Department of Biochemistry, Fujian Education College, Fuzhou 350001 (China); Lin Liping [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Xianzhiqian Street, 36 Zhangzhou, Fujian 363000 (China); Wang Xinxing [Department of Chemistry and Environmental Science, Zhangzhou Normal College, Xianzhiqian Street, 36 Zhangzhou, Fujian 363000 (China); Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou 363000 (China); Li Zhiming [Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou 363000 (China)

    2010-03-24

    8-Quinolineboronic acid phosphorescent molecular switch (8-QBA-PMS) in the 'off' state emitted weak room temperature phosphorescence (RTP) of 8-QBA on the acetylcellulose membrane (ACM) with the perturbation of Pb{sup 2+}. When 8-QBA-PMS was used to label concanavalin agglutinin (Con A) to form 8-QBA-PMS-Con A based on the reaction between -OH of 8-QBA-PMS and -COOH of Con A, 8-QBA-PMS turned 'on' automatically due to its structure change, and RTP of the system increased 2.7 times. Besides, -NH{sub 2} of 8-QBA-PMS-Con A could carry out affinity adsorption (AA) reaction with the -COOH of alpha-fetoprotein variant (AFP-V) to form the product Con A-AFP-V-Con A-8-QBA-PMS containing -NH-CO- bond, causing the RTP of the system to further increase. Moreover, the amount of AFP-V was linear to the {Delta}I{sub p} of the system in the range of 0.012-2.40 (fg spot{sup -1}). Thus, a new affinity sensitive adsorption solid substrate room temperature phosphorimetry using 8-QBA-PMS as labelling reagent (8-QBA-PMS-AASSRTP) for the determination of AFP-V was proposed with the detection limit (LD) of 9 x 10{sup -15} g mL{sup -1}. It had been used to determine AFP-V in human serum with the results agreeing with enzyme-link immunoassay (ELISA), showing promise for the prediction of PHC due to the intimate association between AFP-V and primary hepatocellular carcinoma (PHC). The mechanism of the promethod was also discussed.

  5. Higher Ratio of Serum Alpha-Fetoprotein Could Predict Outcomes in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma and Normal Alanine Aminotransferase.

    Directory of Open Access Journals (Sweden)

    Young-Il Kim

    Full Text Available The role of serum alpha-fetoprotein (AFP levels in the surveillance and diagnosis of hepatocellular carcinoma (HCC is controversial. The aim of this study was to investigate the value of serially measured serum AFP levels in HCC progression or recurrence after initial treatment.A total of 722 consecutive patients newly diagnosed with HCC and treated at the National Cancer Center, Korea, between January 2004 and December 2009 were enrolled. The AFP ratios between 4-8 weeks post-treatment and those at the time of HCC progression or recurrence were obtained. Multivariate logistic regression analysis was performed to correlate the post-treatment AFP ratios with the presence of HCC progression or recurrence.The etiology of HCC was related to chronic hepatitis B virus (HBV infection in 562 patients (77.8%, chronic hepatitis C virus (HCV infection in 74 (10.2%, and non-viral cause in 86 (11.9%. There was a significant decrease in serum AFP levels from the baseline to 4 to 8 weeks after treatment (median AFP, 319.6 ng/mL vs. 49.6 ng/mL; p 1.0 was an independently associated with HCC progression or recurrence. Among the different causes of HCC analyzed, this association was significant only for HCC related to chronic hepatitis B (p< 0.001 and non-viral causes (p<0.05, and limited only to patients who had normal alanine aminotransferase (ALT levels.Serial measurements of serum AFP ratios could be helpful in detecting progression or recurrence in treated patients with HBV-HCC and normal ALT.

  6. Alpha-fetoprotein still is a valuable diagnostic and prognosis predicting biomarker in hepatitis B virus infection-related hepatocellular carcinoma.

    Science.gov (United States)

    Yao, Mingjie; Zhao, Jingmin; Lu, Fengmin

    2016-01-26

    Use of serum alpha-fetoprotein (AFP) in clinical practices has been challenged in recent years, due to the lack of specificity and sensitivity. Here we conducted a retrospective study to evaluate the diagnostic and prognostic value of serum AFP among hepatocellular carcinoma (HCC) patients with their pathogenic features taken into consideration. The cohort for this study comprised 318 cases of hepatitis and 731 cases of cirrhosis, as well as 796 HCC patients. Using 11.62ng/mL as a cut-off value, the positive rate of AFP test among serum hepatitis B surface antigen (HBsAg) positive HCC patients was significantly higher than that in those HBsAg negative HCC patients (79.55% vs 56.49%, P < 0.000). Similarly, the median serum AFP level in HCC patients with serum HBsAg positive was significantly higher than that in those HBsAg negative HCC patients (423.89ng/ml vs 40.82ng/ml, P < 0.000). In addition, Kaplan-Meier curve analysis revealed that lower preoperative AFP level implicated a much higher overall survival rate. Of note, such prognosis predicting value was only seen in those chronic HBV infection-related HCC patients, but not among the HCC patients etiologically irrelevant to HBV infection. We believe that serum AFP is of diagnosis and prognostic predicting value for HCC with chronic HBV infection, and strongly suggest use of serum AFP as a biomarker in China and other HBV infection endemic area like Southeast Asia.

  7. Higher Ratio of Serum Alpha-Fetoprotein Could Predict Outcomes in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma and Normal Alanine Aminotransferase

    Science.gov (United States)

    Park, Joong-Won

    2016-01-01

    Background The role of serum alpha-fetoprotein (AFP) levels in the surveillance and diagnosis of hepatocellular carcinoma (HCC) is controversial. The aim of this study was to investigate the value of serially measured serum AFP levels in HCC progression or recurrence after initial treatment. Methods A total of 722 consecutive patients newly diagnosed with HCC and treated at the National Cancer Center, Korea, between January 2004 and December 2009 were enrolled. The AFP ratios between 4–8 weeks post-treatment and those at the time of HCC progression or recurrence were obtained. Multivariate logistic regression analysis was performed to correlate the post-treatment AFP ratios with the presence of HCC progression or recurrence. Results The etiology of HCC was related to chronic hepatitis B virus (HBV) infection in 562 patients (77.8%), chronic hepatitis C virus (HCV) infection in 74 (10.2%), and non-viral cause in 86 (11.9%). There was a significant decrease in serum AFP levels from the baseline to 4 to 8 weeks after treatment (median AFP, 319.6 ng/mL vs. 49.6 ng/mL; p 1.0 was an independently associated with HCC progression or recurrence. Among the different causes of HCC analyzed, this association was significant only for HCC related to chronic hepatitis B (p< 0.001) and non-viral causes (p<0.05), and limited only to patients who had normal alanine aminotransferase (ALT) levels. Conclusion Serial measurements of serum AFP ratios could be helpful in detecting progression or recurrence in treated patients with HBV-HCC and normal ALT. PMID:27304617

  8. In vitro radionuclide therapy and in vivo scintigraphic imaging of alpha fetoprotein producing hepatocellular carcinoma by targeted sodium iodide symporter gene expression

    International Nuclear Information System (INIS)

    Kim, Kwang Il; Lee, Yong Jin; Lee, Tae Sup; Song, Inho; Cheon, Gi Jeong; Lim, Sang Moo; Kang, Joo Hyun; Chung, June Key

    2012-01-01

    This study aimed to develop a gene expression targeting method for specific imaging and therapy of alpha fetoprotein (AFP) producing hepatocellular carcinoma (HCC) cells, using an adenovirus vector containing the human sodium/iodide symporter (hNIS) gene driven by an AFP enhancer/promoter. The recombinant adenovirus vector, AdAFPhNIS (containing the hNIS gene driven by human AFP enhancer/promoter) was prepared. After in vitro infection by the adenovirus, hNIS gene expression in AFP producing cells and in AFP nonproducing cells was investigated using 125 I uptake assay and semi quantitative reverse transcription polymerase chain reaction (RT-PCR). The killing effect of 131 I vitro clonogenic assay. In addition, tumor bearing mice were intravenously injected with the adenovirus, and scintigraphic images were obtained. The expression of hNIS was efficiently demonstrated by 125 I uptake assay in AFP producing cells, but not in AFP nonproducing cells. AFP producing HCC targeted gene expression was confirmed at the mRNA level. Furthermore, in vitro clonogenic assay showed that hNIS gene expression induced by AdAFPhNIS infection in AFP producing cells caused more sensitivity to 131 I than that in AFP nonproducing cells. Injected intravenously in HuH-7 tumor xenografts mice by adenovirus, the functional hNIS gene expression was confirmed in tumor by in vivo scintigraphic imaging. An AFP producing HCC was targeted with an adenovirus vector containing the hNIS gene using the AFP enhancer/promoter in vitro and in vivo. These findings demonstrate that AFP producing HCC specific molecular imaging and radionuclide gene therapy are feasible using this recombinant adenovirus vector system

  9. 8-Quinolineboronic acid as a potential phosphorescent molecular switch for the determination of alpha-fetoprotein variant for the prediction of primary hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Liu Jiaming; Li Feiming; Liu Zhenbo; Lin Changqing; Lin Shaoqin; Lin Liping; Wang Xinxing; Li Zhiming

    2010-01-01

    8-Quinolineboronic acid phosphorescent molecular switch (8-QBA-PMS) in the 'off' state emitted weak room temperature phosphorescence (RTP) of 8-QBA on the acetylcellulose membrane (ACM) with the perturbation of Pb 2+ . When 8-QBA-PMS was used to label concanavalin agglutinin (Con A) to form 8-QBA-PMS-Con A based on the reaction between -OH of 8-QBA-PMS and -COOH of Con A, 8-QBA-PMS turned 'on' automatically due to its structure change, and RTP of the system increased 2.7 times. Besides, -NH 2 of 8-QBA-PMS-Con A could carry out affinity adsorption (AA) reaction with the -COOH of alpha-fetoprotein variant (AFP-V) to form the product Con A-AFP-V-Con A-8-QBA-PMS containing -NH-CO- bond, causing the RTP of the system to further increase. Moreover, the amount of AFP-V was linear to the ΔI p of the system in the range of 0.012-2.40 (fg spot -1 ). Thus, a new affinity sensitive adsorption solid substrate room temperature phosphorimetry using 8-QBA-PMS as labelling reagent (8-QBA-PMS-AASSRTP) for the determination of AFP-V was proposed with the detection limit (LD) of 9 x 10 -15 g mL -1 . It had been used to determine AFP-V in human serum with the results agreeing with enzyme-link immunoassay (ELISA), showing promise for the prediction of PHC due to the intimate association between AFP-V and primary hepatocellular carcinoma (PHC). The mechanism of the promethod was also discussed.

  10. Maternal and fetal mechanisms of B cell regulation during pregnancy: human Chorionic Gonadotropin stimulates B cells to produce IL-10 while alpha-fetoprotein drives them into apoptosis

    Directory of Open Access Journals (Sweden)

    Franziska Fettke

    2016-12-01

    Full Text Available Maternal immune tolerance towards the fetus is an essential requisite for pregnancy. While T cell functions are well documented, little is known about the participation of B cells. We have previously suggested that IL-10 producing B cells are involved in pregnancy tolerance in mice and humans. By employing murine and human systems, we report now that fetal trophoblasts positively regulate the generation of IL-10 producing B cells. We next studied the participation of hormones produced by the placenta as well as the fetal protein alpha-fetoprotein (AFP in B cell modulation. Human Chorionic Gonadotropin (hCG, but not progesterone, estrogen or a combination of both, was able to promote changes in B cell phenotype and boost their IL-10 production, which was abolished after blocking hCG. The hCG-induced B cell phenotype was not associated with augmented galactosylation, sialylation or fucosylation of IgG subclasses in their Fc. In vitro, hCG induced the synthesis of asymmetrically glycosylated antibodies in their Fab region. Interestingly, AFP had dual effects depending on the concentration. At concentrations corresponding to maternal serum levels, it did not modify the phenotype or IL-10 secretion of B cells. At fetal concentrations, however, AFP was able to drive B cells into apoptosis, which may indicate a protective mechanism to avoid maternal B cells to reach the fetus.Our data suggests that the fetus secrete factors that promote a pregnancy-friendly B cell phenotype, unraveling interesting aspects of B cell function and modulation by pregnancy hormones and fetal proteins.

  11. Alpha-fetoprotein, identified as a novel marker for the antioxidant effect of placental extract, exhibits synergistic antioxidant activity in the presence of estradiol.

    Directory of Open Access Journals (Sweden)

    Hye Yeon Choi

    Full Text Available Placenta, as a reservoir of nutrients, has been widely used in medical and cosmetic materials. Here, we focused on the antioxidant properties of placental extract and attempted to isolate and identify the main antioxidant factors. Porcine placental extracts were prepared through homogenization or acid hydrolysis, and their antioxidant activity was investigated in the human keratinocyte HaCaT cell line. Treatment with homogenized placental extract (H-PE increased the cell viability of H2O2-treated HaCaT cells more than two-fold. H-PE treatment suppressed H2O2-induced apoptotic and necrotic cell death and decreased intracellular ROS levels in H2O2-treated HaCaT cells. The antioxidant factors in H-PE were found to be thermo-unstable and were thus expected to include proteins. The candidate antioxidant proteins were fractionated with cation-exchange, anion-exchange, and size-exclusion chromatography, and the antioxidant properties of the chromatographic fractions were investigated. We obtained specific antioxidant fractions that suppressed ROS generation and ROS-induced DNA strand breaks. From silver staining and MALDI-TOF analyses, alpha-fetoprotein (AFP precursor was identified as a main marker for the antioxidant effect of H-PE. Purified AFP or ectopically expressed AFP exhibited synergistic antioxidant activity in the presence of estradiol. Taken together, our data suggest that AFP, a serum glycoprotein produced at high levels during fetal development, is a novel marker protein for the antioxidant effect of the placenta that exhibits synergistic antioxidant activity in the presence of estradiol.

  12. Clinical and molecular sub-classification of hepatocellular carcinoma relative to alpha-fetoprotein level in an Asia-Pacific island cohort.

    Science.gov (United States)

    Nishioka, Scott T; Sato, Miles M; Wong, Linda L; Tiirikainen, Maarit; Kwee, Sandi A

    2018-01-01

    Increased serum alpha-fetoprotein (AFP) levels are associated with specific molecular sub-classes of hepatocellular carcinoma (HCC), supporting AFP as a predictive or therapeutic biomarker for precision treatment of this disease. Considering recent efforts to validate HCC molecular classification systems across different populations, we applied existing signature-based classification templates to Hawaii cohorts and examined whether associations between HCC molecular sub-class, AFP levels, and clinical features found elsewhere can also be found in Hawaii, a region with a unique demographic and risk factor profile for HCC. Whole-genome expression profiling was performed on HCC tumors collected from 40 patients following partial hepatectomy. Tumors underwent transcriptome-based categorization into 3 molecular sub-classes (S1, S2, and S3). Patient groups based on molecular sub-class and AFP level were then compared with regards to clinical features and survival. Differences associated with AFP level and other clinical parameters were also examined at the gene signature level by gene set enrichment analysis. Statistically confident (false discovery rate 400 ng/mL predicted significant tumor enrichment for genes corresponding to MYC target activation, high cell proliferation, poor clinical prognosis, and the S2 sub-class. AFP > 400 ng/mL and non-S3 tumor classification were found to be significant predictors of overall survival. Distinct sub-classes of HCC associated with different molecular features and survival outcomes can be detected with statistical confidence in a Pacific Island cohort. Molecular classification signatures and other predictive markers for HCC that are valid for all patient populations are needed to support multi-center efforts to develop targeted therapies for HCC.

  13. Rapid and simple half-quantitative measurement alpha-fetoprotein by poly(dimethylsiloxane) microfluidic chip immunochromatographic assay

    Science.gov (United States)

    Tong, Chao; Jin, Qinghui; Zhao, Jianlong

    2008-03-01

    In this article, a kind of microfluidic method based on MEMS technology combined with gold immunochromatographic assay (GICA) is developed and discussed. Compared to the traditional GICA, this method supplies us convenient, multi-channel, in-parallel, low cost and similar efficiency approach in the fields of alpha-fetopro-tei (AFP)detection. Firstly, we improved the adhesion between the model material SU-8 and Silicon wafer, optimized approaches of the fabrication of the SU-8 model systematically, and fabricate the PDMS micro fluid chip with good reproduction successfully. Secondly, Surface modification and antibody immobilization methods with the GICA on the PDMS micro fluid analysis chip are studied, we choose the PDMS material and transfer GICA to the PDMS micro fluid chip successfully after researching the antibody immobilization efficiency of different materials utilized in fabrication of the micro fluid chip. In order to improve the reaction efficiency of the immobilized antibody, we studied the characteristics of micro fluid without the gas drive, and the fluid velocity control in our design; we also design structure of grove to strengthen the ability of immobilizing the antibody. The stimulation of the structure shows that it achieves great improvement and experiments prove the design is feasible.

  14. A cyclized peptide derived from alpha fetoprotein inhibits the proliferation of ER-positive canine mammary cancer cells.

    Science.gov (United States)

    Torres, Cristian Gabriel; Pino, Ana María; Sierralta, Walter Daniel

    2009-06-01

    The effects of estradiol (E2) and of an AFP-derived cyclized peptide (cP) on the proliferation of primary cultures of cancer cells isolated from spontaneous canine mammary tumors were studied. The cellular response to E2 and cP was related to the expression of estradiol receptor (isoforms alpha and beta). In ER-positive cells, 2 nM estradiol increased cell proliferation and the phosphorylation of ERK1/2; 2 microg/ml cP inhibited all these effects. Estradiol also increased HER2 immunoreactivity in ER-positive cells, an effect that was reverted to its basal values by cP. Estradiol stimulated in these cells the release of MMP2 and MMP9 and the shedding of HB-EGF, effects that the cP did not affect. ER-negative cells were refractory to estradiol or cP. All canine mammary tumor cells in culture responded to treatments analogously to human mammary cancer cells. Our results support the proposal of cP as a new, potentially effective therapeutic agent for the management of mammary cancer.

  15. Alpha-fetoprotein and (18)F-FDG positron emission tomography predict tumor recurrence better than Milan criteria in living donor liver transplantation.

    Science.gov (United States)

    Hong, Geun; Suh, Kyung-Suk; Suh, Suk-Won; Yoo, Tae; Kim, Hyeyoung; Park, Min-Su; Choi, YoungRok; Paeng, Jin Chul; Yi, Nam-Joon; Lee, Kwang-Woong

    2016-04-01

    Given the organ shortage for liver transplantation (LT) and the limitations of the current morphology-based selection criteria, improved criteria are needed to achieve the maximum benefit of LT for hepatocellular carcinoma (HCC). We hypothesized that a combination of biological markers may better predict the prognosis than the Milan criteria. HCC patients (n=123) with preoperative data on serum alpha-fetoprotein (AFP) levels and (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG PET) positivity underwent live-donor LT between January 2003 and December 2009. The cut-off values for serum AFP levels (200 ng/ml) and (18)F-FDG PET positivity (1.10) for tumor recurrence were determined by c-statistics using receiver operating characteristic curves. Univariate and multivariate analyses with preoperative variables were performed to find pre-transplant prognostic factors. Disease-free survival rates and overall survival rates were analysed with regard to serum AFP levels and (18)F-FDG PET positivity. The 5-year disease-free survival rates and overall survival rates were 80.3% and 81.6% respectively. (18)F-FDG PET positivity (hazard ratio (HR) 9.766, 95% CI 3.557-26.816; p<0.001) and serum AFP level (HR 6.234, 95% CI 2.643-14.707; p<0.001) were the only significant pre-transplant prognostic factors in the multivariate analysis; tumor number and size were not significant. A combination of criteria showed that the biologically high-risk group (AFP level ⩾200 ng/ml and PET-positive) had an HR of 29.069 (95% CI 8.797-96.053; p<0.001) compared with the double-negative group. Use of the Milan criteria yielded an HR of 1.351 (95% CI 0.500-3.652; p=0.553). The combination of the serum AFP level and (18)F-FDG PET data predicted better outcomes than those using the Milan criteria, improving objectivity when adult-to-adult living donor LT is contemplated. Copyright © 2015 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

  16. The efficacy of modified docetaxel-cisplatin-5-fluorouracil regimen as first-line treatment in patients with alpha-fetoprotein producing gastric carcinoma

    Science.gov (United States)

    Bozkaya, Yakup; Doğan, Mutlu; Yazıcı, Ozan; Erdem, Gökmen Umut; Demirci, Nebi Serkan; Zengin, Nurullah

    2017-01-01

    Alpha-fetoprotein producing gastric carcinoma (AFP-PGC) is a rare cancer for which limited data on the clinicopathological features and treatment modalities exist. The aim of this study was to compare the efficacy of modified docetaxel-cisplatin-5-fluorouracil (mDCF) as the first-line chemotherapy regimen in metastatic AFP-PGC and non-AFP-PGC. The patients diagnosed with metastatic gastric cancer who were given mDCF as first-line therapy were retrospectively reviewed. The patients with a basal serum AFP level over 9 ng/ml were defined as AFP-PGC patients. In total, 169 patients (34 with AFP-PGC and 135 with non-AFP-PGC) were included in this study. AFP-PGC patients had more liver metastases than non-AFP-PGC patients (p < 0.001). A decrease in basal AFP levels after three cycles of chemotherapy was significantly different in AFP-PGC group (p = 0.001). Overall disease control rate was 79.4% (partial response [PR] - 44.1%, stable disease [SD] - 35.3%), and 82.2% (complete response - 3%, PR - 36.2%, SD - 43%) in AFP-PGC and non-AFP-PGC patients, respectively. There was no difference between AFP-PGC and non-AFP-PGC groups in overall and progression-free survival rates (11.3 versus 11.4 months and 7.7 versus 7.1 months, respectively). Rates of grade 3-4 hematologic toxicity were 8.8% and 6.7% for neutropenia in AFP-PGC and non-AFP-PGC group, respectively and 5.9% and 7.4% for anemia. In conclusion, mDCF regimen is well-tolerated with acceptable toxicity outcomes in both AFP-PGC and non-AFP-PGC patients. A statistically significant decrease in AFP levels after mDCF regimen indicate that AFP might be considered as a supplemental marker of response to mDCF chemotherapy in AFP-PGC patients. However, further prospective clinical trials are required in this area. PMID:28273032

  17. Rapid and sensitive lateral flow immunoassay method for determining alpha fetoprotein in serum using europium (III) chelate microparticles-based lateral flow test strips

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Rong-Liang; Xu, Xu-Ping; Liu, Tian-Cai; Zhou, Jian-Wei; Wang, Xian-Guo; Ren, Zhi-Qi [Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong (China); Hao, Fen [DaAn Gene Co. Ltd. of Sun Yat-sen University, 19 Xiangshan Road, Guangzhou 510515 (China); Wu, Ying-Song, E-mail: wg@smu.edu.cn [Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou 510515, Guangdong (China)

    2015-09-03

    Alpha-fetoprotein (AFP), a primary marker for many diseases including various cancers, is important in clinical tumor diagnosis and antenatal screening. Most immunoassays provide high sensitivity and accuracy for determining AFP, but they are expensive, often complex, time-consuming procedures. A simple and rapid point-of-care system that integrates Eu (III) chelate microparticles with lateral flow immunoassay (LFIA) has been developed to determine AFP in serum with an assay time of 15 min. The approach is based on a sandwich immunoassay performed on lateral flow test strips. A fluorescence strip reader was used to measure the fluorescence peak heights of the test line (H{sub T}) and the control line (H{sub C}); the H{sub T}/H{sub C} ratio was used for quantitation. The Eu (III) chelate microparticles-based LFIA assay exhibited a wide linear range (1.0–1000 IU mL{sup −1}) for AFP with a low limit of detection (0.1 IU mL{sup −1}) based on 5ul of serum. Satisfactory specificity and accuracy were demonstrated and the intra- and inter-assay coefficients of variation (CV) for AFP were both <10%. Furthermore, in the analysis of human serum samples, excellent correlation (n = 284, r = 0.9860, p < 0.0001) was obtained between the proposed method and a commercially available CLIA kit. Results indicated that the Eu (III) chelate microparticles-based LFIA system provided a rapid, sensitive and reliable method for determining AFP in serum, indicating that it would be suitable for development in point-of-care testing. - Highlights: • Europium (III) chelate microparticles was used as a label for LIFA. • Quantitative detection by using H{sub T}/H{sub C} ratio was achieved. • LIFA for simple and rapid AFP detection in human serum. • The sensitivity and linearity was more excellent compared with QD-based ICTS. • This method could be developed for rapid point-of-care screening.

  18. Horseradish peroxidase-loaded nanospheres attached to hollow gold nanoparticles as signal enhancers in an ultrasensitive immunoassay for alpha-fetoprotein

    International Nuclear Information System (INIS)

    Li, Ya; Yuan, Ruo; Chai, Yaqin; Zhuo, Ying; Su, Huilan; Zhang, Yuxia

    2014-01-01

    We report on a novel electrochemical signal amplification strategy for use in immunoassays. The highly responsive immunoelectrode was constructed in the following way: (1) The surface of a gold electrode was covered with a layer single-walled carbon nanotubes dispersed in chitosane functionalized with L-cysteine; (2) Gold nanoparticles containing protein A and anti-alpha-fetoprotein (anti-AFP) were then covalently attached to the surface via the thiol groups of the chitosane. The electrode is then exposed to the analyte (AFP) which then is bound by the antibody. In the next step, a gold-conjugated secondary antibody is added that was prepared in the following way: (1) Horseradish peroxidase was crosslinked and the resulting spheres were coated with hollow gold nanoparticles (hollow Au-NPs) to give nanospheres of ∼100 nm in diameter. (2) These were the coated with thionine and, in a last step, with secondary antibody. The use of these materials has several attractive features: The HRP-NPs functionalized with hollow Au-NPs possess a large surface area that can load the large amount of secondary antibody. Thionine (Thi) is highly redox active and improves the intensity of the signal. Carbon nanotubes were used because they possess an excellent electron transfer rate and large surface area. Following incubation of the modified electrode (a) with a sample containing AFP, (b) then with the secondary antibody, and (c) with washing buffer, the electrode is placed in a solution containing H 2 O 2 . The HRP in the smart secondary antibody causes the catalytic decomposition of H 2 O 2 and the results in an electrical current that is linearly related to the concentration of AFP in the 0.025 to 5.0 ng mL −1 concentration range. The detection limit for AFP is as low as 8.3 pg mL −1 . We believe that this novel kind of immunoassay represents a promising tool for use in sensitive clinical assays. (author)

  19. Application of PET-CT in monitoring residual and extrahepatic metastatic lesions for hepatocellular carcinoma with positive alpha fetoproteins after interventional therapy

    International Nuclear Information System (INIS)

    Zhu Guangyu; Teng Gaojun; Guo Jinhe; Deng Gang; He Shicheng; Fang Wen; Li Guozhao; Chen Xiaohui; Wei Xiaoying

    2010-01-01

    Objective: To investigate the value of positron emission tomography-computed tomography (PET-CT) in monitoring the residual lesions in lipiodol sedimentary region and extrahepatic metastastic lesions of hepatocellular carcinoma (HCC) with alpha fetoproteins (AFP) positive after interventional therapy. Methods: The data of 20 cases with primary HCC confirmed by histopathology were retrospectively analyzed. Their AFP levels decreased to normal range after interventional treatments, but rose to abnormal high level during following-up. After the abdominal routine imaging examinations, the definite diagnosis of the residual lesions in lipiodol sedimentary region or extrahepatic lesions can't be made confidently. All cases were scanned by PET-CT, and according to their PET-CT results, the further treatments were given and the therapeutic results were monitored with radiology and AFP tests. Results: In all 20 cases, 10 of them were detected to have the extrahepatic metastastic lesions by PET-CT, including 4 with abdominal wall metastasis upon the liver, 3 with solitary pulmonary metastasis with diameter less than 1 cm, 2 with mesenteric metastasis, 1 with metastasis of operative incisions, but these lesions were neglected by abdominal routine imaging examinations. Eight cases showed the uneven lipiodol sedimentary region in the primary lesion by CT or MRI examination, but can't be diagnosed whether it was residual lesion with other examinations including DSA. A definite diagnosis was obtained by PET-CT. In 2 cases, lymph nodes less than 1.5 cm were found in the hepatic portal area (PHA) and retroperitoneum on CT images, which was determined to be metastatic by PET-CT. All the detected lesions were given further treatments of surgery or interventional therapy. Most patients showed decreased AFP levels except the 2 patients with lymph node metastasis. The imaging examinations also indicated that the treatments had a good effect on lesions. Conclusion: In the patients with

  20. Alpha-fetoprotein level > 1000 ng/mL as an exclusion criterion for liver transplantation in patients with hepatocellular carcinoma meeting the Milan criteria.

    Science.gov (United States)

    Hameed, Bilal; Mehta, Neil; Sapisochin, Gonzalo; Roberts, John P; Yao, Francis Y

    2014-08-01

    Serum alpha-fetoprotein (AFP) has been increasingly recognized as a marker for a poor prognosis after liver transplantation (LT) for hepatocellular carcinoma (HCC). Many published reports, however, have included a large proportion of patients with HCC beyond the Milan criteria, and the effects of incorporating AFP as an exclusion criterion for LT remain unclear. We studied 211 consecutive patients undergoing LT for HCC within the Milan criteria according to imaging under the Model for End-Stage Liver Disease organ allocation system between June 2002 and January 2009. The majority (93.4%) had locoregional therapy before LT. The median follow-up was 4.5 years (minimum = 2 years). The Kaplan-Meier 1- and 5-year patient survival rates were 94.3% and 83.4%, respectively. In a univariate analysis, significant predictors of HCC recurrence included vascular invasion [hazard ratio (HR) = 10, 95% confidence interval (CI) = 3.9-26, P 1000 ng/mL (HR = 4.5, 95% CI = 1.3-15.3, P = 0.02), and an AFP level > 500 ng/mL (HR = 3.1, 95% CI = 1.04-9.4, P = 0.04). In a multivariate analysis, vascular invasion was the only significant predictor of tumor recurrence (HR = 5.6, 95% CI = 1.9-19, P = 0.02). An AFP level > 1000 ng/mL was the strongest pretransplant variable predicting vascular invasion (odds ratio = 6.8, 95% CI = 1.6-19.1, P = 0.006). The 1- and 5-year rates of survival without recurrence were 90% and 52.7%, respectively, for patients with an AFP level > 1000 ng/mL and 95% and 80.3%, respectively, for patients with an AFP level ≤ 1000 ng/mL (P = 0.026). Applying an AFP level > 1000 ng/mL as a cutoff would have resulted in the exclusion of 4.7% of the patients fr m LT and a 20% reduction in HCC recurrence. In conclusion, an AFP level > 1000 ng/mL may be a surrogate for vascular invasion and may be used to predict posttransplant HCC recurrence. Incorporating an AFP level > 1000 ng/mL as an exclusion criterion for LT within the Milan criteria may further improve posttransplant

  1. Cost-effectiveness analysis of hepatocellular carcinoma screening by combinations of ultrasound and alpha-fetoprotein among Alaska Native people, 1983–2012

    Directory of Open Access Journals (Sweden)

    Prabhu P. Gounder

    2016-05-01

    Full Text Available Background: The American Association for the Study of Liver Diseases (AASLD recommends semi-annual hepatocellular carcinoma (HCC screening using ultrasound (US in persons with chronic hepatitis B (CHB virus infection at high risk for HCC such as Asian males aged ≥40 years and Asian females aged ≥50 years. Objective: To analyse the cost-effectiveness of 2 HCC screening methods in the Alaska Native (AN health system: US-alone, or screening by alpha-fetoprotein (AFP initially and switching to US for subsequent screenings if AFP >10 ng/mL (AFP→US. Design: A spreadsheet-based model was developed for accounting the costs of 2 hypothetical HCC screening methods. We used epidemiologic data from a cohort of 839 AN persons with CHB who were offered HCC screening by AFP/US semi-annually during 1983–2012. We assumed that compared with AFP→US, US-alone identifies 33% more tumours at an early stage (defined as a single tumour ≤5 cm or ≤3 tumours ≤3 cm in diameter. Years of life gained (YLG attributed to screening was estimated by comparing additional years of survival among persons with early- compared with late-stage tumours. Screening costs were calculated using Medicare reimbursement rates in 2012. Future screening costs and YLG were projected over a 30-year time horizon using a 3% discount rate. Results: The total cost of screening for the cohort by AFP→US would have been approximately $357,000 ($36,000/early-stage tumour detected compared to $814,000 ($59,000/early-stage tumour detected by US-alone. The AFP→US method would have yielded an additional 27.8 YLG ($13,000/YLG compared with 38.9 YLG ($21,000/YLG for US-alone. Screening by US-alone would incur an additional $114,000 per extra early-tumour detected compared with AFP→US and $41,000 per extra YLG. Conclusions: Although US-alone HCC screening might have yielded more YLG than AFP→US, the reduced costs of the AFP→US method could expand access to HCC screening in resource

  2. The efficacy of modified docetaxel-cisplatin-5-fluorouracil regimen as first-line treatment in patients with alpha-fetoprotein producing gastric carcinoma

    Directory of Open Access Journals (Sweden)

    Yakup Bozkaya

    2017-05-01

    Full Text Available Alpha-fetoprotein producing gastric carcinoma (AFP-PGC is a rare cancer for which limited data on the clinicopathological features and treatment modalities exist. The aim of this study was to compare the efficacy of modified docetaxel-cisplatin-5-fluorouracil (mDCF as the first-line chemotherapy regimen in metastatic AFP-PGC and non-AFP-PGC. The patients diagnosed with metastatic gastric cancer who were given mDCF as first-line therapy were retrospectively reviewed. The patients with a basal serum AFP level over 9 ng/ml were defined as AFP-PGC patients. In total, 169 patients (34 with AFP-PGC and 135 with non-AFP-PGC were included in this study. AFP-PGC patients had more liver metastases than non-AFP-PGC patients (p < 0.001. A decrease in basal AFP levels after three cycles of chemotherapy was significantly different in AFP-PGC group (p = 0.001.Overall disease control rate was 79.4% (partial response [PR] - 44.1%, stable disease [SD] - 35.3%, and 82.2% (complete response - 3%, PR - 36.2%, SD - 43% in AFP-PGC and non-AFP-PGC patients, respectively. There was no difference between AFP-PGC and non-AFP-PGC groups in overall and progression-free survival rates (11.3 versus 11.4 months and 7.7 versus 7.1 months, respectively. Rates of grade 3-4 hematologic toxicity were 8.8% and 6.7% for neutropenia in AFP-PGC and non-AFP-PGC group, respectively and 5.9% and 7.4% for anemia. In conclusion, mDCF regimen is well-tolerated with acceptable toxicity outcomes in both AFP-PGC and non-AFP-PGC patients. A statistically significant decrease in AFP levels after mDCF regimen indicate that AFP might be considered as a supplemental marker of response to mDCF chemotherapy in AFP-PGC patients. However, further prospective clinical trials are required in this area.

  3. Influence of the. cap alpha. -,. beta. -, and. gamma. -subunits of the energy-transducing adenosine triphosphatase from Micrococcus lysodeikticus in the immunochemical properties of the protein and in their reconstitution studied by a radioimmunoassay method

    Energy Technology Data Exchange (ETDEWEB)

    Larraga, V; Mollinedo, F; Rubio, N; Munoz, E [Unidad de Biomembranas, Instituto de Inmunologia y Biologia Microbiana, Madrid (Spain)

    1981-03-01

    A sensitive radioimmunoassay was developed for the energy-transducing adenosine triphosphatase (F/sub 1/-ATPase, EC 3.6.1.3) of Micrococcus lysodeikticus and the assay was extended to the ..cap alpha..-, ..beta..-, and ..gamma..-subunits of the enzyme. These subunits were isolated and cross-reactions studied.

  4. Characterization and application of a radioimmunoassay for reduced, carboxymethylated human luteinizing hormone. cap alpha. -subunit. [/sup 125/I tracer technique

    Energy Technology Data Exchange (ETDEWEB)

    Keutmann, H.T.; Beitins, I.Z.; Johnson, L.; McArthur, J.W.

    1978-12-01

    We have established a double antibody RIA using a rabbit antiserum prepared against reduced, carboxymethylated (RCXM) human LH ..cap alpha..-subunit, with RCXM-..cap alpha.. as tracer and standard. This antiserum did not cross-react with any native gonadotropins or subunit, and reacted only weakly with RCXM-..cap alpha... A tryptic digest of RCXM ..cap alpha..-subunit was completely reactive, while chymotryptic digestion abolished all immunoreactivity. By testing with separate tryptic fragments, the recognition site could be localized to a segment close to the amino-terminus of the peptide chain. When applied to measurement of serum and urine, an immunoreactive species, parallel to RCXM ..cap alpha..-subunit by serial dilution, was found in concentrations of 1-2 ng/ml in serum and 3-4 ng/ml in urine. Similar levels of the immunoreactive component were found in conditions of elevated gonadotropins (e.g. pregnancy) as well as gonadotropin deficiency (panhypopituitarism and Kallmann's syndrome). After stimulation with LHRH, no rise was noted at times up to 6 h despite the fact that both LH and LH-..cap alpha.. were elevated. The data indicate that the sequence-specific antiserum may be detecting an immunoreactive form of ..cap alpha..-subunit of LH whose kinetics of appearance and disappearance differs from those of the native subunit.

  5. A novel anti-alpha-fetoprotein single-chain variable fragment displays anti-tumor effects in HepG2 cells as a single agent or in combination with paclitaxel.

    Science.gov (United States)

    Ji, Xiaonan; Shen, Yanli; Sun, Hao; Gao, Xiangdong

    2016-08-01

    Human hepatocellular carcinoma (HCC) has a high rate of tumor recurrence and metastasis, resulting in shortened survival time. The function of alpha-fetoprotein (AFP) as a regulatory factor in the growth of HCC cells has been well defined. The aim of this study was to investigate the use of a novel AFP-specific single-chain variable fragment that blocked AFP and inhibited HCC cell growth. The results indicated that the anti-AFP single-chain variable fragment (scFv) induced growth inhibition of AFP-expressing HCC cell lines in vitro through induction of G1 cell cycle arrest and apoptosis. The mechanism of apoptosis probably involved with blocking AFP internalization and regulation of the PTEN/PI3K/Akt signaling network. Moreover, the anti-AFP-scFv also effectively sensitized the HepG2 cells to paclitaxel (PTX) at a lower concentration. The combination effect of PTX and anti-AFP-scFv displayed a synergistic effect on HepG2 cells both in vitro and in vivo. Our results demonstrated that targeting AFP by specific antibodies has potential immunotherapeutic efficacy in human HCC.

  6. Development of on-chip fully automated immunoassay system "μTASWako i30" to measure the changes in glycosylation profiles of alpha-fetoprotein in patients with hepatocellular carcinoma.

    Science.gov (United States)

    Kurosawa, Tatsuo; Watanabe, Mitsuo

    2016-12-01

    Glycosylation profiles significantly change during oncogenesis. Aberrant glycosylation can be used as a cancer biomarker in clinical settings. Different glycoforms can be separately detected using lectin affinity electrophoresis and lectin array-based methods. However, most methodologies and procedures need experienced technique to perform the assays and expertise to interpret the results. To apply glycomarkers for clinical practice, a robust assay system with an easy-to-use workflow is required. Wako's μTASWako i30, a fully automated immunoanalyzer, was developed for in vitro diagnostics based on microfluidic technology. It utilizes the principles of liquid-phase binding assay, where immunoreactions are performed in a liquid phase, and electrokinetic analyte transport assay. Capillary electrophoresis on microfluidic chip has enabled the detection of different glycoform types of alpha-fetoprotein (AFP), a serum biomarker for hepatocellular carcinoma. AFP with altered glycosylation can be separated based on the reactivity to Lens culinaris agglutinin on electrophoresis. The glycoform AFP-L3 was reportedly more specific in hepatocellular carcinoma. This assay system can provide a high sensitivity and rapid results in 9 min. The test results for ratio of AFP-L3 to total AFP using μTASWako i30 are correlated with those of conventional methodology. The μTASWako assay system and the technology can be utilized for glycosylation analysis in the postgenomic era. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Homogeneous immunoassay for the cancer marker alpha-fetoprotein using single wavelength excitation fluorescence cross-correlation spectroscopy and CdSe/ZnS quantum dots and fluorescent dyes as labels

    International Nuclear Information System (INIS)

    Wang, Jinjie; Liu, Heng; Huang, Xiangyi; Ren, Jicun

    2016-01-01

    The article describes sensitive and selective homogeneous immunoassays for the liver cancer biomarker alpha-fetoprotein (AFP) in human serum by using single wavelength excitation fluorescence cross-correlation spectroscopy (SW-FCCS). Both competitive and sandwich immunoassay modes were applied, and AFP served as a model analyte. Fluorescent CdSe/ZnS quantum dots (with a 655 nm emission peak) and the fluorophore Alexa Fluor 488 (520 nm emission) were chosen to label the antibodies in the sandwich mode, and the antibody and the antigen in the competitive mode. Under optimized conditions, the sandwich assay has a linear dynamic range that covers the 20 pM to 5.0 nM concentration range. The competitive assay, in turn, extends from 180 pM to 15.0 nM. The respective detection limits are 20 pM and 180 pM. The method was successfully applied to directly determine AFP in (spiked) clinical samples, and results were in good agreement with data obtained via ELISAs. (author)

  8. Characteristic of expression levels of HepPar-1, alpha-fetoprotein, cytokeratin 7 and 20 by the cells of cholangiocellular cancer in trephine biopsy of the liver

    Directory of Open Access Journals (Sweden)

    V. A. Tumanskiy

    2014-10-01

    Full Text Available Aim. Expression level of immunohistochemical markers such as HepPar-1, AFP, CK7, CK20 and the area of immunopositive cells in cholangiocellular liver cancer, and their differences from hepatocellular carcinoma Methods and results. Histopathological, histochemical and immunohistochemical research of trephine was determined in the liver in 90 patients with biopsy. Among them 53 patients had hepatocellular, 36 – сholangiocellular liver cancer, 1 patient had mixed hepato-cholangiocellular carcinoma. Level of expression of immunohistochemical markers of tumor cells and the area of immunopositive tumor cells in the tumor was determined by photo-digital morphometry. It was established that expression of α-fetoprotein is determined in 47.22% of patients with cholangiocellular liver carcinoma in tumor cells, when AFP-immunopositive cells represent 17,25 ± 9,67% of the total area of tumor cells. Positive expression of HepPar-1 cells in cholangiocellular liver cancer wasn’t detected (unlike hepatocellular carcinoma, when cytoplasmic expression of HepPar-1 by tumor hepatocytes is determined in 92.45% of cases. Expression of СK7 by cholangiocellular carcinoma cells was observed in 97.22% of patients, and the expression of CK20 – in 45.29% patients, immunopositive cells represent 43,55 ± 9,93% and 50,28 ± 16,35% of the tumor area, respectively. Medium strength correlation was determined between the level of AFP and CK7 expression by tumor cells in cholangiocellular carcinoma. Direct strong bond was determined between level of AFP and CK20 expression. Negative weak correlation was determined between the level of CK7 and CK20.

  9. The role of unexplained high serum alpha-fetoprotein (AFP and human chorionic gonadotropin (hCG levels in the second trimester to determine poor obstetric outcomes

    Directory of Open Access Journals (Sweden)

    Hümeyra Öztürk

    2014-09-01

    Full Text Available Objective: To investigate the relationship between gestational complications and high levels of maternal serum alfa-fetoprotein (MSAFP and/or beta human chorionic gonadotropin (hCG and to determine whether these markers are effective predictors of poor pregnancy outcomes. Materials and Methods: In this study, we enrolled a total of 679 women at 15-20 gestational weeks with MSAFP and hCG below or above 2.0 multiples of the median (MoM; of those, 200 women with normal MSAFP and hCG MoM formed the control group. Pre-eclampsia, intrauterine growth retardation (IUGR, preterm labor, preterm delivery, placental abruption, placenta previa, placenta accreta, preterm premature rupture of the membranes (PPROM, intrauterine fetal death, as well as neonatal and perinatal morbidity rates were evaluated. Results: A significant relationship was found between adverse pregnancy outcomes and abnormal elevation of hCG and AFP levels in the second trimester. In cases of isolated elevation of hCG, preeclampsia and preterm labor/spontaneous preterm birth rate were slightly higher than in the control group (p=0.043, p=0.015, while IUGR, PPROM, placental abruption, and intrauterine fetal death rates were all similar (p=0.063, p=0.318, p=1.00, p=0.556. In case having an elevation in both markers, increased rate of obstetric complications have been observed. A significant relationship was found between the high levels of maternal serum AFP and hCG MoM and poor pregnancy outcomes like preeclampsia, IUGR, PPROM, intrauterine fetal death (p=0.003, p=0.001, p=0.040, p=0.006. Conclusion: To our knowledge, up to now, no definitive follow-up and treatment protocols have been established for patients at increased risk. In light of these findings, it is recommended to inform and educate patients about the most likely signs and symptoms of complications, to make more often antenatal visits, to perform more frequent ultrasound examination (fetal growth, AFI, etc., NST, arterial

  10. Expression of human α-fetoprotein in yeast

    International Nuclear Information System (INIS)

    Yamamoto, Ritsu; Sakamoto, Takashi; Nishi, Shinzo; Sakai, Masaharu; Morinaga, Tomonori; Tamaoki, Taiki

    1990-01-01

    Human α-fetoprotein (AFP) was expressed in Saccharomyces cerevisiae, with a plasmid containing the cDNA sequence for human AFP fused with the rat AFP signal peptide. The recombinant AFP was purified from the yeast lysate by DEAE-cellulose and immunoaffinity chromatography. The amino acid composition and the molecular weight of the recombinant AFP were similar to those of hepatoma AFP. N-terminal amino acids sequence analysis indicated that the signal peptide had been processed. The recombinant and hepatoma AFP reacted identically in Ouchterlony immunodiffusion and radioimmunoassay tests. These observations indicated that the yeast recombinant protein had the properties of native AFP

  11. New Solid Phases for Estimation of Hormones by Radioimmunoassay Technique

    International Nuclear Information System (INIS)

    Sheha, R.R.; Ayoub, H.S.M.; Shafik, M.

    2013-01-01

    The efforts in this study were initiated to develop and validate new solid phases for estimation of hormones by radioimmunoassay (RIA). The study argued the successful application of different hydroxy apatites (HAP) as new solid phases for estimation of Alpha fetoprotein (AFP), Thyroid Stimulating hormone (TSH) and Luteinizing hormone (LH) in human serum. Hydroxy apatites have different alkali earth elements were successfully prepared by a well-controlled co-precipitation method with stoichiometric ratio value 1.67. The synthesized barium and calcium hydroxy apatites were characterized using XRD and Ftir and data clarified the preparation of pure structures of both BaHAP and CaHAP with no evidence on presence of other additional phases. The prepared solid phases were applied in various radioimmunoassay systems for separation of bound and free antigens of AFP, TSH and LH hormones. The preparation of radiolabeled tracer for these antigens was carried out using chloramine-T as oxidizing agent. The influence of different parameters on the activation and coupling of the used apatite particles with the polyclonal antibodies was systematically investigated and the optimum conditions were determined. The assay was reproducible, specific and sensitive enough for regular estimation of the studied hormones. The intra-and inter-assay variation were satisfactory and also the recovery and dilution tests indicated an accurate calibration. The reliability of these apatite particles had been validated by comparing the results that obtained by using commercial kits. The results finally authenticates that hydroxyapatite particles would have a great potential to address the emerging challenge of accurate quantitation in laboratory medical application

  12. Irregular vascular pattern by contrast-enhanced ultrasonography and high serum Lens culinaris agglutinin-reactive fraction of alpha-fetoprotein level predict poor outcome after successful radiofrequency ablation in patients with early-stage hepatocellular carcinoma.

    Science.gov (United States)

    Takada, Hitomi; Tsuchiya, Kaoru; Yasui, Yutaka; Nakakuki, Natsuko; Tamaki, Nobuharu; Suzuki, Shoko; Nakanishi, Hiroyuki; Itakura, Jun; Takahashi, Yuka; Kurosaki, Masayuki; Asahina, Yasuhiro; Enomoto, Nobuyuki; Izumi, Namiki

    2016-11-01

    Radiofrequency ablation (RFA) is considered the most effective treatment for early-stage hepatocellular carcinoma (HCC) patients unsuitable for resection. However, poor outcome after RFA has occasionally been reported worldwide. To predict such an outcome, we investigated imaging findings using contrast-enhanced ultrasonography (CEUS) with Sonazoid and serum tumor markers before RFA. This study included 176 early-stage HCC patients who had initially achieved successful RFA. Patients were examined using CEUS; their levels of alpha-fetoprotein (AFP), Lens culinaris agglutinin-reactive fraction of AFP (AFP-L3), and des-gamma-carboxy prothrombin before RFA were measured. Sonazoid provided parenchyma-specific contrast imaging and facilitated tumor vascular architecture imaging through maximum intensity projection (MIP). Kaplan-Meier analysis examined cumulative rates of local tumor progression, intrasubsegmental recurrence, and survival; factors associated with these were determined with Cox proportional hazards analysis. Local tumor progression (n = 15), intrasubsegmental recurrence (n = 46), and death (n = 18) were observed. Irregular pattern in MIP classification and serum AFP-L3 level (>10%) before RFA were identified as independent risk factors for local tumor progression and intrasubsegmental recurrence. These two factors were independently associated with poor survival after RFA (irregular pattern in MIP: hazard ratio, (HR) = 8.26; 95% confidence interval, (CI) = 2.24-30.3; P = 0.002 and AFP-L3 > 10%: HR = 2.94; 95% CI = 1.09-7.94; P = 0.033). Irregular MIP pattern by CEUS and high level of serum AFP-L3 were independent risk factors for poor outcome after successful RFA. The Patients with these findings should be considered as special high-risk group in early-stage HCC. © 2016 The Authors. Cancer Medicine published by John Wiley & Sons Ltd.

  13. Radioimmunoassay of plasma progesterone

    Energy Technology Data Exchange (ETDEWEB)

    Langer, L; Veleminsky, J [Institute of Clinical Endocrinology, Lubochna (Czechoslovakia); Hampl, R; Starka, L [Vyzkumny Ustav Endokrinologicky, Prague (Czechoslovakia); Holan, J [Comenius Univ., School of Medicine, Martin (Czechoslovakia). Dept. of Physics and Nuclear Medicine

    1978-06-30

    A simple modification of plasma progesterone radioimmunoassay is described. 11..cap alpha..-Hydroxyprogesterone hemisuccinate - BSA conjugate was used as an immunogen. (1,2,6,7-H-3) Progesterone specific radioactivity 82 Ci.mmol/sup -1/ was purchased from Radiochemical Centre Amersham (England). The method has been applied for the analysis of more than 2000 plasma samples. The typical fluctuation of progesterone in plasma during the menstrual cycle, using data obtained with this method is illustrated. The reliability criteria of the method are given.

  14. Radioimmunoassays for tumor diagnosis

    International Nuclear Information System (INIS)

    Dressler, J.

    1983-01-01

    Aside from imaging techniques several (radio-)immunological analyses are used for tumor diagnosis. Oncofetal antigens, for instance the carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP), have become the most important substances for many malignancies. However, nearly all of the so-called tumor markers are not suitable for early diagnosis or screening either because of low sensitivity or low tumor specifity. On the other hand follow-up measurements give a very sensitive index of the success of treatment and may indicate tumor progression when other signs are still not present. In some carcinomas and under some clinical circumstances tumorspecific markers are available and mandatory for detection and/or staging: AFP in hepatoma, acid phosphatase in metastasizing carcinoma of the prostate and serum thyreoglobulin in differentiated thyroid cancer. (orig.) [de

  15. Preoperative serum levels of ca 72-4, cea, ca 19-9, and Alpha-fetoprotein in patients with gastric cancer Níveis séricos pré-operatórios de CA 72-4, CEA, CA 19-9 e Alfa-fetoproteína em pacientes com câncer gástrico

    Directory of Open Access Journals (Sweden)

    Rejane Mattar

    2002-06-01

    Full Text Available INTRODUCTION: The clinical importance of preoperative serum levels of CA 72-4, carcinoembryonic antigen (CEA, CA 19-9, and alpha-fetoprotein (AFP was prospectively evaluated in 44 patients with gastric cancer. METHOD: The serum tumor marker levels were determined by commercial radioimmunoassay kits. Positivity for CA 72-4 (>4 U/mL, CEA (>5 ng/mL, CA 19-9 (>37 U/mL, and AFP (>10 ng/mL were correlated according to the stage, histology, and lymph node metastasis. RESULTS AND DISCUSSION: CA 72-4 showed a higher positivity rate for gastric cancer (47.7% than CEA (25%, CA 19-9 (25%, and AFP (0%. The combination of CA 72-4 with CEA and CA 19-9 increased the sensitivity to 61.4%. The positivity rates of CA 72-4 in patients at stages I and II (initial disease and in patients at stages III and IV (advanced disease were 9% and 60.6%, respectively (P INTRODUÇÃO: A importância clínica dos níveis séricos pré-operatórios de CA 72-4, antígeno carcinoembrionário (CEA, CA 19-9 e alfa-fetoproteína (AFPfoi avaliada prospectivamente em 44 pacientes com câncer gástrico. MÉTODOS: Os marcadores tumorais foram quantificados com o emprego de kits comerciais de radioimunoensaio. A positividade dos marcadores, CA 72-4 (>4 U/ml, CEA (>5 ng/ml, CA 19-9 (>37 U/ml e AFP (>10 ng/ml, foi correlacionada com o estágio da doença, a histologia do tumor e comprometimento de linfonodo. RESULTADOS E DISCUSSÃO: O marcador CA 72-4 apresentou maior positividade para o câncer gástrico (47,7% que CEA (25%, CA 19-9 (25% e AFP (0%. A associação de CA 72-4, CEA e CA 19-9 aumentou a sensibilidade para 61,4%. A positividade do CA 72-4 nos pacientes com estágios I e II (Doença Inicial e nos pacientes com estágios III e IV (Doença Avançada foi de 9 e 60,6%, respectivamente (p<0,005. Não foi observada correlação entre os níveis séricos de CEA e CA 19-9 com o estágio do câncer gástrico. O CA 72-4 apresentou tendência de sugerir comprometimento de linfonodo, mas n

  16. Radioimmunoassay apparatus

    International Nuclear Information System (INIS)

    1981-01-01

    Apparatus for performing a quantitative radioimmunoassay comprising: a substantially spherical bead for carrying an antibody and a gripper for gripping said bead, said gripper comprising an integrally formed unit having a single elongate handle portion and a plurality of resilient fingers arranged at the base of the handle so that when said bead is secured within said fingers, said bead may be freely rotated about any diametric axis of the bead. In particular the invention relates to an apparatus for a two site immunoradiometric assay for serum ferritin in human blood samples. (author)

  17. Radioimmunoassay for determination of tumor markers in the diagnosis of rectal cancer recurrences

    International Nuclear Information System (INIS)

    Ozhiganov, E.L.; Kuznetsova, L.F.

    1991-01-01

    The levels of tumor markers were determined in patients with rectal cancer recurrences by radioimmunoassay. An increase in a CEA level was observed most frequently. An increase in the levels of α-fetoprotein, ferritin and β 2 -microglobulin was observed. It was shown that the most specific and effective diagnostic test of rectal cancer recurrences was the determination of a CEA level

  18. The sandwich-type electrochemiluminescence immunosensor for {alpha}-fetoprotein based on enrichment by Fe{sub 3}O{sub 4}-Au magnetic nano probes and signal amplification by CdS-Au composite nanoparticles labeled anti-AFP

    Energy Technology Data Exchange (ETDEWEB)

    Zhou Hankun [State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering of Ningbo University, Ningbo 315211 (China); Gan Ning, E-mail: ganning@nbu.edu.cn [State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering of Ningbo University, Ningbo 315211 (China); Li Tianhua; Cao Yuting; Zeng Saolin [State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering of Ningbo University, Ningbo 315211 (China); Zheng Lei, E-mail: nfyyzl@163.com [Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515 (China); Guo Zhiyong [State Key Laboratory Base of Novel Functional Materials and Preparation Science, Faculty of Material Science and Chemical Engineering of Ningbo University, Ningbo 315211 (China)

    2012-10-09

    Highlights: Black-Right-Pointing-Pointer Sandwich immunoreaction, testing a large number of samples simultaneously. Black-Right-Pointing-Pointer The magnetic separation and enrichment by Fe{sub 3}O{sub 4}-Au magnetic nano probes. Black-Right-Pointing-Pointer The amplification of detection signal by CdS-Au composite nanoparticles labeled anti-AFP. Black-Right-Pointing-Pointer Almost no background signal, which greatly improve the sensitivity of detection. - Abstract: A novel and sensitive sandwich-type electrochemiluminescence (ECL) immunosensor was fabricated on a glassy carbon electrode (GCE) for ultra trace levels of {alpha}-fetoprotein (AFP) based on sandwich immunoreaction strategy by enrichment using magnetic capture probes and quantum dots coated with Au shell (CdS-Au) as the signal tag. The capture probe was prepared by immobilizing the primary antibody of AFP (Ab1) on the core/shell Fe{sub 3}O{sub 4}-Au nanoparticles, which was first employed to capture AFP antigens to form Fe{sub 3}O{sub 4}-Au/Ab1/AFP complex from the serum after incubation. The product can be separated from the background solution through the magnetic separation. Then the CdS-Au labeled secondary antibody (Ab2) as signal tag (CdS-Au/Ab2) was conjugated successfully with Fe{sub 3}O{sub 4}-Au/Ab1/AFP complex to form a sandwich-type immunocomplex (Fe{sub 3}O{sub 4}-Au/Ab1/AFP/Ab2/CdS-Au), which can be further separated by an external magnetic field and produce ECL signals at a fixed voltage. The signal was proportional to a certain concentration range of AFP for quantification. Thus, an easy-to-use immunosensor with magnetic probes and a quantum dots signal tag was obtained. The immunosensor performed at a level of high sensitivity and a broad concentration range for AFP between 0.0005 and 5.0 ng mL{sup -1} with a detection limit of 0.2 pg mL{sup -1}. The use of magnetic probes was combined with pre-concentration and separation for trace levels of tumor markers in the serum. Due to the

  19. Radioimmunoassay of thymosin alpha 1

    International Nuclear Information System (INIS)

    Goldstein, A.L.; McClure, J.E.

    1983-01-01

    The present invention relates to an immunoassay for thymosin α 1 , a heat stable, acidic polypeptide composed of 28 amino acid residues. This immunopotentiating hormone is a component of thymosin fraction 5 and also has been found to be present in the blood of mammalian subjects. The immunogen utilized to prepare the antibody for the instant assay is readily obtained by covalently bonding thymosin α 1 to a conventional immunological carrier material. Thymosin α 1 may be radioisotopically labelled with tritium, carbon 14 or iodine 125

  20. Progesterone radioimmunoassay

    International Nuclear Information System (INIS)

    Allen, R.M.; Redshaw, M.R.

    1981-01-01

    This patent claims a radioimmunoassay for progesterone, which comprises contacting, in an acidic medium a sample of liquid with a predetermined amount of antibodies raised against a progesterone-protein complex, the protein being attached to the 11-position of progesterone by means of a bridging group and with a predetermined amount of a progesterone derivative having an iodinatable group attached to its 3-position by means of a bridging group, the iodinatable group being iodinated with one or more atom(s) of a radioisotope of iodine, separating the steroid bound in the resulting antibody-antigen complex from the free steroid and measuring the radioactivity of the free steroid component or of the antibody-antigen complex. Sufficient sensitivity has been achieved to enable a progesterone assay to be carried out directly on a sample of biological fluid, such as serum, plasma, urine or milk. (U.K.)

  1. Radioimmunoassay in clinical practice

    Energy Technology Data Exchange (ETDEWEB)

    Ametov, A S

    1982-01-01

    A wide application of radioimmunoassay in clinical practice is shown. The main theoretical aspects of radioimmunoassay and the fields of application in clinical practice - endocrinology, oncology, allergology, cardiology, pharmacology, pediatrics, hematology, obstetrics and gynecology, are presented.

  2. Steroid radioimmunoassays

    International Nuclear Information System (INIS)

    Shinde, Y.; Hacker, R.R.; Ntunde, B.; Smith, V.G.

    1981-01-01

    An estrogen radioimmunoassay was used to study the problem of blanks in steroid assays. Negligible binding (1.5 percent) in the non-antibody tubes prevailed throughout the study. The assay was validated using accepted procedures. Both water and solvent blanks had estrogen concentrations of 7-9 pg/tube. However, neither water nor solvent blanks showed a dose-related response, indicating that they were 'real' blanks. Exogenous estradiol, when added to water and solvent in quantities less than the estimated blank, was not quantitatively recovered. However, exogenous estradiol added to the water solvent in quantities greater than the blank estimate was quantitatively recovered. The sensitivity of the reference standard curve was 6-10 pg/tube, approximately the same as the blank estimate. These results indicated that the estimates of water and solvent blanks were measures of the assay sensitivity. In such circumstances, it is suggested that blank estimates should not be subtracted from sample values. If the blank estimates are high, attention should be directed towards improving the sensitivity of the assay

  3. Trypsin radioimmunoassay

    International Nuclear Information System (INIS)

    Scheithauer, R.; Wolf, F.; Tympner, F.

    1981-01-01

    In 29 patients with suspicion of pancreatic disease standard secretin-pancreozymin-test was performed parallel to trypsin determination by radioimmunoassay before and after stimulation with secretin. Mean serum trypsin in normal subjects was 175 ng BIT/ml (range 90-250), the maximum after stimulation being 20 minutes after secreting injection (range 110-550). Preliminary normal values are 270 ng BIT/ml for basal concentration and 650 ng BIT/ml for 20 min after secretin stimulation. In the group of normals there was no case of misinterpretation. In patients with several pathological parameters (n = 10) basal trypsin concentration was increased in 7 cases, the stimulated value was concordant with the definite diagnosis in every case. Significant advantage for diagnostics was derived in patients having had pancreatic diseases before, however being actually normal with respect to standard diagnostic parameters. All these patients revealed increased trypsin concentrations after stimulation and 50% of them showed increased basal values. Equivocal results were seen in patients with endstage pancreatitis as well as in case of obstruction during reduced secretion. (orig.) [de

  4. Tumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    Ainsworth, Mark A; Bendtzen, Klaus; Brynskov, Jørn

    2007-01-01

    To investigate if the combined assessment of anti-infliximab antibodies (Ab) and the degree of TNF-alpha binding capacity (TNF-alpha-BC) afforded by infliximab may predict the response to infliximab treatment in patients with Crohn's disease (CD).......To investigate if the combined assessment of anti-infliximab antibodies (Ab) and the degree of TNF-alpha binding capacity (TNF-alpha-BC) afforded by infliximab may predict the response to infliximab treatment in patients with Crohn's disease (CD)....

  5. Handbook of radioimmunoassay

    International Nuclear Information System (INIS)

    Abraham, G.E.

    1977-01-01

    This handbook provides clear, detailed descriptions of ways to set up radioimmunoassay procedures for a variety of polypeptides and low molecular weight compounds. It covers extensively the subjects of statistical evaluation of radioimmunoassay, instrumentation in radioimmunoassay, and radiation safety in the performance of radioimmunoassay. Related nonconventional methods are also discussed. Contributors to this handbook have presented their own procedures for performing the radioimmunoassay and their rationale for choosing particular reagents and conditions. Emphasis is on providing sufficient information to enable relatively inexperienced immunoassayists to set up assay systems with a minimum of difficulty

  6. Carcinoembryonic antigen radioimmunoassay in hepatic tumor

    International Nuclear Information System (INIS)

    Aburano, Tamio; Tonami, Norihisa; Hisada, Kinichi

    1976-01-01

    Carcinoembryonic antigen (CEA) radioimmunoassay with the sandwich method was performed in addition to both α 1 -fetoprotein (AFP) radioimmunoassay and liver scintigraphy to elevate the diagnostic accuracy of hepatic tumor in nuclear medicine. All of the ten healthy controls and 47 of 52 cases with benign disease showed a CEA titer less than 2.5ng/ml. 78 of 188 cases (41%) of malignant disease showed a titer of over 2.5ng/ml; however most positive cases were metastatic, especially to the liver. In metastatic liver cancer, thirtythree out of 46 cases (72%) showed a strongly positive CEA titer. Over 5ng/ml was taken as the lower limit for predicting metastasis to the liver. On the other hand, in primary liver cancer thirty-two out of 35 cases (91%) showed a strongly positive AFP titer over 200ng/ml, although only one case showed a CEA titer over 5ng/ml. Seven cases (15%) of metastatic liver cancer also showed a strongly positive AFP titer; however six of these positive cases showed a CEA titer over 5ng/ml. In metastatic liver cancer, eleven out of 46 cases (24%) showed no clearcut focal defects on liver scintigram. Nine of these negative cases showed a CEA titer over 5ng/ml, and at subsequent operation metastatic liver lesions were found. The overall diagnostic accuracy for detecting metastatic liver cancer with a combination of both methods was 95%. CEA radioimmunoassay was found to be useful for the elucidation of the nature of focal hepatic lesions in addition to AFP radioimmunoassay, and moreover could be used as an adjunct to liver scintigraphy for the detection of metastatic lesions in the liver. (auth.)

  7. Radioimmunoassays in prenatal genetic diagnosis

    International Nuclear Information System (INIS)

    Santavy, J.; Janouskova, M.; Fingerova, H.; Krikal, Z.

    1981-01-01

    Prenatal medicine strives to reveal hereditary disorders and congenital malformations before delivery. The application of RIA significantly widened the spectrum of available diagnostic possibilities. We first focused our attention on determining alpha-1-fetoprotein in the amniotic fluid and the serum. We used the results of 33 examinations of the amniotic fluid and 100 samples of the blood serum to compile a graph of physiological values during pregnancy. The graph is used in assessing clinical samples in suspect congenital disorders of neural tube closure and other malformations. In the last two years we have tested testosterone determination in the amniotic fluid to ascertain prenatally the fetal sex in early pregnancy. The results were satisfactory and agreed in 70.6%. (author)

  8. Determination of α-fetoprotein levels in maternal serum and its application in prenatal diagnosis

    International Nuclear Information System (INIS)

    Rokos, A.; Jedlickova, B.; Hradil, M.

    1989-01-01

    Alpha-fetoprotein determination is described in the screening of neural tube (NT) disorders. The benefits and constraints are described of the AFP-NT screening and the possibilities are assessed of revealing Down's syndrome fetuses. The use is evaluated of the RIA-test-AFP kit in Czechoslovakia. (E.J.). 1 fig., 3 tabs., 10 refs

  9. Radioimmunoassay for human ACTH

    Energy Technology Data Exchange (ETDEWEB)

    Matsumura, S; Oyama, H; Tenku, A; Horino, M [Kawasaki Medicial School, Kurashiki, Okayama (Japan); Kobayashi, K

    1977-03-01

    A radioimmunoassay method for human adrenocorticotropic hormone (ACTH) is described. The antiserum was produced in a guinea pig by multiple injections of a total of 1 mg of porcine ACTH adsorbed with CM-cellulose and mixed with complete Freund's adjuvant. The antiserum used for this study at a final dilution of 1:500,000 was obtained from a guinea pig after 10 immunizations. A highly purified native ACTH (Li, ..cap alpha.. sub(h)sup(1-39) ACTH) was used as an assay standard and a tracer hormone. Separation of free and bound hormone was achieved by dextran coated charcoal. Extraction of ACTH from plasma samples was performed by Donald's method modified with silicic acid. The antibody appeared to be directed against the C-terminal portion of the hormone molecule and showed no significant reaction with other pituitary hormones (GH, TSH, LH, FSH, Hpr, Oxytocin, Arginine-and Lysine-Vasopressin). Plasma ACTH levels of 5 healthy subjects at nine o'clock averaged 32 +- 8.5 pg/ml (M +- SD). Plasma ACTH concentrations in a case of Sheehan's syndrome and of an untreated adrenogenital syndrome at nine o'clock were less than 20 and 194 pg/ml, respectively. A case of Cushing's syndrome accompanied with bilateral nodular hyperplasia of the adrenal cortex showed diminished diurnal variation and normal levels of plasma ACTH. In contrast, elevated plasma ACTH levels and lack of diurnal rhythm of ACTH secretion were observed in a suspected case of Cushing's disease.

  10. Alfa-fetoprotein secreting ovarian sex cord-stromal tumor

    Directory of Open Access Journals (Sweden)

    Kusum D Jashnani

    2013-01-01

    Full Text Available Ovarian sex cord-stromal tumors are relatively infrequent neoplasms that account for approximately 8% of all primary ovarian tumors. They are a heterogeneous group of neoplasms composed of cells derived from gonadal sex cords (granulosa and Sertoli cells, specialized gonadal stroma (theca and Leydig cells, and fibroblasts. They may show androgenic or estrogenic manifestations. We report such a tumor associated with markedly raised serum alpha-fetoprotein (AFP levels in a young female presenting with a mass and defeminising symptoms. Serum AFP levels returned to normal on removal of tumor.

  11. Alpha-feto protein radioimmunoassay and its values for the diagnosis of hepatoma among normal Japanese population and those with chronic schistosomiasis

    International Nuclear Information System (INIS)

    Lio, Masahiro; Yamada, Hideo; Luchi, Masahiko

    1972-01-01

    Since its first report made by Ablevet al., α-feto protein which appears both in fetal serum and serum in the cases with hepatoma drew attention of hepatologist in order to establish an qualitative method to diagnose the presence of hepatoma. Since conventional liver scintigraphy is not able to make the diagnosis of the nature of lesion which appears as space occupying lesion need for the more qualitative method facilitated the establishment of the method to detect the presence of α-feto protein in serum. α-feto protein in the serum of hepatoma cases was reported to be identical to fetal α-feto protein by the immunological method. At first, assay of the α-feto protein was performed using the precipitation reaction in agar gel and became quite popular among clinical hepatologist. Detection sensitivity of this conventional method, however, is approximately 10 μgr/ml and α-feto protein was detected in 80% of hepatoma patient. However detection percentage of the hepatoma cases associated with chronic schistosomiasis was found to be as poor as 30% which was measured by our group at Kofu municipal Hospital. Because of the frequent incidences of deformed and small liver by scintigraphy among cases with chronic schistosomiasis and these abnormal liver scintigraphy easily lead one to middiagnose the presence of hepatoma, establishment of more sensitive assay method is needed. After the success of purification and crystallization of the α-feto protein by the group of Univ. of Hokkaido, the radioimmunoassay technique was developed in Japan and α-feto-125 Kit is now became available commercially. Initial trial of this new promising and this double antibody technique enabled us to measure 2 mμgr/ml of α-feto protein which was almost 500-1,000 times more sensitive than those of the conventional method.

  12. Solid phase radioimmunoassays

    International Nuclear Information System (INIS)

    Wide, L.

    1977-01-01

    Solid phase coupled antibodies were introduced to facilitate the separation of bound and free labelled ligand in the competitive inhibition radioimmunoassay. Originally, the solid matrix used was in the form of small particles and since then a number of different matrices have been used such as very fine powder particles, gels, paper and plastic discs, magnetic particles and the inside surface of plastic tubes. The coupling of antibodies may be that of a covalent chemical binding, a strong physical adsorbtion, or an immunological binding to a solid phase coupled antigen. New principles of radioimmunoassay such as the solid phase sandwich techniques and the immunoradiometric assay were developped from the use of solid phase coupled antigens and antibodies. The solid phase sandwich techniques are reagent excess methods with a very wide applicability. Several of the different variants of solid phase techniques are suitable for automation. Advantages and disadvantages of solid phase radioimmunoassays when compared with those using soluble reagents are discussed. (orig.) [de

  13. Radioimmunoassay for tetrahydrocortisol

    International Nuclear Information System (INIS)

    Mougey, E.H.

    1978-01-01

    A radioimmunoassay for urinary tetrahydrocortisol (THF) is described. Antibodies were produced in rabbits against a THF-monohemisuccinate:bovine serum albumin conjugate. The radioimmunoassay procedure involves enzymic hydrolysis of the urine, extraction with ethyl acetate, radioimmunoassay, and separation of free from bound steroid with Somogyi reagents. The antibody showed some cross-reaction with tetrahydro substance S (36%) which does not occur in urine in appreciable amounts and with tetrahydrocortisone (10%) which does. An analysis of monkey urine extracts before and after tlc purification revealed that the THF antibody was specific enough to permit assay of urines for THF during stress experiments without a purification step. THF values obtained correlated very highly with 17-hydroxycorticosteroid values on the same urines obtained from monkeys during a chair restraint experience (r = 0.97). Also for comparison pruposes these same urines were assayed for free THF, total cortisol, and free cortisol. The free (unconjugated) steroids showed the greatest percentage increase over basal levels

  14. Radioimmunoassay for biopterin

    International Nuclear Information System (INIS)

    Nagatsu, Toshiharu; Yamaguchi, Tokio; Kato, Takeshi; Sugimoto, Takashi; Matsuura, Sadao.

    1979-01-01

    Specific antibodies against biopterin, neopterin, and 6,7-dimethylpterin were prepared and a new type of radioimmunoassay, to which these pterins were subjected was developed. This new type of radioimmunoassay was used to determine biopterin in human urine. Specific antiserum against biopterin did not crossreact significantly with tetrahydro-biopterin, dihydro-biopterin, neopterin, 6,7-dimethylpterin, pterin, or folic acid and showed 30% binding of biopterinyl-caproyl- ( 125 I ) tyramide at a dilution of 1:800. The antisera against neopterin and dimethylpterin also showed a high specificity and did not cross-react significantly with the other pterins. The sensitivity of the radioimmunoassay was increased about ten-fold by pre-incubating the antiserum with 6,7-dimethylpterinyl-caproyl to remove antibodies against the caproic acid moiety of the biopterin conjugate. The recovery of biopterin or tetrahydro-biopterin added human urine was nearly 100% according to this type of radioimmunoassay, and the biopterin concentrations in the urine that were obtained by means of this type of radioimmunoassay showed a fairly good agreement with the values obtained by means of bioassay. The biopterin contents per milliliter of human urine showed considerable variations depending on the subjects, but those per milligram of creatinine were found to be fairly constant in normal subjects. Therefore, biopterin concentrations per milligram of creatinine in human urine may be a good indicator of biopterin metabolism in clinical chemistry. This new type of radioimmunoassay is simple, highly specific, and reproducible. Therefore, it may be very useful for the screening of atypical phenylketonuria due to biopterin deficiency and also for the study of the pathogenesis of various biopterin metabolic diseases. (J.P.N.)

  15. Radioimmunoassay for biopterin

    Energy Technology Data Exchange (ETDEWEB)

    Nagatsu, T; Yamaguchi, T; Kato, T [Tokyo Inst. of Tech. (Japan); Sugimoto, T; Matsuura, S

    1979-06-01

    Specific antibodies against biopterin, neopterin, and 6,7-dimethylpterin were prepared and a new type of radioimmunoassay, to which these pterins were subjected was developed. This new type of radioimmunoassay was used to determine biopterin in human urine. Specific antiserum against biopterin did not crossreact significantly with tetrahydro-biopterin, dihydro-biopterin, neopterin, 6,7-dimethylpterin, pterin, or folic acid and showed 30% binding of biopterinyl-caproyl- ( /sup 125/I ) tyramide at a dilution of 1:800. The antisera against neopterin and dimethylpterin also showed a high specificity and did not cross-react significantly with the other pterins. The sensitivity of the radioimmunoassay was increased about ten-fold by pre-incubating the antiserum with 6,7-dimethylpterinyl-caproyl to remove antibodies against the caproic acid moiety of the biopterin conjugate. The recovery of biopterin or tetrahydro-biopterin added human urine was nearly 100% according to this type of radioimmunoassay, and the biopterin concentrations in the urine that were obtained by means of this type of radioimmunoassay showed a fairly good agreement with the values obtained by means of bioassay. The biopterin contents per milliliter of human urine showed considerable variations depending on the subjects, but those per milligram of creatinine were found to be fairly constant in normal subjects. Therefore, biopterin concentrations per milligram of creatinine in human urine may be a good indicator of biopterin metabolism in clinical chemistry. This new type of radioimmunoassay is simple, highly specific, and reproducible. Therefore, it may be very useful for the screening of atypical phenylketonuria due to biopterin deficiency and also for the study of the pathogenesis of various biopterin metabolic diseases.

  16. Radioimmunoassay of pancreatic glucagon

    International Nuclear Information System (INIS)

    Nooijen, W.J.

    1979-01-01

    The author presents some of the problems and concepts related to the development of a radioimmunoassay of pancreatic glucagon. A specific derivatization of glucagon for raising specific anti-glucagon antisera is introduced, and special procedures for diminishing the non-specific effect are outlined. (G.T.H.)

  17. Erythropoietin radioimmunoassay studies

    International Nuclear Information System (INIS)

    Garcia, J.F.

    1982-01-01

    A highly sensitive radioimmunoassay capable of measuring erythropoietin concentrations in very small amounts of serum as been developed. After establishing normal human values on a large series of serum samples, serum samples from patients with polycythemia vera and patients with secondary polycythemia were obtained and the erythropoietin concentrations measured

  18. Radioimmunoassay in endocpinology

    International Nuclear Information System (INIS)

    Ametov, A.S.; Torjtsina, L.K.

    1983-01-01

    Radioimmunoassay of the level of various hormones in blood for the diagnosis of thyroid gland diseases, diabetes mellitus, pancreatic gland diseases, adrenal cortex, phophorous-calcium exchange, cerebral-hypophysial deseases, peripheric endocrine glands diseases and others are considered. It is shown that CEA methods implantation in clinical practice permits to obtain a number of new data on mechanism of neurohumoral interrelations of various organs and organism systems

  19. Automation systems for radioimmunoassay

    International Nuclear Information System (INIS)

    Yamasaki, Paul

    1974-01-01

    The application of automation systems for radioimmunoassay (RIA) was discussed. Automated systems could be useful in the second step, of the four basic processes in the course of RIA, i.e., preparation of sample for reaction. There were two types of instrumentation, a semi-automatic pipete, and a fully automated pipete station, both providing for fast and accurate dispensing of the reagent or for the diluting of sample with reagent. Illustrations of the instruments were shown. (Mukohata, S.)

  20. Feto portador de síndrome de turner e tetralogia de fallot associadas à elevação de alfafetoproteína materna Fetal turner syndrome and tetralogy of fallot associated with elevated maternal serum alpha-fetoprotein levels

    Directory of Open Access Journals (Sweden)

    Eduardo Vieira Neto

    1998-06-01

    Full Text Available A síndrome de Turner fetal e suas complicações, a hidropisia e o higroma cístico, podem produzir alteração dos marcadores bioquímicos de soro materno inicialmente utilizados no rastreamento de síndrome de Down e de defeitos de tubo neural (DTN. Os autores relatam o caso de uma gestante de 37 anos, que foi rastreada para síndrome de Down e DTN no início do 2º trimestre. Foi constatado aumento da alfafetoproteína de soro materno (MSAFP e o rastreamento foi considerado positivo para DTN. Foi realizado exame ultra-sonográfico tridimensional, que não demonstrou nenhuma anormalidade fetal ou placentária, caracterizando o caso como elevação idiopática de MSAFP. No 3º trimestre, a gravidez evoluiu com acentuada oligoidrâmnia e alteração do fluxo uteroplacentário, obrigando à instituição de terapia com corticosteróides e parto cesáreo na 34ª semana gestacional. O concepto do sexo feminino foi encaminhado à UTI neonatal, onde foram diagnosticadas tetralogia de Fallot e síndrome de Turner. Esse caso incentivou os autores a rever a literatura sobre marcadores bioquímicos de soro materno na síndrome de Turner e nas malformações cardíacas congênitas. Ao final, propõe-se um protocolo para elevação idiopática de MSAFP.Turner syndrome and its complications, hydrops and cystic hygroma, can produce alterations in maternal serum biochemical markers used in screening for Down's syndrome and neural tube defects (NTD. The authors report the case of a 37-year-old pregnant woman, screened for Down's syndrome and NTD in the second trimester of pregnancy. The maternal serum alpha-fetoprotein (MSAFP level was increased and the test was considered screen positive for NTD. A three-dimensional ultrasound investigation was performed, but no fetal or placental anomalies were found, indicating a case of unexplained increased msafp. In the third trimester severe oligohydramnios and disturbances in uteroplacental arterial circulation

  1. Comparative study of radioimmunoassay dates

    International Nuclear Information System (INIS)

    Venegas Sanchez, Ruth.

    1986-01-01

    The radioimmunoassay is frequently used in clinical chemistry for the concentration determination of several substances like hormones as thyrotropine and thyroxine. In this experiment the dates of tyroxine radioimmunoassay are processed by three methods: a) like the recommendation of the IAEA, b) Dr. G. Chase method, c) according to the provider. The best method was Dr. Chase's. (author)

  2. Automation of radioimmunoassay

    International Nuclear Information System (INIS)

    Yamaguchi, Chisato; Yamada, Hideo; Iio, Masahiro

    1974-01-01

    Automation systems for measuring Australian antigen by radioimmunoassay under development were discussed. Samples were processed as follows: blood serum being dispensed by automated sampler to the test tube, and then incubated under controlled time and temperature; first counting being omitted; labelled antibody being dispensed to the serum after washing; samples being incubated and then centrifuged; radioactivities in the precipitate being counted by auto-well counter; measurements being tabulated by automated typewriter. Not only well-type counter but also position counter was studied. (Kanao, N.)

  3. Radioimmunoassay in microtiter plates

    International Nuclear Information System (INIS)

    Hirschberg, T.; Hirschberg, H.

    1987-01-01

    A radioimmunoassay was performed in the wells of casein-coated microplates employing 125 I-labelled sheep anti-human second antibody. The antigen-antibody complexes were thereafter dislodged from the well walls using detergent sodium dodecyl sulfate (SDS) and the entire contents of the wells were simultaneously absorbed into 48 cellulose acetate-absorbing cartridges. All 48 cartridges were transferred to counting vials and the radioactivity determined by standard gamma counting techniques. The particular advantage of the method described here is the ease with which the supernatants can be collected and transferred to counting vials with minimal handling of radioactive samples. 6 refs.; 1 figure; 1 table

  4. Radioimmunoassay for Melatonin

    International Nuclear Information System (INIS)

    Tapp, E.; Skinner, R.G.; Phillips, V.

    1980-01-01

    A radioimmunoassay for melatonin has been developed and used to measure the level of melatonin of male and post-menopausal female patients coming to operation for benign and malignant conditions. The amount of melatonin in the serum of the females was considerably lower than that in males. No difference could be found between patients suffering from benign and malignant conditions. A patient with a non-parenchymatous pineal tumour had considerably lower levels in the serum at three months after surgery and radiotherapy. A further month later melatonin could not be found in samples of serum taken over a 24-hour period. (author)

  5. Radioimmunoassay of steroid hormone

    International Nuclear Information System (INIS)

    Murakami, Tadashi

    1975-01-01

    Low acid pepsin treated gamma-globulin was applied to ammonium sulfate salting out method, which was a method to separate bound fraction from free one in radioimmunoassay of steroid hormone, and the effect of the separation and the standard curve were examined. Pepsin treated gamma-globulin was prepared in pH 1.5 to 5.5 and then the pepsin was completely removed. It had an effect to accelerate the precipitation in radioimmunoassay of steroid hormone labelled with 3 H. The effect of pepsin treated gamma-globulin to adhere free steroid hormone and to slat out bound one was compared with that of human gamma-globulin. Pepsin treated gamma-globulin, which was water soluble, could easier reach its optimal concentration, and the separation effect was better than human gamma-globulin. The standard curve of it was steeper, particularly in a small dose, and the reproducibility was also better. It could be applied not only to aldosterone and DOC, but also to the steroid hormones, such as progesterone and DHEA, and it seemed suitable for routine measurement method. (Kanao, N.)

  6. Radioimmunoassay of oxytocin

    International Nuclear Information System (INIS)

    Dawood, M.Y.; Raghavan, K.S.; Pociask, C.

    1978-01-01

    The evaluation of a radioimmunoassay of oxytocin is described. The method involved careful collection and transportation of blood at 4 0 C, acidification of the plasma, extraction with Fuller's earth and radioimmunoassay using antisera raised in rabbits immunized against oxytocin conjugated to bovine serum albumin and 125 I-labelled oxytocin. The antisera showed insignificant cross-reaction with a variety of small peptides including vasopressin and vasotocin. The limit of detection of the assay was 2.5 pg with intra-assay and interassay coefficients of variation of 7 to 15% and 12 to 18% respectively. Seventy-seven per cent (88 out of 116) of the pregnant women tested had detect-able maternal plasma oxytocin. Serial samples of maternal plasma showed a significant increase in oxytocin from the first to the second stage of labour and a significant decrease in the third stage. Oxytocin concentrations in the umbilical arterial plasma were significantly higher in patients in labour. The significance of these findings is discussed. (author)

  7. Radioimmunoassay of platelet proteins

    International Nuclear Information System (INIS)

    Pepper, D.S.

    1987-01-01

    The radioimmunoassay of platelet-specific proteins has proven to be an excellent way of monitoring platelet activation in vivo. In contrast to earlier methods such as aggregometry, which has been the major tool used in the evaluation of antiplatelet drugs, the RIAs are capable of working with samples which have been subjected to physiological conditions such as haematocrit, oxygen tension, shear rate and ionized calcium concentration. Also, in contrast to aggregometry, no choice of agonist is necessary. Thus, for the first time it has been possible to monitor the effects of therapeutic intervention with drugs upon the platelet release reaction in vivo. It seems reasonable to equate the release reaction in vivo with activation in vivo, though the stimuli necessarily remain unknown. Nevertheless, the fact that a significant number of the compounds mentioned in Table 3 are indeed capable of reducing platelet activation in vivo and that this effect can be measured objectively is a major step forward in our understanding of platelet pharmacology. Two important goals remain to be achieved, however, the establishment of nonhuman animal models for the evaluation of newer compounds in vivo and longer-term goal of proving in the clinical setting the relevance or otherwise of platelet activation per se to the clinical outcome of a particular disease. In this respect, the availability of accurate, reliable and specific radioimmunoassays has a central role

  8. Diagnostic Utility of Alpha-fetoprotein and Des-gamma ...

    African Journals Online (AJOL)

    2018-02-07

    ] High DCP levels also reflect .... using the multiple logistic regression models. P < 0.05 was considered statistically significant. .... that C-reactive protein has significant diagnostic power in AFP-negative, HBV-related HCC.[23].

  9. Diagnostic Utility of Alpha-fetoprotein and Des-gamma ...

    African Journals Online (AJOL)

    2017-12-05

    Dec 5, 2017 ... reports on the clinical characteristics of hepatocellular carcinoma in relation to the combination of the ... This is an open access article distributed under the terms of the Creative Commons .... Microsoft Excel and then transferred to STATA 10 software ..... Sohn W, Paik Y, Kim J, Kwon C, Job J, Cho J, et al.

  10. Radioimmunoassay of anabolic steroids

    International Nuclear Information System (INIS)

    Hampl, R.; Stranska, I.; Starka, L.; Picha, J.; Chundela, B.

    1978-01-01

    Alternative antisera against 17 α-methyltestosterone and 19-nortestosterone were raised and used for radioimmunoassay of anabolic steroids. Tritiated compounds were used as radioligands. The RIA method suitable for doping control is proposed for 17 α-alkylated anabolic steroids in both plasma and urine, using qoat antiserum against methyltestosterone-3-carboxymethyloxime-BSA. Sensitivity of the method was expressed as least amount of nonradioactive methandienone which, when added to normal urine or plasma, caused statistically significant decrease of measured supernatant radioactivity at 99% level. The amounts from 50 to 500 pg were tested, each in eight parallel determinations. The amounts of 100 pg for plasma and 200 pg for urine met these criteria. The respective coefficients of variation did not depend on the amount of steroid added in this range. They averaged 4.60% for plasma and 4.95% for urine, respectively. (T.I.)

  11. Radioimmunoassay for jasmonic acid

    Energy Technology Data Exchange (ETDEWEB)

    Knoefel, H.D.; Brueckner, C.; Kramell, R.; Sembdner, G.; Schreiber, K. (Akademie der Wissenschaften der DDR, Halle/Saale. Inst. fuer Biochemie der Pflanzen)

    1984-01-01

    A radioimmunoassay (RIA) for the natural plant growth regulator jasmonic acid (JA) was developed. The antiserum was raised in rabbits against (+-)-JA linked to bovine serum albumin. As tracer tritium labelled (+-)-JA (spec. act. 7.4 x 10/sup 9/ Bq x mmol/sup -1/) was used. Cross-reactivity studies with compounds structurally related to JA demonstrated the antiserum to be specific for JA, abscisic acid normally present in the same extract does not interfer. The RIA has a detection limit of 2 ng (-)-JA methylester, a measuring range 2-200 ng, and no extensive purification is required prior to estimation. Therefore, in JA analysis the RIA described is superior to GC, HPLC, and bioassay. This new method has been employed for studies on the distribution of JA in different plant organs of the broad bean, Vicia faba L.

  12. Radioimmunoassay of synthetic steroids

    Energy Technology Data Exchange (ETDEWEB)

    Raynaud, J -P; Bucourt, R; Salmon, J

    1975-12-01

    The sensitivity of a radioimmunoassay depends on the intrinsic association constant of the interaction between ligand and antibody. Its specificity depends on the position of the chain which forms the link with the antigen. Thus, an antibody specific of estradiol has been obtained by coupling estradiol to albumin via a chain at position 7. For synthetic steroids the structure of which is sufficiency different from that of natural hormones, the requirements for a sensitive assay method not involving chromatography are simply maximum affinity and positioning of the couple at a site which does not undergo metabolic attack. These criteria were used to develop assays for R 2858 and R 2453 which obviate the need to administer radioactive product in clinical pharmacology. Cross-reaction with structural analogs may be used to assay competitors. Thus, R 2323 antibody, highly specific for endogenous steroids, may be used to assay other trienes such as R 1697 (trenbolone) and R 2010 (norgestrienone).

  13. Delta antibody radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Kselikova, M; Urbankova, J

    1985-11-15

    The principle and procedure are described of the radioimmunoassay of delta antibody (delta-Ab) using the ABBOTT ANTI-DELTA kit by Abbott Co. A description is given of the kit, the working procedure and the method of evaluation. The results are reported of the incidence of delta-Ab in sera of patients with viral hepatitis B, in haemophiliacs, carriers of the hepatitis B virus surface antigen (HBsAg) and blood donors. The presence was detected of delta-Ab in one HBsAg carrier. The necessity is emphasized of delta-Ab determinations in the blood of donors in view of the antibody transfer with blood and blood preparations.

  14. Radioimmunoassay of erythropoietin

    Energy Technology Data Exchange (ETDEWEB)

    Goldwasser, E; Sherwood, J B [Chicago Univ., IL (USA). Dept. of Biochemistry

    1981-07-01

    A brief review of the historical development of the radioimmunoassay (RIA) for serum erythropoietin is given. It has been shown that there is reasonable agreement between the results obtained by RIA and those obtained by the previously used bioassay. By RIA, a mean normal serum titre of 18 mu/ml erythropoietin has been determined in a study of 445 individuals. Serum erythropoietin results for patients with polycythaemia vera have not been shown to be significantly different from normal values but in patients with secondary polycythaemia, serum titres averaging 94 mu/ml have been found. The predicted physiological changes in erythropoietin titre have also been demonstrated in humans using the RIA; increasing after bleeding and decreasing after red cell administration. Studies of erythropoietin levels in experimental animals have shown that, with the particular antiserum used, the sensitivity of the RIA is markedly reduced.

  15. A radioimmunoassay for chicken avidin

    International Nuclear Information System (INIS)

    Kulomaa, M.S.; Elo, H.A.; Tuohimaa, P.J.

    1978-01-01

    A double-antibody solid-phase radioimmunoassay for chicken avidin is reported. Avidin was labelled with 125 I by the chloramine-T method. The bound and free avidin were separated with a second antibody bound to a solid matrix. In the logit-log scale the standard curve was linear from 1-2 to 100-200ng of avidin/ml. Cross-reaction of ovalbumin was less than 0.015%. Saturation of biotin-binding sites of avidin with an excess of biotin decreased radioimmunoassay values by about 15%. Recovery studies indicated that avidin can be assayed from all chicken tissues studied with radioimmunoassay, whereas the [ 14 C]biotin/bentonite method gave poor recoveries for avidin in the liver and kidney. Radioimmunoassay and the [ 14 C]biotin/bentonite method gave similar concentrations for oviduct avidin. (author)

  16. Clinical indications of prolactin radioimmunoassay

    International Nuclear Information System (INIS)

    Lengyel, A.M.J.; Vieira, J.G.H.; Zanella, M.R.; Zampieri, M.; Chacra, A.R.

    1980-01-01

    Is a review is presented of the main clinical uses of prolactin measurements, including the galactorrhea-amenorrhea syndrome, an experiment employing the prolactin radioimmunoassay is related. (Author) [pt

  17. Automatic computation of radioimmunoassay data

    International Nuclear Information System (INIS)

    Toyota, Takayoshi; Kudo, Mikihiko; Abe, Kanji; Kawamata, Fumiaki; Uehata, Shigeru.

    1975-01-01

    Radioimmunoassay provided dose response curves which showed linearity by the use of logistic transformation (Rodbard). This transformation which was applicable to radioimmunoassay should be useful for the computer processing of insulin and C-peptide assay. In the present studies, standard curves were analysed by testing the fit of analytic functions to radioimmunoassay of insulin and C-peptides. A program for use in combination with the double antibody technique was made by Dr. Kawamata. This approach was evidenced to be useful in order to allow automatic computation of data derived from the double antibody assays of insulin and C-peptides. Automatic corrected calculations of radioimmunoassay data of insulin was found to be satisfactory. (auth.)

  18. Radioimmunoassay of polypeptide hormones and enzymes

    International Nuclear Information System (INIS)

    Felber, J.P.

    1974-01-01

    General principles of radioimmunoassay are reviewed. Detailed procedures are reviewed for the following hormones: insulin, pituitary hormones, gonadotropins, parathyroid hormone, ACTH, glucagon, gastrin, and peptide hormones. Radioimmunoassay of enzymes is also discussed. (U.S.)

  19. Radioimmunoassay for urinary albumin

    International Nuclear Information System (INIS)

    Woo, J.; Floyd, M.; Cannon, D.C.; Kahan, B.

    1978-01-01

    We describe a rapid, sensitive, and precise radioimmunoassay for urinary albumin (U/sub alb/). Aliquots of diluted urine were incubated at room temperature for 1 h with 125 I-labelled albumin and a rabbit antiserum monospecifid for human albumin. Phase separation was effected by the double-antibody technique. The dose-response curve was linear in the range of 15.6 to 10,000 ng, equivalent to 4 to 3000 mg/liter of urine. The limit of sensitivity was 16 ng of albumin. The coefficient of assay variation was 4.8%, both at 44 mg/liter and at 1304 mg/liter. A displacement curve obtained with a serially diluted urine sample of high albumin concentration was completely superimposable with the curve for which human albumin was used as a standard. In 26 normal individuals the range for U/sub alb/ was 2.2 to 12.6 mg/24 h, and for albumin clearance (C/sub alb/), 1.8 x 10 -5 --19.6 x 10 -5 ml/min. After renal homografts in 25 patients, U/sub alb/ ranged from 16.9 to 9928 mg/24 h, and C/sub alb/ from 2.7 x 10 -4 to 1.7 x 10 -1 ml/min. Both increased U/sub alb/ and C/sub alb/ correlated well with the severity of renal homograft rejection

  20. Radioimmunoassay for a human prostate specific antigen

    International Nuclear Information System (INIS)

    Machida, T.; Miki, M.; Ohishi, Y.; Kido, A.; Morikawa, J.; Ogawa, Y.

    1983-01-01

    As a marker for prostatic cancer, a prostate-specific antigen was purified from human prostatic tissues. Double antibody radioimmunoassay utilizing immune reaction was developed on the basis of the purified prostatic antigen (PA). Measurement results have revealed that PA radioimmunoassay is much better than prostatic acid phosphatase (PAP) radioimmunoassay in the diagnosis of prostatic cancer

  1. Radioimmunoassay for human myoglobin

    International Nuclear Information System (INIS)

    Kondo, Akira

    1981-01-01

    1 A new radioimmunoassay (RIA) for human myoglobin (Mb) using two antibody technique was developed as a microassay of Mb. This RIA is characterized as follows; 1) Radioiodination of human Mb was successfully done for the first time by chloramine T method, which yielded Iabelled Mb with high specific activities ranging 40 - 60 μCi/μg. 2) Affinity chromatography was used to obtain purified anti-human Mb antibody, which improved the sensitivity of the RIA remarkably, and excluded the effects of serum and urine. 2 The sensitivity of the RIA was 1 ng/ml in sera and 2 ng/ml in urine. The average recovery was 92.7%. Both intra-assay and inter-assay coefficients of variation were below 10%. 3 With this method, Mb purified from skeletal and cardiac muscles was shown to have same immunological nature. 4 Serum concentrations of Mb in normal adults were 13.1 +- 6.1 ng/ml (mean +- S.D.) with a range of 1 - 28 ng/ml. No sex difference was observed. Urinary Mb Ievels in normal adults were less than 4 ng/ml and 24-hour urinary excretions were less than 6 μg. With this method, serum and urinary Mb concentrations were determined in patients with various disorders such as myopathies and myocardial disorders and with anesthesia. Serum Mb concentrations were revealed to be elevated to 50000 ng/ml in malignant hyperthermia, 4000 ng/ml in acute myocardial infarction, 1000 ng/ml in Duchenne muscular dystrophy and 4000 ng/ml in polymyositis. Urinary Mb level was elevated when serum Mb rose to 2000 - 3000 ng/ml. 5 In order to apply RIA of Mb in daily clinical practices, time needed for the assay was shortened by separation of free and antibody-bound Mb by the polyethylene glycol method immediately after incubation was performed at 37 0 C for 2 hours. (author)

  2. Radioimmunoassay for human myoglobin

    Energy Technology Data Exchange (ETDEWEB)

    Kondo, A. (Tokushima Univ. (Japan). School of Medicine)

    1981-04-01

    1 A new radioimmunoassay (RIA) for human myoglobin (Mb) using two antibody technique was developed as a microassay of Mb. This RIA is characterized as follows; 1) Radioiodination of human Mb was successfully done for the first time by chloramine T method, which yielded Iabelled Mb with high specific activities ranging 40 - 60 ..mu..Ci/..mu..g. 2) Affinity chromatography was used to obtain purified anti-human Mb antibody, which improved the sensitivity of the RIA remarkably, and excluded the effects of serum and urine. 2 The sensitivity of the RIA was 1 ng/ml in sera and 2 ng/ml in urine. The average recovery was 92.7%. Both intra-assay and inter-assay coefficients of variation were below 10%. 3 With this method, Mb purified from skeletal and cardiac muscles was shown to have same immunological nature. 4 Serum concentrations of Mb in normal adults were 13.1 +- 6.1 ng/ml (mean +- S.D.) with a range of 1 - 28 ng/ml. No sex difference was observed. Urinary Mb Ievels in normal adults were less than 4 ng/ml and 24-hour urinary excretions were less than 6 ..mu..g. With this method, serum and urinary Mb concentrations were determined in patients with various disorders such as myopathies and myocardial disorders and with anesthesia. Serum Mb concentrations were revealed to be elevated to 50000 ng/ml in malignant hyperthermia, 4000 ng/ml in acute myocardial infarction, 1000 ng/ml in Duchenne muscular dystrophy and 4000 ng/ml in polymyositis. Urinary Mb level was elevated when serum Mb rose to 2000 - 3000 ng/ml. 5 In order to apply RIA of Mb in daily clinical practices, time needed for the assay was shortened by separation of free and antibody-bound Mb by the polyethylene glycol method immediately after incubation was performed at 37/sup 0/C for 2 hours.

  3. Radioimmunoassay for pyridostigmine

    Energy Technology Data Exchange (ETDEWEB)

    Meyer, H.G.; Lukey, B.J.; Gepp, R.T.; Corpuz, R.P.; Lieske, C.N.

    1988-01-01

    Pyridostigmine is a cholinergic drug used for the treatment of myasthenia gravis and for antagonizing the effects of non-depolarizing muscle relaxants. In addition, military organizations in several countries have an active interest in pyridostigmine as a pretreatment compound for nerve agent poisoning. Anti-pyridostigmine antibodies were produced in rabbits using a pyridostigmine analog conjugated to keyhole limpet hemocyanin. These antibodies were used for development of a radioimmunoassay that has a linear standard curve (r2=0.986) ranging from 0.5 to 10.0 ng/ml of pyridostigmine bromide in a 0.1-ml plasma sample. This assay measures pyridostigmine in plasma with better sensitivity and much greater through-put than do current state-of-the-art high-performance liquid chromatography techniques. In addition, only small volumes (100ml) of the plasma samples are required. Plasma levels of pyridostigmine were determined in the rat after intramuscular administration (0.056mg/kg) of pyridostigmine bromide. Estimates of the various pharmacokinetic parameters were calculated using the computer program NONLIN84. The results were as follows: apparent volume of distribution = 1.97 1/kg, absorption rate constant = 0.277 min-1, elimination rate constant = 0.0273 min-1, area under the curve = 1010 ng x min/ml, absorption rate half-life = 2.41 min, elimination rate half-life = 24.8 min, maximal plasma concentration (Cmax) = 21.3 ng/ml and time to Cmax = 9.02 min.

  4. Peptide radioimmunoassays in clinical medicine

    International Nuclear Information System (INIS)

    Geokas, M.C.; Yalow, R.S.; Straus, E.W.; Gold, E.M.

    1982-01-01

    The radioimmunoassay technique, first developed for the determination of hormones, has been applied to many substances of biologic interest by clinical and research laboratories around the world. It has had an enormous effect in medicine and biology as a diagnostic tool, a guide to therapy, and a probe for the fine structure of biologic systems. For instance, the assays of insulin, gastrin, secretin, prolactin, and certain tissue-specific enzymes have been invaluable in patient care. Further refinements of current methods, as well as the emergence of new immunoassay techniques, are expected to enhance precision, specificity, reliability, and convenience of the radioimmunoassay in both clinical and research laboratories

  5. Radioimmunoassay reagent and its use in a radioimmunoassay

    International Nuclear Information System (INIS)

    Polito, A.J.; Knight, W.S.

    1977-01-01

    A radioimmunoassay to detect or determine a steroid of the cortisone and aldosterone group has been developed. The invention particularly concerns a steroid derivative containing imidazole group which is radioactively labelled. The invented reagents are labelled with iodine 125. (VJ) [de

  6. Recent advances in steroid radioimmunoassay

    International Nuclear Information System (INIS)

    Jeffcoate, S.L.

    1977-01-01

    The advances since 1974 in the techniques of measuring steroid molecules by radioimmunoassay are reviewed in this paper. They are considered under the following headings: preparation and use of antisera; preparation and use of tracers; preparation of biological samples before assay; dispensing of the reagents in the assay; separation of free and bound radioactivity; counting and data processing; quality control and standardization. (orig.) [de

  7. In vitro inhibition of enterobacteria-reactive CD4+Tumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    Mangano, K.; Sardesai, N.; D'Alcamo, M.

    2008-01-01

    VGX-1027 is an isozaxoline compound that has recently been found to primarily target the function of murine macrophages but not of T cells, inhibiting secretion of tumor necrosis factor (TNF)-alpha in response to different Toll-like receptor agonists in vitro and in vivo. The well-defined role...

  8. B-lymfocytdepletring og andre biologiske behandlingsmuligheder ved Graves' oftalmopatiTumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    El, Fassi D.; Hegedus, L.; Nielsen, Claus Henrik

    2008-01-01

    The current medical treatment options for Graves' ophthalmopathy (GO) are unsatisfactory. Recent treatment of GO patients with the B-lymphocyte depleting monoclonal antibody rituximab or with the anti-tumor necrosis factor-alpha agents etanercept and infliximab has shown promising results. We...

  9. A sensitive radioimmunoassay for fentanyl

    International Nuclear Information System (INIS)

    Michiels, M.; Hendriks, R.; Heykants, J.

    1977-01-01

    Antiserum to fentanyl was obtained in rabbits repeatedly injected with carboxyfentanyl conjugated to bovine serum albumin. Using the antiserum, a highly sensitive radioimmunoassay has been developed, based on the dextran-coated charcoal method. It proved possible to assay the drug directly in plasma, in amounts as small as 30 picogram in 0.5 ml. The antibody was highly specific for fentanyl and no cross-reaction was observed with its major metabolites. This sensitive and specific radioimmunoassay method was employed to determine fentanyl in plasma from six volunteers after an intravenous bolus of 0.2 mg, and in plasma from dogs treated both intravenously and subcutaneously with 0.02 mg/kg. The plasma level of fentanyl could be followed for up to 6 h after a therapeutic dose in dogs and man. (orig.) [de

  10. Infections and treatment of patients with rheumatic diseasesTumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    Atzeni, F.; Bendtzen, K.; Bobbio-Pallavicini, F.

    2008-01-01

    /inflammatory conditions, and current therapies have the aim of providing adequate (low) compensatory doses, the timing of GC administration, such as during the nocturnal turning-on phase of tumour necrosis factor (TNF) secretion, can be extremely important. The use of the lowest possible GC dose, at night......, and for the shortest possible time should therefore greatly reduce the risk of infections. Infection is a major co-morbidity in rheumatoid arthritis (RA), and conventional disease-modifying anti-rheumatic drugs (DMARDs) can increase the risk of their occurrence, including tuberculosis. TNF-alpha plays a key role...

  11. Radioimmunoassay for plasma renin activity

    International Nuclear Information System (INIS)

    1975-01-01

    A radioimmunoassay for the determination of renin activity in blood plasma is described. The plasma sample is mixed with a generator buffer solution also containing an inhibitor for enzymes which convert angiotensin I into other substances. The renin in the plasma sample converts angiotensinogen into angiotensin I. The amount of angiotensin I is then measured with a competitive binding method using 125 I-labelled angiotensin I and antibodies to angiotensin I

  12. Determination of phenobarbital by radioimmunoassay

    International Nuclear Information System (INIS)

    Yamaoka, A.; Takatori, T.

    1979-01-01

    A radioimmunoassay for phenobarbital has been studied. Antiphenobarbital antisera were obtained by repeated immunization of rabbits with p-succinamidophenobarbital conjugated to bovine serum albumin. Less than 0.2 pmol of phenobarbital could be measured by this procedure. The specificity of the antibodies was directed to substituents on the nitrogen atoms of the barbituric rings as well as to substituents at the carbon 5-position of the ring. (Auth.)

  13. Radioimmunoassays of papain in beer

    International Nuclear Information System (INIS)

    Rauch, P.; Fukal, L.; Strejcek, F.; Kas, J.

    1984-01-01

    A radioimmunoassay (RIA) of papain is described which is capable of detecting 0.2 μg of papain/ml in beer, a level approximately 1% of that normally used for chillproofing. Changes in incubation conditions significantly accelerate the papain determination with only slight loss of accuracy. Apart from the high sensitivity it has been shown that the method is highly specific and distinguishes papain from the other chillproofing enzymes used. (author)

  14. The promotion of radioimmunoassay in human health

    International Nuclear Information System (INIS)

    Dudley, R.A.

    1983-01-01

    Radioimmunoassay is an analytical technique which makes use of highly specific and sensitive antibodies to segregate particular substances of interest and radioactive tracers to permit quantification of minute amounts. Some procedures use specific biological ''reagents'' other than antibodies and tracers other than radionuclides. Radioimmunoassay plays an enormous role in medical diagnosis and research. Depending on the services to be performed, the radioimmunoassay laboratories are classified into 4 categories. The laboratory of each category is staffed and equipped with facilities according to its scope and quantity of work. From 1980-1982, nearly US$ 2 million had been used under the Agency's Technical Cooperation Programme for the promotion of radioimmunoassay in human health

  15. Promotion of radioimmunoassay in human health

    Energy Technology Data Exchange (ETDEWEB)

    Dudley, R A [International Atomic Energy Agency, Vienna (Austria). Div. of Life Sciences

    1983-06-01

    Radioimmunoassay is an analytical technique which makes use of highly specific and sensitive antibodies to segregate particular substances of interest and radioactive tracers to permit quantification of minute amounts. Some procedures use specific biological ''reagents'' other than antibodies and tracers other than radionuclides. Radioimmunoassay plays an enormous role in medical diagnosis and research. Depending on the services to be performed, the radioimmunoassay laboratories are classified into 4 categories. The laboratory of each category is staffed and equipped with facilities according to its scope and quantity of work. From 1980-1982, nearly US $2 million had been used under the Agency's Technical Cooperation Programme for the promotion of radioimmunoassay in human health.

  16. Optimization of AFP-radioimmunoassay using Antibody Capture Technique

    International Nuclear Information System (INIS)

    Moustafa, K.A.

    2003-01-01

    Alpha-fetoprotein (AFP) is a substance produced by the unborn baby. When the neural tube is not properly formed large amounts of AFP pass into the amniotic fluid and reach the mother's blood. By measuring AFP in the mother's blood and amniotic fluid, it is possible to tell whether or not there is a chance that the unborn baby has a neural tube defect. AFP also used as a tumor marker for hepatocellular carcinoma. There are many different techniques for measuring AFP in blood, but the most accurate one is the immunoassay technique. The immunoassays can be classified on the basis of methodology into three classes; (1) the antibody capture assays, (2) the antigen capture assay, (3)the two-antibody sandwich assays. In this present study, the antibody capture assay in which the antigen is attached to a solid support, and labeled antibody is allowed to bind, will be optimized

  17. Radioimmunoassay of plasma corticotrophin

    International Nuclear Information System (INIS)

    Hummer, L.

    1977-01-01

    An assay has been established, based on antibodies against the N-terminal part of the ACTH molecule with a high affinity, a detection limit of 2 pg ACTH when assaying 200 ul unextracted plasma, and implying a total incubation time of three days. The antibody has been obtained by immunizing guinea-pigs with synthetic human 1-24 ACTH coupled to bovine serum albumin. The selected antibody has an equilibrium constant of 4 x 10 11 litres/mole in a final dilution of 1/320.000. The antiserum reacts with synthetic human 1-39 ACTH as well as with synthetic human 1-24 ACTH, and the hormonally inactive synthetic human 11-24 ACTH fragment as well as alpha- and beta-melanocyte-stimulating hormones do not cross-react in the assay. The interassay coefficient of variation of replicate estimates was 11-13%. The reproducibility of the standard curve have been evaluated by calculating the amount of ACTH corresponding to 5% of the (B/T)o value, 1.2+-0.4 pg ACTH/tube+-SD and 50% of (B/T)o value, 15.7+-2.6 pg ACTH/tube+-SD. Validation of the assay has been obtained by assaying samples from patients with verified adrenal disorders, and the accuracy is supported by ACTH determinations in test, where metyrapon had been administered intravenously. A stimulation of ACTH production by insulin-induced hypoglycaemia has been found as well. Special attention is always paid as to the conditions for the blood sampling. A reference interval of 10-76 ng/l has been found (115 normal subjects). (orig.) [de

  18. Radioimmunoassay method for detection of gonorrhea antibodies

    International Nuclear Information System (INIS)

    1975-01-01

    A novel radioimmunoassay for the detection of gonorrhea antibodies in serum is described. A radionuclide is bound to gonorrhea antigens produced by a growth culture. In the presence of gonorrhea antibodies in the serum, an antigen-antibody conjugate is formed, the concentration of which can be measured with conventional radiometric methods. The radioimmunoassay is highly specific

  19. Pancreatic elastase in human serum. Determination by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Geokas, M.C. (Univ. of California, Davis); Brodrick, J.W.; Johnson, J.H.; Largman, C.

    1977-01-10

    This study demonstrates that a serine endopeptidase of pancreatic origin (elastase 2) circulates in human blood. A specific and highly sensitive radioimmunoassay has been developed for pancreatic elastase 2 in human serum. The inactivation of elastase 2 employed as radioiodinated tracer with an active site-specific reagent (phenylmethanesulfonyl fluoride) was necessary to prevent its binding by serum ..cap alpha../sub 1/-antitrypsin and ..cap alpha../sub 2/-macroglobulin while maintaining its immunoreactivity. The assay is based upon competition of standard human pancreatic elastase 2 with /sup 125/I-labeled phenylmethanesulfonyl elastase 2 for specific antibody binding sites, after which a second antibody precipitation step is used to separate bound from free /sup 125/I-labeled phenylmethanesulfonyl elastase 2. The minimum detectable concentration of elastase 2 was 0.9 ng/ml. The average normal fasting serum level determined was 71 ng/ml, approximately 80-fold greater than the minimum detectable amount.

  20. A radioimmunoassay for abscisic acid

    International Nuclear Information System (INIS)

    Walton, D.; Dashek, W.; Galson, E.

    1979-01-01

    We have developed a radioimmunoassay (RIA) for abscisic acid (ABA) in the 0.1 ng to 2.5 ng range. Antibodies were obtained from rabbits immunized with ABA bound via its carboxyl group to bovine serum albumin. Cross-reactivity studies indicate that ABA esters are completely cross-reactive with ABA, while trans, trans abscisic acid (t-ABA) phaseic acid (PA) and dihydrophaseic acid (DPA) have much lower but significant cross-reactivities. Purification methods which reduce the levels of cross-reacting substances are described. (orig.) 891 AJ/orig. 892 MKO [de

  1. Radioimmunoassay and other related techniques

    International Nuclear Information System (INIS)

    Zarkawi, Moutaz

    1993-04-01

    The article reviews principles, requirements and reliability criteria of radioimmunoassay (RIA). Since basic reactions involved in RIA and related techniques are derived from reactions which take place in the immune system (IS) of humans and animals, the IS and the way it works will be described. In addition to RIA which involves the use of isotopes as tracers (labels), other non-radioisotopic and recent immunoassay techniques i.e. enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (CLIA) and fluoroimmunoassay (FIA) will be dealt with. Some important and related terms will be defined and explained. (author). 59 refs., 4 figs

  2. Radioimmunoassay of cholylglycine in serum

    International Nuclear Information System (INIS)

    Wakushima, Tadashi; Yamanishi, Yasuhito; Hirayama, Chisato

    1979-01-01

    Serum levels of cholylglycines (CG) were determined by radioimmunoassay and that of total bile acids (TBA) by enzymatic method. In normal subjects, serum levels of CG, TBA and CG/TBA ratio were 0.6 +- 0.4 μM, 7 +- 2 μM, and 0.08 +- 0.06, respectively. They were increased markedly in acute hepatitis and moderately in chronic hepatitis and cirrhosis. Thus, measurement of serum CG as compared with serum TBA appears to be a sensitive liver test. (author)

  3. Radioimmunoassay in children with glomerulonephritis

    International Nuclear Information System (INIS)

    Ametov, A.S.; Gavryushova, L.P.; Shashinka, M.; Mumladze, Eh.B.; Tvorogova, T.M.; Dakhuk, B.; Dronova, V.I.; Toritsina, D.K.; Petrova, T.V.

    1985-01-01

    Proceeding from a radioimmunoassay of various biologically active substances in the blood and urine (ACTH, cortisol, FSH, LH, prolatin, progesterone, estradiol, plasma renin activity and β 2 -microglobulin) of 220 children with glomerulonephritis change of all indices with relation to the type and gravity of glomerulonephritis as well as renal function was revealed. The nature of influence on corticosteroid and immunosuppressive therapy was shown. The authors consider it appropriate to use the determination of biologically active substances in the blood and urine for a more profound estimation of a child's status

  4. Heterologous radioimmunoassay for arginine vasopressin

    International Nuclear Information System (INIS)

    Shimamoto, K.; Murase, T.; Yamaji, T.

    1976-01-01

    A sensitive and specific radioimmunoassay for arginine vasopressin (AVP) was developed utilizing the antisera against lysine vasopressin (LVP) in combination with a labeled AVP. The assay employs an acetone extraction procedure and detects as little as 0.8 pg. per millimeter of AVP in human plasma. In normal subjects, the mean (+- S.D.) plasma concentration of AVP was 4.9 +- 1.2 pg. per milliliter after fluid deprivation and 1.2 +- 0.4 pg. per milliliter after water loading. Plasma AVP levels correlated significantly with plasma osmolalities. In four patients with diabetes insipidus, plasma AVP concentrations ranged from less than 0.8 to 1.2 pg. per milliliter, whereas six patients with the syndrome of inappropriate ADH secretion showed plasma levels of AVP which correspond to those of the dehydrated state in normal subjects or greater, although plasma osmolalities were low in all cases. It was concluded that the present radioimmunoassay method for AVP provides a useful way of assessing neurohypophyseal function in man

  5. Apparatus for use in radioimmunoassays

    International Nuclear Information System (INIS)

    Chen, C-H.; Tsay, H-M.; Heyer, R.E.

    1979-01-01

    Apparatus for solid-phase antibody separation techniques used in radioimmunoassays is described in this invention. It consists of a rectangular prism tray with multiple wells protruding into its interior from one side. Near the base of the tray is an orifice used for creating evacuated condition within the structure. At the base of each well there is an orifice of such size and shape as to retain an aqueous liquid under given pressure conditions but permit the evacuation of this liquid at reduced pressure. The outlet of these orifices is in the shape of an inverted conical frustrum. Each of the wells contains an antibody coated disc of porous cellulose paper surrounded by a plastic support. The porous nature of the cellulose paper ensures contact between the antibody coating and the antigen. The use of antibody coated porous cellulose paper in combination with the vacuum operated apparatus simplifies the manipulative steps whilst still maintaining the sensitivity of the radioimmunoassay. It also obviates the need for aspiration and thus lessens the risk of contamination from one sample to another. (UK)

  6. A sensitive radioimmunoassay for colchicine

    International Nuclear Information System (INIS)

    Scherrmann, J.M.; Boudet, L.; Pontikis, R.; Nguyen-Hoang-Nam; Fournier, E.

    1980-01-01

    A sensitive and rapid radioimmunoassay for colchicine was developed using an antibody raised in goats after injection of N-desacetylthiocolchicine conjugated with bovine serum albumin. The incubation time is short and dextran-coated charcoal was used for the separation. The sensitivity of the assay was 0.35 ng/ml urine or serum. A low cross-reactivity was observed with colchicine derivatives and the other drugs tested. The RIA was applied to the measurement of colchicine in the serum of a patient given a 1mg oral dose of the drug. Peak levels of colchicine were attained at 2 hrs. following administration and the drug was rapidly distributed, the concentration decreasing to < 1 ng/ml by 8 hrs. The 24 and 48 hr. concentrations indicated a prolonged excretion of colchicine and a long elimination half-time. (U.K.)

  7. Radioimmunoassay for nortriptyline and amitriptyline

    International Nuclear Information System (INIS)

    Turner, P.

    1977-01-01

    A description has been given (Aherne, G.W.; Marks, V.; Mould, G.; Stout, G.; Lancet; 1214, June 4 (1977)) of a radioimmunoassay (RIA) technique having advantages for monitoring patient response by estimation of plasma concentrations of nortriptyline and amitriptyline. A wide range of monoamine-reuptake-inhibiting drugs is now used in the treatment of depressive illness. There are other analytical techniques such as gas-liquid chromatography which can be used in the estimation of most of these drugs and their major metabolites, as well as for the identification of some other centrally-acting drugs such as benzodiazepines. The chromatographic technique was able to detect the administration of the wrong drug in one comparative study of two antidepressive drugs. Chromatographic and RIA methods therefore both have their own analytical roles according to the clinical or research problem being investigated. (U.K.)

  8. Improved radioimmunoassay of plasma cortisol

    International Nuclear Information System (INIS)

    Donohue, J.; Sgoutas, D.

    1975-01-01

    We report a simplified radioimmunoassay procedure for determination of plasma cortisol. The method offers the advantage that tracer, antibody, and assay buffer are added in a single step with the use of semi-automated equipment. Thus, critical pipetting procedures are minimized, and assay time is reduced to 4 h. No prior extraction or chromatographic purification of cortisol is required. The procedure is simple, reliable, and accurate. For either between-assay or within-assay determinations, the coefficients of variation are about 6 percent. Our results compare well with those obtained from a procedure that included extraction and chromatographic purification of cortisol in several plasma samples. Conditions for optimizing the assay are also discussed. (auth)

  9. Preparation of albumin radioimmunoassay kit

    International Nuclear Information System (INIS)

    Chen Suoshu; Wang Yanzhen; Wang Zhenshan

    1990-01-01

    This paper presents the preparation of Albumin (Alb) radioimmunoassay kit and its preliminary clinical application. The kit is mainly applied to the measurement of Alb concentration in human urine, adopting second antibody-PEG method. The measurement range is (1-50 μg/ml). The curve obtained with a serially diluted urine sample of high Alb concentration was a straight line. Recovery, detectability, intra- and inter-batch variation coefficients were 95.7%-103.6%, 0.1 μg/ml, 5.8% and 6.4% respectively. The kit was tried out clinically for measuring Alb in urine samples in about 1200 normal individuals and 600 various patients of related renal diseases. The preliminary clinical results show that Alb RIA is conducive to the early diagnosis of kidney function abnormalities

  10. Methadone radioimmunoassay: two simple methods

    International Nuclear Information System (INIS)

    Robinson, K.; Smith, R.N.

    1983-01-01

    Two simple and economical radioimmunoassays for methadone in blood or urine are described. Haemolysis, decomposition, common anticoagulants and sodium fluoride do not affect the results. One assay used commercially-available [1- 3 H](-)-methadone hydrobromide as the label, while the other uses a radioiodinated conjugate of 4-dimethylamino-2,2-diphenylpentanoic acid and L-tyrosine methyl ester. A commercially-available antiserum is used in both assays. Normethadone and α-methadol cross-react to a small extent with the antiserum while methadone metabolites, dextropropoxyphene, dipipanone and phenadoxone have negligible cross-reactivities. The 'cut-offs' of the two assays as described are 30 and 33 ng ml -1 for blood, and 24 and 21 ng ml -1 for urine. The assay using the radioiodinated conjugate can be made more sensitive if required by increasing the specific activity of the label. (author)

  11. Radioimmunoassay method for triiodothyronine and thyroxine

    International Nuclear Information System (INIS)

    Hollander, C.S.

    1975-01-01

    This invention relates to a radioimmunoassay method for triiodothyronine or thyroxine or triiodothyronine and thyroxine present in unextracted serum containing thyroxine binding prealbumin and thyroxine binding globulin. Procedures using 125 I and 131 I are described

  12. Measurement of antibodies to tubulin by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Mead, G M; Cowin, P; Whitehouse, J M.A. [CRC Medical Oncology Unit, Southampton General Hospital, Southampton, U.K.

    1979-07-24

    A solid-phase double antibody radioimmunoassay capable of measuring antibody to tubulin, the principal component of microtubules, is described. This assay is simple, combining sensitivity with specificity and also allowing determination of antibody subclasses.

  13. Validation of a radioimmunoassay for rat insulin

    International Nuclear Information System (INIS)

    Delattre, E.; Boschero, A.C.

    1984-01-01

    A methodological approach that permits, in a radioimmunoassay, the evaluation of rat insulin by using bovine insulin as reference is presented. As in general the technics for radioimmunoassay of different substances follow the same principles (competitive inhibition), we believe that the methodology presented here could be used for evaluation of other hormones when the adequated referential, of know biological activity, is not available. (Author) [pt

  14. The measurement of platelet activation by radioimmunoassay in asthma

    International Nuclear Information System (INIS)

    Wu Guoxin; Sun Jian; Li Jianyong; Ruan Changgeng

    1992-02-01

    Radioimmunoassay with specific monoclonal antibody was used to evaluate the platelet activation in 14 cases of acute bronchial asthma. The result showed that the number of molecules of alpha-granule membrane protein (GMP-140) which was exposed on the surface of platelet following secretion significantly increased on the surface of platelet and in plasma, while the number of molecules of glycoprotein (GP) I b and GPIII a did not change significantly; the concentration of thromboxane B 2 in plasma was raised, while the concentration of 6-keto-PGF 1a was within the normal limits; the concentrations of β-thromboglobulin (β-TG) and platelet factor 4(PF 4 ) in plasma increased significantly; the number of platelets decreased. These results strongly confirmed that the degree of platelet activation was enhanced during acute asthmatic attack. The significance of platelet activation in the pathogenesis of asthma should be further investigated

  15. Gastrin radioimmunoassay. Description and application of a novel method

    International Nuclear Information System (INIS)

    Nemeth, J.; Jakab, B.; Schweibert, I.; Szolcsanyi, J.; Oroszi, G.; Szilvassy, Z.

    2002-01-01

    Development and application of a novel gastrin radioimmunoassay (RIA) are described. 125 I-labeling of non-sulphated human gastrin-17 (nshG-17) was performed by the iodogen method and the mono-iodinated hormone, as RIA tracer, was separated by reversed-phase high performance liquid chromatography (HPLC). Serum gastrin levels were measured in response to intravenous application of isoproterenol, a non-selective beta and phenylephrine, a selective alpha-1 receptor agonist using a newly developed method specific for the C-terminal part of the hormone in rats. Isoproterenol at clinically relevant doses elicited a significant increase in serum gastrin concentration in a dose-dependent fashion, whereas phenylephrine was without effect. (author)

  16. Automated, computer interpreted radioimmunoassay results

    International Nuclear Information System (INIS)

    Hill, J.C.; Nagle, C.E.; Dworkin, H.J.; Fink-Bennett, D.; Freitas, J.E.; Wetzel, R.; Sawyer, N.; Ferry, D.; Hershberger, D.

    1984-01-01

    90,000 Radioimmunoassay results have been interpreted and transcribed automatically using software developed for use on a Hewlett Packard Model 1000 mini-computer system with conventional dot matrix printers. The computer program correlates the results of a combination of assays, interprets them and prints a report ready for physician review and signature within minutes of completion of the assay. The authors designed and wrote a computer program to query their patient data base for radioassay laboratory results and to produce a computer generated interpretation of these results using an algorithm that produces normal and abnormal interpretives. Their laboratory assays 50,000 patient samples each year using 28 different radioassays. Of these 85% have been interpreted using our computer program. Allowances are made for drug and patient history and individualized reports are generated with regard to the patients age and sex. Finalization of reports is still subject to change by the nuclear physician at the time of final review. Automated, computerized interpretations have realized cost savings through reduced personnel and personnel time and provided uniformity of the interpretations among the five physicians. Prior to computerization of interpretations, all radioassay results had to be dictated and reviewed for signing by one of the resident or staff physicians. Turn around times for reports prior to the automated computer program generally were two to three days. Whereas, the computerized interpret system allows reports to generally be issued the day assays are completed

  17. Radioimmunoassay of human urinary kallikrein

    International Nuclear Information System (INIS)

    Goering, W.

    1980-01-01

    Using a human urinary kallikrein, purified by means of Trasylol sepharose, it has been possible to develop a radioimmunoassay of kallikrein capable of detecting the substance down to a concentration of 0.5 ng/ml. The specific activity of the tracer labelled with 125-iodine using the Chloramine-T method was 30-70 nCi/ng of kallikrein. The antiserum titres for the antikallikrein serum were 1:20.000 up to 1:50.000. Human urine, submandibular and parotid salivae as well as pancreatic secretion in this RIA reacted in the same manner as the kallikrein standard solution. The kallikrein content in urine, as determined by the RIA was between 0 and 300 ng/ml, in the saliva between 400 and 2.000 ng/ml, and in the pancreatic juice between 300 and 12.000 ng/ml. Using human serum, only an incomplete immunological cross-reaction could be achieved. In human liquor as well as in animal preparation, no cross-reacting substances could be detected. (orig.) [de

  18. Radioimmunoassay of triiodothyronine in urine

    International Nuclear Information System (INIS)

    Kosowicz, J.; Gembicki, M.; Schneider, E.; Eder, M.

    1977-01-01

    In 21 cases of hypothyroidism, in 39 cases of hyperthyroidism, in 54 healthy subjects, in 23 pregnant women, and in certain internal diseases determinations of triiodothyronine were carried out in urine by radioimmunoassay. Anti-T 3 antibodies were obtained in rabbits and sheep immunized with a complex of bovine albumin with triiodothyronine ester. Labelled triiodothyronine of high specific activity was obtained by iodinating triodothyronine by the chloramine method. Determinations of triiodothyronine were performed in morning urine and the obtained values were calculated for one-hour excretion. In healthy subjects the excretion of T 3 was from 20 to 95 ng/hour, in hyperthyroidism it was significantly raised to from 120 to over 600 ng/hour, while in most cases of hypothyroidism it was decreased. In pregnancy the urinary excretion of T 3 was normal amounting to from 34 to 87 ng/hour, although in most cases the serum T 3 concentration was raised. In cases of anorexia nervosa and in obese starving subjects the excretion of T 3 fell significantly, and similarly low excretion was found in some cases of debilitating diseases and myocardial infarction. (author)

  19. Radioimmunoassay of canine growth hormone

    International Nuclear Information System (INIS)

    Eigenmann, J.E.; Eigenmann, R.Y.

    1981-01-01

    A sensitive radioimmunoassay (RIA) for canine growth hormone (GH) was developed. Antibodies were elicited in rhesus monkeys. One antiserum exhibited a working titer at a dilution of 1:500 000. Radioiodination was performed enzymatically employing lactoperoxidase. Logit-log transformation and least squares fitting resulted in straight line fitting of the standard curve between 0.39 and 50 ng/ml. Formation of large-molecular [ 125 I]GH during storage caused diminished assay sensitivity. Therefore [ 125 I]GH was re-purified by gel chromatography. Using this procedure, high and reproducible assay sensitivity was obtained. Tracer preparations were used for as long as 3 months after iodination. Diluted plasma from normal and acromegalic dogs resulted in a dose-response curve parallel to the standard curve. Canine prolactin exhibited a cross-reactivity of 2%. The within-assay coefficient of variation (CV) was 3.8 and the between-assay CV was 7.2%. Mean plasma GH concentration in normal dogs was 1.92 +- 0.14 ng/ml (mean +- SEM.) GH levels in acromegalic dogs were appreciably higher. Insulin-induced hypoglycaemia, arginine and ornithine administration resulted in inconsistent and sluggish GH increment. A better response was obtained by injecting a low dose of clonidine. Clonidine administration to hypopituitary dogs resulted in absent or poor GH increment. (author)

  20. Improved radioimmunoassay for human TSH

    International Nuclear Information System (INIS)

    Spencer, C.A.; Nicoloff, J.T.

    1980-01-01

    This study concerns the optimization of the human TSH (h-TSH) radioimmunoassay with special emphasis on reducing the heterogeneity of the 125 I h-TSH tracer. Enzymatic iodination of h-TSH with glucose oxidase/lactoperoxidase was shown to be superior to either low or high dose chloramine-T procedures, producing a high specific activity reagent (70-150 μCi/μg) with minimal evidence of damage. Tracer purification procedures not only affected initial immunoactivity but also storage stability and heterogeneity of the resulting 125 I h-TSH. The assay developed using these technical approaches shows a sensitivity limit of 0.005+-0.001 (S.E.M.) μU/tube; 50% displacement at 0.18+-0.08 (S.E.M.) μU/tube and complete delineation between euthyroid (n=49, 2.44+-0.18 (S.E.M.) mU/l, range 1.00-6.08) and hyperthyroid (n=62, 0.34+-0.02 (S.E.M.) mU/l, range 0.10-0.85), serum h-TSH levels. (Auth.)

  1. Theoretical and practical application of radioimmunoassays

    International Nuclear Information System (INIS)

    Sokolowski, G.; Wood, W.G.

    1981-01-01

    This book describes experiences made not only in developing and producing commercial reagent kits, but also those made in university research and diagnostic laboratories. The fundamental principles (radioactivity, immunoassay) are explained briefly and tersely. Then the components of the radioimmunoassay are described in detail and illustrated by selected practical examples (production of antibodies, separation of bound and free portions, standards). This chapter is followed by systematic instructions for composing a radioimmunoassay. The actual conditions of the complex problem of quality safety is explained, a field in which particularly the authors are to be regarded as pioneers. Finally an outlook indicates that radioactive alternatives, incorporating the principle of immunologic determinations methods, are available or will be developed for at least a few radioimmunoassays. (orig./MG) [de

  2. Construction of Expression Vector for Anti-Alpha-Fetoprotein Gene and Its Inhibition Effects on Alpha-Fetoprotein Positive Hepg2 Cells

    Science.gov (United States)

    Wang, Ze; Zhang, Hui

    As research previously demonstrated, suppression of AFP expression or its biological activities might inhibit the proliferation of AFP positive human hepatocellular carcinoma cells. In this study, we constructed an anti-AFP gene vector and transfected it to HepG2 cells. RT-PCR showed AFP gene expression in the transfected cells was reduced. MTT assay suggested the proliferation of the transfected cells was also inhibited comparing with the untransfected cells. This result provides a new insight into AFP as the target for preventing and treating hepatocellular carcinoma.

  3. Assessment of the specificity of norethisterone radioimmunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Bedolla-Tovar, N; Rahman, S A; Cekan, S Z; Diczfalusy, E [Swedish Medical Research Council, Karolinska Hospital, Stockholm

    1978-06-01

    It is concluded that (1) the significance of cross- reaction studies as well as that of the parallelism test for the assessment of the over-all specificity of the assay is limited, (2) a single chromatography prior to the radioimmunoassay proper improves the assay specificity, but may not be sufficient to remove all interfering compounds, (3) a comparison of the direct and chromatographic assay procedures using several antisera is useful for the selection of the relatively most specific radioimmunoassay procedure. In the present study, this is the technique employing either antiserum C or antiserum D, the latter, however, only after chromatography.

  4. Solid phase tube radioimmunoassay for digoxin detection

    International Nuclear Information System (INIS)

    Stellner, K.; Glatz, C.; Linke, R.

    1975-01-01

    A solid phase radioimmunoassay with 125 I is described for cardiac patients. The test for the digoxin determination and the poisoning due to cardiac glycosides can be measured very accurately and carried out easily. In addition, the test determination can be automatically performed in connection with other tests. (GSE/LH) [de

  5. Adsorption columns for use in radioimmunoassays

    International Nuclear Information System (INIS)

    1976-01-01

    Adsorption columns are provided which can be utilized in radioimmunoassay systems such as those involving the separation of antibody-antigen complexes from free antigens. The preparation of the columns includes the treatment of retaining substrate material to render it hydrophilic, preparation and degassing of the separation material and loading the column

  6. Rationale for the development of radioimmunoassays

    Energy Technology Data Exchange (ETDEWEB)

    Roulston, J E; Adam, T [Charing Cross Group of Hospitals, London (UK)

    1978-02-01

    A theoretical study of the principles of radioimmunoassay and allied techniques is presented with a view to the application in practical developmental research. Both equilibrium and sequential saturation methods are discussed and their respective advantages noted. Simple methods for the estimation of the affinity constant by saturation techniques and use of the Scatchard plot are outlined.

  7. Determination of serum IgD radioimmunoassay

    International Nuclear Information System (INIS)

    Fayol, V.; Hartmann, D.J.; Sabbagh, I.; Ville, G.

    1986-01-01

    We describe a sensitive liquid phase radioimmunoassay for serum IgD. Extreme values obtained from 85 control patients sera are 0.2 and 121 mg/l with an arithmetic mean of 25 mg/l. In atopic patients (with high serum IgE levels), arithmetic mean is 47 mg/l [fr

  8. Determination of serum IgD radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Fayol, V.; Hartmann, D.J.; Sabbagh, I.; Ville, G.

    1986-01-01

    We describe a sensitive liquid phase radioimmunoassay for serum IgD. Extreme values obtained from 85 control patients sera are 0.2 and 121 mg/l with an arithmetic mean of 25 mg/l. In atopic patients (with high serum IgE levels), arithmetic mean is 47 mg/l.

  9. Improved radioimmunoassay for thyroid hormone and reagent

    International Nuclear Information System (INIS)

    1980-01-01

    Improvements in the radioimmunoassay of the thyroid hormones, thyroxine or triiodothyronine, are described. Hydrolyzed cross-linked polyacrylamide particles covalently bonded against the thyroid hormone are employed as solid phase substrates for the thyroid hormone antibodies. The polyacrylamide particles are dyed yellow or blue to facilitate the various manipulative steps during the radioimmunoassay. The particles are characterized by their ability to form stable hydrophilic suspensions. As a result the reaction mixture, during which thyroid hormone is separated from serum proteins and competitive binding in the presence of radioactive tracer with the antibody occurs, requires no agitation to maintain the desired homogeneous condition. This is in contrast to the settling problems experienced with cellulose, dextran and glass particles. In addition, the non-specific binding property of the polyacrylamide particles is so low that the initially separated solid phase particles following incubation can be directly measured for radioactivity levels without any initial washings thus increasing the speed and convenience of the assay procedure. Details of the preparation of the dyed, hydrolyzed polyacrylamide particles, the coupling of antiserum to these particles and the radioimmunoassay procedure are given. Data obtained from the radioimmunoassays of hypothyroid, euthyroid and hyperthyroid sera demonstrated the satisfactory performance of the assay. (U.K.)

  10. Evaluation of radioimmunoassay data by a minicomputer

    International Nuclear Information System (INIS)

    Venot, A.; Ingrand, J.; Roucayrol, J.C.; Valeyre, J.; Deltour, G.

    1977-01-01

    A program is proposed for routine use in every radioimmunoassay laboratory part of a bigger unit such as a department of nuclear medicine. Its main features are maniability and ability to avoid undue effects of outlying experimental values. A non specialized technician can operate himself, the data being fed off-line by a paper tape into a Multi 8 Intertechnique minicomputer [fr

  11. Radioimmunological determination of. cap alpha. -MSH and ACTH in the rat

    Energy Technology Data Exchange (ETDEWEB)

    Usategui Echeverria, R; Oliver, C; Vaudry, H; Lombardi, G; Rozenberg, I; Vague, J [Centre Hospitalier Universitaire de la Timone, 13 - Marseille (France)

    1975-09-01

    Specific and sensitive radioimmunoassay methods for ..cap alpha..-MSH and ACTH are reported. They make possible specific measurements of each hormone in rat plasma or pituitary extracts. Endogenous ..cap alpha..-MSH and ACTH extracted from plasma or pituitary show the same immunoreactivity than synthetic ..cap alpha..-MSH and ACTH. ..cap alpha..-MSH and ACTH levels vary independently from each other in the following conditions: circadian rythm, corticoid treatment, adrenalectomy, ether stress, haloperidol injection.

  12. Enhancement of human adaptive immune responses by administration of a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensisTumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors

    DEFF Research Database (Denmark)

    Lobner, M.; Walsted, A.; Larsen, R.

    2008-01-01

    tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, and interferon (IFN)-gamma was increased after Immunlina...... administration for 3 days (P alpha, IFN-gamma, and IL-6 responses to TT were enhanced after 8 and 14 days (P ...) and fell below baseline levels after 14 days (P alpha...

  13. Determination of a glucose-containing tetrasaccharide in urine by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Zopf, D.A.; Levinson, R.E. (National Cancer Inst., Bethesda, MD (USA)); Lundblad, A. (University Hospital, Lund (Sweden). Dept. of Clinical Chemistry)

    1982-01-15

    A radioimmunoassay is described that allows rapid determination of a urinary oligosaccharide - Glc..cap alpha..1-6Glc..cap alpha..1-4Glc..cap alpha..1-4Glc ((Glc)/sub 4/) - at concentrations >2pmol/..mu..l. Antibodies produced in rabbits immunized with the phenethylamine derivative of (Glc)/sub 4/ coupled to keyhole limpet hemocyanin bind tritiated (Glc)/sub 4/-alditol. Studies comparing the activities of (Glc)/sub 4/ and several of its derivatives and analogues as inhibitors of binding of tritiated (Glc)/sub 4/-alditol show that (Glc)/sub 4/ is detected with maximum specificity and sensitivity only after reduction to (Glc)/sub 4/-alditol. Quantitation of (Glc)/sub 4/ in urine samples reduced with sodium borohydride gives excellent agreement with a previously used but more laborious method that employs gas-liquid chromatography-mass spectrometry (GLC/MS) (correlation coefficient = 0.98). Studies of the excretion rate of (Glc)/sub 4/ in urine of women during pregnancy using the radioimmunoassay method confirm and extend previous results using the GLC/MS method.

  14. Immunochemical heterogeneity of calcitonin in man: effect on radioimmunoassay

    International Nuclear Information System (INIS)

    Snider, R.H.; Moore, C.F.

    1977-01-01

    Determinations of blood levels of human calcitonin by radioimmunoassay have varied considerably in different laboratories. Much of the controversy over calcitonin levels can be attributed to the multiplicity of immunoreactive forms of the hormone (iCT), the differing region specificities of the antisera utilized for measurement by radioimmunoassay, protein effects, different rates of degradation of the various iCT fractions and the specific methodology of the radioimmunoassay

  15. Programs for radioimmunoassays and radioreceptorassays using SPSS-X.

    Science.gov (United States)

    Amador, A G; Hodges, S L

    1989-01-01

    Two sets of programs written using SPSS-X, and designed for the analysis of data obtained by radioimmunoassay or radioreceptorassay are presented. The main advantages of the programs are that they are easily edited, their commands are simple and logical, and they can be used for any type of radioimmunoassay or radioreceptorassay, respectively. The programs for radioimmunoassay can also be used for fluorescenceimmunoassay and luminescenceimmunoassay.

  16. Radioimmunoassay of deoxynivalenol in wheat and corn

    International Nuclear Information System (INIS)

    Xu, Y.C.; Zhang, G.S.; Chu, F.S.

    1986-01-01

    With the availability of antibody against deoxynivalenol triacetate (DON-triacetate), a radioimmunoassay (RIA) for DON in wheat was developed. DON is extracted from the sample with acetonitrile-water defatted with hexane, and then reacted with acetic anhydride to form DON-triacetate. The reaction mixture is loaded onto a C-18 cartridge to remove excess reagents and impurities. Acetylated DON is eluted from the cartridge with 50% methanol in water, and then analyzed by radioimmunoassay utilizing antiserum against DON-triacetate and tritiated DON-triacetate. Overall recovery for DON added to wheat between 50 and 5000 ppb was 86% with a standard deviation of 7% and coefficient of variation of 8%. The limit of detection for DON was about 20 ppb. Analysis of 12 naturally contaminated wheat, corn, and mixed feed samples for DON revealed that RIA results agreed well with thin layer chromatographic analyses performed by other laboratories

  17. 125I Radioimmunoassay of serum ursodeoxycholyl conjugates

    International Nuclear Information System (INIS)

    Hill, A.; Ross, P.E.; Bouchier, I.A.D.

    1983-01-01

    A radioimmunoassay for serum ursodeoxycholic conjugates using an iodine-125 ligand has been developed. The bile acid was present in normal fasting serum (0.19 +- SD 0.19 μmol/l, n=24) and 2-hour post-prandial serum (0.8 +- SD 0.8 μmol/l, n=16). Gallstone patients undergoing oral ursodeoxycholic acid therapy had significantly higher post-prandial serum levels (21.5 +- SD 14.0 μmol/l, n=15) by radioimmunoassay. Gas liquid chromatography analysis indicated that in normal serum ursodeoxycholic acid was totally conjugated, whereas sera from gallstone patients contained a proportion as the free bile acid (10.2 +- SD 8.1 μmol/l, n=15). Following an oral dose of ursodeoxycholic acid, both unconjugated and conjugated forms of the bile acid appeared in the serum of healthy individuals. (Auth.)

  18. Radioimmunoassay analysis of baculovirus granulins and polyhedrins

    International Nuclear Information System (INIS)

    Summers, M.D.; Hoops, P.

    1980-01-01

    Granulin and polyhedrin proteins were purified by preparative sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis from the baculoviruses Autographa californica, Rachiplusia ou, Heliothis zea, Heliothis armigera. Trichoplusia ni, and Spodoptera frugiperda. Antisera were raised against Autographa californica (Ac) polyhedrin and Trichoplusia ni (Tn) granulin and analyzed for homologous and heterologous immunoreactivity by immunodiffusion and radioimmunoassay (RIA). Ac polyhedrin and Tn granulin antisera recognized antigenic determinants on several baculovirus polyhedrin and granulin proteins even though the heterologous proteins had different immunoreactivities when compared by competition radioimmunoassay. Antigenic differences among granulin and polyhedrin proteins were also detected by altered slopes of the competition reaction curves. Antiserum raised against Ac polyhedrin which was purified in the presence of SDS was tested by competition RIA for its ability to detect and react with native polyhedrin produced in the infected TN-368 cells. Ac polyhedrin antiserum had similar if not identical ability to bind to native polyhedrin and to polyhedrin purified in the presence of SDS

  19. Anamnesis of the radioimmunoassay: A historical reminiscence

    Energy Technology Data Exchange (ETDEWEB)

    Kallee, E

    1984-01-27

    Anamnesis of the Radioimmunoassay - A Historical Reminiscence: Several requirements have to be met before the first radioimmunoassay of a proteohormone (i.e., insulin) could be developed. One of these requirements was that iodination ought not substantially impair the immunologic properties of the insulin molecule. Since the reversibility of antigen adsorption to antibodies is the most important basis of all immunoassays, the adsorptive behavior of proteins first had to be studied at extremely low protein concentrations. Only after these requirements had been investigated was direct electrophoretic detection of a nonprecipitating anti-hormone antibody possible by desorption of insulin on filter paper. Later, the inaccuracy of the species specificity of insulin-binding antibodies proved to be particularly useful for the determination of insulin in human sera.

  20. Radioimmunoassay of human Hageman factor (factor XII)

    International Nuclear Information System (INIS)

    Saito, H.; Ratnoff, O.D.; Pensky, J.

    1976-01-01

    A specific, sensitive, and reproducible radioimmunoassay for human Hageman factor (HF, factor XII) has been developed with purified human HF and monospecific rabbit antibody. Precise measurements of HF antigen were possible for concentrations as low as 0.1 percent of that in normal pooled plasma. A good correlation (correlation coefficient = 0.82) existed between the titers of HF measured by clot-promoting assays and radioimmunoassays among 42 normal adults. Confirming earlier studies, HF antigen was absent in Hageman trait plasma, but other congenital deficient plasmas, including those of individuals with Fletcher trait and Fitzgerald trait, contained normal amounts of HF antigen. HF antigen was reduced in the plasmas of patients with disseminated intravascular coagulation or advanced liver cirrhosis, but it was normal in those of patients with chronic renal failure or patients under treatment with warfarin. HF antigen was detected by this assay in plasmas of primates, but not detectable in plasmas of 11 nonprimate mammalian and one avian species

  1. Folic acid derivatives for use in radioimmunoassay

    International Nuclear Information System (INIS)

    Ali, A.

    1981-01-01

    The chemical preparation of two folic acid derivatives, labelled with 125 I or 131 I, is described for use in radioimmunoassay of folic acid and its metabolites in biological fluids such as blood serum. Labelled compounds of the present invention more closely resemble folic acid in that they have glutamic acid in the terminal position. Examples of the use of these compounds in three different assays are given. (U.K.)

  2. Significance and radioimmunoassay of gastric inhibitory polypeptide

    International Nuclear Information System (INIS)

    Zheng Ping; Zeng Minde; Yuan Jimin

    1995-01-01

    We have established the GIP Radioimmunoassay which has high sensitivity and specificity by labelling with iodogen and purified with HPLC. Using this method, the plasma GIP level was measured in 64 cases of which there are 10 normal individuals, 25 cases of diabetes and 29 cases of liver cirrhosis . The results showed that the plasma GIP level was significantly increased in patients with liver cirrhosis and correlated to degree of glucose tolerance damage

  3. Status of radioimmunoassay services in the Philippines

    International Nuclear Information System (INIS)

    Valencia, I.G.; Bonoan, L.S.; Medina, V.F.O.

    1986-01-01

    This paper discusses the participation of the Philippines as a member state of the International Atomic Energy Agency (IAEA) in the Regional Cooperation Agreement (RCA) project on radioimmunoassay of thyroid related hormones in Asia and the Pacific. Seven of the twenty-three radioassay laboratories engaged in RIA work have signified their agreement to participate in the program. The proposed organization and function structure of the project, the projected RIA use and strategy to popularize use of RIA are given. (ELC)

  4. Radioimmunoassay of measles virus antibodies in SSPE

    International Nuclear Information System (INIS)

    Jankowski, M.A.; Gut, W.; Kantoch, M.

    1982-01-01

    A sensitive radioimmunoassay (RIA) was introduced for detecting measles virus IgG and IgM antibodies. The hyperimmune response to the measles virus could be demonstrated more accurately by RIA than by haemagglutination inhibition (HI). The ratio between RIA and HI antibody titres was decidedly higher in sera and cerebrospinal fluids of patients with subacute sclerosing panencephalitis than in those of other groups tested. (author)

  5. Radioimmunoassay to quantitatively measure cell surface immunoglobulins

    International Nuclear Information System (INIS)

    Krishman, E.C.; Jewell, W.R.

    1975-01-01

    A radioimmunoassay techniques developed to quantitatively measure the presence of immunoglobulins on the surface of cells, is described. The amount of immunoglobulins found on different tumor cells varied from 200 to 1140 ng/10 6 cells. Determination of immunoglobulins on the peripheral lymphocytes obtained from different cancer patients varied between 340 to 1040 ng/10 6 cells. Cultured tumor cells, on the other hand, were found to contain negligible quantities of human IgG [pt

  6. Radioimmunoassays - what is their diagnostic value

    International Nuclear Information System (INIS)

    Panitz, N.

    1983-01-01

    A selection is made of hormones and enzymes which are measurable by radioimmunoassay and their organ-specific function is described. Endocrine function tests together with the stimulating or suppressing substances, reaction in the organism and the examined regulation are listed. Non-radioactive immunological determination methods and the use of monoclonal antibodies in vitro and in vivo with ''tumor imaging'' are indicated. (orig.) [de

  7. Radioimmunoassay of parathyroid hormone: past and future

    International Nuclear Information System (INIS)

    Yalow, R.S.

    1986-01-01

    In this report on radioimmunoassay of parathyroid hormone (iPTH) it was shown that the rate of disappearance of iPTH from plasma differed markedly in patients with primary hyperthyroidism or those with uremia and secondary hyperparathyroidism and that for each patient the rate of disappearance depended on the antiserum used for assay. The heterogeneity of iPTH in plasma was soon rapidly confirmed in many laboratories. (Auth.)

  8. Qualitative determination of tubulin by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Joniau, M [Louvain Univ. (Belgium). Interdisciplinary Research Centre; Brabander, M de [Janssen Pharmaceutica, Beerse (Belgium). Lab. of Oncology; Mey, J de [Brussels Univ. (Belgium). Medische Stichting Koningin Elisabeth; Hoebeke, J [Brussels Univ. (Belgium). Lab. of Biochemical Pathology

    1977-06-15

    The use of an antiserum specific for tubulin in cytochemical studies was described previously. Here a report is presented on its application in a radioimmunoassay for tubulin useful in the concentration range of 0.5 to 20 ..mu..g tubulin/ml. The advantages of this technique over the classical colchicine binding assay are discussed in terms of sensitivity and nonsusceptibility to the degree of polymerization and thermal denaturation of tubulin allowing its application to cell culture homogenates.

  9. Simultaneous radioimmunoassay for luteinizing hormone and prolactin

    International Nuclear Information System (INIS)

    Steele, M.K.; Deschepper, C.F.

    1985-01-01

    A combined radioimmunoassay (RIA) for the measurement of the anterior pituitary proteins luteinizing hormone (LH) and prolactin (PRL) is described and compared with individual RIAs for these hormones. The standard curves and the sample values for LH and PRL were identical when determined in a combined or in an individual RIA. This technique may prove useful to a number of laboratories where it is desirable to determine levels of more than one hormone in limited sample volumes

  10. Studies on the radioimmunoassay of thyroid hormones

    International Nuclear Information System (INIS)

    Kim, J.R.; Awh, O.D.; Park, K.B.; Kim, Y.S.

    1980-01-01

    To establish radioimmunoassay (RIA) systems of 3,5,3'-triiodo-L-thyronine (T 3 ) and thyroxine (T 4 ), various experiments such as 125 I labelling, antibody raising, preparation of hormone-free sera and efficient separations of the free hormones from those of antibody bound etc. were conducted. By optimizing many factors, assay systems were successfully established. Some detailed methodological aspects were described. (author)

  11. Human aldolase B subunit-specific radioimmunoassay

    International Nuclear Information System (INIS)

    Asaka, M.; Alpert, E.

    1983-01-01

    A radioimmunoassay was developed for the direct quantification of aldolase B in human serum and tissues. The method is a double-antibody radioimmunoassay technique using radioiodinated aldolase B homopolymer as ligand, chicken antibodies to aldolase B and rabbit antibodies to chicken IgG. This radioimmunoassay was shown to be specific for the aldolase B subunit, with no cross-reactivity with either human aldolase A subunit or homopolymeric human aldolase C (C 4 ). The lowest measurable amount by this method was 2 ng/ml. Aldolase B is predominantly found in normal liver tissue, with relatively-high aldolase B levels also observed in kidney. Aldolase B levels in the serum obtained from 11 normal subjects ranged from 23 to 38 ng/ml, with a mean of 28.5 +- 9.2 (S.D.) ng/ml. Almost all of patients with hepatitis had serum aldolase B levels greater than 30 ng/ml. In cancer patients, serum aldolase B was slightly elevated in patients with metastatic liver cancer and primary lever cell carcinoma, whereas no elevation of serum aldolase B was shown in patients without liver metastasis. (Auth.)

  12. The production of antibodies for radioimmunoassay

    International Nuclear Information System (INIS)

    Court, G.

    1975-01-01

    Three factors which affect the outcome of any immunisation schedule designed to produce antisera for radio-immunoassay, the antigen, the method of immunisation and the choice of animal are considered. Several factors concerning the nature of the antigen are dealt with, for example, the molecular size and immunogenicity of the antigen. It is noted that the larger polypeptide and proteins are sufficiently immunogenic to elicit a useful antibody response alone and that whilst substances with molecular weights of less than 2000 may produce a response alone they will probably produce a better one if they are conjugated (chemically coupled) to a much larger molecule. The method of immunisation is discussed including a consideration of the use of adjuvant and the route and timing of injections. It is noted that antisera showing the relevant properties for radio-immunoassay are rarely produced without emulsification of the immunogen in Freund's adjuvant although this is not an absolute requirement for antibody production. Data are presented comparing the intramuscular and multiple intradermal routes of injection. The results, however, fail to demonstrate any major advantage for either method although the latter may be more economical, producing high titre antisera with relatively small amounts of immunogen. Because of their convenience rabbits are generally the first choice of animal for raising antisera for radioimmunoassay although guinea pigs, chickens and sheep have been used successfully in many cases

  13. Radioimmunoassay of measles virus hemagglutinin protein G

    International Nuclear Information System (INIS)

    Lund, G.A.; Salmi, A.A.

    1982-01-01

    Guinea pig and rabbit antisera from animals immunized with purified measles virus hemagglutinin (G) protein were used to establish a solid-phase four-layer radioimmunoassay for quantitative measurement of the G protein. The sensitivity of the assay was 2 ng of purified G protein, and 200 μg of protein from uninfected Vero cells neither decreased the sensitivity nor reacted non-specifically in the assay. Radioimmunoassay standard dose-response curves were established and unknown values interpolated from these using the logit program of a desktop computer. Using this procedure, a measles virus growth curve in infected Vero cells was determined by measurement of G protein production. Under these same conditions, hemagglutination was not sensitive enough to detect early hemagglutinin production. Viral antigens in canine distemper virus, Newcastle disease virus, parainfluenza viruses 1-4, simian virus 5, and respiratory syncytial virus-infected cell lysates did not cross-react in the radioimmunoassay. A small degree of cross-reactivity was detected with mumps viral antigens, both with Vero cell-derived (wild-type strain) and egg-derived (Enders strain) purified virus preparations and with a cell lysate antigen prepared from wild-type mumps virus-infected Vero cells. (Auth.)

  14. Radioimmunoassay of measles virus hemagglutinin protein G

    Energy Technology Data Exchange (ETDEWEB)

    Lund, G A; Salmi, A A [Turku Univ. (Finland)

    1982-08-01

    Guinea pig and rabbit antisera from animals immunized with purified measles virus hemagglutinin (G) protein were used to establish a solid-phase four-layer radioimmunoassay for quantitative measurement of the G protein. The sensitivity of the assay was 2 ng of purified G protein, and 200 ..mu..g of protein from uninfected Vero cells neither decreased the sensitivity nor reacted non-specifically in the assay. Radioimmunoassay standard dose-response curves were established and unknown values interpolated from these using the logit program of a desktop computer. Using this procedure, a measles virus growth curve in infected Vero cells was determined by measurement of G protein production. Under these same conditions, hemagglutination was not sensitive enough to detect early hemagglutinin production. Viral antigens in canine distemper virus, Newcastle disease virus, parainfluenza viruses 1-4, simian virus 5, and respiratory syncytial virus-infected cell lysates did not cross-react in the radioimmunoassay. A small degree of cross-reactivity was detected with mumps viral antigens, both with Vero cell-derived (wild-type strain) and egg-derived (Enders strain) purified virus preparations and with a cell lysate antigen prepared from wild-type mumps virus-infected Vero cells.

  15. Localization of radioiodinated antibody to alpha-fetoprotein in rats with transplanted hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Koji, T; Ishii, N; Kono, K [Nagasaki Univ. (Japan)

    1980-11-01

    The specific localization of radiolabeled antibody to AFP was investigated in rats bearing subcutaneous transplants of AH-7974 ascites hepatoma. Total body scintigrams 48 to 168 hours after injection of /sup 125/I-anti rat AFP horse antibody showed remarkable localization of radioactivity on the tumors, as a control, a faint outline of tumor was observed by /sup 125/I-normal horse IgG. The tumor/blood radioactivity ratio in the antibody group exceeded 1.0 and was approximately four times higher than that in the control group 7 days after injection, and suggested an active accumulation of radioactive antibody in the tumor tissue. In the subcellular distribution of radioactivity in the tumor tissue, about 30 to 60% of the total radioactivity were found in a fraction of the cell membrane plus nucleus. Furthermore autoradiograms of the fixed tissue sections in the antibody group revealed the specific localization of the radioactivity on the tumor cell surface as black grains.

  16. Alpha-fetoprotein, HCV and HBV infections in Nigerian patients with ...

    African Journals Online (AJOL)

    The aim of this study was to determine the relationship of serum AFP with HBV and HCV infections among Nigerian patients with, Primary Hepato-Cellular Carcinoma . Forty-two Nigerian adults comprising of 3 groups – PHCC, Liver cirrhosis (LC) and Controls were recruited into the study. The findings showed that only nine ...

  17. Maternal seric alpha-fetoprotein: determination of the medium values in Costa Rican pregnant women

    International Nuclear Information System (INIS)

    Salas Chaves, P.; Villalobos Castro, A.

    1999-01-01

    The present study focuses on the normal mean values of AFP in 124 Costa rican pregnant women. In the women 554 determinations were done between the fifteenth and twentieth week of gestation. For this purpose, and Irma test and normal values were established. The concentration values of AFP were expressed as multiples of the median, considering a value of 2.5 Mm as the normal superior limit and of 0.25 as the inferior limit. The confidence limits of the median were 24-57 Ku/I and the 2.5MM value located between 60-143 Ku/I. (author)

  18. Effects of alpha fetoprotein on escape of Bel 7402 cells from attack of lymphocytes

    International Nuclear Information System (INIS)

    Li, Mengsen; Liu, Xinhua; Zhou, Sheng; Li, Pingfeng; Li, Gang

    2005-01-01

    Involvement of AFP against apoptosis of tumor cell has been implicated in its evasion of immune surveillance. However, the molecular events of immune escape mechanisms are still unknown. The major observations reported here relate to a possible mechanism by which heptoloma Bel 7402 cells escape immune surveillance in vitro. Western blotting and a well-characterized cofocal scanning image were performed to analyze the expression of Fas/FasL and caspase-3 in co-cultured Bel 7402 and Jurkat cells. After co-culture with Jurkat cells, up-regulated Fas and reduced FasL expression could be observed. Treatment with AFP could remarkably inhibit the elevated Fas and, whereas, induce the FasL expression in co-cultured Bel 7402 cells. Cells co-culture could induce the expression of caspase-3 in both cells line. The elevated caspase-3 in Bel 7402 cells was abolished following the treatment of AFP. The expression of caspase-3 was elevated in co-cultured Jurkat cells treated with AFP. No detectable change on the expression of survivin was examined in both cells line. Monoclonal antibody against AFP treatment alone did not obviously influence the growth of cells, as well as the expression of Fas/FasL and caspase-3. However, the effect of AFP could be blocked by antibody. our results provide evidence that AFP could promote the escape of liver cancer cells from immune surveillance through blocking the caspase signal pathway of tumor cells and triggering the Fas/FasL interaction between tumor cells and lymphocytes

  19. Solid phase separation technique for use in radioimmunoassays

    International Nuclear Information System (INIS)

    Tu, J.I.

    1979-01-01

    A radioimmunoassay procedure, and article of manufacture for carrying out that procedure, are disclosed herein. The solid phase separation technique utilized in the radioimmunoassay of this invention utilizes a test tube, the internal surface of which has been coated with two antibody layers

  20. Quality control of radioiodinated gastrin for radioimmunoassay

    International Nuclear Information System (INIS)

    Ginabreda, M.G.P.; Borghi, V.C.; Bettarello, A.

    1988-07-01

    Radioiodinated human gastrin has been prepared at IPEN laboratory for radioimmunoassay use. This work developed the quality control of this tracer analyzing parameters of the labelling reaction, chromatographic purification and radioimmunoassay. The radioiodination yield obtained in five experiments was reproducible and similar when analyzed on 7% polyaraylamide gel eletrophoresis - PAGE - (mean + - SD of 51.70 + - 10.76%) and by1 25 I incorporation checked through thrichloroacetic acid precipitation - TCA - (57-36 + - 9.69%). Similary, after purification the labelled gastrin revaled high and reproducible purity degree when submitted to PAGE (96.57 + - 1.06%) and CA (94.82 + - 4.20%) analysis. The respective specific activities varied from 62 to 307 uCi/ug, being determined by the self-displacement method, which is based on the immunoactivity of the tracer. In this way, the antibody titers required to bind 50% of the tracer ranged from 1:32.000 to 1:180.000. Consequently, the respective doses producing 50% fall in the maximum response of the radioimmunoassays ranged from 155.0 to 24.0 pmol/1, but remained unchanged for each tracer even after three months of its preparations. The tracers presented very low non-specific binding values (1.78 + - 0.79%), stablespecific binding values (46.49 + - 5.65%) and a good between-assay precision, evaluated by an internal quality control sample (25.71 + - 4.30%) with coefficient of variation of 16.74%). The PAGE analysis of the unlabeled gastrin used in the first and last radioiodination revealed an unique and unaltered component, confirming the quality of the tracers. (author) [pt

  1. A linear graph for digoxin radioimmunoassay

    International Nuclear Information System (INIS)

    Smith, S.E.; Richter, A.

    1975-01-01

    The determination of drug or hormone concentrations by radio-immunoassay involves interpolation of values for radioisotope counts within standard curves, a technique which requires some dexterity in curve drawing and which results in some inaccuracy in practice. Most of the procedures designed to overcome these difficulties are complex and time-consuming. In radioimmunoassays involving saturation of the antibody-binding sites a special case exists in that the bound radioactivity is directly proportional to the specific activity of the ligand in the system. Thus a graph of the ratio of radioactivity bound in the absence to that in the presence of added non-radioactive ligand is linear against the concentration of added ligand (Hales,C.N., and Randle, P.J., 1963, Biochem. J., vol. 88, 137). A description is given of a simple and convenient modification of their method, and its application to the routine clinical determination of digoxin using a commercial kit (Lanoxitest β digoxin radioimmunoassay kit, Wellcome Reagents Ltd.). Specially constructed graph paper, which yields linearity with standard solutions, was designed so that it could be used directly without data transmission. The specific activity function appears as the upper arithmetical horizontal scale; corresponding values of the concentration of non-radioactive ligand in the solution added were individually calculated and appear on the lower scale opposite the appropriate values of the upper scale. The linearity of the graphs obtained confirmed that binding of digoxin was approximately constant through the range of clinical concentrations tested (0.5 to 8ng/ml), although binding declined slightly at higher concentrations. (U.K.)

  2. Diagnostic value of α-fetoprotein in hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Hernández Perera, Julio César

    2009-01-01

    In the world the hepatocellular carcinoma (HCC) is the fifth to sixth most common malignancy. An increase in its incidence is looming in Cuba as a result of the population aging and the increase of morbi-mortality due to liver cirrhosis (CH). α-fetoprotein (AFP) by SUMA®, was used in this work as a tumor marker technique in 189 Cirrhotic patients evaluated at the CIMEQ, between January 1999 and September 2005. The main factors associated with an elevation of AFP were HCC, viral cirrhosis, and the age of 50 years or more. Presented CHC 22 (11.64%) patients, who had an average age of 55 years and the predominant sex was male (86,36%). The most important cause associated with this malignant disease was viral CH, mainly the virus of hepatitis C. This tumor marker showed a sensitivity and specificity of 68.18% and 92.17%, respectively. When combined with abdominal ultrasound was increase 86.36% sensitivity and 100% specificity. Elevated AFP levels associated with tumor prognosis factors. It can be concluded that the AFP had value in the diagnosis of HCC. This work is the first of its kind carried out in Cuba, and allows the design of a workable strategy for the research, monitoring and prognosis of HCC, having a wide network of laboratories SUMA®,, with a low cost per each determination. (author)

  3. Radioimmunoassay of nortestosterone and related steroids

    International Nuclear Information System (INIS)

    Hampl, R.; Starka, L.; Picha, J.; Chundela, B.

    1979-01-01

    A radioimmunoassay of nortestosterone and related steroids, including its principal metabolites, is described and evaluated. Antisera against nortestosterone-17β-hemisuccinate- and nortestosterone-3-carboxymethyloxime-bovine serum albumin were raised in goats. By using a mixture of such antisera with different selectivity, the cross-reactions of several naturally occuring steroids can be reduced. The method can be applied for the detection of nortestosterone in both unprocessed or hydrolyzed urine extracts and also in plasma. It has been used as a screening test for anabolics in doping control. (orig.) [de

  4. Radioimmunoassay for thyroid-stimulating hormone (TSH)

    International Nuclear Information System (INIS)

    Blakemore, J.I.; Lewin, N.; Burgett, M.W.

    1978-01-01

    This invention provides a method for the radioimmunoassay of thyroid-stimulating hormone which utilizes a rapid and convenient version of a double antibody procedure. Highly purified second antibody is bound, by means of covalent bonds, to hydrolyzed polyacrylamide particles to produce a two-phase system. The solid phase comprises immobilized second antibody bound to the reaction product of labeled and unlabeled thyroid-stimulating hormone with the first antibody (first antibody-antigen complex) and the liquid phase comprises free (unbound) labeled and unlabeled thyroid-stimulating hormone. The two phases are separated and the radioactivity of either phase is measured

  5. Application of radioimmunoassay in food industry

    International Nuclear Information System (INIS)

    Rauch, P.; Fukal, L.; Kas, J.

    1983-01-01

    Radioimmunoassay can be used in evaluating the quality and wholesomeness of food. The method is advantageous for its speed, specificity, high sensitivity, relative ease of performance, and the possibility of performing a great number of parallel determinations using automation and computer evaluation. It can be used in determining macromolecules of proteins and enzymes. The determination of papain in beer is shown by way of example. Other possibilities of the method include: the determination of microbial toxins of the peptide nature, vitamins, hormones, antibiotics, pesticides and their residues, alkaloids, and carcinogenic materials. (M.D.)

  6. Aflatoxin B1 use in radioimmunoassay

    International Nuclear Information System (INIS)

    Liu Yinkun; Zhang Xiaying; Chen Ruiqun; Gu Tianjue

    1987-01-01

    Antibodies against Aflatoxin B 1 (AFB 1 ) were obtained after multiple-site injections of bovine serum albumin-AFB 1 conjugate into rabbits. The greatest specific binding effciency of antibody for AFB 1 is 70∼80%. The sensitivity of radioimmunoassay (RIA) for AFB 1 is between 0.46∼2.3 ng. The retrievel rate of AFB 1 by RIA from intentionally contaminated serum and rice is between 70∼80%. Detailed methods for the preparation of conjugate, immune serum, and methods for antibody titer determination are described

  7. Radioimmunoassay of serum myoglobin in neuromuscular diseases

    Energy Technology Data Exchange (ETDEWEB)

    Askmark, H; Osterman, P O; Roxin, L E; Venge, P [University Hospital, Uppsala (Sweden)

    1981-01-01

    Radioimmunoassay of serum myoglobin was performed in 85 patients with muscular symptoms. Elevated levels were found in 93% of patients with myogenic myopathy, in 54% with myasthenia gravis and in 50% with neurogenic myopathy. All 11 patients with polymyositis had elevated myoglobin concentrations. In six of seven patients with polymyositis, who were followed up with repeated determinations, a clear relationship between myoglobin levels and clinical course was found. In general serum myoglobin seemed to be a more sensitive indicator of muscle disease than creatine kinase.

  8. Determination of carbofuran in water by radioimmunoassay

    International Nuclear Information System (INIS)

    Zhu Guonian; Wu Huiming; Yang Ting; Hu Xiuqing

    2004-01-01

    A competitive radioimmunoassay (RIA) method was established to detect carbofuran in water samples. Compared with traditional analytical methods RIA provided a easy procedure with higher sensitivity. The detective limitation of RIA for carbofuran was proved to be 0.175 ng/ml. A study was performed to test validation of the RIA. In this study carbofuran residues in water samples were detected simultaneously by RIA and by high performance liquid chromatograph (HPLC). The linear correlation coefficient of the test results was measure to be 0.9985. (authors)

  9. Radioimmunoassay. A revolution in the analytic procedure

    Energy Technology Data Exchange (ETDEWEB)

    Strecker, H; Eckert, H G [Farbwerke Hoechst A.G., Frankfurt am Main (Germany, F.R.). Radiochemisches Lab.

    1978-04-01

    Radioimmunoassay is an analytic method which combines the sensitivity of radioactive measurement and the specificity of the antigen-antibody reaction Substances down to a concentration of some picograms per ml serum (or biological material) can be measured in the presence of a millionfold excess of otherwise interfering substances. The method is easy to carry out (test tube chemistry). The main field of application at the moment is in endocrinology; further possibilities of application are in pharmaceutical research, environmental protection, forensic medicine, and for general analytic purposes. Radioactive sources are used only in vitro in the nanocurie range, i.e. radiation exposure is negligible.

  10. Transfer of estradiol to human milk. [Radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Nilsson, S.; Nygren, K.G.; Johansson, E.D.B.

    1978-11-15

    A radioimmunoassay for the measurement of estradiol in human milk is evaluated. The detection limit was found to be 25 pg of estradiol per milliliter of milk. In milk samples collected from four lactating women during three to four months and from one pregnant and lactating woman, the concentration of estradiol was found to be below the detection limit of the assay. When six lactating women were given vaginal suppositories containing 50 or 100 mg of estradiol, it was possible to estimate the estradiol concentration in milk. A ratio of transfer of estradiol from plasma to milk during physiologic conditions is calculated to be less than 100 : 10.

  11. Radioimmunoassay in the diagnosis of fascioliasis

    Energy Technology Data Exchange (ETDEWEB)

    Tomanek, J; Willomitzer, J; Chroustova, E [Veterinary Research Inst., Brno (Czechoslovakia)

    1978-06-30

    The objective of the study was to develop a sensitive and specific radioimmunoassay (RIA) method for the diagnosis of F. hepatica infection that would be convenient to use in examining large numbers of samples. The main problem was the preparation of a suitable antigen or specific fraction, since the crude extract of adult Fasciola hepatica is a heterogeneous mixture of somatic, metabolic and host substances with possible nonspecific antigenic properties. The antigen was a protein fraction separated from crude extract of adult F. hepatica by gel filtration and labelled with /sup 125/iodine.

  12. Solid phase double-antibody radioimmunoassay procedure

    International Nuclear Information System (INIS)

    Niswender, G.D.

    1977-01-01

    The present invention is concerned with the radioimmunoassay (RIA) procedure for assaying body fluid content of an antigenic substance which may either be an antigen itself or a hapten capable of being converted, such as by means of reaction with a protein, to an antigenic material. The present invention is concerned with a novel and improved modification of a double-antibody RIA technique in which there is a first antibody that is specific to the antigenic substance suspected to be present in a body fluid from which the assay is intended. The second antibody, however, is not specific to the antigenic substance or analyte, but is an antibody against the first antibody

  13. Radioimmunoassay procedure using a stabilized complex

    International Nuclear Information System (INIS)

    Sultanian, I.V.; Irani, J.H.

    1978-01-01

    An improved radioimmunoassay procedure involves the use of a stabilized complex of labelled antigen and antibody which has an extended shelf life as compared to the same complex absent the stabilizers. Since the time needed to incubate the mixture of labelled antigen and antibody is eliminated, the time for completing the assay is considerably shortened and simplified. The components for carrying out the procedure are packaged in a kit basically including standard antigen for generation of a standard curve, a stabilized labelled antigen-antibody complex and reference serum, if used. A plurality of stabilizers are used in the complex to provide a shelf life of six weeks or more. 10 claims

  14. Radioimmunoassay of serum myoglobin in neuromuscular diseases

    International Nuclear Information System (INIS)

    Askmark, H.; Osterman, P.O.; Roxin, L.-E.; Venge, P.

    1981-01-01

    Radioimmunoassay of serum myoglobin was performed in 85 patients with muscular symptoms. Elevated levels were found in 93% of patients with myogenic myopathy, in 54% with myasthenia gravis and in 50% with neurogenic myopathy. All 11 patients with polymyositis had elevated myoglobin concentrations. In six of seven patients with polymyositis, who were followed up with repeated determinations, a clear relationship between myoglobin levels and clinical course was found. In general serum myoglobin seemed to be a more sensitive indicator of muscle disease than creatine kinase. (author)

  15. Radioimmunoassay in the diagnosis of fascioliasis

    International Nuclear Information System (INIS)

    Tomanek, J.; Willomitzer, J.; Chroustova, E.

    1978-01-01

    The objective of the study was to develop a sensitive and specific radioimmunoassay (RIA) method for the diagnosis of F. hepatica infection that would be convenient to use in examining large numbers of samples. The main problem was the preparation of a suitable antigen or specific fraction, since the crude extract of adult Fasciola hepatica is a heterogeneous mixture of somatic, metabolic and host substances with possible nonspecific antigenic properties. The antigen was a protein fraction separated from crude extract of adult F. hepatica by gel filtration and labelled with 125 iodine. (T.I.)

  16. Radioimmunoassay of sodium cromoglycate in human plasma

    Energy Technology Data Exchange (ETDEWEB)

    Brown, K; Gardner, J J; Lockley, W J.S.; Preston, J R; Wilkinson, D J [Fisons plc, Loughborough (UK). Pharmaceutical Div.

    1983-01-01

    A sensitive radioimmunoassay method for sodium cromoglycate in human plasma is described. The lowest quantifiable concentration of sodium cromoglycate is 0.93 nmol/l when 0.1 ml plasma samples are analysed. The range of the method is limited; both 0.01 and 0.1 ml volumes of plasma must be analysed to encompass the concentration range 0.93-139 nmol/l which may be encountered in plasma samples from patients and human volunteers. The method is specific for sodium cromoglycate as indicated by a low cross-reactivity of the anti-cromoglycate antiserum with a number of drugs.

  17. HLA-DR typing by radioimmunoassay

    International Nuclear Information System (INIS)

    Tosi, R.; Tanigaki, N.; Centis, D.; Rossi, P.L.; Alfano, G.; Ferrara, G.B.; Pressman, D.

    1980-01-01

    A radioimmunoassay procedure is described by which peripheral blood lymphocytes can be typed for HLA-DR specificities. The major advantages of this method are the following: simple and reproducible procedure, no need for B lymphocyte separation, no need for optimal viability, and no need for preabsorption of antisera with platelets. This method will find an application in the genetic and biochemical analysis of the HLA complex, and in the clinical tests of Ia antigens for diagnostic or prognostic purposes and in retrospective transplant studies

  18. Development of a radioimmunoassay for papain

    International Nuclear Information System (INIS)

    Rauch, P.; Fukal, L.; Marek, M.; Strejcek, F.; Kas, J.

    1984-01-01

    Various well-tried radioimmunoassay (RIA) techniques were compared for the quantitation of papain. The evaluation of individual assays was performed by logit-log analysis. The most compatible analytical steps were combined in order to obtain the optimal analytical conditions of the assay. The preferred RIA involves papain labelling with lactoperoxidase, a double antibody method as the separation step and a 24 h incubation period at 2 0 C. It permits the detection of 16 ng of papain per tube. In contrast, a method using immobilized antibody was satisfactory for rapid quantitation of papain with quite acceptable accuracy. (Auth.)

  19. Determination of hCG-alpha subunit in threatened pregnancy

    International Nuclear Information System (INIS)

    Talas, M.; Pohanka, J.; Fingerova, H.; Janouskova, M.; Krikal, Z.; Prasilova, J.; Zupkova, H.

    1987-01-01

    Radioimmunoassay of the hCG-alpha subunit was made using an antibody anti hCG-alpha serum, highly purified hCG-alpha for 125 I-labelling and the standard hCG-alpha. Sera of healthy pregnant women sampled throughout the whole pregnancies were used to determine x-bar±S.D. of hCG-alpha for 14-day intervals. Included in the study were groups of women with high risk of premature labor, late toxemia of pregnancy, twins and fetal hypotrophy. It was shown that increased hCG-alpha is found in pregnant women in whom signs of late toxemia of pregnancy are combined with high risk of premature labor, or with twin pregnancies, while in those with fetal hypotrophy hCG-alpha is within normal limits. (author). 3 figs., 7 refs

  20. Radioimmunoassay of bleomycin in plasma and urine

    International Nuclear Information System (INIS)

    Teale, J.D.; Clough, J.M.; Marks, V.

    1977-01-01

    Antibodies to bleomycin were raised by immunization of sheep and rabbits with bleomycin-albumin conjugates. The combination of a high-titre, high-avidity sheep antiserum and iodinated bleomycin produced a radioimmunoassay sensitive to 8 ng of bleomycin per ml of plasma or urine. Untreated specimens (100 μl) of plasma or urine could be added directly to the assay tubes. The anti-serum was specific for bleomycin and showed no cross-reaction with other anti-cancer agents used in combination chemotherapy. Over a concentration range of 20 to 100 ng/ml. recovery of bleomycin from plasma was 110% and from urine, 93%. Repeated assay of plasma samples showed a decrease in bleomycin levels unless the samples were kept at 4 0 C or below. Assay of bleomycin levels in plasma and urine from patients under treatment with bleomycin showed similarities with results reported using a microbiological assay. The radioimmunoassay offers a more reliable, rapid and sensitive method for the measurement of bleomycin. (author)

  1. Radioimmunoassay in the diagnosis of arterial hypertension

    Energy Technology Data Exchange (ETDEWEB)

    Slavnov, V N; Olejnik, V A; Yakovlev, A A; Yugrinov, O G; Markov, V V [Kievskij Nauchno-Issledovatel' skij Inst. Ehndokrinologii i Obmena Veshchestv (Ukrainian SSR)

    1984-11-01

    The paper is concerned with the results of a study of the aldosterone concentration and renin activity, the general level of catecholamines and their fractions in the peripheral blood and blood taken at selective venography from the vena cava inferior, renal and adrenal veins of 108 patients with hypertension, aldosteroma and idiopathic hyperaldosteronism, adrenal and extraadrenal pheochromocytoma, renovascular and renoparenchymatous arterial hypertension. The aldosterone concentration and renin activity were determined with radioimmunoassay, and the general content of catecholamines and their fractions with a radioenzymatic method using standard kits. It has been shown that the radioimmunoassay to determine the aldosterone concentration and renin activity makes possible differential diagnosis of hypertension, aldosteroma, idiopathic and secondary hyperaldosteronism. A considerable increase in the blood plasma renin activity on the affected side was revealed in the patients with renovascular hypertension, and in renoparenchymatous hypertension it was equal in both renal veins. The study of the total content of catecholamines and their fractions in the blood from different parts of the venous system can be used for topical diagnosis of adrenal and extraadrenal pheochromocytoma.

  2. Radioimmunoassay in the diagnosis of arterial hypertension

    International Nuclear Information System (INIS)

    Slavnov, V.N.; Olejnik, V.A.; Yakovlev, A.A.; Yugrinov, O.G.; Markov, V.V.

    1984-01-01

    The paper is concerned with the results of a study of the aldosterone concentration and renin activity, the general level of catecholamines and their fractions in the peripheral blood and blood taken at selective venography from the vena cava inferior, renal and adrenal veins of 108 patients with hypertension, aldosteroma and idiopathic hyperaldosteronism, adrenal and extraadrenal pheochromocytoma, renovascular and renoparenchymatous arterial hypertension. The aldosterone concentration and renin activity were determined with radioimmunoassay, and the general content of catecholamines and their fractions with a radioenzymatic method using standard kits. It has been shown that the radioimmunoassay to determine the aldosterone concentration and renin activity makes it possible to resort to differential diagnosis of hypertension, aldosteroma, idiopathic and secondary hyperaldosteronism. A considerable increase in the blood plasma renin activity on the affected side was revealed in the patients with renovascular hypertension, and in renoparenchymatous hypertension it was equal in both renal veins. The study of the total content of catecholamines and their fractions in the blood from different parts of the venous system can be used for topical diagnosis of adrenal and extraadrenal pheochromocytoma

  3. Radioimmunoassay for hepatitis B core antigen

    International Nuclear Information System (INIS)

    Sagnelli, E.; Pereira, C.; Triolo, G.; Vernace, S.; Paronetto, F.

    1982-01-01

    Serum hepatitis B core antigen (HBcAg) is an important marker of hepatitis B virus replication. We describe an easy, sensitive radioimmunoassay for determination of HBcAg in detergent-treated serum pellets containing Dane particles. Components of a commercial kit for anticore determination are used, and HBcAG is measured by competitive inhibition of binding of 125 I-labeled antibodies to HBcAg with HBcAg-coated beads. We assayed for HBcAG in the sera of 49 patients with hepatitis B surface antigen (HBsAg)-positive chronic hepatitis, 50 patients with HBsAg-negative chronic hepatitis, and 30 healthy volunteers. HBcAg was detected in 41% of patients with HBsAg-positive chronic hepatitis but not in patients with HBsAg-negative chronic hepatitis. Hepatitis Be antigen (an antigen closely associated with the core of Dane particles) determined in the same sera by radioimmunoassay, was not detected in 50% of HBcAg-positive sera

  4. Human C-peptide. Pt. 1. Radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Beischer, W; Keller, L; Maas, M; Schiefer, E; Pfeiffer, E F [Ulm Univ. (Germany, F.R.). Abt. Innere Medizin, Endokrinologie und Stoffwechsel

    1976-08-01

    Synthetic human C-peptide bearing a tyrosine group at its amino end is labelled with /sup 125/iodine using chloramin T or hydrogen peroxide and lactoperoxidase. The results of the two methods are compared. Antiserum to synthetic human C-peptide (without tyrosine), which was partially coupled to rabbit albumin, is raised in guinea pigs and goats. Goats show to be superior to guinea pips concerning antibody production. The so-called 'hook effect' phenomenon is observed when setting up the standard curves for the radioimmunoassay. Monotonically decreasing standard curves are obtained on dilution of antiserum with a high antibody titer which was produced by repeated immunization in goats. Free C-peptide and C-peptide bound to antiserum are separated using the anion exchange resin amberlite. Using this separation technique we excluded unspecific binding of labelled C-peptide to protein fractions in serum of diabetics. The sensitivity of our radioimmunoassay is approx. 0.3 ng C-peptide/ml serum. Intra- and interassay variability are below 10%. Human proinsulin is the only substance found to crossreact with the antiserum.

  5. Steroid radioimmunoassay: contribution of standards to blank values, ch. 4

    International Nuclear Information System (INIS)

    Froelich, M.; Termorshuizen, W.; Kenter, E.; Molenaar, A.J.

    1977-01-01

    The sensitivity of the radioimmunoassay of steroids is considerably reduced by high blank values which may be derived in part from co-chromatographed standards. Blank levels approach the detection limit of the radioimmunoassay of aldosterone, testosterone and androstenedione when 10,000 dpm (30-35 pg) labelled steroids are used as reference standard. When 20 μg aldosterone, testosterone, or androstenedione is used as standard, blank levels of up to 12,800 pg were measured in the radioimmunoassay. Application of the standards on a separate strip does not improve the results. From the experiments it appeared that contamination took place by transport by the solvent

  6. A competitive-inhibiton radioimmunoassay for influenza virus envelope antigens

    International Nuclear Information System (INIS)

    Russ, G.; Styk, B.; Vareckova, E.; Polakova, K.

    1976-01-01

    A double-antibody competitive-inhibition radioimmunoassay for influenza virus envelope antigens is described. A viral antigen preparation from influenza A virus recombinant MRC11 [antigenically identical to A/Port Chalmers/1/73 (H3N2)] consisting of haemagglutinin and neuraminidase was labelled with radioiodine. Rabbit antisera were allowed to react with the labelled antigen and the resultant antigen-antibody complexes were precipitated with the appropriate antiglobulin. The competitive-inhibition radioimmunoassay very sensitively elucidated differences even among closely related influenza virus strains. Attempts have been made to eliminate neuraminidase from radioimmunoprecipitation to obtain a competitive-inhibition radioimmunoassay system for haemagglutinin alone. (author)

  7. Radioiodinated derivatives for steroid radioimmunoassay. Application to the radioimmunoassay of cortisol

    International Nuclear Information System (INIS)

    Gomez-Sanchez, C.; Milewich, L.; Holland, O.B.

    1977-01-01

    Gamma-emitting steroid tracers for use in the radioimmunoassay of steroids have a number of advantages over the more common tritiated tracers. The steroid derivatives aldosterone-3-(p-hydroxybenzoyl) hydrazone, aldosterone-3-(p-hydroxyphenylpropionyl)hydrazone, deoxycorticosterone-3-(p-hydroxyphenylpropionyl)hydrazone, and cortisol-3-(p-hydroxyphenylpropionyl)hydrazone were synthesized by a one-step procedure and iodinated ([ 125 I]). To illustrate the usefulness of these derivatives, we describe the details of a cortisol radioimmunoassay. The use of the radioiodinated tracer appeared to increase the specificity of the antigen-antibody reaction when compared with [ 3 H]cortisol. The methodology involved in the preparation of the steroid derivatives described above can be extended to other 3-oxo-4-ene-containing steroids, with the advantages of economy, simplicity, and versatility

  8. Quantification of the adrenal cortex hormones with radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Badillo A, V.; Carrera D, A. A.; Ibarra M, C. M., E-mail: vbadillocren@hotmail.co [Universidad Autonoma de Zacatecas, Unidad Academica de Estudios Nucleares, Calle Cipres No. 10, Fracc. La Penuela, 98068 Zacatecas (Mexico)

    2010-10-15

    The pathologies of the adrenal cortex -adrenal insufficiency and Cushing syndrome- have their origin on the deficit or hypersecretion of some of the hormones that are secreted by the adrenal cortex, which is divided in three zones anatomically defined: the external zone, also called the zona glomerulosa, which is the main production site of aldosterone and mineralocorticoids; the internal zone, or zona reticularis, that produces androgens; and the external zone, or zone 1 orticotrop, which is responsible for producing glucocorticoids. In this work, a quantitative analysis of those hormones and their pathologic trigger was made; the quantification was made in the laboratory by means of highly sensitive and specific techniques, in this case, the radioimmunoassay, in which a radioisotope I-125 is used. This technique is based on the biochemical bond-type reaction, because it requires of a substance called the linker, which bonds to another called ligand. This reaction is also known as antigen-antibody (Ag-Ab), where the results of the reaction will depend on the quantity of antigen in the sample and on its affinity for the antibody. In this work, a 56 patients (of which 13 were men and 43 women) study was made. The cortisol, the ACTH, the androsterone and the DHEA values were very elevated in the majority of the cases corresponding to women, predominating cortisol; while in men, a notorious elevation of the 17 {alpha}-OH-PRG and of the DHEA-SO{sub 4} was observed. Based on that, we can conclude that 51 of them did not have mayor complications, because they just went to the laboratory once, while the remaining 5 had a medical monitoring, and they visited the laboratory more than one occasion, tell about a difficulty on their improvement. According to the results, an approximate relation of 8:2 women:men, respectively, becomes clear to the hormonal pathologies of the adrenal cortex. (Author)

  9. Simultaneous use of two prostaglandin radioimmunoassays employing two antisera of differing specificity. II. Relative stability of prostaglandins E1, E2, and F1alpha in cell cultures of BALB/c 3T3 and SV3T3 mouse fibroblasts

    International Nuclear Information System (INIS)

    Ritzi, E.M.; Stylos, W.A.

    1976-01-01

    The relative stability of Prostaglandins (PGs) E1, E2 and F1α in cultures of BALB/c 3T3 and SV3T3 cells has been evaluated using 3 different approaches. First, total recovery of tritium in the ethyl acetate phase following incubation and extraction of PGF1α and PGE1 demonstrated greater stability for PGF1α (88.8 percent) than PGE1 (65.9 percent). Second, analysis of incubated, extracted, tritiated PGs by thin layer chromatography revealed decreases of up to 23 percent in the PGE zone following incubation of 3H-PGE1. With increasing time of incubation, decreases in the PGE zone were accompanied by increase in PGA-like compounds. 3H-PGF1α demonstrated greater stability, having greater than 90 percent recovery of the tritium in the PGF zone. A third approach to the assessment of PG stability in culture was the comparison of the production of individual PGs by radioimmunoassay (RIA). The data obtained by RIA indicated a lag in the increase of PGA and PGB, until an initial rise in PGE was noted, suggesting that PGA and PGB may be secondary products arising from PGE which exhibits only partial stability in culture. By employing two RIAs, one for total PGE and one for PGA and PGB, the composite determination PG [E + (A + B)] can be used to provide a more meaningful determination of PG production because of the instability of the PGs. On the other hand, individual determinations are helpful in assessing the stability of PGEs in cell cultures

  10. Report on the national seminar on radioimmunoassays

    International Nuclear Information System (INIS)

    1979-01-01

    Deliberations of the National Seminar on Radioimmunoassays (RIA) held at Bombay during 16-20 January 1978 are reported. Various aspects of the application of RIA techniques in India were discussed in the seminar. They included the basic requirements of RIA in India, the state-of-the-art of RIA in India, radiation protection in RIA, quality control of RIA, usefulness and limitations of RIA in clinical diagnosis and use of RIA in tropical infectious diseases. Difficulties encountered in the practice of RIA techniques in India were found to be mainly related to availability of 125 I, RIA kits, antisera and hormones. The need for establishing a centralised assay service for RIA was examined and it was concluded that such a step would be premature in the present situation. Recommendations made and guidelines spelt out deal with resources for RIA, research, referral assay service, training and information exchange and dissemination. (M.G.B.)

  11. Radioimmunoassay for thyroid stimulating hormone (TSH)

    International Nuclear Information System (INIS)

    1980-01-01

    An improved double antibody radioimmunoassay method is described for the determination of thyroid stimulating hormone (TSH) in biological and other fluids. Highly purified second antibody is immobilised on to hydrophilic, hydrolyzed polyacrylamide particles of a suspendable size to form a solid phase second antibody reagent. The immobilised second antibody reagent is used to precipitate the reaction product of the first antibody with labelled and unlabelled thyroid stimulating hormone (TSH-anti-TSH-complex) so as to produce a two-phase system which permits rapid and efficient separation of bound TSH in the solid phase from free TSH in the liquid phase. Details of the preparation of this novel second antibody-polyacrylamide reagent and of the assay procedure for human TSH are described. (U.K.)

  12. Radioimmunoassay of luteinizing hormone in hypothyroidism

    International Nuclear Information System (INIS)

    Sobieszczyk, S.

    1975-01-01

    Radioimmunoassay of luteinizing hormone was performed in 18 women with primary hypothyroidism and 15 women with secondary hypothyroidism. The results of determinations were compared with LH values found in healthy women at reproductive age and after menopause. It was observed that in primary hypothyroidism the level of LH is normal, in young women it was from 6 to 25 m IU/ml, while in the postmenopausal period it increased to 70 to 200 m IU/ml. In secondary hypothyroidism due to pituitary hypofunction the LH level is undetectable or lies in the range of lowest values observed in healthy subjects, not exceeding 8 m IU/ml. Determinations of serum LH may be useful for differential diagnosis of primary and secondary hypothyroidism. (author)

  13. A radioimmunoassay for neurotensin in human plasma

    International Nuclear Information System (INIS)

    Blackburn, A.M.; Bloom, S.R.

    1979-01-01

    A radioimmunoassay was developed for detecting the neurotensin peptide in human plasma. The plasma was specific for neurotensin as no cross-reaction was found with any of the other gut hormones tested. Changes of 5 pmol/l could be detected with 95% confidence. Neurotensin was unstable in both blood and plasma but considerable protection was afforded by addition of aprotinin, rapid separation of plasma and immediate deep freezing. Neurotensin-like immunoreactivity was detected in human plasma in both a small and large molecular form. The mean fasting level of plasma neurotensin-like immunoreactivity in 36 healthy volunteers was 29 +- 3 pmol/l. A significant increment of 27 +- 8 pmol/l plasma neurotensin immunoreactivity was detected after a large meal in nine healthy men. In view of the present results in man and also of neurotensin's potent pharmacological actions in experimental animals, neurotensin appears to fulfil some of the criteria needed for a hormone. (UK)

  14. Sensitive radioimmunoassay for plasma arginine vasopressin

    International Nuclear Information System (INIS)

    Thibonnier, M.; Soto, M.E.; Corvol, P.; Milliez, P.; Marchetti, J.; Menard, J.

    1980-01-01

    Using an ion exchange resin, a sensitive radioimmunoassay for plasma arginine vasopressin (AVP) was developed. This assay was characterized by the absence of blank values, an excellent recovery rate, great sensitivity (0.1 pg of AVP was significantly detected) and reproducibility. In 8 normal men, plasma AVP after overnight dehydration was 1.57+-0.17 pg/ml, and dropped to 0.58+-0.11 pg/ml after 20 ml/kg oral water loading. Significant correlations between plasma AVP levels and plasma or urinary osmolality confirm the validity of this assay. In complete pituitary diabetes insipidus (n=4) plasma AVP was undetectable whereas it was frankly increased in Schwartz-Bartter syndrome (3 to 33 pg/ml, n=8) [fr

  15. Standardised multicentre procedure for plasma gonadotrophin radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Ferguson, K M; Hayes, M; Jeffcoate, S L [Chelsea Hospital for Women, London (UK)

    1982-09-01

    A radioimmunoassay method for the assay of luteinising hormone (LH) and follicle-stimulating hormone (FSH) in serum/plasma has been designed for use in laboratories of varying expertise in the United Kingdom. The major sources of experimental error leading to poor within-laboratory performance and between-laboratory comparability were identified: quality of tracer, use of calibration standards, and separation procedure. A simple rugged kit was designed which was extensively tested first in our laboratory and then in a small multi-centre field trial before being made available. It is now used routinely by 26 health service and research laboratories. The working range of the assays is 1-50 IU/l (LH) and 0.3-16 IU/l (FSH). The between-batch reproducibility was 5-11% (CV) over the dose range 4.8-18 IU/l (LH) and 1.6-15 IU/l (FSH).

  16. Radioimmunoassay of 17-beta - estradiol in plasma

    International Nuclear Information System (INIS)

    Kyian, T.S.; Wajchenberg, B.L.

    1980-01-01

    A radioimmunoassay technique for the measurement of plasma E 2 was standardized utilizing a highly specific antisera against E 2 [-6(-0-carboximetil)-oxime] BSA without the need of previous chromatographic purification. The anti-E 2 serum was highly specificic, showing high affinity with affinity constants:K 1 =1.62 x 10 12 M -1 and K 2 =2.94x10 11 M -1 , calculated by Scatchard plot. The standard-curve sensitivity was 2 picograms. The method was specific and accurate, showing an intra-assay precision with a mean C.V. of 2.9%, with the inter-assay evaluation showing a mean C.V. of 5.0%. This method was employed to evaluate E 2 secretion during the menstrual cycle in 6 normal female. (Author) [pt

  17. Radioimmunoassay in basic and clinical pharmacology

    International Nuclear Information System (INIS)

    Patrono, C.; Peskar, B.A.

    1987-01-01

    The subject of the book is the development, validation and application of radioimmunoassay (RIA) techniques for the measurement of a variety of substances in animal and human body fluids. The book discusses methodological and conceptual issues related to the main classes of mediators of drug action and to drugs themselves, as assayed by this particular analytical technique. A number of introductory chapters provide basic information concerning production and characterization of antibodies, labeling techniques, statistical aspects and validation criteria, insight into problems related to the development and validation of RIA for the newly discovered mediator(s). In the following chapters, the emphasis is placed on the technical details relevant to each class of compounds and on specific aspects of their applications to basic and/or clinical pharmacological studies. New developments in this area, such as monoclonal antibodies and non-radioactive labeling techniques, are also covered

  18. Beta-endorphin radioimmunoassay: specificity studies

    Energy Technology Data Exchange (ETDEWEB)

    Colas-Linhart, N.; Perdrisot, R.; Petiet, A.; Bok, B.

    1986-01-01

    This note describes the technical details of a cerebrospinal fluid (CSF) B-Endorphin (B-End) radioimmunoassay (RIA). We used an antiserum raised in rabbits against human B-End which cross-reacs 100% with B-Lipotrophin (B-Lph). Thus, filtration chromatography is used to separate both peptides. The assay is sensitive (limit detection=14 pmoles/l), reproducible (the intra and inter assay coefficients of variation are 5 and 6% respectively). Specificity studies are performed to verify the cross-reactions with other opioid peptides and the non specific reactions with the biological fluid (CSF). In order to evaluate the effects of iodine-containing contrast media on the RIA, additional standard curves were analyzed in the presence of varying concentrations of two contrast materials.

  19. Quantitation of secretory protein levels by radioimmunoassay

    International Nuclear Information System (INIS)

    Klein, J.L.; Dawson, J.R.

    1978-01-01

    A radioimmunoassay was designed for the detection of secretory protein, a component of secretory immunoglobulin A, in human serum. The assay uses free secretory protein isolated from human colostrum, and antisera raised in rabbits to be purified antigen. The mean level of secretory protein in the control group was 2.34+-0.41 μg/ml (mean+-S.E.M.). The level in cord blood was slightly lower (0.74+-0.26 μg/ml), while the level in patients with ovarian carcinoma was significantly increased (12.67+-1.43 μg/ml). Pregnant women have increasingly secretory protein levels with increasing length of gestation (5.86+-2.02, 11.55+-1.30 and 17.00+-1.16 μg/ml for the first, second and third trimesters, respectively. (Auth.)

  20. Radioimmunoassay for progesterone in bovine milk

    International Nuclear Information System (INIS)

    Ruiz, Miriam; Figueredo, Nancy; Castillo, Sonia; Pizarro

    2002-01-01

    A system for the measurement of progesterone in bovine milk by radioimmunoassay has been developed and validated. This assay includes an iodine tracer purified by HPLC, the standard prepared in fat-free milk and an antibody anti-progesterone combined with second antibody. The detection limit of the assay is at 0.2 nmol/L calculated from the maximum binding menus two standard deviations and the precision is satisfactory. In the recovery assay was used 4 milk different samples and the result was 98% of recuperation. The progesterone was determinate in milk samples from post-partum animals taking samples three times per week for 40 days. The assay is simple, rapid and possibility the progesterone measurement without sample dilution, distinguish the cyclic changes of this hormone that reflect the ovarian activity in the animals. (author)

  1. Radioimmunoassay for GRF and CRF in humans

    International Nuclear Information System (INIS)

    Stalla, G.K.; Losa, M.; Kaliebe, T.; Stalla, J.; Schopohl, J.; Muller, O.A.; Von Werder, K.

    1987-01-01

    In 1981 the structure of ovine CRF was established. One year later others isolated a 44 amino acid peptide with GH releasing activity from a pancreatic islet cell carcinoma of an acromegalic patient (hp GRF/sup 1-44/). In 1983 the gene of human CRF was cloned and the amino acid sequence of hCRF could be elucidated. It differs in 7 amino acids from oCRF. Many investigators demonstrated the biological activity of these peptides in vitro and in vivo. The aim of the authors study was to establish radioimmunoassays for GRF and hCRF with the synthetic derivates, measure endogenous GFR and CRF and circulating GRF- and CRF-levels after intravenous injection and calculate metabolic clearance rate and half-time of disappearance from serum for both releasing hormones

  2. Radioimmunoassay of human plasma protein C

    International Nuclear Information System (INIS)

    Wang Bocheng; Li Jinquan; Jing Jian; Zhang Manda

    1995-01-01

    A radioimmunoassay method for the measurement of human plasma protein C (PC) is established. PC was isolated and purified from human plasma. The antisera against PC was obtained by immunizing rabbits. Iodination of PC was carried out with chroramine-T. The sensitivity was 3.94% μg/L, and the assay covered 6.25∼1024 μg/L for PC. The intra-assay and inter-assay CV were 4.4% and 9.68% respectively, with a recovery rate of 104.28%. There was no cross reaction with factor II. The normal value was 3.84 +- 0.34 mg/L in 36 normal persons. Value of 1.03 +-0.41 mg/L was found in 16 patients with fulminating hepatitis complicated with coagulation disturbance. It is an effective approach for the diagnosis of hereditary or acquired PC deficiency and also for the study of thrombotic diseases

  3. Radioimmunoassay for medical diagnosis and research

    Energy Technology Data Exchange (ETDEWEB)

    Dudley, R A; Vavrejn, B [International Atomic Energy Agency, Vienna (Austria). Div. of Life Sciences

    1982-12-01

    Health and disease in living systems depend on the dynamic interplay of thousands of biochemical substances occurring in living systems in concentrations ranging from parts per hundred to parts per billion or trillion. Radioimmunoassay (RIA) is a highly specific and sensitive technique for measuring the concentration of such biochemical substances. It represents one of the most dramatically expanding areas of medical diagnosis and research. Reviewed here is the recent progress on RIA, with emphasis on methodology and on its adaptation and application in developing countries. The number of biological substances (ligands) being assayed by RIA continues to expand. RIA is central to diagnosis, epidemiology and research. It has been successfully applied in the study of parasitic and infectious diseases. Introduction of RIA into developing countries, for which the Agency's help is sought and given, presents numerous problems: personnel, equipment, adaptability of techniques to local needs, and public support.

  4. Radioimmunoassay of urine oxytocin in man

    International Nuclear Information System (INIS)

    Zebidi, A.; Geelen, G.; Allevard, A.M.; Sempore, B.; Jarsaillon, E.; Meunier, C.; Gharib, C.

    1978-01-01

    A radioimmunoassay (RIA) for oxytocin (OT) in urine is described. 125 I-OT was prepared, and antibodies were raised in rabbits against OT coupled to bovine serumalbumine. This allowed us to set up a RIA for OT which limit of detection is 1.25 pg/tube (0.6 μU). The use of an extraction procedure using CG 50 Amberlite is essential. The recovery after extraction reaches 70.5 %. pH 5 is the optimum pH were urine samples must be stored. The superposition of the elution peak of endogenous OT on that of exogenous hormone is an argument in favour of the validity of such an extraction procedure. Daily urinary excretion of OT reaches 9.58 mU +- 3.48 in 18 healthy young men [fr

  5. Radioimmunoassay of cholecystokinin in tissue and plasma

    International Nuclear Information System (INIS)

    Jansen, J.B.M.J.

    1984-01-01

    The physiological and pathophysiological role of the pancreas hormone, the polypeptide 'cholecystokinin' (CCK) is not well-established yet. This is due to the lack of specific and reliable radioimmunoassays for CCK. The aim of this thesis is to develop such an assay meeting the requirements of high specificity and sensitivity. Several problems were faced, such as (1) the cross-reactivity of existing antibodies with the stomach hormone gastrin and (2) changes in immunoreactivity caused by the introduction of the labelling isotope 125 I and various labels (prepared according to the Bolton-Hunter method) into the polypeptide. The reliability of the assay for the measurement in human tissue and blood is extensively evaluated, inter alia, in patients with pancreas insufficiency (alcohol, cystic fibrosis) and with coeliac disease. (Auth.)

  6. Radioimmunoassays for catalase and glutathion peroxidase

    International Nuclear Information System (INIS)

    Baret, A.; Courtiere, A.; Lorry, D.; Puget, K.; Michelson, A.M.

    1982-01-01

    Specific and sensitive radioimmunoassays for human, bovine and rat catalase (CAT) and glutathion Peroxidase (GPX) are described. The obtained values are expressed as enzymatic units per μg of immunoreactive protein. They appear to closely correspond to specific activities of the purified enzymes determined by colorimetric protein-assay. Indeed, the values of the specific activities of purified human CAT is 57.9 k/mg and that of purified rat GPX is 180 units/mg. This result validates the present RIAs and the association of the two techniques allows the determination of a further parameter. In conclusion, RIAs for CAT and GPX can be applied with great specificity and sensitivity to a wide variety of human, rat and bovine medias

  7. Radioimmunoassay of progesterone in unextracted serum

    International Nuclear Information System (INIS)

    Haynes, S.P.; Corcoran, J.M.; Eastman, C.J.; Doy, F.A.

    1980-01-01

    A rapid, precise radioimmunoassay for progesterone in 25 μL of unextracted serum is described. Progesterone is released from its binding protein by adding an optimal amount of cortisol, which binds to the same protein (cortisol binding globulin) as progesterone. The amount of cortisol required does not cross react with the specific progesterone antibody used. This approach considerably shortens assay time and removes a tedious and imprecise stage in the conventional assay of serum progesterone. Results correlated well (r = 0.97) with a method involving organic solvent extraction of progesterone from serum. During the two years we have used this mehod in a busy diagnostic endocrine laboratory, the between-assay precision (CV) for low-, medium-, and high-concentration quality control sera was 12, 7, and 9%, respectively. Data from participation in an independent external quality-control program verified the adequacies of the method

  8. A beta-endorphin radioimmunoassay: specificity studies

    International Nuclear Information System (INIS)

    Colas-Linhart, N.; Perdrisot, R.; Petiet, A.; Bok, B.

    1986-01-01

    This note describes the technical details of a cerebrospinal fluid (CSF) B-Endorphin (B-End) radioimmunoassay (RIA). We used an antiserum raised in rabbits against human B-End which cross-reacs 100% with B-Lipotrophin (B-Lph). Thus, filtration chromatography is used to separate both peptides. The assay is sensitive (limit detection=14 pmoles/l), reproducible (the intra and inter assay coefficients of variation are 5 and 6% respectively). Specificity studies are performed to verify the cross-reactions with other opioid peptides and the non specific reactions with the biological fluid (CSF). In order to evaluate the effects of iodine-containing contrast media on the RIA, additional standard curves were analyzed in the presence of varying concentrations of two contrast materials [fr

  9. Radioimmunoassay of cholecystokinin in human plasma

    International Nuclear Information System (INIS)

    Byrnes, D.J.; Henderson, L.; Borody, T.; Rehfeld, J.F.

    1981-01-01

    A sensitive radioimmunoassay for cholecystokinin (CCK) has been developed. Porcine CCK-33 was labelled by conjugation with 125 I-hydroxyphenyl-propionic acid succinimide ester. Antibodies were raised against porcine CCK-33 covalently coupled to egg albumin. Plasma samples were extracted with 96% ethanol prior to assay. Free and bound hormone were separated by dextran-coated charcoal. The antibodies bound CCK-8 and CCK-33 with equimolar potency. The assay detection limit was 1 pmol/l plasma. Within and between assay coefficients of variation were +-12.7 and 13.0% at mean plasma CCK concentrations of 13.2 and 13.6 pmol/l. The concentration of CCK in 47 normal fasting subjects ranged from undetectable to 22 pmol/l. Ingestion of a mixed meal in 9 normal subjects increased the plasma concentration from 8.3 +- 2.5 S.E. to 24.4 +- 6.5 pmol/l. (Auth.)

  10. Radioimmunoassay of influenza A virus haemagglutinin. I

    International Nuclear Information System (INIS)

    Russ, G.; Styk, B.; Polakova, K.

    1978-01-01

    Haemagglutinin released from influenza A virus recombinant MRC11 [antigenically identical to the strain A/Port Chalmers/1/73 (H3N2)] by bromelain treatment and purified by rate zonal centrifugation (further on B-HA) was examined for possible contamination by neuraminidase. Specific enzymatic activities of the MRC11 virus and the B-HA respectively showed that B-HA contained less than 0.1% of enzymatically active neuraminidase originally present in the virus. Gel double diffusion tests, specificities of rabbit antisera induced by B-HA as well as radioimmunoprecipitation experiments demonstrated that B-HA was devoid of any antigenically active neuraminidase. Precipitation of 125 I-labelled B-HA with antisera to influenza virus recombinants with N2 neuraminidase was evidently caused by antibodies to host antigenic determinant(s) present in these sera. As for purity and radioimmunoprecipitation properties, B-HA is quite suitable for radioimmunoassay experiments. (author)

  11. Automatic computation of radioimmunoassay data. Insulin and C-peptide

    Energy Technology Data Exchange (ETDEWEB)

    Toyota, T; Kudo, M; Abe, K [Hirosaki Univ., Aomori (Japan). School of Medicine; Kawamata, F; Uehata, S

    1975-09-01

    Radioimmunoassay provided dose response curves which showed linearity by the use of logistic transformation (Rodbard). This transformation which was applicable to radioimmunoassay should be useful for the computer processing of insulin and C-peptide assay. In the present studies, standard curves were analysed by testing the fit of analytic functions to radioimmunoassay of insulin and C-peptides. A program for use in combination with the double antibody technique was made by Dr. Kawamata. This approach was evidenced to be useful in order to allow automatic computation of data derived from the double antibody assays of insulin and C-peptides. Automatic corrected calculations of radioimmunoassay data of insulin was found to be satisfactory.

  12. Synthesis of tritium or deuterium labelled 19-nor-3. cap alpha. -hydroxy-5. cap alpha. -androstan-17-one from nortestosterone

    Energy Technology Data Exchange (ETDEWEB)

    Protiva, J; Klinotova, E [Karlova Univ., Prague (Czechoslovakia). Prirodovedecka Fakulta; Filip, J [Ustav pro Vyzkum, Vyrobu a Vyuziti Radioisotopu, Prague (Czechoslovakia); Hampl, R [Research Inst. of Endocrinology, Praha (Czechoslovakia)

    1982-10-20

    Tritium and/or deuterium (5-H) labelled 19-nor-3..cap alpha..-hydroxy-5..cap alpha..-androstan-17-one (norandrosterone) was prepared from nortestosterone in view to use it as a radioligand for radioimmunoassay of the main nortestosterone metabolites. Based upon model experiments using testosterone and deuterium labelling, the following four step procedure was established: nortestosterone was oxidized with pyridine chlorochromate and the resulting 19-nor-4-androsten-3,17-dione was tritiated with tritium gas under catalysis with tris(triphenylphosphine)rhodium chloride to give (4,5..cap alpha..-/sup 3/H)19-nor-5..cap alpha..-androstan-3,17-dione. A selective reduction of the latter compound yielded (5-/sup 3/H)19-nor-3..cap alpha..-hydroxy-5..cap alpha..-androstan-17-one of the molar radioactivity 0.3 TBq (8.15 Ci)/mmol.

  13. Determination of plasma oxytocin by radioimmunoassay

    International Nuclear Information System (INIS)

    Ogawa, Satsuki; Fukuchi, Soitsu; Miura, Tadashi

    1978-01-01

    A simple radioimmunoassay was applied to the measurement of oxytocin in human plasma. A high specificity of immunoassay was demonstrated by the fact that large excess of angiotensin I and II, and ACTH did not displace labelled oxytocin from the antibody. Lysine-8-vasopressin and arginine-8-vasopressin showed very little cross-reaction in the assay, possessing only 0.002% of the immunological potency of oxytocin. The specific activity of 125 I-oxytocin was 166 μCi/μg. Adsorption and extraction capacities of Florisil were 96.6 +- 2.1% and 85.7 +- 2.5%, respectively. Intra- and inter-assay variability were 7.2 +- 4.9% and 4.3 +- 2.2%, respectively. The sensitivity of the assay was below 1 pg/tube. Normal levels of plasma oxytocin were 0 - 2.2 pg/ml (n=13) in males and 0 - 10.4 pg/ml (n=10) in females. Plasma oxytocin levels in the 39th and 40th weeks of pregnancy were 27.9 +- 4.14 pg/ml (n=4) and 29.8 +- 17.1 pg/ml (n=13), respectively. The levels increased to 33.1 +- 12.1 pg/ml (n=7) and 37.1 +- 17.5 pg/ml (n=7) in the first and third stages of labor, and decreased to 13.6 +- 5.25 pg/ml (n=6) on the 2nd to 8th day after labor. The radioimmunoassay for oxytocin in plasma is considered to be sufficiently applicable for clinical use. (auth.)

  14. Autism Spectrum Disorders and Maternal Serum α-Fetoprotein Levels During Pregnancy

    DEFF Research Database (Denmark)

    Abdallah, Morsi; Grove, Jakob; Hougaard, David M

    2011-01-01

    Objective: Numerous studies have been trying to disentangle the complex pathophysiology of autism spectrum disorders (ASD). In our study, we explored the potential role of maternal serum (MS) α-fetoprotein (AFP) in the prediction and the pathophysiology of ASD. Methods: A total of 112 patients...

  15. Examination of illegal, non declared injection preparations on anabolic hormones and development of a radioimmunoassay for 19-nortestosterone

    Energy Technology Data Exchange (ETDEWEB)

    Rapp, M.; Meyer, H.H.D.

    Procedures are described for the identification and quantification of anabolic hormones in preparations for injection. To perform radioreceptorassays for the most important groups of anabolics, estrogens and androgens, hydrolysis of esters is absolutely necessary in order to release the active substances from depot preparations. After moderate hydrolysis (30 min; 45/sup 0/C) it was possible to identify 19-nortestosterone-17..beta.. as active substance in three various illegal solutions for injection and one of them contained in addition estradiol-17..beta... After more drastic hydrolysis (2 h; 70/sup 0/C) there was a strong disintegration of 19-nortestosterone-17..beta.. and new more polar compounds were found, e.g. in small amounts 6-ketoestradiol, with estrogenic activity. Among the anabolic steroids only trenbolone-17..beta.. behaved in similar manner, and the disintegration during drastic hydrolytic conditions gave an indication for the existence of 19-norsteroids. Specific radioimmunoassays and GC/MS were used for final identification of the anabolic agents. Quantitation was carried out with HPLC/UV and radioimmunoassays after moderate hydrolysis. Since 19-nortestosterone-17..beta.. was identified a specific radioimmunoassay for this hormone was developed. Antibodies were raised against 19-nortestosterone-17..beta..-HS-BSA in order to be able to measure 19-nortestosterone-17..cap alpha.., 19-nortestosterone-17..beta.., and 19-norandrostenedione which were expected as the main bovine metabolites.

  16. Radioimmunoassay of methaqualone in human urine compared with chromatographic methods

    International Nuclear Information System (INIS)

    Mule, S.J.; Kogan, M.; Jukofsky, D.

    1978-01-01

    The 125 I-radioimmunoassay for methaqualone in human urine was evaluated by a comparison with newly modified gas-liquid chromatographic and thin-layer chromatographic methods. The statistically significant sensitivity value for the radioimmunoassay was at 2 μg of methaqualone per liter of urine. The coefficient of variation was 2.88 -+ 0.16% intraassay. There was cross-reactivity only with metabolites of methaqualone, 4'-hydroxymethaqualone being twice as sensitively measured as methaqualone. There was complete agreement between results by radioimmunoassay and by gas-liquid chromatography in 96.7% of the samples analyzed. Only 1.2% of the radioimmunoassay values were false positives, and 2.1% false negatives (phi = 0.8917, P < 0.001). Comparisons between the thin-layer chromatographic data and the gas--liquid chromatographic or radioimmunoassay data showed less agreement because of the 50- to 200-fold higher sensitivity of the latter techniques. Gas--liquid chromatography therefore appears to represent the best reference method for the evaluation of the radioimmunoassay, which appears to be a very sensitive and reliable technique for detecting methaqualone and its metabolites in human urine

  17. Radioimmunoassay for platelet activation specific protein GMP-140 on the platelet surface and in plasma

    International Nuclear Information System (INIS)

    Wu Guoxin; Li Jianyong; Ruan Changgeng

    1991-08-01

    Using monoclonal antibody (McAb) SZ-51 which is specific for an alpha-granule membrane protein (GMP-140) on the surface of human activated platelets, the platelet GMP-140 expression in fixed whole blood was measured by direct radioimmunoassay and GMP-140 microparticles in plasma was measured by sandwich method. The GMP-140 molecules per platelet or milliliter (mL) were calculated for the following subjects; acute myocardial infarction; cerebro thrombosis; diabetic mellitus; asthma attack; epidemic hemorrhagic fever etc.. By comparing with the concentration of thromboxane B 2 (TXB 2 ) and von Willebrand factor (vWF) in plasma, it is confirmed that the measurement of GMP-140 molecules is better than that of TXB 2 and vWF. It is a sensitive and specific method for evaluating the platelet activation degree in vivo. The establishment of this method will be useful to diagnosing the thrombotic disorders and studying the pathogenesis of some other diseases

  18. Radioimmunoassay and related procedures in medicine

    International Nuclear Information System (INIS)

    1978-01-01

    Full text: Radioassay procedures for the measurement of substances such as hormones, vitamins and drugs in the body fluids and tissues, above all in the blood, are now in the front rank of medical applications of radioactive materials. These procedures, which are carried out on specimens in the medical laboratory and do not involve the administration of any radioactive material to the patient, are now widely employed in the routine diagnosis and investigation of disease, whilst their use in research has led to important advances in many branches of medicine. Typical of radioassay methods is radioimmunoassay, which depends on the antigen-antibody reaction between the substance to be measured and the antibodies in an antiserum against that substance produced in a guinea-pig, rabbit, sheep or other animal. The importance of radioimmunoassay was recently underlined by the award of the Nobel Prize in medicine for 1977 to Dr. Rosalyn Yalow of the United States of America for her pioneer work on the method over the past two decades, particularly in relation to the measurement of protein hormones. This symposium was the third on the subject to have been sponsored by the IAEA. The first took place in Vienna in 1969 and the second in Istanbul in 1973. During the four years from 1973 to 1977, the growing commercial availability of reagents and kits for radioassays brought them into routine use. This in turn led to an increasing awareness of the need for assay standardization and quality control and to an increasing attention to techniques of assay data analysis. The burgeoning demands made on assay services stimulated interest in the possibilities for automation of assay procedures. Promising new methods were developed, notably solid-phase radioassay and radioreceptor assay. At the same time there was a resurgence of interest in alternative assay methods not based on the use of radioactive materials, which made a critical re-examination of the entire subject desirable. The

  19. Radioimmunoassay and related procedures in medicine

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1978-02-15

    Full text: Radioassay procedures for the measurement of substances such as hormones, vitamins and drugs in the body fluids and tissues, above all in the blood, are now in the front rank of medical applications of radioactive materials. These procedures, which are carried out on specimens in the medical laboratory and do not involve the administration of any radioactive material to the patient, are now widely employed in the routine diagnosis and investigation of disease, whilst their use in research has led to important advances in many branches of medicine. Typical of radioassay methods is radioimmunoassay, which depends on the antigen-antibody reaction between the substance to be measured and the antibodies in an antiserum against that substance produced in a guinea-pig, rabbit, sheep or other animal. The importance of radioimmunoassay was recently underlined by the award of the Nobel Prize in medicine for 1977 to Dr. Rosalyn Yalow of the United States of America for her pioneer work on the method over the past two decades, particularly in relation to the measurement of protein hormones. This symposium was the third on the subject to have been sponsored by the IAEA. The first took place in Vienna in 1969 and the second in Istanbul in 1973. During the four years from 1973 to 1977, the growing commercial availability of reagents and kits for radioassays brought them into routine use. This in turn led to an increasing awareness of the need for assay standardization and quality control and to an increasing attention to techniques of assay data analysis. The burgeoning demands made on assay services stimulated interest in the possibilities for automation of assay procedures. Promising new methods were developed, notably solid-phase radioassay and radioreceptor assay. At the same time there was a resurgence of interest in alternative assay methods not based on the use of radioactive materials, which made a critical re-examination of the entire subject desirable. The

  20. Approaches to a markedly increased sensitivity of the radioimmunoassay for thyrotropin-releasing hormone by derivatization

    Energy Technology Data Exchange (ETDEWEB)

    Visser, T J; Klootwijk, W [Erasmus Universiteit, Rotterdam (Netherlands). Dept. of Internal Medicine 3 and Clinical Endocrinology

    1981-04-03

    Studies on the specificity of the antisera obtained suggested that the sensitivity of the radioimmunoassay for TRH may be increased substantially by prior conversion of the hormone into dinitrophenylene derivatives. To test this possibility, several TRH-Dnp derivatives were prepared by reaction of TRH with equimolar amounts of 1,5-difluoro-2,4-dinitrobenzene yielding N/sup im/-(5-fluoro-2,4-dinitrophenyl)TRH. This intermediate was reacted with ammonia, histamine, tyramine or N/sup ..cap alpha../-acetyl-lysine methyl ester (N/sup ..cap alpha../Ac-LysOMe) to yield the respective unsubstituted and N-substituted N/sup im/-(5-amino-2,4-dinitrophenyl)TRH derivatives: TRH-Dnp-NH/sub 2/, TRH-Dnp-histamine, TRH-Dnp-tyramine and TRH-Dnp-N/sup ..cap alpha../Ac-Lys-OMe. N/sup im/-(2,4-Dinitrophenyl)TRH was prepared similarly by reaction of TRH with 1-fluoro-2,4-dinitrobenzene. The products were isolated by means of high-performance liquid chromatography (HPLC) and were found to be pure by HPLC and thin-layer chromatography using several solvent systems. TRH-Dnp-histamine and TRH-Dnp-tyramine were labelled with /sup 125/I using the chloramine-T method. The labelled products were purified to homogeneity by ion-exchange chromography on SP-Sephadex and adsorption chromatography on Sephadex LH-20, respectively, and were found by HPLC to be pure.

  1. Radioimmunoassay and the hormones of thyroid function

    International Nuclear Information System (INIS)

    Stahl, R.J.

    1975-01-01

    Radioimmunoassay (RIA) has provided the tools for wide-reaching investigations that have changed and continue to change many important concepts of thyroid physiology and pathophysiology. The RIA for human thyrotropin (TSH) was developed in 1965; development of the RIA for triiodothyronine (T 3 ), thyroxine(T 4 ), thyroxine-binding globulin (TBG), and recently, thyrotropin-releasing hormone (TRH) and thyroglobulin (Tg) followed. The capacity to measure nanogram and picogram concentrations with relative ease and speed has permitted the demonstration of dynamic relationships of the intrathyroidal and circulating thyroid hormones to each other and to the pituitary and hypothalamic regulating hormones. Evidence for the presence of cross-influences between TRH and other hypothalamic regulating hormones on the secretion of pituitary hormones has accumulated. The impact of the new information on clinical practice is now becoming evident. There is new appreciation of the value of assaying serum T 3 and TSH concentrations in the clinical management of patients with disturbed function of the thyroid, pituitary, or hypothalamus. The necessary components for RIA performance can be purchased separately or in kit form from commercial sources. With appropriate quality-control procedures, precise, sensitive, and reliable data can be generated. Awareness of the specific technical problems relating to the RIA of these hormones is absolutely necessary to assure reliable results. The availability of kits or their components permits the performance of these studies in the community hospital and in reliable commercial-service laboratories. (U.S.)

  2. Chemical Kinetics of Progesterone Radioimmunoassay System

    International Nuclear Information System (INIS)

    Abdel-Fattah, A.A.; Moustsfs, K.A.; El-Kolally, M.T.

    2004-01-01

    Progesterone is one of the steroids secreted by the corpus Iuteum in females during the menstrual cycle, and in a much higher amount by the placenta during pregnancy. It is also secreted in a minor quantities by the adrenal cortex in both males and females. Measurement of serum progesterone represents one of diagnostic values in menstrual disorders and infertility. The progesterone radioimmunoassay is based on the competition between unlabelled progesterone and a fixed quantity of 125 I-labeled progesterone for a limited number of binding sites on progesterone specific antibody. Allowing for a fixed amount of magnetizable immunosorbent to react, the antigen-antibody complex is bound on solid particles which are then separated by magnetic rack, and the radioactivity of the solid phase was counted using gamma counter. In this work, the chemical kinetics of the assay was followed, where the specific rate constant (K) was calculated at 4 degree and 37 degree and the activation energy (E act ) were calculated and the reaction rate was deduced

  3. Radioimmunoassay in developing countries: General principles

    Energy Technology Data Exchange (ETDEWEB)

    Piyasena, R D

    1993-12-31

    Radioimmunoassay (RIA) is probably the most commonly performed nuclear medicine technique. It is an in vitro procedure, where no radioactivity is administered to the patient. But this alone is not the reason for its widespread use. It provides the basis for extremely sensitive and specific diagnostic tests, and its use in present day medicine has brought a virtual information explosion in terms of understanding the pathophysiology of many diseases. The fact that the technology involved is within the technical and economic capabilities of the developing world is evident from the increasing demand for its introduction or expansion of existing services. RIA facilities need not be restricted to urban hospitals, as in the case of in vivo nuclear medicine techniques, but may be extended to smaller district hospitals and other laboratories in peripheral areas. It is also possible to send blood samples to a central laboratory so that a single centre can serve a wide geographical area. There are many laboratories in the industrialized world that receive a major proportion of samples for assay by mail. In recent years, substantial RIA services have been established in many of the developing countries in Asia and Latin America. The International Atomic Energy Agency (IAEA) and World Health Organisation (WHO) have made vital contributions to these activities and have played a catalytic role in assisting member states to achieve realistic goals. In the past five years, more than 250 individual RIA laboratories in developing member states have been beneficiaries of IAEA projects

  4. Radioimmunoassay study of neurophysins in human plasma

    International Nuclear Information System (INIS)

    Reinharz, A.C.; Tissot-Berthet, M.-C.; Vallotton, M.B.

    1978-01-01

    Using a homologous system we have developed a specific and sensitive radioimmunoassay for the measurement of one of the human neurophysins in unextracted human plasma. This neurophysin is specifically secreted in response to oestrogen and has therefore been referred to as human oestrogen-stimulated neurophysin (h-OeSN). The plasma concentration was 0.57 plus minus 0.17 ng/ml (SD) in females and 0.88 plus minus 0.76 ng/ml (SD) in males. This difference is not significant. In women on oral contraceptives, plasma h-OeSN was 2.0 plus minus 1.1 ng/ml. During pregnancy h-OeSN increased progressively to 3.7 plus minus 2.9 ng/ml at the end of the first trimester, and 5.2 plus minus 2.8 ng/ml at term. Plasma h-OeSN concentrations increased rapidly and markedly in men treated with ethinyl-oestradiol. We have also demonstrated the presence of h-OeSN in amniotic and cerebrospinal fluids. A second human neurophysin, which is stimulated by nicotine but not by oestrogen, was also measured. This neurophysin was monitored by a heterologous system using antiserum raised against bovine neurophysin II (b-NII), and b-NII as the standard and tracer. (author)

  5. Radioimmunoassay of creatine kinase BB isoenzyme

    Energy Technology Data Exchange (ETDEWEB)

    Jianguo, Geng [Shanghai Medical Univ. (China). Zhongshan Hospital; and others

    1988-11-01

    A radioimmunoassay of creatine kinase BB isoenzyme (CK-BB) was developed by using CK-BB purified from human brain. The CK-BB antiserum was raised by immunizing rabbite and {sup 125}I-CK-BB iodinated with Bolton-Hunter reagent. The affinity constant was 3.0 x 10{sup 9} mol/L. No cross reactions with creatine kinase MM isoenzyme and neuron-specific enolase were found. The measuring range was 3.5 x 10{sup -8} {approx} 1.2 x 10{sup -5} mmol/L, the average recovery rate 97.5%, with the inter and intrassay CV 3.1% and 12%, respectively. The average serum CK-BB concentration in 83 normal persons was 1.5 x 10{sup -7} +- 8.1 x 10{sup -8} mmol/L, quite different from the values of acute myocardial infarction (5.2 x 10{sup -6} +- 1.2 x 10{sup -4} mmol/L, n = 28) and cerebral vascular accident (8.4 x 10{sup -4} +- 5.0 x 10{sup -4} mmol/L, n = 10).

  6. Radioimmunoassay of bovine heart protein kinase

    International Nuclear Information System (INIS)

    Fleischer, N.; Rosen, O.M.; Reichlin, M.

    1976-01-01

    Immunization of guinea pigs with bovine cardiac cAMP-dependent protein kinase (ATP : protein phosphotransferase, EC 2.7.1.37) resulted in the development of precipitating antibodies to the cAMP-binding subunit of the enzyme. Both the phosphorylated and nonphosphorylated cAMP-binding protein of the protein kinase reacted with the antiserum. A radioimmunoassay was developed that detects 10 ng of holoenzyme and permits measurement of enzyme concentrations in bovine cardiac muscle. Bovine liver, kidney, brain, and skeletal muscle contain protein kinases which are immunologically identical to those found in bovine cardiac muscle. However, the proportion of immunoreactive enzyme activity differed for each tissue. All of the immunologically nonreactive enzyme in skeletal muscle and heart was separable from immunoreactive enzyme by chromatography on DEAE-cellulose. Rat tissues and pig heart contained protein kinase activity that cross reacted immunologically in a nonparallel fashion with bovine cardiac enzyme. These results indicate that cAMP-dependent protein kinases within and between species are immunologically heterogeneous

  7. Radioimmunoassay for 2,4-dichlorophenoxyacetic acid

    International Nuclear Information System (INIS)

    Knopp, D.; Dobberkau, H.J.; Nuhn, P.

    1985-01-01

    Antisera to 2,4-dichlorophenoxyacetic acid (2,4-D), a widely used herbicide, have been obtained from rabbits following immunization with various 2,4-D-protein conjugates. Employing [ 125 I] 2,4-D-tyramine as the radioligand for the antisera, very poor assay sensitivity was achieved because of a much higher affinity of the antibodies to the tracer. When using [6- 3 H] 2,4-D (specific radioactivity 465 GBq/mmol) a sensitive and specific radioimmunoassay (RIA) for 2,4-D could be developed, which allows determination directly in water, plasma and urine samples. Levels as low as approximately 100 pg (450 femtomoles) of 2,4-D can be detected. The antiserum is fairly specific for 2,4-D. Other related phenoxycarboxylic acids and dichlorophenol showed a cross-reactivity smaller than 10%. After a single administration of 2,4-D (0.91 mg/100 g body weight, orally) to rats, plasma and urine levels were determined at different times. Results correspond to those found in the literature, thus indicating the utility of the RIA. Further applications and limitations are discussed. (orig.)

  8. Radioimmunoassay of arginine vasopressin in human plasma

    International Nuclear Information System (INIS)

    Uhlich, E.; Weber, P.; Groeschel-Stewart, U.; Roeschlau, T.; Wuerzburg Univ.

    1975-01-01

    Antibodies for the radioimmunoassay of arginine vasopressin (AVP) described here were produced in rabbits using synthetic AVP coupled to rabbit γ-globulin with carbodiimide. In three out of six rabbits, significant antibody titres were obtained. Using the best antisera produced, 40% of labelled AVP was bound at a final dilution of 1 : 50,000. After iodination of synthetic AVP with 125 I using the chloramin-T method, a gel filtration on Sephadex G-25 was performed to purify the iodinated AVP. For separation of antibody bound and free hormone, a second antibody precipitation was used. There was no crossreactivity with oxytocin. AVP was extracted from plasma after ammoniumsulfate precipitation of the proteins by adsorption to Florisil. The recovery of AVP added to plasma in amounts of 5-25 pg/ml was 60 +- 15% (n = 6). The minimum amount of AVP detectable was 1 pg per ml plasma. The plasma level in normal adults under standard conditions was 3.4 +- 2.2 pg/ml. This is in agreement with data recently published by other researchers. The applicability and reproducibility was further tested in measurements of samples taken hourly during the entire day under water diuresis and after hormonal stimulation of AVP. (orig.) [de

  9. Falsely elevated triiodothyronine values in radioimmunoassay

    International Nuclear Information System (INIS)

    Skovsted, L.; Moelholm Hansen, J.E.; Nygaard, B.

    1995-01-01

    Five patients with falsely elevated serum triiodothyronine (T 3 ) concentrations (>9 nmol/l) in a radioimmunoassay are reported. The high T 3 -values disagreed with the other thyroid variables investigated as well as with the clinical observations. In sera from all patients a normal non-specific binding of T 3 was found, thus excluding abnormal serum-protein-binding of the hormone. An ethanol extraction of T 3 from serum before RIA reduced the T 3 content in serum from all patients to normal levels (2.0-2.4 nmol/l). These findings indicate the presence in the sera of substances, probably of protein nature, that were interfering with the assay by binding the reagent-antibody and not the antigen. Addition of non-immune rabbit serum prevented this interference and normalized the T 3 -values (1.8-2.4 nmol/l). Thus the interfering substance in T 3 -RIA could be an anti-rabbit antibody, the interaction of which can be eliminated by a minor modification of the assay making it possible to differentiate true from false T 3 -values. (au) 16 refs

  10. Combined radioimmunoassay for triiodothyronine and thyroxine

    International Nuclear Information System (INIS)

    Denning, C.E.; Schick, L.A.

    1980-01-01

    A method for the combined radioimmunoassay for the hormones triiodothyronine (T-3) and thyroxine (T-4) in serum or plasma, wherein the radiolabeled t-3 and t-4 reagents are t-3 and t-4 labeled with the same radioisotope. The hormones are extracted from the serum or plasma sample and separated from their binding proteins by adsorption to a crosslinked dextran gel at a highly alkaline ph. After washing away the dissociated proteins, a predetermined limiting amount of t-3 antibody is incubated with the gel and thereafter radiolabeled t-3 which has become extracted from the gel by antibody binding is washed away. Excess t-3 antibody and a predetermined limiting amount of t-4 antibody are then incubated with the gel and radiolabeled t-3 and t-4 which has become extracted from the gel by antibody binding are thereafter washed away. The respective relative amounts of radiolabeled t-3 and t-4 retained by the gel after the incubations with the respective limiting amounts of t-3 and t-4 antibody are compared with standard results to quantitate the hormones in the sample. The use of excess t-3 antibody in the t-4 antibody incubation is critical to the accuracy of the method, enabling the use of the same radioisotope for radiolabeling t-3 and t-4

  11. Radioimmunoassay of renin in human renal tissues

    International Nuclear Information System (INIS)

    Wowra, B.

    1981-01-01

    A method has been developed to quantitatively determine renin in human kidney tissue. The angiotensin I split off angiotensinogs by renin was radioimmunologically determined. The renin-renin substrate reaction rate followed a saturation kinetics, as it increased the larger the substrate content in the incubation medium until it acquired a maximum value; the reaction rate decreased with substrate concentrations over 40 mg/ml incubation medium. The discontinuance of the renin reaction after incubation by adding acid, boiling and neutralizing again, gave highest renin values. The RIA scattering was 8.3% for double determination of the same sample, for the determination in different RIA additions 7.0%. The detection limit was 20 pg angiotensin I. A direct comparison of radioimmunoassay and bioassay exhibited a very significant agreement of both methods, where the radioimmunologically measured renin values were on average four times larger than those obtained using biological technique. The definition of the so-called normal values for absolute and specific renin concentration in human kidney tissue enabled one to assess the renin values in various syndromes. (orig./MG) [de

  12. Radioimmunoassay in developing countries: General principles

    International Nuclear Information System (INIS)

    Piyasena, R.D.

    1992-01-01

    Radioimmunoassay (RIA) is probably the most commonly performed nuclear medicine technique. It is an in vitro procedure, where no radioactivity is administered to the patient. But this alone is not the reason for its widespread use. It provides the basis for extremely sensitive and specific diagnostic tests, and its use in present day medicine has brought a virtual information explosion in terms of understanding the pathophysiology of many diseases. The fact that the technology involved is within the technical and economic capabilities of the developing world is evident from the increasing demand for its introduction or expansion of existing services. RIA facilities need not be restricted to urban hospitals, as in the case of in vivo nuclear medicine techniques, but may be extended to smaller district hospitals and other laboratories in peripheral areas. It is also possible to send blood samples to a central laboratory so that a single centre can serve a wide geographical area. There are many laboratories in the industrialized world that receive a major proportion of samples for assay by mail. In recent years, substantial RIA services have been established in many of the developing countries in Asia and Latin America. The International Atomic Energy Agency (IAEA) and World Health Organisation (WHO) have made vital contributions to these activities and have played a catalytic role in assisting member states to achieve realistic goals. In the past five years, more than 250 individual RIA laboratories in developing member states have been beneficiaries of IAEA projects

  13. Estimation of urinary angiotensin II by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Fukuchi, S [Tohoku Univ., Sendai (Japan). School of Medicine

    1974-11-01

    Urine samples were collected from fasting subjects after maintaining posture for 2 hr in early morning. Urinary angiotensin II was extracted with SE-Sephadex. The extracts, after being dissolved in phosphate buffer, pH 7.5, were measured by radioimmunoassay. Recovery, sensitivity and accuracy were found to be satisfactory. The normal values obtained from 6 subjects were 52-280 pg/2 hr. The values were almost normal in essential hypertension and in chronic glomerulonephritis. They were high in 3 out of 6 cases with renovascular hypertension and subsequently dropped after surgery. In 6 cases with primary aldosteronism, very low levels were found. These increased after the removal of adrenal adenomas. No positive correlation between simultaneous plasma and urinary angiotensin samples was apparent. Also no positive correlation between urinary angiotensin and urine volume was found. In renovascular hypertention, during glucose infusion, lower values in urine volume and angiotensin excretion were observed on the stenotic side as compared to the intact side. Thus, the angiotensin excretion rate does not appear to be regulated by arterial angiotensin concentration, but rather by the angiotensin perfusion rate.

  14. Serum erythropoietin levels by radioimmunoassay in polycythaemia

    Energy Technology Data Exchange (ETDEWEB)

    Birgegaard, G.; Miller, O.; Caro, J.; Erslev, A. (Cardeza Foundation for Hematological Research, Jefferson University, Philadelphia, Pa.)

    1982-01-01

    A radioimmunoassay (RIA) method for erythropoietin (Epo) was developed and validated against the polycythaemic mouse assay. The correlation was good, with a r=0.94. Several other criteria of specificity were also filled by the RIA, which had a lower detection limit of 5 mU/ml. The mean serum-Epo level in 6 patients with secondary polycythaemia, 50.2 +- 26.2 mU/ml, was significantly higher than in a group of 11 normal subjects, 28.7 +- 7.2 mU/ml (P<0.0002). However, the Epo level in 31 polycythaemia vera (PV) patients, M = 21.9 +- 6.6 mU/ml, was not significantly different from normal (P = 0.006). Since previous studies with bioassay of heat-treated and concentrated plasma samples have shown a decreased serum-Epo level in PV, Epo levels were measured before and after heat treatment and concentration of samples from normals and polycythaemics. It was found that the levels of immunoreactive material increased after heat treatment and 40 times concentration in samples from normals and patients with secondary polycythaemias, but decreased in PV. We conclude that the Epo levels in serum in the low range measured by our and previous RIA:s probably are not true Epo levels but are partly due to an unspecific serum effect, that was removed by heat treatment.

  15. Radioimmunoassay of creatine kinase BB isoenzyme

    International Nuclear Information System (INIS)

    Geng Jianguo

    1988-01-01

    A radioimmunoassay of creatine kinase BB isoenzyme (CK-BB) was developed by using CK-BB purified from human brain. The CK-BB antiserum was raised by immunizing rabbite and 125 I-CK-BB iodinated with Bolton-Hunter reagent. The affinity constant was 3.0 x 10 9 mol/L. No cross reactions with creatine kinase MM isoenzyme and neuron-specific enolase were found. The measuring range was 3.5 x 10 -8 ∼ 1.2 x 10 -5 mmol/L, the average recovery rate 97.5%, with the inter and intrassay CV 3.1% and 12%, respectively. The average serum CK-BB concentration in 83 normal persons was 1.5 x 10 -7 +- 8.1 x 10 -8 mmol/L, quite different from the values of acute myocardial infarction (5.2 x 10 -6 +- 1.2 x 10 -4 mmol/L, n = 28) and cerebral vascular accident (8.4 x 10 -4 +- 5.0 x 10 -4 mmol/L, n = 10)

  16. Radioimmunoassay of penicilloyl groups in biological fluids

    International Nuclear Information System (INIS)

    Wal, J-M.; Bories, G.

    1975-01-01

    Penicilloyl-protein conjugates may be formed in man or animals after therapeutic treatment but cannot be detected by the classical microbiological assay methods used for penicillin. Allergies have been noted in consumers of products coming from animals treated with penicillin (milk particularly). Although these compounds present a risk to public health, they passed unnoticed through hygienic food inspection. Therefore a specific and sensitive method was devised for the assay of these derivatives in biological fluids where they can be present in trace amounts. An iodine-125 labelled conjugate has been prepared. Already usable for the detection of antipenicilloyl antibodies in sera of penicillin allergic patients, it has been used for the development of a radioimmunoassay of penicilloyl groups. Assay is done directly on milk, urine and serum without previous extraction and with a detection limit of a few ppb. It permits a rapid, specific and easy to handle determination in the sera of hospital patients as well as in the inspection of animal products at the slaughter house and at the dairy

  17. Development of radioimmunoassay for prolactin binding protein

    Energy Technology Data Exchange (ETDEWEB)

    Raikar, R.S.; Sheth, A.R. (Institute for Research in Reproduction, Bombay (India))

    1982-01-01

    Using a homogenous prolactin binding protein (PBP) preparations from rat seminal vesicle secretion, a sensitive and specific radioimmunoassay (RIA) for PBP has been developed. The assay was highly specific and showed no cross-reaction with other protein hormones from various species. The antiserum had an affinity constant (Ka) of 2.66 x 10/sup 10/ M/sup -1/. The assay sensitivity was in the range of 0.5-1.0 ng of pure PBP per assay tube and the intra- and inter-assay coefficients of variations were 6-8% and 12-14.5% respectively. The overall recovery of PBP to the rat seminal vesicle secretion was 96.8%. Using this RIA, PBP levels in various biological fluids and reproductive tissues were measured. Azoospermic human semen contained significantly higher levels of PBP than normospermic semen. The seminal vesicle of rat exhibited the highest concentration of PBP. Administration of antiserum to PBP to mature male rats resulted in a significant reduction in the weight of ventral prostrate and serum prolactin levels were significantly elevated in these animals suggesting that the antibody raised against the PBP was capable of blocking prolactin receptors.

  18. Radioimmunoassay for chlorinated dibenzo-p-dioxins

    International Nuclear Information System (INIS)

    Albro, P.W.; Chae, K.; Luster, M.I.; Mckinney, J.D.

    1980-01-01

    The invention provides a double-antibody radioimmunoassay method for the determination of chlorinated dibenzo-p-dioxins, particularly, 2,3,7,8-tetrachlorodibenzo-p-dioxin, in environmental samples including animal tissues such as monkey liver and adipose tissues. The limit of detection is approximately 25 picograms for 2,3,7,8-tetrachlorodibenzo-pdioxin. Assuming an appropriate cleanup procedure is used, chlorinated dibenzofurans are the only likely interferences, and these can be distinguished through the use of two antisers of different dibenzo-furan/dibenzodioxin selectivities. The invention includes the preparation of a reproducible antigen, an appropriate radiolabeled hapten, and effective sample extracts. A feature of the assay method is the use of a nonionic detergent (e.g., ''cutscum'' or ''triton x-305'') to solubilize the extremely hydrophobic dibenzo-p-dioxins in a manner permitting their binding by antibodies. The immunoassay is applicable to screening samples in order to minimize the demand for mass spectrometric screening, and to routine monitoring for exposure to known chlorinated dibenzo-p-dioxins in potentially contaminated environments

  19. Radioimmunoassay for human health in developing countries

    International Nuclear Information System (INIS)

    Piyasena, R.D.; Airey, P.L.; Ganatra, R.D.; Nofal, M.

    1989-01-01

    Since first introduced in the early 1960s, radioimmunoassay (RIA) has gained wide acceptance as an analytical method adopted by an increasing number of developing countries as an appropriate technology that can be managed within the capabilities of local infrastructures. An informed estimate would be that there are, at present, more than 500 hospitals, university, or other laboratories in the developing world engaged in RIA on some scale. In the developing world, RIA is used primarily for patient management, but research activity is also increasing as expertise and resources improve. The majority of patient samples processed are in relation to thyroid disorders. However, the technique also is used widely in the investigation of other endocrine conditions and public health problems. Some developing countries have gained the capability to perform radioisotopic microassays in areas of clinical and research importance such as steroid receptor quantification in breast tissue; diagnosis of bacterial and parasitic disorders; investigation of infertility and sterility; narcotic drug abuse; and organ transplantation. 1 fig

  20. A simple radioimmunoassay for plasma cortisol

    International Nuclear Information System (INIS)

    Seth, J.; Brown, L.M.

    1978-01-01

    A simple radioimmunoassay (RIA) for plasma cortisol is described which combines the advantages of (i) direct analysis of untreated plasma samples, (ii) use of solid-coupled anti-cortisol antibodies and (iii) use of a gamma-labelled radioligand. The reagents are relatively easily prepared and stable, and the analysis can be completed in 4 h. Inter-assay precision (C.V.) is 8-11%. Critical examination of specificity using high pressure liquid chromatography showed that 23-35% of the immunoassayable material in plasma was not cortisol. RIA results on samples collected under basal conditions were an average 40 nmol/l lower than fluorimetric results, while in insulin hypoglycaemia and synacthen (ACTH) stimulation tests, this difference increased to over 100 nmol/l. The RIA is technically more simple than fluorimetric, competitive-protein-binding, and many RIA methods, and can be used with advantage in the routine investigation of adrenocortical function. However, using the present antiserum, the RIA is not applicable to investigations on patients receiving metyrapone, nor in suspected cases of congenital adrenal hyperplasia. (Auth.)

  1. Steroid radioimmunoassays. The problems of blanks

    Energy Technology Data Exchange (ETDEWEB)

    Shinde, Y; Hacker, R R; Ntunde, B; Smith, V G [Guelph Univ., Ontario (Canada). Dept. of Animal and Poultry Science

    1981-06-01

    An estrogen radioimmunoassay was used to study the problem of blanks in steroid assays. Negligible binding (1.5 percent) in the non-antibody tubes prevailed throughout the study. The assay was validated using accepted procedures. Both water and solvent blanks had estrogen concentrations of 7-9 pg/tube. However, neither water nor solvent blanks showed a dose-related response, indicating that they were 'real' blanks. Exogenous estradiol, when added to water and solvent in quantities less than the estimated blank, was not quantitatively recovered. However, exogenous estradiol added to the water solvent in quantities greater than the blank estimate was quantitatively recovered. The sensitivity of the reference standard curve was 6-10 pg/tube, approximately the same as the blank estimate. These results indicated that the estimates of water and solvent blanks were measures of the assay sensitivity. In such circumstances, it is suggested that blank estimates should not be subtracted from sample values. If the blank estimates are high, attention should be directed towards improving the sensitivity of the assay.

  2. Parathyroid hormone: radioimmunoassay and clinical interpretation

    International Nuclear Information System (INIS)

    Hawker, C.D.

    1975-01-01

    A radioimmunoassay for serum immunoreactive parathyroid hormone (iPTH), which has had widespread clinical use for five years, is described in detail. The iPTH results in large groups of patients are reported, and are discussed in relation to the specificity of the assay and in relation to other assays. The assay has excellent precision and is highly proficient in discrimination of groups of patients. Ninety-three percent of 412 patients with surgically proven primary hyperparathyroidism were confidently separated from normal subjects or patients with hypercalcemia owing to other causes, while 86 percent of 160 patients with chronic renal failure and secondary hyperparathyroidism had iPTH values more than 2 S.D. above the normal mean. Results in patients with ectopic hyperparathyroidism were lower than in primary hyperparathyroidism although these groups showed considerable overlap. The antiserum used in this assay for iPTH appears to be specific for the carboxy-terminal region of the secreted or intact form of PTH but recognizes predominantly the secreted form rather than carboxy-terminal fragments believed to be in the circulation. It does not recognize amino terminal fragments. The assay is useful in selective venous catheterization for preoperative localization of hyperfunctioning parathyroid tissue

  3. Immunodiagnosis of opportunistic mycoses: detection of fungal antigenemia by radioimmunoassays in systemic candidiasis and aspergillosis

    International Nuclear Information System (INIS)

    Weiner, M.H.

    1980-01-01

    The authors have developed radioimmunoassays to the Candida carbohydrate, mannan, and to an Aspergillus cell wall carbohydrate. They evaluate these radioimmunoassays with sera from rabbit models of disseminated mycoses, and further evaluate the radioimmunoassays for their diagnostic usefulness in a panel of patient sera. (Auth.)

  4. Buffett's Alpha

    DEFF Research Database (Denmark)

    Frazzini, Andrea; Kabiller, David; Heje Pedersen, Lasse

    Berkshire Hathaway has realized a Sharpe ratio of 0.76, higher than any other stock or mutual fund with a history of more than 30 years, and Berkshire has a significant alpha to traditional risk factors. However, we find that the alpha becomes insignificant when controlling for exposures to Betting...

  5. Alpha Blockers

    Science.gov (United States)

    ... quickly, but their effects last only a few hours. Long-acting medications take longer to work, but their effects last longer. Which alpha blocker is best for you depends on your health and the condition being treated. Alpha blockers are ...

  6. Infusion of hypertonic saline before elective hysterectomy: effects on cytokines and stress hormonesTumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    Kolsen-Petersen, J.A.; Bendtzen, K.; Tonnesen, E.

    2008-01-01

    factor-alpha. Serum cortisol and vasopressin were measured at these time points and 48 h after operation. Epinephrine and norepinephrine (n=26) were quantified at baseline, after infusion, 25 min after incision, 1, and 4 h after surgery. Finally, C-reactive protein was measured at baseline, 24, and 48 h...... in a double-blind study to infusion of NaCl 7.5% (HS), NaCl 0.9% (NS4), both 4 ml kg(-1), or NaCl 0.9% 32 ml kg(-1) (NS32) over 20 min. Blood was collected at baseline, 1, 4, and 24 h after surgery (n=34) for the determination of interleukin (IL)-1beta, IL-6, IL-8, IL-10, IL-12, IL-1ra, and tumour necrosis...

  7. Radioimmunoassay for tumor antigen of human cervical squamous cell carcinoma

    International Nuclear Information System (INIS)

    Kato, H.; Torigoe, T.

    1977-01-01

    A heterologous antiserum for human cervical squamous cell carcinoma was prepared and specificity determined by Ouchterlony immunodiffusion and immunofluorescence studies. With this antiserum, a tumor antigen was purified from human cervical squamous cell carcinoma tissue. The specificities of the antigen and the antiserum were then re-examined by a radioimmunoassay method using 125 I-labeled purified antigen. Although normal cervical tissue extract showed a moderate cross-reactivity in the radioimmunoassay, the circulating antigen activity could not be detected in normal women or in several patients with other carcinomas, whereas 27 of 35 patients with cervical squamous cell carcinoma showed detectable serum antigen activity. All patients with advanced stages of cervical squamous cell carcinoma showed detectable antigen levels. These results indicate that there is a quantitative abnormality, at least, of this tumor antigen in patients with cervical squamous cell carcinoma and that the radioimmunoassay for the antigen is a potentially useful tool in clinical care

  8. Determination of triiodothyronine in serum by enzyme- and radioimmunoassay

    International Nuclear Information System (INIS)

    Oellerich, M.; Haindl, H.; Medizinische Hochschule Hannover

    1981-01-01

    An evaluation of a heterogeneous enzyme immunoassay for determination of triiodothyronine in serum (Enzymun-Test T 3 , Boehringer Mannheim) is presented. The enzyme immunoassay was compared with the laboratory routine radioimmunoassay. The precision of both assays was satisfactory at triiodothyronine concentrations between 1.0 and 8.0 nmol/l (coefficients of variation from day to day 3 from 96-104% and with the radioimmunoassay from 88-111%. A comparison of the results obtained by Enzymun-Test T 3 and the radioimmunoassay in a series of 103 patients showed a good correlation between both methods. L-thyroxine did not cause a relevant cross-reaction in the enzyme immunoassay. About 20 unknown samples can be analyzed in triplicate by Enzymun-Test T 3 within 260 minutes. (orig.) [de

  9. Estimation of antibodies to human cytomegalovirus by immunofluorescence and radioimmunoassay

    International Nuclear Information System (INIS)

    Jankowski, M.; Gut, W.; Nawrocka, E.

    1980-01-01

    The 125 I labelled IgG fraction against rabbit IgG of goat origin was employed for the detection of CMV IgG and IgM antibodies in the double indirect radioimmunoassay. The results were compared with those obtained in complement fixation, indirect immunofluorescence and anti-complement immunofluorescence tests. The application of labelled anti-fc antisera, instead of antisera against whole IgG in the tests for detection of specific CMV IgG antibody resulted in increased sensitivity of radioimmunoassay and reduction of non-specific cytoplasmatic reactions in preparations stained by indirect immunofluorescence. The absorption of sera with protein A rich staphylococci and aggregates to immunoglobulin eliminated unwanted secondary IgM staining caused by rheumatoid factors both in indirect immunofluorescence and radioimmunoassay tests for CMV antibodies. (author)

  10. Radioimmunoassay for pantothenic acid in blood and other tissues

    International Nuclear Information System (INIS)

    Wyse, B.W.; Wittwer, C.; Hansen, R.G.

    1979-01-01

    We described a radioimmunoassay for pantothenic acid in biological tissues. D-Pantothenic acid was conjugated with bovine serum albumin by use of a bromoacetyl derivative of pantothenic acid, and antibody to this antigen was raised by injecting it into the foot pads of rabbits. For the radioimmunoassay, a 100-fold dilution of the resulting antiserum was incubated with radiolabeled panthothentic acid. The antibodies were precipitated and dissolved, and the radioactivity of the solution was measured in a liquid scintillation counter. Between 5 and 125 ng of pantothenic acid can be detected in 75 μL of tissue extract. Validation included recovery and precision studies, parallelism with tissue extracts, and competitive binding studies. Results of the radioimmunoassay and those of microbiological assay with use of Lactobacillus plantarum correlated well

  11. TRH radioimmunoassay for unextracted human urine

    International Nuclear Information System (INIS)

    Mitsuma, Terunori; Hirooka, Yoshibumi; Nihei, Noriyuki

    1975-01-01

    The authors developed a TRH radioimmunoassay for unextracted human urine using anti-TRH antibody produced by immunization of rabbits with a TRH-bis-diazotized-bovine serum albumin conjugate. The antibody had no crossreactivity with TRH analogues, amino acids or pituitary hormones, but with L or DL-Aze3-TRH. TRH was radioiodinized by Greenwood-Hunter's method, followed by purification on Sephadex G-10. Inactivation of TRH by serum was well documented. The authors found however that this inactivation of TRH could be prevented by adjusting the pH to 3.0 or by keeping the temperature between 4 0 C and -20 0 C. All assay procedures were performed in 0.01 M phosphate buffer with 0.15 M NaCl (pH 7.5) at 4 0 C. Free and bound forms were separated with a second antibody system. In this system, sensitivity was 0.01 ng/tube, recovery was approximately 100%, intrassay reproducibility was 3.2% and interassay variation was 9.8%. TRH levels in urine measured with this system were undetectable to 9.0 ng/ml in normal subjects, undetectable in hyperthyroid patients or a tertiary hypothyroid patient and 13 to 24 ng/ml in primary hypothyroid patients. Approximately 6 percent of the intravenously administered TRH was excreted into the urine within 12 hours following administration in a normal subject. As a result this assay system is quite attractive for clinical determination as well as research application. (Evans, J.)

  12. Oxytocin determination by radioimmunoassay in cattle. 1

    International Nuclear Information System (INIS)

    Schams, D.; Schmidt-Polex, B.; Kruse, V.

    1979-01-01

    A radioimmunoassay for oxytocin in cow plasma is described. Antisera were raised in rabbits against synthetic oxytocin coupled to bovine thyroglobulin. Iodinated oxytocin free of unlabelled oxytocin and most likely also free of diiodo-oxytocin was used as radioactive tracer. The tracer showed a high degree of purity, and was stable on storage. It could be used in the assay for 2-3 months. The assay showed very little cross-reactivity with vasopressin. Acetone was used for the extraction of oxytocin from plasma as well as from standards made of synthetic exytocin in pooled cow plasma. Inhibition curves obtained with plasma collected from cows at parturition were parallel to those obtained with the oxytocin standard preparation. The mean recovery of oxytocin added to cow plasma was 106% (SD=14). The within-assay coefficient of variation (CV) varied from 5.2 to 10.9%, and the between-assay CV was in the order of 13%. The assay sensitivity was 1 pg (0.5 μU) per tube, corresponding to 3 pg/ml plasma. Around the time of milking the plasma oxytocin profile showed a strong response to the preparation for milking, and a further effect related to the attachment of the teat cups of the milking machine. Peak concentrations were in the range of 15-50 pg/ml. During parturition there was a peak of oxytocin (65 pg/ml) coinciding with the expulsion phase. After this peak levels decreased by remained measurably elevated until the expulsion of the placenta. The plasma disappearance curve for immunoreactive oxutocin after the infusion of 100 IU oxytocin over a period of 1 h showed two components with apparent half-lives of 7-9 and 25 min, respectively. (author)

  13. Radioimmunoassay of thyrotropin concentrated from serum

    International Nuclear Information System (INIS)

    Nisula, B.C.; Louvet, J.P.

    1978-01-01

    A method for concentrating human TSH (hTSH) from serum for use in RIAs is described. The method takes advantage of the affinity of the plant lectin, concanavalin A, for the carbohydrate portion of the hTSH molecule. The hTSH from 2.5 ml serum was adsorbed to concanavalin A covalently linked to sepharose and then radioimmunoassayed using the hTSH antiserum and hTSH for iodination distributed by the National Pituitary Agency. For the RIA standard curve, the hTSH reference preparation was concentrated from a serum wwith undetectable hTSH in order to correct for recovery and to control for nonspecific effects. The percentage of serum hTSH extracted from 2.5 ml serum with the concentration procedure was 76.6 +- 3.4% (mean +- SD). The coefficient of correlation between serum hTSH, determined with the concentration procedure, and serum hTSH determined without was 0.979 (P < 0.001). Over 95% of normal adult men and women had detectable levels of serum hTSH, ranging from < 0.56 to 4.0 μU/ml. The mean of detectable serum hTSH levels in normal adult women (n = 11) was 1.54 +- 1.03 μU/ml (mean +- SD) and in normal men (n = 9) was 2.02 +- 1.15 μU/ml (mean +- SD). Clinically hyperthyroid patients with diffuse and nodular toxic goiters (n = 8) and patients with hypothyroidism secondary to pituitary disease (n = 6), four of whom were taking replacement doses of thyroid hormone, had undetectable serum hTSH levels. Serum hTSH in patients with primary hypothyroidism uniformly exceeded the normal range. This hTSH concentrating procedure enhances the effective sensitivity and, therefore, the clinical utility of the RIA for hTSH in serum

  14. Problems in radioimmunoassay of human lutropin with commercially available regents

    International Nuclear Information System (INIS)

    Hammond, J.E.; Phillips, J.C.; Straight, C.B.; Hammond, M.G.

    1980-01-01

    To evaluate five commercially available reagent sets supplied for the radioimmunoassay of lutropin, we determined whether there was parallelism between the curve given by dilutions of the standards supplied by the manufacturers, by dilutions of a serum pool, and by dilutions of a standard preparation from human pituitaries, LER-907. These studies demonstrated significant analytical problems with three of the five sets. We conclude that each user should carefully evaluate all commercially available radioimmunoassays for lutropin (and, by inference, for other peptide hormones) before use

  15. A sensitive radioimmunoassay for a component of mouse casein

    International Nuclear Information System (INIS)

    Enami, Jumpei; Nandi, S.; California Univ. Berkeley

    1977-01-01

    Mouse casein (m.w. 22,000 daltons) has been purified by employing Sephadex G-100 and DEAE-cellulose column chromatographies. A sensitive radioimmunoassay method has been developed by using [ 125 I]-labelled casein and antiserum elicited in rabbits after injection of glutaraldehyde-treated casein. The assay method is capable of detecting as little as 0.1 ng of casein. The use of the present radioimmunoassay method in detecting casein production in cultured mouse mammary explants has also been demonstrated

  16. Radioimmunoassay of type D oncovirus from continuous J-96 cells

    International Nuclear Information System (INIS)

    Vlasenkova, N.K.; Altshtejn, A.D.; Zhdanov, V.M.; Kitsak, V.Ya.

    1978-01-01

    The radioimmunoassay of the J-96 virus and an extract of J-96 cells in the homologous and heterologous systems aimed at detecting antigenic determinants of p25 of Mason-Pfizer virus and group-specific and interspecies antigenic determinants p30 of Rauscher leukaemia virus demonstrated that (1) J-96 virus contains a major internal protein immunologically identical with p25 protein of Mason-Pfizer virus based on the antigenic determinants detectable by the radioimmunoassay used; and (2) no interspecies antigenic determinants characteristic of the major internal protein of mammalian type C viruses were detectable in the J-96 virus or the J-96 cell extract. (author)

  17. A double antibody radioimmunoassay specific for placental alkaline phosphatase

    International Nuclear Information System (INIS)

    Dass, S.; Bagshawe, K.D.

    1984-01-01

    Placental alkaline phosphatase (PLAP) is normally found in enzymically measurable amounts in second and third trimester pregnancy serum. Its occurrence in sera and tumours from patients with malignant disease has led to the development of methods to specifically identify and quantitate the enzyme. Recently immunological techniques have been used, employing antibodies raised to purified PLAP; these include solid phase radioimmunoassays and enzyme-immunoassay. The development of a sensitive, specific, automated double-antibody radioimmunoassay for the measurement of PLAP in serum is reported. (Auth.)

  18. Development and application of radioimmunoassays for vitamin D metabolites

    International Nuclear Information System (INIS)

    Clemens, T.L.

    1980-04-01

    A sensitive radioimmunoassay for 1,25-dihydroxycholecalciferol, the most potent derivative of vitamin D 3 is described and has been used to measure its concentration in human serum from normal subjects, patients with clinical osteomalacia,, anephric patients and those with chronic renal failure. The absorption and clearance of 1,25-dihydroxycholecalciferol have been studied in healthy and uremic patients by following the changes of its circulating concentration after administration of a single (2ug) dose of synthetic 1,25-dihydroxycholecalciferol orally. The radioimmunoassay was also used to investigate the underlying cause of the hypercalcaemia of sarcoidosis. (author)

  19. Logistics of national radioimmunoassay services and radioimmunoassay trouble shooting in developing countries

    International Nuclear Information System (INIS)

    Hazra, D.K.; Shukla, A.K.; Arvind, B.; Khandelwal, S.; Singh, R.; Lahiri, V.L.

    1986-01-01

    The logistics of national radioimmunoassay services are analysed in regard to: applications of immunoassays in developing countries; provision of matched reagents; collection, storage and transport of patient samples, particularly from rural areas; development of robust new immunoassay systems, especially those using non-isotopic labels; quality control; and interrelation with international agencies and commercial suppliers. The effect of using different methods and temperatures of sample storage on representative glycoprotein, steroid, thyroid and communicable disease analytes is examined. The need for stratification of assay services (district, state and national/regional levels) for various tests is discussed in relation to their clinical urgency. A trouble shooting guide suited to developing countries has been written. Starting from various symptoms of assay failure, it describes the differential diagnosis of likely causes, lists appropriate tests to confirm the diagnosis and finally suggests remedial action to prevent future similar mishaps or to salvage the assay. (author)

  20. Radioimmunoassay of glicagon secretion in diabetes mellitus

    International Nuclear Information System (INIS)

    Milanova, St.; Koparanova, O.; Milkov, V.; Visheva, N.; Kurtev, I.; Maleeva, A.

    1988-01-01

    Radioimunoassay of glucagon, lh, fsh, aldosterone, cortisol, acth, sth and lth was performed using preparations from the firms SORIN, AMERSHAM and HOECHST. The basal glucagon secretion was estimated in 403 diabetic patients and 84 normal subjects. It was transitorily suppressed by glucose. The alpha-cellular reactivity in diabetic patients was stimulated by insulin-induced hypoglycemia and with arginine and vasopressin. The tested group of diabetic patients had absolute hyperglucagonemia, despite the hyperglycemia, which is an evidence of abnormal alpha-cellular function. The insulin-dependent nature of hyperglucagonemia in diabetes mellitus and the hyperreactivity of glucagon secretion after protein stimulation was demonstrated. The correlation of these results leads to essential diagnostic inferences