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Sample records for alpha fetoprotein radioimmunoassay

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

    International Nuclear Information System (INIS)

    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.)

  2. RADIOCHEMICAL STUDIES ON ALPHA FETOPROTEIN RADIOIMMUNOASSAY SOLID PHASE COATED BEADS

    International Nuclear Information System (INIS)

    Alpha fetoprotein (AFP) is a marker for hepatocellular and germ cell carcinoma. There are many different techniques used for measuring AFP in blood where the most accurate one is the immunoassay technique. The aim of the present study was to evaluate, optimize and prepare anti-AFP solid phase coated beads and use it for the determination of AFP in serum. The anti-AFP polyclonal antibodies were prepared by immunization of five rats with a highly purified AFP antigen and the anti-sera obtained were used for coating polystyrene beads to obtain the solid phase coated beads. Also, the AFP antigen was labelled with 125 I using chloramin-T (Ch-T) as oxidizing agent and the tracer obtained was purified using sephadex G-25 (PD-10) chromatography. The assay was performed using a set of AFP standards prepared by diluting the cord blood. The suitable conditions for coating process were obtained which include pH 8, molarity of coating buffer 0.05 M, volume of coating buffer 100 ml and dilution of antibody 1:1000 for coating 1000 beads. Because the coated beads prepared using borate buffer is more suitable than prepared using carbonate or phosphate buffers, they were chosen to complete the optimization and validation study. The optimization and characterization of the assay were performed to evaluate the quality of the proposed system. The system prepared proved a low cost, simple, sensitive and accurate results. The prepared system can be used to evaluate AFP in the blood and this will be helpful in diagnosing some diseases such as hepatocellular carcinoma and neural tube defects

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

    International Nuclear Information System (INIS)

    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 125I 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)

  4. Alpha fetoprotein

    Science.gov (United States)

    Fetal alpha globulin; AFP ... Greater than normal levels of AFP may be due to: Cancer in testes , ovaries, biliary (liver secretion) tract, stomach, or pancreas Cirrhosis of the liver Liver cancer ...

  5. 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)

    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 125I-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

  6. 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)

    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.)

  7. Use of human chorionic gonadtropin and alpha-fetoprotein radioimmunoassays: specificity and apparent half-life determination after delivery and in patients with germ cell tumors

    International Nuclear Information System (INIS)

    The specificity of commercially available hCG and hCG-β antibodies (anti-hCG-β from N.I.A.M.D.D. (SB6), Serono (S), Biosigma (B), Union Carbide (UC) (I.R.E.) and anti-hCG from Union Carbide) were compared. Using 125I-hCG (CR 115), the crossreactivity with LH (LER 960) was 5.5% for SB6, 0.25% for S and 0.3% for B. In the homologous hCG system UC, crossreactivity was 0.06% with hCG-β and 1.2% with LH, in the hCG-β system UC, it was 2.2% with hCG and 0.01% with LH. Parallel standard curves for hCG, the 2nd I.R.P. hCG and hCG-β were found exclusively with the 125I-hCG, anti-hCG-β system S. Consequently accurate estimates of the total hCG or hCG-β content in serum and standardization with the 2nd I.R.P. is possible. This system is the most useful for clinical purposes. The serum half-life of hCG was calculated in 10 pregnant women after delivery and was found to be 10 to 34h. AFP half-life time, determined by the double antibody radioimmunoassay was 4.0+-1.8 (+- SD) days in pregnant women after delivery (n=60) and 3.8+-0.9 days in neonates during the first 16 days of life (n=26). Altogether, apparent half-life determinations were carried out in 29 patients with nonseminomatous germ cell tumours. The results indicate that half-life determinations of hCG and AFP may have a predictive value with respect to the monitoring of therapy. (Auth.)

  8. 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...... 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...... (high or low) and the tests were later found to be false positives. There was a strong association between anxiety experienced in conjunction with the alpha-fetoprotein screening programme and the alpha-fetoprotein test result. Two percent of the women with a normal test result reported severe anxiety...

  9. 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...

  10. Maternal serum alpha fetoprotein in monozygotic and dizygotic twin pregnancies.

    Science.gov (United States)

    Thom, H; Buckland, C M; Campbell, A G; Thompson, B; Farr, V

    1984-01-01

    Maternal serum alpha-fetoprotein (MSAFP) values in the second trimester have been related to pregnancy outcome for 100 normal twin pairs, 42 monozygous (MZ) and 58 dizygous (DZ), liveborn after 28 weeks gestation. The median MSAFP value was 1.9 multiples of the median value (MOM) for uncomplicated singleton pregnancies. Both very low and very high MSAFP values were associated with twins of low birthweight. MSAFP values were higher in MZ than DZ twin pregnancies particularly those with dizygotes of like-sex. This effect was even more marked when only dichorionic like-sex twin pairs were compared. PMID:6209699

  11. Hereditary persistence of alpha-fetoprotein (HPAFP) : review of the literature

    NARCIS (Netherlands)

    Houwert, A. C.; Giltay, J. C.; Lentjes, E. G. W. M.; Lock, M. T. W. T.

    2010-01-01

    Alpha-fetoprotein (AFP) serum levels are raised in several clinical conditions, ranging from non-pathological conditions to malignancies. Hereditary persistence of alpha-fetoprotein (HPAFP) is a rare benign disorder with elevated AFP levels. HPAFP is described as a benign autosomal dominantly inheri

  12. Maternal serum alpha-fetoprotein and fetal triploidy.

    Science.gov (United States)

    Pircon, R A; Towers, C V; Porto, M; Gocke, S E; Garite, T J

    1989-10-01

    Fetal triploidy is commonly found in early pregnancy. The majority of these pregnancies spontaneously abort in the first trimester. Occasionally, the pregnancy progresses to the second and third trimesters. We reviewed the maternal serum alpha-fetoprotein (MSAFP), amniotic fluid alpha-fetoprotein (AFP), amniotic fluid acetylcholinesterase (ACHE), fetal pathology, and placental pathology in sex second-trimester pregnancies complicated by fetal triploidy. Four of these patients had MSAFP values greater than 7.5 multiples of the median (MoM). Five of six pregnancies had MSAFP values greater than 2.25 MoM. All five of these patients had a partial mole. Four patients had amniotic fluid AFP values greater than 2.0 MoM. Two fetuses had associated neural tube defects. These were the only patients with positive amniotic fluid ACHE. None of the other patients had fetuses with anomalies that are known to be associated with an elevated MSAFP. The elevated MSAFP appeared to be related to the presence of a partial mole. Two of the five cases with an MSAFP greater than 2.25 MoM did not have sonographic evidence of a significant anomaly. Therefore, karyotyping can be of benefit in evaluating patients with elevated MSAFP. PMID:2480590

  13. Diagnostic value of alpha-fetoprotein in liver cancer

    International Nuclear Information System (INIS)

    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)

  14. Maternal alpha-fetoprotein screening: two years' experience in a low-risk district

    OpenAIRE

    1981-01-01

    Over a two-year period, 3479 pregnant women in the Kings' Lynn Health District were screened for neural tube defects by estimation of maternal serum alpha-fetoprotein. Most pregnancies were scanned by sonar for fetal maturity. Eight women had fetuses with open neural tube defects; four with anencephaly were associated with very high alpha-fetoprotein values. Of the four with open neural tube defects without anencephaly, only one was detected by screening and confirmed after amniocentesis. One...

  15. Abnormal maternal serum alpha fetoprotein and pregnancy outcome.

    Science.gov (United States)

    Zarzour, S J; Gabert, H A; Diket, A L; St Amant, M; Miller, J M

    1998-01-01

    The objective was to assess the occurrence of miscarriages, low birth weight, and karyotype abnormalities found with low and elevated maternal serum alpha-fetoprotein (MSAFP) among women who had genetic amniocentesis performed. A retrospective study of 2,159 women who had MSAFP analysis prior to amniocentesis was conducted. Pregnancy outcomes were obtained from record review and physicians follow-up. Limits of MSAFP used in analysis were MOM) (lower levels) and >2.0 MOM (upper levels). Autosomal trisomy was found in 1.6% with low, 0.9% normal, and 0.6% with elevated MSAFP values. Sex chromosome abnormalities were present only in patients with normal MSAFP, [45X (n = 6), 47XXY (n = 2), 69XXX]. Of five open neural tube defects, four had elevated MSAFP and one had a normal value. Omphalocele was identified in four patients, two with normal and two with elevated MSAFP. Gastroschisis was found in one low and one elevated MSAFP. Amniotic fluid alpha-fetoprotein (AFAFP) values did not correlate with MSAFP values. Patients with low MSAFP levels had a greater prevalence of abnormal karyotype (19 of 249, prevalence = 0.076) than patients with an elevated MSAFP level (2 or 166, prevalence = 0.012 OR (odds ratio) = 0.20 (P value = 0.024) when unadjusted for maternal age, and OR = 0.09 (P value = 0.001) when adjusted for maternal age. Spontaneous abortion occurred more often in patients with elevated (4 of 166, or 4%) than normal or low (20 of 1948, or 1%) values of MSAFP (odds ratio 4.32, P = 0.020 when adjusted for maternal age). Birth weight below 2,500 g was present less frequently with low or normal MSAFP (136 of 1,760, or 7.7%) than in elevated MSAFP (21 of 144 or 14.6%) (odds ratio 2.04, P = 0.005, unadjusted; and odds ratio = 2.32, P = 0.003, adjusted for maternal age). Female fetuses were present more often with low MSAFP (136 of 249, or 55%) than elevated levels 43% (71 of 164, or 43%; P = 0.024). We conclude that patients undergoing genetic amniocentesis with MSAFP

  16. Unexplained elevations of maternal serum alpha-fetoprotein.

    Science.gov (United States)

    Katz, V L; Chescheir, N C; Cefalo, R C

    1990-11-01

    Alpha-fetoprotein (AFP) is a commonly used prenatal screening test for congenital anomalies. However, when anomalies are excluded after high resolution ultrasound and/or amniocentesis, an elevated maternal serum AFP (MSAFP) has been found to be associated with a 2- to 4-fold increase in low birthweight resulting from both preterm delivery and intrauterine growth retardation. Unexplained MSAFP elevations are also associated with up to 10-fold increase of placental abruption and a 10-fold increase in perinatal mortality. Results from studies of over 225,000 screened pregnancies indicate that 20 and 38 per cent of women with an unexplained MSAFP elevation may have an adverse pregnancy outcome. Twin gestations with MSAFP elevations greater than four multiples of the median are associated with similar constellations of pregnancy complications. Maternal serum AFP elevations in women with pregnancy complications are most likely the result of a leak of AFP across the placenta. Optimum management of women with unexplained elevations has not yet been established; however, evaluation of fetal growth throughout gestation is important in these patients. PMID:1700347

  17. The alpha-fetoprotein enhancer region activates the albumin and alpha-fetoprotein promoters during liver development

    OpenAIRE

    Jin, Lin; Long, Lingyun; Green, Michael A.; Spear, Brett T.

    2009-01-01

    The four members of the albumin gene family encode the serum transport proteins albumin, α-fetoprotein, α-albumin, and vitamin D-binding protein. These genes are transcribed primarily in the liver with each having a different pattern of developmental expression. The tight linkage of these genes, particularly that of albumin, α-fetoprotein and α-albumin, and their liver-specific expression, has led to the suggestion that these genes share common regulatory elements. To directly examine whether...

  18. 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

    1992-01-01

    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 no

  19. 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

    1991-01-01

    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 compar

  20. 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 pr

  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. PMID:26359849

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

    International Nuclear Information System (INIS)

    An own modification of the double antibody radioimmunoassay for AFP using 125I-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)

  3. 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

  4. Dexamethasone inhibits alpha-fetoprotein gene transcription in neonatal rat liver and isolated nuclei.

    OpenAIRE

    Huang, D. P.; Cote, G J; Massari, R J; Chiu, J F

    1985-01-01

    The effect of dexamethasone on rat alpha-fetoprotein (AFP) expression has been further examined. Quantitation of serum AFP levels from newborns treated with dexamethasone showed a dose-response relationship between the quantity of dexamethasone administered and the reduction in AFP serum level. RNA blots, utilizing cloned AFP cDNA as probe, showed a marked reduction in AFP mRNA in dexamethasone treated livers. The extent of AFP mRNA depletion was correlated with dexamethasone dosage. The effe...

  5. Electrophoresis and electro-affinity transfer with specific antibodies to alpha-fetoprotein for detection of circulating immune complexes of alpha-fetoprotein.

    Directory of Open Access Journals (Sweden)

    Taketa,Kazuhisa

    1984-08-01

    Full Text Available A combination of agarose gel electrophoresis and a newly developed technique of electro-affinity transfer was applied to the detection of circulating immune complexes of human alpha-fetoprotein (AFP and anti-AFP. After electrophoretic transfer to nitrocellulose membrane, to which affinity-purified polyclonal horse antibodies to human AFP were bound, the membranes were treated with or without rabbit immunoglobulins to human AFP, followed by overlaying with horseradish peroxidase-labeled goat anti-rabbit IgG for color development. Artificial complexes formed in vitro from human AFP and rabbit anti-AFP were clearly separated from free AFP by the agarose electrophoresis. The complexes were stained 20-40% as dark as the equivalent amount of free AFP by treatment with rabbit anti-AFP, and 10-20% as dark without the antibody treatment over a wide range of antigen-antibody ratios.

  6. Elevated maternal serum alpha-fetoprotein levels in patients with subchorionic hematoma.

    Science.gov (United States)

    Kumbak, Banu; Sahin, Levent

    2010-07-01

    Subchorionic hematoma might be associated with poor pregnancy outcome. Two intra cytoplasmic sperm injection pregnancies complicated with subchorionic hematoma were found to have elevated mid-trimester maternal serum alpha-fetoprotein levels. One of them had miscarriage at 16 weeks' gestation and the other delivered a healthy baby by cesarean section. The valid interpretation of triple test result might be complicated by subchorionic hematoma. Therefore, it is better not to order triple test in such cases to avoid unnecessarily provoking the anxiety of the couple. PMID:19883262

  7. Alpha fetoprotein antagonizes apoptosis induced by paclitaxel in hepatoma cells in vitro

    OpenAIRE

    Mingyue Zhu; Wei Li; Yan Lu; Xu Dong; Yi Chen; Bo Lin; Xieju Xie; Junli Guo; Mengsen Li

    2016-01-01

    Hepatocellular carcinoma (HCC) cell resistance to the effects of paclitaxel has not been adequately addressed. In this study, we found that paclitaxel significantly inhibited the viability of HLE, Bel 7402 and L-02 cells in a dose- and time-dependent manner. HLE cells and L-02 cells resisted the cytotoxicity of paclitaxel when transfected with pcDNA3.1-afp vectors. However, Bel 7402 cell sensitivity to paclitaxel was increased when transfected with alpha fetoprotein (AFP)-siRNA. Bel 7402 cell...

  8. Prussian blue modified amperometric FIA biosensor: one-step immunoassay for alpha-fetoprotein.

    Science.gov (United States)

    Guan, Jian-Guo; Miao, Yu-Qing; Chen, Jian-Rong

    2004-03-15

    The aim of this study was to develop a rapid method to measure alpha-fetoprotein (AFP) in human serum by use of one-step sandwich enzyme-linked immunosorbent assay (ELISA)-based immunobiosensor with disposable screen-printed carbon electrode technology. Prussian blue (PB) was deposited using cyclic voltammetry (CV) on the surface of electrode to catalyze H202 from the reaction of glucose oxidase. It took only about 30 min to complete the whole experimental procedure through flow injection analysis (FIA). A detection range obtained is in the range from 5 to 500 ng/ml AFP. This detection seems to be quick, reliable and practical. PMID:15128097

  9. Radioimmunoassay

    International Nuclear Information System (INIS)

    Consideration is given to radioimmunoassay technique. Referring to radioimmunoassay reagents, orientation is given to the choice of an antibody that fulfills the method requirements and hormone labelling. A table with theoretical specific activities for some iodinated hormones is shown. An analysis is made of the techniques of separation of the free phase of the hormone from that is bound to the antibody in complex form, based in general separation principles such as differential migration, free hormone adsorption and antibody-bound hormones separation. A table summarizing many substances that can be assayed by radioimmunoassay or competitive protein - binding assay is presented. Radioimmunological assays used in medical routine by Brazilian physicians are described such as those of renin, insulin, pituitary gonadotropins, human chorionic gonadotropin, progesterone, human growth and cortisol

  10. 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 Mendelian-in

  11. The immunosuppression role of alpha-fetoprotein in human hepatocellular carcinoma.

    Science.gov (United States)

    Meng, Wenbo; Bai, Bing; Bai, Zhongtian; Li, Yan; Yue, Ping; Li, Xun; Qiao, Liang

    2016-06-01

    Human hepatocellular carcinoma (HCC) is the third most common cause of cancer-related death. Alpha-fetoprotein (AFP) is perhaps the best-defined tumor marker for HCC, and as such, it is widely used in clinical settings as an adjuvant diagnostic and prognostic tool. Up to 70% of HCC cases exhibit elevated serum level of AFP, but its pathophysiological functions in HCC are poorly defined. It is now known that AFP is not just a fetal form of carrier protein and a tumor marker, it is also critically involved in the regulation of several important cellular functions, such as cell growth, differentiation, apoptosis, angiogenesis, and immune regulation. In this mini-review, we summarize the recent development of AFP in hepatocellular carcinoma. PMID:27448785

  12. 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...... with ASD and 243 control subjects were included in a case-control study, using a historic birth cohort maintained at Statens Serum Institute. Measurements of MS-AFP were obtained from a multicentre screening program, whereas clinical data were obtained from nationwide registers. Association between MS-AFP...... and ASD status was analyzed using logistic regression models and nonparametric tests. Results: Crude, but not adjusted, estimates showed that MS-AFP levels were slightly, but significantly, higher in mothers of children with ASD, compared with their control subject counterparts. People with ASD had an...

  13. Alpha-fetoprotein, the major fetal serum protein, is not essential for embryonic development but is required for female fertility

    OpenAIRE

    Gabant, Philippe; Forrester, Lesley; Nichols, Jennifer; Van Reeth, Thierry; De Mees, Christelle; Pajack, Bernard; Watt, Alistair; Smitz, Johan; Alexandre, Henri; Szpirer, Claude; Szpirer, Josiane

    2002-01-01

    The alpha-fetoprotein gene (Afp) is a member of a multigenic family that comprises the related genes encoding albumin, alpha-albumin, and vitamin D binding protein. The biological role of this major embryonic serum protein is unknown although numerous speculations have been made. We have used gene targeting to show that AFP is not required for embryonic development. AFP null embryos develop normally, and individually transplanted homozygous embryos can develop in an AFP-deficient microenviron...

  14. 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

  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

    OpenAIRE

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

    2014-01-01

    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 t...

  16. Elevated Serum Levels of Alpha-Fetoprotein in Patients with Infantile Hemangioma Are Not Derived from within the Tumor

    OpenAIRE

    Itinteang, Tinte; Chibnall, Alice M.; Marsh, Reginald; Jonathan C. Dunne; de Jong, Sophie; Davis, Paul F.; Leadbitter, Philip; Tan, Swee T.

    2016-01-01

    Aims The embryonic-like stem cell origin of infantile hemangioma (IH) and the observed elevated serum levels of alpha-fetoprotein (AFP) in patients with hepatic IH led us to investigate if this tumor was the source of AFP. Materials and methods We measured serial serum levels of AFP in patients with problematic proliferating IH treated with surgical excision or propranolol treatment. We also investigated the expression of AFP in extrahepatic IH samples using immunohistochemical sta...

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

    OpenAIRE

    Olivier Brock; Matthieu Keller; Quentin Douhard; Julie Bakker

    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 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 e...

  18. Telomerase Inhibition Decreases Alpha-Fetoprotein Expression and Secretion by Hepatocellular Carcinoma Cell Lines: In Vitro and In Vivo Study

    OpenAIRE

    Roula Tahtouh; Anne-Sophie Azzi; Nada Alaaeddine; Soulaima Chamat; Hasnaa Bouharoun-Tayoun; Layal Wardi; Issam Raad; Riad Sarkis; Najibe Abou Antoun; George Hilal

    2015-01-01

    Alpha-fetoprotein (AFP) is a diagnostic marker for hepatocellular carcinoma (HCC). A direct relationship between poor prognosis and the concentration of serum AFP has been observed. Telomerase, an enzyme that stabilizes the telomere length, is expressed by 90% of HCC. The aim of this study was to investigate the effect of telomerase inhibition on AFP secretion and the involvement of the PI3K/Akt/mTOR signaling pathway. Proliferation and viability tests were performed using tetrazolium salt. A...

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

    International Nuclear Information System (INIS)

    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.)

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

    International Nuclear Information System (INIS)

    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 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 5/mm3 (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.

  1. 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.

  2. Alpha-fetoprotein expression is a potential prognostic marker in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Dénes G(o)r(o)g; János Reg(o)ly-Mérei; Sándor Paku; László Kopper; Péter Nagy

    2005-01-01

    AIM: To characterize the alpha-fetoprotein (AFP) positive and negative hepatocellular carcinoma (HCC) samples.METHODS: Thirty-seven paraffin-embedded human HCC samples were analyzed by immunohistochemistry for the following antigens: AFP, β-catenin, p53, CD44, MSH-2,MLH-1, and HNF-4. The tumors were divided into two groups based on the AFP expression. The immunophenotypic data and important clinical parameters were studied between the two groups.RESULTS: Twenty-one of the thirty-seven examined HCCs were AFP positive. Seven with nuclear p53 staining were AFP positive, while seven tumors with nuclear β-catenin staining were AFP negative. CD44 staining and high histological tumor grade were more frequent among the AFP-positive HCCs. The other immunophenotypical and dinical parameters did not show statistically significant difference in their distribution between the AFP positive and negative samples.CONCLUSION: AFP expression in HCC correlates with unfavorable prognostic factors, while nuclear β-catenin positivity is more common among the AFP-negative liver tumors. This observation supports the microarray data onin vivo human tumors.

  3. 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. PMID:27468205

  4. 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

  5. Alpha fetoprotein antagonizes apoptosis induced by paclitaxel in hepatoma cells in vitro.

    Science.gov (United States)

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

    2016-01-01

    Hepatocellular carcinoma (HCC) cell resistance to the effects of paclitaxel has not been adequately addressed. In this study, we found that paclitaxel significantly inhibited the viability of HLE, Bel 7402 and L-02 cells in a dose- and time-dependent manner. HLE cells and L-02 cells resisted the cytotoxicity of paclitaxel when transfected with pcDNA3.1-afp vectors. However, Bel 7402 cell sensitivity to paclitaxel was increased when transfected with alpha fetoprotein (AFP)-siRNA. Bel 7402 cell resistance to paclitaxel was associated with the expression of the "stemness" markers CD44 and CD133. Paclitaxel significantly inhibited growth and promoted apoptosis in HLE cells and L-02 cells by inducing fragmentation of caspase-3 and inhibiting the expression of Ras and Survivin, but pcDNA3.1-afp vectors prevented these effects. However, paclitaxel could not significantly promote the cleavage of caspase-3 or suppress the expression of Ras and Survivin in Bel 7402 cells. Silenced expression of AFP may be synergistic with paclitaxel to restrain proliferation and induce apoptosis, enhance cleavage of caspase-3, and suppress the expression of Ras and Survivin. Taken together, AFP may be an important molecule acting against paclitaxel-inhibited proliferation and induced apoptosis in HCC cells via repressing the activity of caspase-3 and stimulating the expression of Ras and Survivin. Targeted inhibition of AFP expression after treatment with paclitaxel is an available strategy for the therapy of patients with HCC. PMID:27255186

  6. Contribution of alpha-fetoprotein in liver transplantation for hepatocellular carcinoma

    Science.gov (United States)

    Charrière, Bérénice; Maulat, Charlotte; Suc, Bertrand; Muscari, Fabrice

    2016-01-01

    Alpha-fetoprotein (AFP) is the main tumor biomarker available for the management of hepatocellular carcinoma (HCC). Although it is neither a good screening test nor an accurate diagnostic tool for HCC, it seems to be a possible prognostic marker. However, its contribution in liver transplantation for HCC has not been fully determined, although its use to predict recurrence after liver transplantation has been underlined by international societies. In an era of organ shortages, it could also have a key role in the selection of patients eligible for liver transplantation. Yet unanswered questions remain. First, the cut-off value of serum AFP above which liver transplantation should not be performed is still a subject of debate. We show that a concentration of 1000 ng/mL could be an exclusion criterion, whereas values of liver transplantation. These recent tools have associated AFP values with morphological criteria, thus refining pre-existing criteria, such as Milan, University of California, San Francisco, or “up-to-seven”. We provide a review of the different criteria submitted within the past years. Finally, AFP can be used to monitor recurrence after transplantation, although there is little evidence to support this claim. Future challenges will be to draft new international guidelines to implement the use of AFP as a selection tool, and to determine a clear cut-off value above which liver transplantation should not be performed.

  7. Alpha-fetoprotein-producing early gastric cancer of the remnant stomach: report of a case.

    Science.gov (United States)

    Shibata, Yutaka; Sato, Kouhei; Kodama, Masashi; Nanjyo, Hiroshi

    2007-01-01

    A 67-year-old man initially underwent a distal gastrectomy for early gastric cancer (T1, N0, M0; Stage IA) in March 1995. During the follow-up period, an elevation of the serum alpha-fetoprotein (AFP) level (98.8 ng/ml) and a liver tumor (S4) were detected. A left hepatectomy was performed in December 1996. Immunohistochemically, AFP-positive cells were present in both the primary gastric tumor and metastasized liver tumor. The serum AFP level normalized immediately, but it elevated again to 22.4 ng/ml. An endoscopic examination revealed a protruding lesion in the remnant stomach. A total resection of the remnant stomach was performed in February 2005. The tumor was evaluated T1, N0, M0; Stage IA, with positive staining for AFP. The patient has survived without any sign of recurrence for more than 11 years after the first diagnosis of cancer. To the best of our knowledge, this is the first case of a long-term survival of AFP-producing gastric cancer with successfully resected metachronous liver metastasis and gastric remnant carcinoma. PMID:17952534

  8. Radioimmunoassay for determination of alpha subunit of pituitary glycoprotein hormones in patients with pituitary tumors

    International Nuclear Information System (INIS)

    A radioimmunoassay method for alpha subunit has been described and applied for serum alpha subunit determinations in normal subjects and 71 patients with pituitary tumors /45 acromegalic and 26 non-acromegalic/. The labelling of alpha subunit by the chloramine T technique yielded 125I-alpha subunit of high specific activity and high immuno-reactivity. Three purification methods of labelled 125I-alpha subunit were compared; the best separation of undamaged 125I-alpha subunit from impurities was achieved by gel filtration on Ultrogel AcA54 column, whereas gel filtration on Sephadex G-100 and adsorption chromatography on CF-11 cellulose gave less satisfactory results. Microheterogenity of 125I-alpha subunit was disclosed by chromatofocusing on PBE 94; the fractions of high immunoreactivitiy had isoelectric points of 6.0, 5.5 and 4.8. In normal subjects, radioimmunoassay of alpha subunit gave the following results /mean and SD/: 0.75 ng/ml +- 0.41 in males and 0.80 ng/ml +- 0.39 in females in reproductive age. In 9 acromegalic serum alpha subunit concentration were elevated up to 21 ng/ml, and in 8 non-acromegalic up to 30 ng/ml. One woman with acromegaly and high serum alpha subunit concentration had also elevated serum TSH associated with hyperthyroids. Our results disclosed that high serum alpha subunit concentration occurs in 25 % of patients with pituitary adenomas. (Author)

  9. 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.

  10. Elevated serum alpha fetoprotein levels promote pathological progression of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Peng Li; Shan-Shan Wang; Hui Liu; Ning Li; Michael A McNutt; Gang Li; Hui-Guo Ding

    2011-01-01

    AIM: To investigate the biological role of alpha fetoprotein (AFP) and its clinical significance in carcinogenesis of hepatocellular carcinoma (HCC).METHODS: Clinical analysis of HCC patients and immunohistochemical examination were conducted to eva-luate the relationship between serum AFP level and patient mortality. Confocal microscopy, Western blotting, dimethylthiahzolyl-2,5-diphenyl-tetrazolium bromide, Cell Counting Kit-8 assays and flow cytometry were performed to explore the possible mechanism.RESULTS: Among the 160 HCC patients enrolled in this study, 130 patients survived 2 years (81.25%), with a survival rate of 86.8% in AFP 250 μg/L group, demonstrating a higher mortality rate in HCC patients with higher AFP levels. Surgical treatment was beneficial only in patients with low AFP levels. The mortality rate of HCC patients with high AFP levels who were treated surgically was apparently higher than those treated with conservative management. The results of immunohistochemistry showed that AFP and AFP receptor were merely expressed in tissues of HCC patients with positive serum AFP. Consistently, in vitro analysis showed that AFP and AFPS were expressed in HepG2 but not in HLE cells. AFP showed a capability to promote cell growth, and this was more apparent in HepG2 cells, in which the proliferation was increased by 3.5 folds. Cell cycle analysis showed that the percentage of HepG2 cells in S phase after exposure to AFP was modestly increased.CONCLUSION: HCC patients with higher AFP levels show a higher mortality rate, which appears to be attributable to the growth promoting properties of AFP.

  11. A case of an alpha-fetoprotein-producing intrahepatic cholangiocarcinoma suggests probable cancer stem cell origin.

    Science.gov (United States)

    Ishikawa, Kenji; Sasaki, Atsushi; Haraguchi, Naotsugu; Yoshikawa, Yasuji; Mori, Masaki

    2007-03-01

    Recent evidence suggests that some cancers may originate from cancer stem cells, which may derive from carcinogenesis of normal stem cells. A hepatic progenitor cell population, which gives rise to hepatocytes and cholangiocytes, has been suggested in humans, though whether these cells can give rise to malignant tumors has not been confirmed. We report here a case of an alpha-fetoprotein (AFP)-producing intrahepatic cholangiocarcinoma (ICC) in an 81-year-old woman with chronic hepatitis C viral infection, suggesting malignant transformation of hepatic stem cells as a mechanism for hepatic neoplasia. Abdominal computed tomography revealed a low-density mass with surrounding enhancement measuring 5 cm x 5 cm in segments IV and VIII of the liver. The preoperative serum levels of tumor markers were 1.7 ng/ml of carcinoembryonic antigen, 22 mAU/ml of protein induced by vitamin K absence or antagonist II, 43.4 U/ml of carbohydrate antigen 19-9, and 1,560 ng/ml of AFP. Following central bisegmentectomy of the liver, serum AFP levels decreased dramatically. Histologically, the tumor cells showed indistinct glandular structures with abundant fibrous stroma. Immunohistochemical analysis demonstrated that the neoplastic cells reacted strongly to antibodies against AFP and cytokeratin (CK) 7. In addition, cancer cells showed partially positive reaction to anti-CK14, a liver stem cell marker, and to anticluster designation (CD) 133, a hematopoietic stem cell marker, and negative reaction to antihepatocyte paraffin (HepPar) 1. These data may indicate that the tumor was derived from a normal liver stem cell that underwent oncogenic transformation. PMID:17405896

  12. Contribution of alpha-fetoprotein in liver transplantation for hepatocellular carcinoma.

    Science.gov (United States)

    Charrière, Bérénice; Maulat, Charlotte; Suc, Bertrand; Muscari, Fabrice

    2016-07-28

    Alpha-fetoprotein (AFP) is the main tumor biomarker available for the management of hepatocellular carcinoma (HCC). Although it is neither a good screening test nor an accurate diagnostic tool for HCC, it seems to be a possible prognostic marker. However, its contribution in liver transplantation for HCC has not been fully determined, although its use to predict recurrence after liver transplantation has been underlined by international societies. In an era of organ shortages, it could also have a key role in the selection of patients eligible for liver transplantation. Yet unanswered questions remain. First, the cut-off value of serum AFP above which liver transplantation should not be performed is still a subject of debate. We show that a concentration of 1000 ng/mL could be an exclusion criterion, whereas values of AFP could also prove useful. However, evidence is lacking regarding the values that should be used. Today, the real input of AFP seems to be its integration into new criteria to select patients eligible for a liver transplantation. These recent tools have associated AFP values with morphological criteria, thus refining pre-existing criteria, such as Milan, University of California, San Francisco, or "up-to-seven". We provide a review of the different criteria submitted within the past years. Finally, AFP can be used to monitor recurrence after transplantation, although there is little evidence to support this claim. Future challenges will be to draft new international guidelines to implement the use of AFP as a selection tool, and to determine a clear cut-off value above which liver transplantation should not be performed. PMID:27478538

  13. The intracellular mechanism of alpha-fetoprotein promoting the proliferation of NIH 3T3 cells

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    AIM The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS The effect of AFP on the proliferation of NIH 3T3 cells was measured by incorporation of 3H-TdR. Receptor-binding assay of 125I-AFP was performed to detect the properties of AFP receptor in NIH 3T3 cells. The influences of AFP on the [cAMP]i and the activities of protein kinase A (PKA) were determined. Western blot was used to detect the change of K-ras P21 protein expression. RESULTS The proliferation of NIH 3T3 cells treated with 0-80 mg/L of AFP was significantly enhanced. The Scatchard analysis indicated that there were two classes of binding sites with KD of 2.722×10-9M (Bmax=12810 sites per cell) and 8.931× 10-SM (Bmax=l19700 sites per cell) respectively. In the presence of AFP (20 mg/L), the content of cAMP and activities of PKA were significantly elevated . The level of K-ras P21 protein was upregulated by AFP at the concentration of 20 mg/L. The monoclonal antibody against AFP could reverse the effects of AFP on the cAMP content, PKA activity and the expression of K-ras p21 gene. CONCLUSION The effect of AFP on the cell proliferation was achieved by binding its receptor to trigger the signal transduction pathway of cAMP-PKA and alter the expression of K- ras p21 gene.

  14. Clinical Value of Hepatoma-Specific Alpha-Fetoprotein in the Diagnosis of Hepatocellular Carcinoma

    Institute of Scientific and Technical Information of China (English)

    Runzhou Ni; Mingbing Xiao; Fei Jin; Cuihua Lu; Jiefei Huang; Xianyong Meng

    2006-01-01

    OBJECTIVE To study the clinical value of hepatoma-specific alpha-fetoprotein (HS-AFP) in the diagnosis and differential diagnosis of hepatocellular carcinoma (HCC).METHODS A method of vertical slab polyacrylamide gel electrophoresis with discontinuous buffer system was developed to separate AFP subtypes. After separation, the AFP subtypes were transferred to nitrocellulose and reacted first with rabbit anti-human AFP and then with goat anti-rabbit IgG-HRP. Finally, AFP subtypes were visualized by reacting with 3,3'-diaminobenzidine tetrahydrochloride. A HS-AFP band was determined in 82 cases with HCC and 95 cases with benign liver diseases.The correlations between the positive rates of HS-AFP and serum AFP concentration, tumor size as well as portal vein metastasis were analyzed.RESULTS Serum AFP in the cases with various liver diseases was separated into one to several bands. The fastest band on electrophoresis (FAFP) was found in all patients, while the band at the cathodal site (HSAFP) was detected predominantly in HCC but rarely in benign liver diseases. The positive rate of HS-AFP in HCC was 74.4%, which was significantly higher than that in benign liver diseases (9.1%, 7.3% and 10.0% in liver cirrhosis, chronic hepatitis and acute hepatitis respectively). HSAFP was detected in 3 out of 9 HCC cases with AFP<50 μg/L , but in none of 22 cases of benign liver diseases with the same AFP concentration. HS-AFP correlated with serum AFP concentration and tumor size to some extent, but not with portal vein metastasis.CONCLUSION HS-AFP increases the sensitivity of diagnosing HCC in patients with negative AFP, and is useful in distinguishing high AFP due to HCC from that caused by benign liver diseases.

  15. 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

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

    International Nuclear Information System (INIS)

    Highlights: ► QDs-based homogeneous time-resolved fluoroimmunoassay was developed to detect AFP. ► The conjugates were prepared with QDs-doped microspheres and anti-AFP McAb. ► The conjugates were prepared with LTCs and another anti-AFP McAb. ► Excess amounts of conjugates were used for detecting AFP without rinsing. ► 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 Förster resonance transfer (TR-FRET) at 67.3% and the spatial distance of the donor to acceptor calculated to be 66.1 Å. Signals from TR-FRET were found to be proportional to AFP concentrations. The resulting standard curve was log Y = 3.65786 + 0.43863·log X (R = 0.996) with Y the QPs fluorescence intensity and X the AFP concentration; the calculated sensitivity was 0.4 ng mL−1. By assaying test samples against the standard curve, the coefficient of variations was <5%, indicating that QDs were suitable for

  17. 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.

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

    Directory of Open Access Journals (Sweden)

    Zhu Jian

    2010-01-01

    Full Text Available Abstract 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.

  19. 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)

    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)

  20. A meaningful appraisal of cholestasis in serum total bilirubin, cholyglyine, alpha-fetoprotein and carbohydrate antigen19-9

    International Nuclear Information System (INIS)

    Objective: Appraise the clinical significant how the serum total bilirubin (TB), cholylglycine (CG), α-fetoprotein (AFP), carbohydrate antigen 19-9 (CA 19-9) have their concentration changes in the pathological changes of intrahepatic cholestasis through a combined detection to them. Methods: The serum samples from 96 cases of chronic virus hepatitis, 26 cases of liver cirrhosis and 50 cases of normal people were detected by biochemistry for TB, by radioimmunoassay for CG, by electro chemiluminescence for AFP and CA19-9. Results: There is no obvious deference of serum TB in the group without intrahepatic cholestasis, the group of cholestasis without clinical symptoms and the control groups. There is also a marked deference (P<0.01) in the group of cholestasis with clinical symptoms, the group of liver cirrhosis, the group without intrahepatic cholestasis, the control group and the group of cholestasis without clinical symptoms. the serum CG from the groups of intrahepatic cholestasis, the group without intrahepatic cholestasis, and the control group all show a very obvious deference (P<0.01). The serum CA19-9 from the groups of intrahepatic cholestasis and the group without intrahepatic cholestasis show an obvious deference. The serum AFP, CA19-9 from the group of liver cancer show a very obvious deference (P<0.01). Conclusions: In clinc bilirubin is a rough index to reflect cholestasis. It has its own limit in deciding patterns of deferent bile obstruction. In the early stage of intrahepatic cholestasis, that the index of CG is high obviously points out existence of intrahepatic cholestasis. CG and the liver impairment are well interrelated and they are comparatively sensitive indexes of liver function. AFP reflects the regeneration of the liver cell necrosis and it means alarm to the seriousness of intrahepatic cholestasis. CA19-9 is a marker of tumor of biliary tract. The index increase through an initial observation is interrelated to the seriousness of

  1. Heat shock protein 70 chaperoned alpha-fetoprotein in human hepatocellular carcinoma cell line BEL-7402

    Institute of Scientific and Technical Information of China (English)

    Xiao-Ping Wang; Qiao-Xia Wang; Hai-Yan Li; Rui-Fen Chen

    2005-01-01

    AIM: To investigate the interaction between heat shock protein 70 (HSP70) and α-fetoprotein (AFP) in human hepatocellular carcinoma (HCC) cell line BEL7402.METHODS: The expression and localization of HSP70 and AFP in human HCC cell line BEL-7402 were determined by immunocytochemistry and indirect immunofluorescence cytochemical staining. The interaction between HSP70 and AFP in HCC cells was analyzed by immunoprecipitation and Western blot.RESULTS: Immunocytochemical staining detection showed that HCC cell BEL-7402 expressed a high level of HSP70 and AFP synchronously. Both were stained in cell plasma.AFP existed in the immunoprecipitate of anti-HSP70 mAb,while there was HSP70 in the immunoprecipitate of antiAFP mAb.CONCLUSION: HSP70 chaperones AFP in human HCCcell BEL-7402. The interaction between HSP70 and AFP in human HCC cell can be a new route to study the pathogenesis and immunotherapy of HCC.

  2. Downregulation of alpha-fetoprotein siRNA inhibits proliferation of SMMC-7721 cells

    Institute of Scientific and Technical Information of China (English)

    Yun-Shan Wang; Xiao-Li Ma; Tong-Gang Qi; Xiang-Dong Liu; Yue-Sheng Meng; Guang-Ju Guan

    2005-01-01

    AIM: To study the function of α-fetoprotein (AFP) in SMMC-7721 hepatoma cells.METHODS: A hairpin siRNA expressing plasmid pSilencer3.0-H1-afp was constructed and transfected into SMMC-7721 cells with Lipofectamine 2000. The expression of AFP was monitored by real-time RT-PCR and immunoassays, its effect on SMMC-7721 cell proliferation and cell death was detected by MTT and fluorescenceactivated cell sorter (FACS).RESULTS: The AFP-siRNA expressing plasmid downregulated the expression of AFP obviously (about 34%), and inhibited SMMC-7721 cell proliferation, but did not induce apoptosis.CONCLUSION: Downregulation of AFP siRNA inhibits proliferation of SMMC-7721 cells, but cannot cause apoptosis.

  3. Characteristics of magnetic labeling on liver tumors with anti-alpha-fetoprotein-mediated Fe3O4 magnetic nanoparticles

    Directory of Open Access Journals (Sweden)

    Huang KW

    2012-06-01

    Full Text Available Kai-Wen Huang,1 Jen-Jie Chieh,2 Herng-Er Horng,2 Chin-Yih Hong,3 Hong-Chang Yang41Department of Surgery and Hepatitis Research Center, National Taiwan University Hospital, Taipei, 2Institute of Electro-optical Science and Technology, National Taiwan Normal University, Taipei, 3Graduate Institute of Biomedical Engineering, National Chung Hsing University, Taichung, 4Department of Physics, National Taiwan University, Taipei, TaiwanAbstract: For preoperative and intraoperative detection of tumor distribution, numerous multimodal contrast agents, such as magnetic nanoparticles (MNPs with several examination indicators, are currently in development. However, complex materials, configuration, and cost are required for multimodal contrast agents, accompanied by a high possibility of toxicity and low popularity in clinics. Nevertheless, the magnetic labeling of MNPs using bioprobes should be feasible not only in preoperative magnetic resonance imaging (MRI, but also in intraoperative examination based on other magnetic properties. In this study, anti-alpha-fetoprotein (AFP-mediated Fe3O4 MNPs, injected into mice with liver tumors, were used to examine the characteristics of magnetic labeling. Using MRI and scanning superconducting-quantum-interference-device biosusceptometry (SSB, based on alternating current (AC susceptibility, the magnetic labeling occurred significantly on the first day post-injection of anti-AFP magnetic fluid (MF, and then decreased over time. However, for both MF without antibodies and an anti-carcinoembryonic antigen MF, no magnetic labeling occured on the first day of their respective post-injection. The favorable agreement indicates that magnetic labeling possesses two magnetic characteristics: distortion of the imaging field and AC susceptibility. In addition, the results of the biopsy tests, anti-AFP staining, and Prussian blue staining show the same dynamics as those of magnetic methodologies and prove that bound MNPs on

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

    OpenAIRE

    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 ...

  5. SETX mutations are a frequent genetic cause of juvenile and adult onset cerebellar ataxia with neuropathy and elevated serum alpha-fetoprotein

    OpenAIRE

    Nanetti, Lorenzo; Cavalieri, Simona; Pensato, Viviana; Erbetta, Alessandra; Pareyson, Davide; Panzeri, Marta; Zorzi, Giovanna; Antozzi, Carlo; Moroni, Isabella; Gellera, Cinzia; Brusco, Alfredo; Mariotti, Caterina

    2013-01-01

    Objectives/background Ataxia with oculomotor apraxia defines a group of genetically distinct recessive ataxias including ataxia-telangectasia (A-T, ATM gene), ataxia with oculomotor apraxia type 1 (AOA1, APTX gene) and type 2 (AOA2, SETX gene). Although, a few unique clinical features differentiate each of these forms, the patients also share common clinical signs, such as the presence of cerebellar atrophy, sensorimotor axonal neuropathy, and elevated alpha-fetoprotein (AFP) serum level. Mat...

  6. Determination of maternal serum α-fetoprotein (AFP) by a Polish AFP radioimmunoassay - a suitable screening method for the detection of neural tube defects

    International Nuclear Information System (INIS)

    230 double determinations of maternal serum α-fetoprotein (AFP) have been made using a Polish AFP-RIA. On this basis the applicability of this method as screening for neural tube defects has been proved. An additional comparison with a French RIA and an enzyme immunoassay (EIA) from the Serum-Werke Dessau confirmed equal accuracy. There were three pathological increased values of AFP. In one case an anencephalus was found and in the other a spina bifida. In the third case the value was normal at the control examination. General AFP screening in pregnant women is recommended. (author)

  7. A highly sensitive biosensor for tumor maker alpha fetoprotein based on poly(ethylene glycol) doped conducting polymer PEDOT.

    Science.gov (United States)

    Cui, Min; Song, Zhiling; Wu, Yumin; Guo, Bing; Fan, Xiaojian; Luo, Xiliang

    2016-05-15

    Biocompatible polymers, such as poly(ethylene glycol) (PEG), are of great significance in the development of bio-interfaces and biosensors, as they possess excellent biocompatibility and are easy for modification. A novel highly biocompatible polymer composite was synthesized herein through electrochemical polymerization of the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) and a PEG derivative, 4-arm PEG terminated with thiol groups. The electrodeposited conducting polymer composite of PEG doped PEDOT (PEDOT/PEG) exhibited flake-like nanostructure, large surface area and outstanding stability. In order to further immobilize antibodies, gold nanoparticles (AuNPs) were introduced to the PEDOT/PEG composite surface through their unique interaction with the thiol groups. AuNPs modified PEDOT/PEG provided a desirable support for the immobilization of various biomolecules such as antibodies for alpha fetoprotein (AFP), a vital tumor biomarker for liver cancer. The fabricated AFP biosensor demonstrated favorable selectivity, high sensitivity, and ultralow detection limit. Furthermore, owing to the presence of PEG polymers that are highly hydrophilic, such AuNPs/PEDOT/PEG based AFP biosensor also exhibited good anti-fouling ability, and it was capable of assaying target AFP in 10% (V/V) human serum samples, indicating highly feasible potential for clinical diagnosis. PMID:26774088

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

    International Nuclear Information System (INIS)

    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

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

    Directory of Open Access Journals (Sweden)

    Zhou Ying

    2012-01-01

    Full Text Available Abstract Backgrounds 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. Methods 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. Results 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. Conclusions 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.

  10. Relationship between Expression of the Human Alpha-Fetoprotein Gene and DNA Methylation Status of the Promoter Region

    Institute of Scientific and Technical Information of China (English)

    Lijun Chen; Wei Wang; Qiuyue Jin; Ruimin Wang; Wenliang Hu

    2006-01-01

    OBJECTIVE DNA methylation has been regarded as an important epigenetic signature reflecting the transcription state of DNA in cells. This study was to conducted to assess the relationship between human alpha-fetoprotein (AFP) gene expression and the DNA methylation status of the promoter region in three different cells, namely two human hepatocellular carcinoma (HCC) cell lines and normal human fibroblasts.METHODS Transcription of the AFP gene was verified by RT-PCR. After bisulphate treatment of DNA, the methods of MSP and BSP were used to analyze the methylation density and status within single DNA strands of two closely spaced CpG dinucleotides of the promoter region in the different cells.RESULTS RT-PCR analysis indicated that the expression of the AFP gene in HepG2 cells was significantly higher than in SMMC-7721 cells,and that the AFP gene was not expressed in normal human fibroblasts.By MSP and BSP we observed that the promoter region was demethylated in the AFP-high-expressing cell lines, and that the sites of -2,494 bp and -2,431 bp in the AFP genomic sequence can be used as detection sites for early tumorous diagnosis.CONCLUSION These results indicate that the DNA methylation state of the promoter region has a negative correlation with AFP gene expression.

  11. Developmental alterations in the alpha-fetoprotein sugar chain in maternal serum analyzed by lectin affinity electrophoresis.

    Directory of Open Access Journals (Sweden)

    Kawahara N

    1999-06-01

    Full Text Available Our purpose was to investigate developmental alterations of human alpha-fetoprotein (AFP oligosaccharides in maternal serum by lectin affinity electrophoresis and to compare the AFP glycoforms in maternal serum with those in umbilical cord serum and amniotic fluid. AFP glycoforms were separated by affinity electrophoresis with concanavalin A (Con A, lentil lectin (LCA, erythroagglutinating phytohemagglutinin (E-PHA and Allomyrina dichotoma lectin (allo A and detected by sensitive antibody affinity blotting. In maternal serum, increased proportions of Con A-nonreactive AFP (AFP-C1, LCA strongly-reactive AFP (AFP-L3 and E-PHA-reactive AFP (AFP-P4 and AFP-P5 decreased gradually during the early gestational weeks. Allo A-nonreactive AFP (AFP-A1 and asialo-AFP were found only in amniotic fluids during early gestational weeks. The percentages of these glycoforms at full term were almost the same among those body fluids. Since the glycoforms of maternal serum AFP were close to those of umbilical cord serum AFP, lectin-affinity electrophoretic analysis of maternal serum AFP may be useful for evaluating the developmental state of fetus by examining the nature of AFP sugar chain.

  12. In vivo screening of hepatocellular carcinoma using AC susceptibility of anti-alpha fetoprotein-activated magnetic nanoparticles.

    Directory of Open Access Journals (Sweden)

    Jen-Jie Chieh

    Full Text Available With antibody-mediated magnetic nanoparticles (MNPs applied in cancer examinations, patients must pay at least twice for MNP reagents in immunomagnetic reduction (IMR of in vitro screening and magnetic resonance imaging (MRI of in vivo tests. This is because the high maintenance costs and complex analysis of MRI have limited the possibility of in vivo screening. Therefore, this study proposes novel methods for in vivo screening of tumors by examining the AC susceptibility of bound MNPs using scanning superconducting-quantum-interference-device (SQUID biosusceptometry (SSB, thereby demonstrating high portability and improved economy. The favorable agreement between in vivo tests using SSB and MRI demonstrated the feasibility of in vivo screening using SSB for hepatocellular carcinoma (HCC targeted by anti-alpha fetoprotein (AFP-mediated MNPs. The magnetic labeling was also proved by in vitro tests using SSB and biopsy assays. Therefore, patients receiving bioprobe-mediated MNPs only once can undergo in vivo screening using SSB in the future.

  13. 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. PMID:26386330

  14. Enhancement of plasma α-fetoprotein, as measured by sandwich-type radioimmunoassay, and induction of γ-glutamyl transpeptidase-positive hepatic cell foci in rats fed benzo[a]pyrene

    International Nuclear Information System (INIS)

    Male Fischer rats were fed semipurified diets containing 0, 1, 100, and 1000 ppM benzo[a]pyrene (BaP) for 6 or 13 wk. Plasma samples were assayed for α-fetoprotein (AFP) by a new sandwich-type radioimmunoassay (RIA) utilizing a special controlled porous-glass solid phase. This procedure is described in detail. Significant AFP elevation (p less than or equal to 0.01) was observed in the highest BaP treatment group after 5 wk of treatment. The 1 and 100 ppM BaP groups exhibited no AFP elevation throughout the study. Liver sections from the 1000 ppM groups had discrete γ-glutamyl transpeptidase (GGT)-positive foci 10 to 20 cells in diameter by the sixth wk. GGT-positive foci were not evident in liver sections from the other treatment groups. Thus a high level of dietary BaP appears to rapidly alter rat liver cells, indicating hepatic neoplasia

  15. Characteristics of magnetic labeling on liver tumors with anti-alpha-fetoprotein-mediated Fe3O4 magnetic nanoparticles.

    Science.gov (United States)

    Huang, Kai-Wen; Chieh, Jen-Jie; Horng, Herng-Er; Hong, Chin-Yih; Yang, Hong-Chang

    2012-01-01

    For preoperative and intraoperative detection of tumor distribution, numerous multimodal contrast agents, such as magnetic nanoparticles (MNPs) with several examination indicators, are currently in development. However, complex materials, configuration, and cost are required for multimodal contrast agents, accompanied by a high possibility of toxicity and low popularity in clinics. Nevertheless, the magnetic labeling of MNPs using bioprobes should be feasible not only in preoperative magnetic resonance imaging (MRI), but also in intraoperative examination based on other magnetic properties. In this study, anti-alpha-fetoprotein (AFP)-mediated Fe(3)O(4) MNPs, injected into mice with liver tumors, were used to examine the characteristics of magnetic labeling. Using MRI and scanning superconducting-quantum-interference-device biosusceptometry (SSB), based on alternating current (AC) susceptibility, the magnetic labeling occurred significantly on the first day post-injection of anti-AFP magnetic fluid (MF), and then decreased over time. However, for both MF without antibodies and an anti-carcinoembryonic antigen MF, no magnetic labeling occured on the first day of their respective post-injection. The favorable agreement indicates that magnetic labeling possesses two magnetic characteristics: distortion of the imaging field and AC susceptibility. In addition, the results of the biopsy tests, anti-AFP staining, and Prussian blue staining show the same dynamics as those of magnetic methodologies and prove that bound MNPs on tumor tissue are rotatable by an AC magnetic field to express AC susceptibility. Therefore, with the simple configuration of antibody-mediated MNPs, magnetic labeling is also feasible for intraoperative examinations using SSB with high mobility and sensitivity. PMID:22787394

  16. 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.

  17. Alpha-Fetoprotein Detection of Hepatocellular Carcinoma Leads to a Standardized Analysis of Dynamic AFP to Improve Screening Based Detection.

    Science.gov (United States)

    Bird, Thomas G; Dimitropoulou, Polyxeni; Turner, Rebecca M; Jenks, Sara J; Cusack, Pearce; Hey, Shiying; Blunsum, Andrew; Kelly, Sarah; Sturgeon, Catharine; Hayes, Peter C; Bird, Sheila M

    2016-01-01

    Detection of hepatocellular carcinoma (HCC) through screening can improve outcomes. However, HCC surveillance remains costly, cumbersome and suboptimal. We tested whether and how serum Alpha-Fetoprotein (AFP) should be used in HCC surveillance. Record linkage, dedicated pathways for management and AFP data-storage identified i) consecutive highly characterised cases of HCC diagnosed in 2009-14 and ii) a cohort of ongoing HCC-free patients undergoing regular HCC surveillance from 2009. These two well-defined Scottish patient cohorts enabled us to test the utility of AFP surveillance. Of 304 cases of HCC diagnosed over 6 years, 42% (129) were identified by a dedicated HCC surveillance programme. Of these 129, 47% (61) had a detectable lesion first identified by screening ultrasound (US) but 38% (49) were prompted by elevated AFP. Despite pre-HCC diagnosis AFP >20kU/L being associated with poor outcome, 'AFP-detected' tumours were offered potentially curative management as frequently as 'US-detected' HCCs; and had comparable survival. Linearity of serial log10-transformed AFPs in HCC cases and in the screening 'HCC-free' cohort (n = 1509) provided indicators of high-risk AFP behaviour in HCC cases. An algorithm was devised in static mode, then tested dynamically. A case/control series in hepatitis C related disease demonstrated highly significant detection (pAFP in HCC surveillance. We show proof-of-principle that an automated and further refine-able algorithmic interpretation of AFP can identify patients at higher risk of HCC. This approach could provide a cost-effective, user-friendly and much needed addition to US surveillance. PMID:27308823

  18. Diagnostic Performance of Alpha-Fetoprotein, Protein Induced by Vitamin K Absence, Osteopontin, Dickkopf-1 and Its Combinations for Hepatocellular Carcinoma

    OpenAIRE

    Eun Sun Jang; Sook-Hyang Jeong; Jin-Wook Kim; Yun Suk Choi; Philippe Leissner; Christian Brechot

    2016-01-01

    Background & Aims Alpha-fetoprotein (AFP) is the most widely used serum biomarker for hepatocellular carcinoma (HCC), despite its limitations. As complementary biomarkers, protein induced by vitamin K absence (PIVKA-II), osteopontin (OPN), and Dickkopf-1 (DKK-1) have been proposed. This study aimed to perform a head-to-head comparison of the diagnostic performance of AFP, PIVKA-II, OPN and DKK-1 as single or in combination to seek the best biomarker or panel, and to investigate the clinical f...

  19. Prolonged Survival in a Case of Chemotherapy-Sensitive Gastric Cancer That Produced Alpha-Fetoprotein and Protein Induced by Vitamin K Antagonist-II

    OpenAIRE

    Ogasawara, Naotaka; Takahashi, Emiko; Matsumoto, Tomoko; Amaike, Manami; Nohara, Mako; Nagao, Kazuhiro; Ebi, Masahide; Funaki, Yasushi; Sasaki, Makoto; Kasugai, Kunio

    2015-01-01

    The number of reported cases of alpha-fetoprotein (AFP)-producing gastric cancer has gradually increased, with a reported prevalence of 1.3–1.5% of all gastric cancer cases. However, reports of gastric cancer accompanied by elevated serum levels of both AFP and protein induced by vitamin K antagonist-II (PIVKA-II) are rare. The prognosis of AFP- and PIVKA-II-producing gastric cancer has been reported to be very poor because the tumor cells were considered to have a high malignant potential an...

  20. Radioimmunoassay of inhibin based on synthetic human inhibin alpha-chain peptide

    International Nuclear Information System (INIS)

    Polyclonal rabbit antisera were produced against cyclic human inhibin [(Cys6, Tyr7) alpha-(6-30)NH2] peptide, covalently conjugated to bovine serum albumin. The tyrosine residue introduced at position 7 facilitated the oxidative incorporation of radiolabel (125I) to yield a tracer with specific activity of 73.9 Ci/g. These reagents were used to develop a homologous equilibrium radioimmunoassay for human inhibin, with polyethylene glycol, 200 g/L, serving as the separation phase. At a detection limit of 2 micrograms/L (n = 7), immunoactive inhibin was detectable in human pre-ovulatory follicular fluid (128 micrograms/L), seminal plasma (2374 micrograms/L), amniotic fluid (66 micrograms/L), and placental extract (347 micrograms/L). We also demonstrated inhibin immunoreactivity in biological fluids from other mammalian species: macaque, chimpanzee, porcine, and bovine, but not rodent (guinea pig). Although the antisera were raised against a nonbioactive inhibin peptide, immunoglobulins fractionated on Protein A-Sepharose neutralized the bioactivity of human ovarian inhibin. Further characterization of inhibin immuno- and bioactivity was undertaken with immobilized heparin, divalent metal cations, and dye ligands. Only heparin-Sepharose distinguished between immuno- and bioactive inhibin

  1. Hepatitis B Virus X Protein Driven Alpha Fetoprotein Expression to Promote Malignant Behaviors of Normal Liver Cells and Hepatoma Cells

    Science.gov (United States)

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

    2016-01-01

    Background: The infection of Hepatitis B virus (HBV) is closely associated with the development of hepatocellular carcinoma(HCC), HBV-X protein(HBx) is able to induce expression of alpha-fetoprotein(AFP) in normal liver cells, and AFP harbors a function to promote malignant transformation of normal liver cells, but the role AFP playing in malignant behaviors of HCC cells is still unclear. Methods: Fifty-six liver tissue samples were collected from the clinical patients through hepatectomy(include normal liver tissues, HBV-related hepatitis liver tissues and HBV-related HCC tissues), and diagnosis of these tissues by pathology section, expression of AFP, Ras and CXCR4 were evidenced by immunohisochemical staining and Western blotting; The proliferation of human normal liver cells line L-02 cells and human hepatoma cells line, HLE cells(non AFP-producing) were performed by MTT method; Repaired capacity of L-02 and HLE cells were compared by wound healing assay; Migration and invasion of these cells were analyzed by Transwell chamber assay; HBx expressed vectors(pcDNA3.1-HBx) were constructed and transfected into L-02 and HLE cells, effects of pcDNA3.1-HBx on the malignant behaviors were also detected by MTT, Transwell chamber assay and the expression of AFP, Ras and CXCR4 were evidenced by Western blotting. Results: we found that expression of AFP, Ras and CXCR4 in HBV-related HCC and lymph nodes metastasis tissues were significantly elevated compared with HBV-related HCC, non metastasis tissues and HBV-related hepatitis tissues; Expression of AFP, Ras and CXCR4 in HBV-related hepatitis tissues were significantly enhanced compared with normal liver tissues; The growth ratio, migratory and invasive ability, expression of AFP, Ras and CXCR4 of the cells were outstanding promoted while L-02 and HLE cells were transfected with pcDNA3.1-HBx vectors. The proliferation ratio, migration and invasion ability, and expression of Ras and CXCR4 were significantly inhibited while

  2. Telomerase inhibition decreases alpha-fetoprotein expression and secretion by hepatocellular carcinoma cell lines: in vitro and in vivo study.

    Science.gov (United States)

    Tahtouh, Roula; Azzi, Anne-Sophie; Alaaeddine, Nada; Chamat, Soulaima; Bouharoun-Tayoun, Hasnaa; Wardi, Layal; Raad, Issam; Sarkis, Riad; Antoun, Najibe Abou; Hilal, George

    2015-01-01

    Alpha-fetoprotein (AFP) is a diagnostic marker for hepatocellular carcinoma (HCC). A direct relationship between poor prognosis and the concentration of serum AFP has been observed. Telomerase, an enzyme that stabilizes the telomere length, is expressed by 90% of HCC. The aim of this study was to investigate the effect of telomerase inhibition on AFP secretion and the involvement of the PI3K/Akt/mTOR signaling pathway. Proliferation and viability tests were performed using tetrazolium salt. Apoptosis was determined through the Annexin V assay using flow cytometry. The concentrations of AFP were measured using ELISA kits. The AFP mRNA expression was evaluated using RT-PCR, and cell migration was evaluated using a Boyden chamber assay. The in vivo effect of costunolide on AFP production was tested in NSG mice. Telomerase inhibition by costunolide and BIBR 1532 at 5 and 10 μM decreased AFP mRNA expression and protein secretion by HepG2/C3A cells. The same pattern was obtained with cells treated with hTERT siRNA. This treatment exhibited no apoptotic effect. The AFP mRNA expression and protein secretion by PLC/PRF/5 was decreased after treatment with BIBR1532 at 10 μM. In contrast, no effect was obtained for PLC/PRF/5 cells treated with costunolide at 5 or 10 μM. Inhibition of the PI3K/Akt/mTOR signaling pathway decreased the AFP concentration. In contrast, the MAPK/ERK pathway appeared to not be involved in HepG2/C3A cells, whereas ERK inhibition decreased the AFP concentration in PLC/PRF/5 cells. Modulation of the AFP concentration was also obtained after the inhibition or activation of PKC. Costunolide (30 mg/kg) significantly decreased the AFP serum concentration of NSG mice bearing HepG2/C3A cells. Both the inhibition of telomerase and the inhibition of the PI3K/Akt/mTOR signaling pathway decreased the AFP production of HepG2/C3A and PLC/PRF/5 cells, suggesting a relationship between telomerase and AFP expression through the PI3K/Akt/mTOR pathway. PMID:25822740

  3. Telomerase inhibition decreases alpha-fetoprotein expression and secretion by hepatocellular carcinoma cell lines: in vitro and in vivo study.

    Directory of Open Access Journals (Sweden)

    Roula Tahtouh

    Full Text Available Alpha-fetoprotein (AFP is a diagnostic marker for hepatocellular carcinoma (HCC. A direct relationship between poor prognosis and the concentration of serum AFP has been observed. Telomerase, an enzyme that stabilizes the telomere length, is expressed by 90% of HCC. The aim of this study was to investigate the effect of telomerase inhibition on AFP secretion and the involvement of the PI3K/Akt/mTOR signaling pathway. Proliferation and viability tests were performed using tetrazolium salt. Apoptosis was determined through the Annexin V assay using flow cytometry. The concentrations of AFP were measured using ELISA kits. The AFP mRNA expression was evaluated using RT-PCR, and cell migration was evaluated using a Boyden chamber assay. The in vivo effect of costunolide on AFP production was tested in NSG mice. Telomerase inhibition by costunolide and BIBR 1532 at 5 and 10 μM decreased AFP mRNA expression and protein secretion by HepG2/C3A cells. The same pattern was obtained with cells treated with hTERT siRNA. This treatment exhibited no apoptotic effect. The AFP mRNA expression and protein secretion by PLC/PRF/5 was decreased after treatment with BIBR1532 at 10 μM. In contrast, no effect was obtained for PLC/PRF/5 cells treated with costunolide at 5 or 10 μM. Inhibition of the PI3K/Akt/mTOR signaling pathway decreased the AFP concentration. In contrast, the MAPK/ERK pathway appeared to not be involved in HepG2/C3A cells, whereas ERK inhibition decreased the AFP concentration in PLC/PRF/5 cells. Modulation of the AFP concentration was also obtained after the inhibition or activation of PKC. Costunolide (30 mg/kg significantly decreased the AFP serum concentration of NSG mice bearing HepG2/C3A cells. Both the inhibition of telomerase and the inhibition of the PI3K/Akt/mTOR signaling pathway decreased the AFP production of HepG2/C3A and PLC/PRF/5 cells, suggesting a relationship between telomerase and AFP expression through the PI3K

  4. Intrahepatic cholangiocarcinoma detected by elevated levels of alpha-fetoprotein-L3 after hepatectomy for hepatocellular carcinoma in a patient with Budd-Chiari syndrome.

    Science.gov (United States)

    Yamamoto, Masakazu; Otsubo, Takehito; Ariizumi, Shunichi; Nakano, Masayuki; Takasaki, Ken

    2005-01-01

    We report the case of a 57-year-old woman with Budd-Chiari syndrome, hepatocellular carcinoma (HCC), and intrahepatic cholangiocarcinoma (ICC). She underwent partial hepatectomy for HCC in April 2000. After surgery, alpha-fetoprotein (AFP) and protein induced by vitamin K absence II (PIVKA-II) returned to normal levels, but lens culinaris agglutinin-reactive alpha-fetoprotein (AFP-L3) increased, and ultrasonography showed a nodule 2 cm in greatest dimension in the left lateral segment of the liver. We diagnosed this nodule as recurrence from HCC and performed a partial hepatectomy in October 2001. Microscopic examination showed that tubular adenocarcinoma and immunohistochemical staining was focally positive for AFP. AFP-L3 was 0% and AFP was 5 ng/ml 3 months after re-operation. This case was interesting in that ICC was detected by elevated levels of AFP-L3, and ICC produced AFP from the time it was minute in size. PMID:16119710

  5. Isolation characterization and radioimmunoassay of rat alpha-macrofetoprotein (acute phase A2 macroglobulin)

    International Nuclear Information System (INIS)

    Rat alpha-macrofetoprotein (AMF) has been isolated and purified by antibody affinity chromatography and quantitated using a double antibody radioimmunoassay (RIA) with a sensitivity of 20 ng. Two forms of AMF, differing in reference mobilities (R) in 5% polyacrylamide gels and in isoelectric focusing points (pI), are present in serum. The Rsub(f0.10)AMF has a pI of 4.35 and Rsub(f0.25) AMF a pI of 4.65. Rsub(f0.25) AMF competes 2.2 times better than Rsub(f0.10) AMF in an RIA with 125I Rsub(f0.25) AMF as the predominant antigen. The two forms do not differ substantially in mol. wt. AMF has a mol. wt of 700.000 +- 70.000 determined by SDS polyacrylamide gel electrophoresis. Dithiothreitol reduction of AMF resulted in six different chains, the largest mol. wt 175.000. It is suggested that the five smaller chains are the result of interaction of AMF with endoproteases. The E280sup(1%) of AMF is 9.6 +- 0.3 by the biuret method using bovine serum albumin as a standard and 10.6 +- 0.3 by microKjeldahl N analysis. Amino-acid analysis of AMF is compared with the analyses recently reported by others. Normal adult rat serum AMF concentrations determined for two groups of rats are 17 +- 2 and 32 +- 6 μg/ml. (author)

  6. The diagnostic value of combined detection of alpha-fetoprotein, AFP, lentil lectin-reactive alpha-fetoprotein(AFP-L3) and alpha L fucosidase (AFU) level for the patients with primary liver cancer in early stage%联合检测血清AFP、AFP异质体和AFU对早期肝癌诊断价值的研究

    Institute of Scientific and Technical Information of China (English)

    王杰; 赵业民; 陈增银; 史光军

    2013-01-01

    Objective To explore the diagnostic value of combined detection of serum alpha-fetoprotein ( AFP), lentil lectin-reactive alpha-fetoprotein( AFP-L3 ) and alpha L fucosidase ( AFU ) level for the patients with primary liver cancer in early stage. Methods The serum level of AFP, AFP-L3 and AFU were detected in 56 patients with primary liver cancer (PLC), 158 patients with benign liver disease, 40 patients with other malignant tumor without PLC and 40 healthy persons.Results The positive rate of AFP, AFP-L3 and AFU in the group of PLC were signiifcantly higher than that of in the group of benign liver diseases and in the group of other malignant tumor without PLC ( P<0.01 ). Combined detection have a complementary function for the diagnosis of patients PLC in early stage and the sensitivity rate was 94.60%. The sensitivity in the group of combined detection is higher than that of in the group of single detection ( P<0.05 ). Conclusion The sensitivity rate of single detection for AFP, AFP-L3 and AFU for the diagnosis of PLC in the early stage is similar and combined detection could signiifcantly improve the sensitivity rate for the diagnosis of PLC in the early stage and decrease the misdiagnosis rate. The combined detection of AFP, AFP-L3 and AFU can be used as a newly routine detection method for the high population of the patients with PLC in the early stage especially.%目的:探讨血清甲胎蛋白(alpha-fetoprotein ,AFP)、扁豆凝集素亲和型甲胎蛋白异质体(lentil lectin-reactive alpha-fetoprotein, AFP-L3)、α-L-岩藻糖苷酶(alpha L fucosidase, AFU)水平联合检测对原发性肝癌(primary liver cancer, PLC)的诊断价值。方法对56例PLC、158例良性肝病患者、40例非PLC恶性肿瘤患者以及40例健康查体者进行AFP、AFP-L3和AFU进行联合检测。结果 PLC组的AFP、AFP-L3、AFU的阳性率明显高于良性肝病和非PLC恶性肿瘤组( P<0.01),PLC患者血清AFP、AFP-L3、AFU三者之间无相关性,联

  7. Aplasia cutis congenita in a setting of fetus papyraceus associated with small fetal abdominal circumference and high alpha-fetoprotein and amniotic acetylcholinesterase.

    Science.gov (United States)

    Mazza, Joni M; Klein, Janice F; Christopher, Kurt; Silverberg, Nanette B

    2015-01-01

    Fetus papyraceus is the fetal death of one or more fetuses in a multiparous pregnancy. The surviving infants can experience extensive aplasia cutis in an H-shaped distribution over the flanks and abdomen as a consequence of the loss of their fetal sibling. We report the case of a monochorionic, diamniotic pregnancy complicated by a single fetal death at 13 weeks of gestational age. Aplasia cutis of the surviving twin was suggested in utero by three criteria: high amniotic and maternal alpha-fetoprotein, detectable acetylcholinesterase, and small abdominal circumference on prenatal ultrasound. This constellation of findings in the setting of fetus papyraceus can be an indicator of aplasia cutis in the surviving fetus. PMID:24118049

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

    International Nuclear Information System (INIS)

    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-SiO2) 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 NaBH4, 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-SiO2 (HRP-anti-AFP-Ag-SiO2) as secondary antibodies. Compared with pure silver nanoparticles, Ag-SiO2 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-H2O2 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.

  9. 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)

    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

  10. 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)

    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 (uE3) 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 uE3 were well correlated with gestational age, respectively (r=0.94, p=0.003; r=-0.97, p=0.029; r=0.99, p3 weren't (r=-0.17, p=0.22; r=0.36, p=0.09, respectively). The values of aFP, CG and uE3 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 E3 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, 1996

  11. Development and application of radioimmunoassays for interleuking-1 alpha and interleukin-1 beta

    International Nuclear Information System (INIS)

    Specific and sensitive radioimmunoassay (RIA) systems for human IL-1α and IL-1β, using sheep polyclonal antisera have been developed. The assays, which do not require prior extraction of Il-1, have been used to quantify Il-1α and Il-1β in synovial fluid and plasma and to quantify intracellular and extracellular IL-1α and Il-1β produced by human monocytes stimulated with endotoxin. (author). 16 refs.; 3 figs.; 1 tab

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

    Directory of Open Access Journals (Sweden)

    Lei Zheng

    2012-12-01

    Full Text Available 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.

  13. 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)

    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)

  14. 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.

  15. Prolonged Survival in a Case of Chemotherapy-Sensitive Gastric Cancer That Produced Alpha-Fetoprotein and Protein Induced by Vitamin K Antagonist-II.

    Science.gov (United States)

    Ogasawara, Naotaka; Takahashi, Emiko; Matsumoto, Tomoko; Amaike, Manami; Nohara, Mako; Nagao, Kazuhiro; Ebi, Masahide; Funaki, Yasushi; Sasaki, Makoto; Kasugai, Kunio

    2015-01-01

    The number of reported cases of alpha-fetoprotein (AFP)-producing gastric cancer has gradually increased, with a reported prevalence of 1.3-1.5% of all gastric cancer cases. However, reports of gastric cancer accompanied by elevated serum levels of both AFP and protein induced by vitamin K antagonist-II (PIVKA-II) are rare. The prognosis of AFP- and PIVKA-II-producing gastric cancer has been reported to be very poor because the tumor cells were considered to have a high malignant potential and the cancer progressed rapidly. We described a case of gastric cancer producing AFP and PIVKA-II in which chemotherapy was effective and resulted in prolonged survival, and these two tumor markers were useful for monitoring the treatment response. Routine health screening using upper abdominal ultrasonography revealed hepatic tumors in an apparently healthy 65-year-old man. Whole-body computed tomography (CT) revealed multiple hepatic tumors, and an esophagogastroduodenoscopy (EGD) revealed a Bormann type 3 tumor in the lower stomach. A biopsy specimen confirmed that the tumor was immunohistochemically positive for AFP, PIVKA-II, and human epidermal growth factor receptor 2. After chemotherapy, the gastric tumor appeared as a small elevated lesion on EGD, and CT revealed a remarkable reduction in the size of the metastatic liver tumors. The patient is still alive, 35 months after the initial chemotherapy. PMID:26034473

  16. Prolonged Survival in a Case of Chemotherapy-Sensitive Gastric Cancer That Produced Alpha-Fetoprotein and Protein Induced by Vitamin K Antagonist-II

    Directory of Open Access Journals (Sweden)

    Naotaka Ogasawara

    2015-04-01

    Full Text Available The number of reported cases of alpha-fetoprotein (AFP-producing gastric cancer has gradually increased, with a reported prevalence of 1.3-1.5% of all gastric cancer cases. However, reports of gastric cancer accompanied by elevated serum levels of both AFP and protein induced by vitamin K antagonist-II (PIVKA-II are rare. The prognosis of AFP- and PIVKA-II-producing gastric cancer has been reported to be very poor because the tumor cells were considered to have a high malignant potential and the cancer progressed rapidly. We described a case of gastric cancer producing AFP and PIVKA-II in which chemotherapy was effective and resulted in prolonged survival, and these two tumor markers were useful for monitoring the treatment response. Routine health screening using upper abdominal ultrasonography revealed hepatic tumors in an apparently healthy 65-year-old man. Whole-body computed tomography (CT revealed multiple hepatic tumors, and an esophagogastroduodenoscopy (EGD revealed a Bormann type 3 tumor in the lower stomach. A biopsy specimen confirmed that the tumor was immunohistochemically positive for AFP, PIVKA-II, and human epidermal growth factor receptor 2. After chemotherapy, the gastric tumor appeared as a small elevated lesion on EGD, and CT revealed a remarkable reduction in the size of the metastatic liver tumors. The patient is still alive, 35 months after the initial chemotherapy.

  17. 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.

  18. Expression of alpha-fetoprotein messenger RNA in BEL-7404 human hepatoma cells and effect of L-4-oxalysine on the expression

    Institute of Scientific and Technical Information of China (English)

    1998-01-01

    AIM To investigate alpha-fetoprotein (AFP) mRNA expression in BEL-7404 human hepatoma cells and the effect of L-4-oxalysine (OXL) on the expression.METHODS BEl-7404 human hepatoma cells were maintained in RPMI 1640 media. Human AFP cDNA probe was labelled with digoxigenin-11-dUTP by the random primer labelling method. The expression of AFP mRNA in Bel-7404 cells was determined by an in situ hybridization technique with digoxigenin-labelled human AFP cDNA probe. The positive intensities of AFP mRNA in cells were analyzed by microspectrophotometer and expressed as absorbance at 470nm. For the experiment with OXL, cells were incubated with various concentrations of the agent for 72h.RESULTS Essentially all the hepatoma cells contained AFP mRNA in the cytoplasm, although in various amounts. The specificity of the hybridization reaction was confirmed by control experiments in which the use of RNase-treated BEL-7404 cells, non-AFP-producing cells (HL-60 human leukemia cells) or a nonspecific cDNA probe resulted in negative hybridization. When the cells were treated with OXL (25, 50mg/L), the content of AFP mRNA in the cytoplasm was decreased with the inhibition percentages of 34.3% and 70.1%, respectively (P<0.05).CONCLUSION AFP mRNA was expressed in BEL-7404 human hepatoma cells and OXL suppressed AFP mRNA expression in the cells.

  19. 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.

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

    International Nuclear Information System (INIS)

    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+ 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

  1. First-trimester maternal serum alpha-fetoprotein as a marker for fetal chromosomal disorders. Dutch Working Party on Prenatal Diagnosis.

    Science.gov (United States)

    Van Lith, J M

    1994-10-01

    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 pregnancies. The median multiple of the normal median (MOM) in 32 Down's syndrome pregnancies was 0.83 with a 95 per cent confidence interval ranging from 0.60 to 1.04. The difference between the distributions of first-trimester MS-AFP in normal and Down's syndrome pregnancies was statistically significant (t-test: t = 2.34, P MOM = 1.26; seven cases with sex chromosome abnormalities, MOM = 1.07; 22 cases with a chromosomal mosaic pattern in chorionic villi, MOM = 1.08). We conclude that first-trimester MS-AFP can discriminate between normal and Down's syndrome pregnancies, but is not an effective marker. First-trimester MS-AFP has no value as a marker for other fetal chromosomal disorders. PMID:7534926

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

    International Nuclear Information System (INIS)

    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 125I, 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

  3. 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

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

    International Nuclear Information System (INIS)

    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. - 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

  5. 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)

    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 Pb2+. 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, -NH2 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 ΔIp 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.

  6. Diagnostic Performance of Alpha-Fetoprotein, Protein Induced by Vitamin K Absence, Osteopontin, Dickkopf-1 and Its Combinations for Hepatocellular Carcinoma.

    Directory of Open Access Journals (Sweden)

    Eun Sun Jang

    Full Text Available Alpha-fetoprotein (AFP is the most widely used serum biomarker for hepatocellular carcinoma (HCC, despite its limitations. As complementary biomarkers, protein induced by vitamin K absence (PIVKA-II, osteopontin (OPN, and Dickkopf-1 (DKK-1 have been proposed. This study aimed to perform a head-to-head comparison of the diagnostic performance of AFP, PIVKA-II, OPN and DKK-1 as single or in combination to seek the best biomarker or panel, and to investigate the clinical factors affecting their performance.Using 401 stored plasma samples obtained from 208 HCC patients and 193 liver cirrhosis control patients, plasma AFP, PIVKA-II, OPN and DKK-1 levels were measured by ELISA, and receiver operating characteristic curve analyses were performed for each biomarker and for every combination of two to four markers.Of the four biomarkers, AFP showed the highest area under the curve (0.786. The sensitivity and specificity for each single biomarker was 62% and 90.2% (AFP>20 ng/mL, 51.0% and 91.2% (PIVKA-II>10 ng/mL, 46.2% and 80.3% (OPN>100 ng/mL, and 50.0% and 80.8% (DKK-1>500 pg/mL, respectively. Among the combinations of two biomarkers, AFP>20 ng/mL or DKK-1>500 pg/mL showed the best diagnostic performance (sensitivity 78.4%, specificity 72.5%. Triple or quadruple combination did not improve the diagnostic performance further. The patient's age, etiology and tumor invasiveness of HCC affected the performance of each marker.AFP was the most useful single biomarker for HCC diagnosis, and the combined measurement of AFP and DKK-1 could maximize the diagnostic yield. Clinical decision should be based on the consideration of various factors affecting the diagnostic performance of each biomarker. Efforts to seek novel HCC biomarkers should be continued.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  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. Application of Latex particles in detecting alpha- fetoprotein (AFP)%乳胶粒子法检测甲胎蛋白的应用

    Institute of Scientific and Technical Information of China (English)

    陈浩全; 曾嫚妮

    2012-01-01

    Objective:To evaluate the application value of Latex method in detection of alpha - fetoprotein ( AFP). Methods:Repetitive determination tests within and between batches were done on serum specimens, then the determination results were compared and analyzed by Latex method and electrochemiluminescence assay. Results; The results had no significant difference between Latex method and electrochemiluminescence assay, the recoveries were 95.5% for Latex method and 96.7% for electrochemiluminescence assay. Conclusion; Though electrochemiluminescence assay had high sensitivity and precision, it was not suitable for the wide use due to its high cost in instrument and reagent. Latex method had good correlation with electrochemiluminescence assay, which can be applied widely for its rapidness, stable results and low cost.%目的:评价Latex法检测甲胎蛋白(AFP)的临床应用价值.方法:对血清标本做批内批间重复性测定试验及Latex法与电化学发光法进行对比测定.结果:Latex法与电化学发光法检测AFP结果无显著差异,Latex法与电化学发光法平均回收率分别为95.5%和96.7%.结论:电化学发光法灵敏度、精密度高,但其仪器、试剂价格高,不适于全面推广;Latex法与电化学发光法检测AFP结果无显著差异,相关性好,快速、结果稳定,试剂便宜在全自动生化仪上检测,可广泛应用.

  13. The significance of serum 17 alpha-hydroxy progesterone as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    17 α-hydroxy progesterone (17 OH-P), which exists in minute quantities in blood, has been considered to be a difficult substance to determine, and its dynamics and significance have been largely unclarified. We produced a sensitive, highly specific antiserum to 17 OH-P and used it to quantitatively measure by radioimmunoassay the serum 17 OH-P levels in the peripheral blood of both non gravid and gravid females and in umbilical blood. In women with normal menstrual cycles, peripheral serum 17 OH-P levels in the preovulatory period were 309 +- 77 pg/ml, in the luteal phase 757 +- 268 pg/ml and in menopause 158 +- 71 pg/ml. In gravid women, 6 - 20 wk levels were 1.8 +- 0.3 ng/ml, at 26 wk 3.4 +- 0.9 ng/ml, at 30 wks 3.3 +- 0.5 ng/ml, at 34 wks 4.6 +- 0.3 ng/ml, and at 40 wks or term 5.2 +- 0.6 ng/ml. In cases of threatened abortion and toxemia gravidarum, levels were slightly lower than those listed above. At parturition, maternal peripheral levels were 5.9 +- 0.9 ng/ml, umbilical venous levels 16.4 +- 5.2 ng/ml, umbilical arterial levels 10.5 +- 5.8 ng/ml, and retroplacental blood levels 13.2 +- 4.8 ng/ml. From these findings, it is possible to hypothesize that in normal women, the preovulatory 17 OH-P is produced and released by the adrenal cortex, in the luteal phase by the corpus luteum, and in late pregnancy by the placenta. Further, it is possible to identify the ovulatory period, and in pregnancy the placental function. (auth.)

  14. Cytological alteration of cultured rat liver cells by 3'-methyl-4-dimethylaminoazobenzene with special reference to chromosome changes, changes of growth patterns at a colony level and alpha-fetoprotein production.

    Directory of Open Access Journals (Sweden)

    Tokiwa,Takayoshi

    1980-06-01

    Full Text Available A near diploid clone derived from a rat liver cell line was continuously treated with various concentrations of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB in culture. By treatment with 2.8 micrograms/ml, cells with 41 chromosomes formed a mode and which then shifted to 39. The chromosome numbers of cells treated with 5.4 micrograms/ml were widely distributed at early stages, but later the mode shifted to hypotetraploid region. Untreated control cells were confirmed as near diploid. Increased plating efficiency by 3'-Me-DAB as well as the appearance of large sized colonies was obtained. The production of alpha-fetoprotein (AFP by the cells was slightly enhanced by treatment with 3'-Me-DAB. The cells treated with and without 3'-Me-DAB did not produce any tumor in rats 6 months after their intraperitoneal injection.

  15. AFP、APF-L3、GP73、AFP-mRNA联合检测在肝细胞癌诊断中的应用价值%The application value of combined detection for serum alpha-fetoprotein, alpha-fetoprotein-L3, Golgi protein 73 and AFP-mRNA in the diagnosis of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    李永勤; 陈浩; 耿坤静; 杜博勋; 裘银久

    2012-01-01

    Objective To investigate the application value of combined detection for alpha fetoprotein (AFP), alpha-fetoprotein-L3 ( AFP-L3 ) , Golgi protein 73 ( GP73 ) and AFP-mRNA in the diagnosis of hepatocellular carcinoma ( HCC). Methods Enzyme-linked immunosorbent assay ( ELISA) was used to detect the serum levels of GP73 in 73 cases of HCC,41 cases of liver cirrhosis,36 cases of chronic hepatitis B and 25 healthy subjects. AFP was detected by chemiluminescent immunoassay, and AFP-L3 was determined by adsorption centrifuging. AFP-mRNA was tested by fluorescent quantitative RT-PCR. Results There were significant differences in the levels of AFP, AFP-L3, GP73, AFP-mRNA between HCC group and cirrhosis group, normal control group ( P <0.01). The areas evaluated according to ROC curves were 0.691 ( AFP) ,0.859 ( APF-L3), 0.816 (GP73),0.762 (AFP-mRNA) respectively in HCC group,and the sensitivities were 54. 79% ,76. 71% , 80.82% ,53. 42%, respectively, and specificities were 79. 41% ,95. 10% ,82. 35% ,99. 02% , respectively. The areas under ROC curves for the four tumor-related markers were 0.958 by combined detection. The sensitivity of combined detection was significantly increased, which reached 95. 8% , and its specificity was 95. 0% in the diagnosis of HCC. Conclusion The combination detection for AFP, AFP-L3 ,GP73 and AFP-mRNA can obviously enhance the sensitivity and specificity in the diagnosis of HCC,which is superior to individual detection of AFP, AFP-L3.GP73 or AFP-mRNA.%目的 探讨甲胎蛋白(AFP)、甲胎蛋白异质体( AFP-L3)、高尔基体蛋白(GP73)及AFP-mR-NA联合检测在HCC诊断中的价值.方法 收集175例血清、其中肝细胞癌患者73例(HCC组)、肝硬化患者41例(肝硬化组)、慢乙肝患者36例(慢性肝炎组)、正常人25例(正常人组),采用酶联免疫法检测GP73,用化学发光法检测AFP,用亲和吸附离心法检测AFP-L3,采用荧光定量逆转录聚合酶链反应(RTPCR)法检测患者AFP

  16. Effects of curcumin in pediatric epithelial liver tumors: inhibition of tumor growth and alpha-fetoprotein in vitro and in vivo involving the NFkappaB- and the beta-catenin pathways

    Science.gov (United States)

    Bortel, Nicola; Armeanu-Ebinger, Sorin; Schmid, Evi; Kirchner, Bettina; Frank, Jan; Kocher, Alexa; Schiborr, Christina; Warmann, Steven; Fuchs, Jörg; Ellerkamp, Verena

    2015-01-01

    In children with hepatocellular carcinoma (pHCC) the 5-year overall survival rate is poor. Effects of cytostatic therapies such as cisplatin and doxorubicin are limited due to chemoresistance and tumor relapse. In adult HCC, several antitumor properties are described for the use of curcumin. Curcumin is one of the best-investigated phytochemicals in complementary oncology without relevant side effects. Its use is limited by low bioavailability. Little is known about the influence of curcumin on pediatric epithelial hepatic malignancies. We investigated the effects of curcumin in combination with cisplatin on two pediatric epithelial liver tumor cell lines. As mechanisms of action inhibition of NFkappaB, beta-catenin, and decrease of cyclin D were identified. Using a mouse xenograft model we could show a significant decrease of alpha-fetoprotein after combination therapy of oral micellar curcumin and cisplatin. Significant concentrations of curcuminoids were found in blood samples, organ lysates, and tumor tissue after oral micellar curcumin administration. Micellar curcumin in combination with cisplatin can be a promising strategy for treatment of pediatric HCC. PMID:26515460

  17. Combined radioimmunoassay of 17-alpha-hydroxyprogesterone and 11-desoxycortisol, and of dehydroepiandrosterone-sulphate: Methods and some novel clinical findings

    International Nuclear Information System (INIS)

    The radioimmunoassay (RIA) methods for determining 17-alpha-hydroxyprogesterone (17HOP), 11-desoxycortisol (Cpd.-S), and dehydroepiandrosterone-sulphate, are reported. The plasma levels of these hormones were measured in a series of 15 diagnostic groups including adrenal diseases, fertility-related problems and human growth hormone (HGH) deficiency states. The relations of DHEA-S and growth hormone were studied in a group of children with growth retardation who presented an absent response to HGH stimulation tests. The DHEA-S level found in these patients was significantly lower than that of their age-matched controls. Furthermore, in an etiologically different entity, i.e. head-injury patients, similar findings were also made. In a single case with HGH deficiency due to antibodies against HGH, DHEA-S levels were not detectable. It is suggested that the determination of DHEA-S plasma levels can provide further insight into the classification of patients with fertility-related problems. Furthermore, it appears that DHEA-S can be taken as an indicator for the availability of bioactive human growth hormone. The various forms of congenital adrenal hyperplasia (CAH) can be successfully screened and detected by the determination of the plasma levels of 17-alpha-hydroxyprogesterone (17HOP), of 11-desoxycortisol (Cpd.-S), and of dehydroepiandrosterone-sulphate (DHEA-S). These tests are also relevant in the investigation of patients with hirsutism, hyperandrogenism, and amenorrhea. RIA procedures for the measurement of these hormones are described and new clinical data on DHEA-S are presented in relation to fertility problems both in men and women, such as oligozoospermia, oligomenorrhea and aspermia and, in cases of human growth hormone (HGH), deficiency states such as retarded growth, head-injury patients and, in one case, HGH deficiency due to antibodies directed against HGH. The relations between normo- and hyperprolactinaemia and DHEA-S are also examined

  18. Radioimmunoassay apparatus

    International Nuclear Information System (INIS)

    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)

  19. Electrochemical immunosensor for alpha-fetoprotein determination based on ZnSe quantum dots/Azure I/gold nanoparticles/poly (3,4-ethylenedioxythiophene) modified Pt electrode

    International Nuclear Information System (INIS)

    A novel amperometric enzyme immunosensor with amplified sensitivity for the determination of alpha-fetoprotein (AFP) was constructed with layer-by-layer assembly of ZnSe quantum dots (ZnSe QDs)/Azure I/gold nanoparticles (nanoAu)/poly (3,4-ethylenedioxythiophene) (PEDOT) on Pt electrode. Firstly, citrate coated nanoAu was immobilized on the PEDOT polymer film, which was electrochemically synthesized in ionic liquid electrolyte of 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim][BF4]). Then, Azure I was immobilized on nanoAu/PEDOT composite matrix as a redox probe and was used to immobilize ZnSe QDs. Subsequently, AFP antibody (anti-AFP) was adsorbed onto the surface of ZnSe layer. Finally, horseradish peroxidase (HRP) was employed to block sites against nonspecific binding and amplify the current signal of the antigen–antibody reaction. The modification processes were characterized by cyclic voltammetry, scanning electron microscopy. The factors influenced the performances of the proposed immunosensors were studied in detail. Because of the synergism between Azure I and nanoAu/PEDOT to facilitate electron-transfer process, and the small diameter of ZnSe QDs favorable for stabilization of biological activity to a large extent, the immunosensor displayed a high sensitivity, fast analytical time, a relatively low detection limit of 1.1 fg/mL at 3 times of signal-to-noise ratio (S/N = 3), and a especially broad linear response to AFP in a ranges from 5 × 10−5 to 250 ng/mL. Moreover, the selectivity, repeatability, and stability of the proposed immunosensor were acceptable

  20. Progesterone radioimmunoassay

    International Nuclear Information System (INIS)

    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. Trypsin radioimmunoassay

    International Nuclear Information System (INIS)

    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.)

  2. 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

  3. New adjuvant design using layered double hydroxide for production of polyclonal antibodies in radioimmunoassay techniques

    International Nuclear Information System (INIS)

    Various adjuvants have been used to enhance the immune response against specific antigens. So the objective of this work describes the immune stimulating activity of layered double hydroxide (LDH) particles incorporate with mineral oil as a new formulation of adjuvant as compared to known Freund's adjuvant for production of alpha-fetoprotein polyclonal antibody (anti-AFP) for estimation of alpha fetoprotein (AFP) in human serum by radioimmunoassay technique. In this concern, the study comprised two groups of white New Zealand rabbits, 2-2.5 kg body weight and each group comprised three rabbits. The first group vaccinated with AFP antigen emulsified with Freund's adjuvant and the second group vaccinated with AFP antigen emulsified with LDH formulation. The obtained data show that the highest displacement using LDH adjuvant reached (74.2, 61.7 and 66.5 %) while the corresponding values with Freund's adjuvant recorded (64.8, 60.3 and 54.6 %) which indicates that the use of LDH adjuvant as a cellular vehicle is a more suitable choice. Also, the preparation of AFP tracer using lactoperoxidase oxidation method and its purification using gel chromatography on PD-10 column were carried out. Different factors affecting the optimization of the assay process were studied. Validation testes of the assay were carried out. The reproducibility as measured by the intra- and inter- assay variations is satisfactory. The recovery and dilution testes indicated accurate calibration and appropriate matrix. The present technique agreed well with the currently used commercial kit (Siemens, IRMA kit). In conclusion, the liquid phase double antibody RIA technique proved to be sensitive, specific, precis and accurate for routine laboratory use. (author)

  4. Radioimmunoassay of thymosin alpha 1

    International Nuclear Information System (INIS)

    The present invention relates to an immunoassay for thymosin α1, 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 α1 to a conventional immunological carrier material. Thymosin α1 may be radioisotopically labelled with tritium, carbon 14 or iodine 125

  5. Correlação do nível de alfa-feto proteína, índice de sobrevida e recidiva tumoral em pacientes submetidosa transplante hepático Survival and tumor relapse rate according to alpha-fetoprotein level in patients submitted to liver transplantation

    Directory of Open Access Journals (Sweden)

    Elaine Cristina Ataide

    2011-03-01

    Full Text Available RACIONAL: O transplante hepático para carcinoma hepatocelular pode resultar em potencial cura e melhora da sobrevida comparado com operações conservadoras. OBJETIVO: Analisar os índices de recorrência e sobrevida em pacientes transplantados hepáticos por carcinoma hepatocelular e com níveis séricos de alfa-fetoproteína maiores que 200 ng/ml. MÉTODO: Foram analisados retrospectivamente 90 pacientes cirróticos com carcinoma hepatocelular submetidos à transplante hepático ortotópico entre 1997 e 2009. As lesões hepáticas foram diagnosticadas no pré-operatório por ultrassonografia com Doppler, tomografia computadorizada e níveis séricos de alfa fetoproteína. Os pacientes foram divididos em dois grupos de acordo com o nível de alfa-fetoproteína (menor ou maior que 200 ng/ml. O método de Kaplan-Meier foi usado para calcular a taxa de sobrevida. A análise de regressão Cox estudou os fatores preditivos de sobrevida. RESULTADOS: Pacientes com alfa-fetoproteína maior que 200 ng/ml (n=6 apresentaram menor taxa de sobrevida em um e cinco anos e na média de meses comparados com o grupo com alfa-fetoproteína menor que 200 ng/ml (n=84; respectivamente 35%, 18% e 11,8 meses contra 68%, 43% e 28,1 meses. Além disso, a taxa de recidiva foi 16,6% no primeiro grupo, e de 5,6% no outro. Observouse risco de óbito de 1% para cada 10 u de alfa-fetoproteína>200 ng/ml e para cada mm da maior medida de tumor acima de 28 mm. CONCLUSÃO: Os pacientes com valores séricos de alfa-fetoproteína maiores que 200 ng/ml demonstraram menores taxas de sobrevida, porém não foi preditivo de recidiva tumoral.BACKGROUND: Liver transplantation for hepatocellular carcinoma (HCC can result in a potential cure and greater survival than other less radical techniques. AIM: To analyze the survival and recurrence rate in liver transplant recipients with hepatocellular carcinoma and alpha-fetoprotein over 200 ng/ml. METHOD: Analysis, in this retrospective study

  6. 慢性乙型肝炎免疫清除期甲胎蛋白与HBV DNA清除的相关性研究%Correlation between serum alpha-fetoprotein level and HBV DNA clearance in immune clearance phase of patients with chronic hepatitis B

    Institute of Scientific and Technical Information of China (English)

    庞晓鹰; 徐洪涛; 杨秀珍; 咸建春; 沈美龙

    2014-01-01

    目的:探讨乙型肝炎免疫清除期,甲胎蛋白(AFP)与HBV DNA清除之间的相关性。方法收集处于免疫清除期发病2周内AFP升高5倍以上患者共58例,分为非抗病毒治疗组(下称观察组)31例和核苷(酸)类似物抗病毒治疗组(下称阳性对照组)27例,另取同期住院AFP阴性首次发病的慢性乙型肝炎核苷(酸)类似物抗病毒患者30例作为阴性对照组。分析影响HBV DNA清除率的相关因素。结果阳性对照组及阴性对照组患者均行抗病毒治疗,HBV DNA定量与各临床数据之间均无相关性,观察组HBV DNA定量与各指标之间的关系结果显示,AFP与HBV DNA清除具有显著相关性(r=0.8420,P=0.018),以下依次为ALT(r=0.7888,P=0.027)和总胆红素(TBil)(r=0.7816,P=0.032)。HBsAg(r=0.0480,P=0.413)和HBeAg(r=0.3356,P=0.191)与HBV DNA清除无显著相关性。结论对乙型肝炎免疫清除期AFP升高5倍以上患者,可先进行密切病情观察,根据病情需要进行抗病毒治疗。%Objectives To investigate the relationship between serum alpha-fetoprotein level and HBV DNA clearance in immune clearance phase of patients with chronic hepatitis B (CHB). Methods Total of 58 cases with CHB, who were in immune clearance phase and the serum alpha-fetoprotein level>5 upper limits of normal in two weeks since disease onset were enrolled. Among which 31 cases received non-antiviral therapy as experimental group, and 27 cases received antiviral therapy as positive control group. While other 30 cases with CHB who were in immune clearance phase and the serum alpha-fetoprotein level were normal in two weeks since disease onset, were enrolled as negative control group. And the correlation factors of HBV DNA clearance were analyzed, respectively. Results There were no signiifcant correlation between HBV DNA clearance and other data in patients who received antiviral therapy in positive and negative

  7. Clinical application of alpha-fetoprotein variant to screening benign and mlaignant liver dis-eases%甲胎蛋白异质体在鉴别良恶性肝病中的临床应用

    Institute of Scientific and Technical Information of China (English)

    陶绍能; 周建国; 程光华; 戴云海; 张新路; 阮昊

    2014-01-01

    目的:探讨小扁豆凝集素结合型甲胎蛋白异质体( AFP-L3)在良恶性肝病鉴别诊断中的临床价值。方法:应用装有耦联了小扁豆凝集素( LCA)的微量离心柱分离131例肝病患者的AFP-L3,用化学发光免疫检测血清AFP和AFP-L3含量,计算AFP-L3占AFP总量的百分比( AFP-L3%)。结果:与肝硬化( LC)和慢性肝炎( CH)组相比,肝癌( HCC)组中患者血清AFP、AFP-L3与AFP-L3%均升高,经Kruskal-Wallis 秩和检验有统计学意义(χ2=31.457,41.983,413.77;P=0.000,0.000,0.000)。AFP、AFP-L3和AFP-L3%相应ROC曲线下面积(AUC)分别为0.628、0.718和0.795,其差异均有统计学意义(P<0.05)。以AFP-L3%≥6.90%判断为阳性,AFP-L3%诊断肝细胞癌敏感性和特异性分别为66.7%和82.4%。结论:AFP-L3对肝细胞癌诊断准确度明显高于AFP,微量离心柱法检测AFP-L3在良恶性肝脏病变鉴别诊断中具有重要临床价值。%Objective:To verify the clinical value of applying the isoform of alpha -fetoprotein variant(AFP-L3) to screening benign and malignant liver diseases.Methods:AFP-L3 was isolated 131 patients with liver disease using the micro-spin column coupled with lens culinaris agglutinin ( LCA) .Chemi-luminescent immunoassay was performed to determine the content of serum AFP and AFP-L3,and AFP-L3% was calculated.Results:The serum levels of AFP and AFP-L3 as well as AFP-L3% were significantly higher in patients with hepatocellular carcinoma (HCC) compared to patients with liver cirrhosis (LC) and chronic hepatitis (CH),and Kruskal-Wallis one-way analysis of the variance showed statistical difference (χ2 =31.457,41.983,41.377;P=0.000,respectively).The area under the ROC curve (AUC) of AFP,AFP-L3 and AFP-L3 % was 0.628,0.718 and 0.795,respectively,and the differ-ences were significant (P<0.05).Sensitivity and specificity by AFP-L3 %(≥6.90% cut-off value) for diagnosis of HCC were66 .7

  8. 孕中期异常甲胎蛋白与不良妊娠结局的相关性%The association between second - trimester maternal serum abnormal alpha - fetoprotein and adverse pregnancy outcome

    Institute of Scientific and Technical Information of China (English)

    吴满武; 俞信忠; 杨志浩; 张涛; 罗婷婷

    2012-01-01

    目的 探讨孕中期甲胎蛋白(AFP)与不良妊娠结局的相关性,为妊娠管理提供建议.方法 对孕中期14 008例孕妇进行唐氏筛查.研究组:①AFP MoM>2.0孕妇338例.②AFP MoM<0.5孕妇224例.依据AFP MoM值不同分组.对照组:AFP MoM值都在正常范围内的孕妇318例.回顾分析妊娠结局资料.结果 ①AFP MoM>2.0的孕妇发生出生缺陷(ρ<0.01)的机会明显比正常孕妇高.②AFP MoM> 3.5的孕妇发生出生缺陷(P<0.001)和早产(P<0.05)的机会明显比正常孕妇高.③AFP MoM> 5.0的孕妇发生出生缺陷(P<0.001)、早产(P<0.01)和胎儿窘迫(P<0.05)的机会明显比正常孕妇高.④AFP MoM<0.5和<0.25的孕妇均未发现与不良妊娠结局相关.结论 异常水平的标志物除了能够提供开放性神经管缺损风险外,还能提供不良妊娠结局的风险信息,而这些信息对产前遗传咨询和妊娠管理是非常重要.%Objective; Aim of this study is to determine the association between risk of adverse pregnancy outcome and abnormal maternal serum alpha - fetoprotein in second trimester screening. Abnormal levels of maternal serum markers provide information which is important for counseling and pregnancy management. Methods: A total of 14 008 pregnant women underwent maternal serum Down' s syndrome screening. Study groups include:①338 cases were found to have a serum AFP of > 2.0 multiples of the median ( MoM). ②cases serum were found to have a serum AFP 2.0 MoM have higher occurrence of birth defects (P 3. 5 MoM have higher occurrence of birth defects ( P 5.0 MoM have higher occurrence of birth defects (P < 0. 001) and preterm labor ( P < 0. 01) and fetal stress ( P < 0. 05). ④Pregnant women with a serum AFP of < 0. 5 M < 0. 25 MoM were found to have no association with adverse outcomes. Conclusion: Abnormal levels of maternal serum markers can provide risk information of adverse outcomes in addition to the risks for open neural tube defects

  9. Radioimmunoassays in prenatal genetic diagnosis

    International Nuclear Information System (INIS)

    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)

  10. Diagnosis value of alpha-fetoprotein heterogeneity in primary hepatic carcinoma%甲胎蛋白异质体在原发性肝癌中的诊断价值

    Institute of Scientific and Technical Information of China (English)

    陈俊; 王利健; 骆剑明; 吴荣辉; 楼跃民; 朱笑频

    2012-01-01

    目的:探讨甲胎蛋白异质体在肝脏良恶性疾病诊断中的意义.方法:采用微量离心柱法对240例肝病患者(原发性肝癌70例,肝癌术后10例,肝硬化63例,慢性肝炎97例),采用化学发光法检测AFP及AFP-L3水平,计算AFP-L3和AFP比例.结果:原发性肝癌组患者血清中的AFP,AFP-L3%阳性率显著高于肝癌术后组,肝硬化组和慢性肝炎组.AFP-L3%的灵敏度为86.96%,特异性为88.30%,Youden指数为0.75.AFP的灵敏度为75.68%,特异性为85.54%,Youden指数为0.61.从Youden指数可以知道AFP-L3%是比AFP更好的诊断肝癌的指标.原发性肝癌组患者血清中的AFP、AFP-L3%含量显著高于肝癌术后组、肝硬化组和慢性肝炎组(F值分别为74.45、54.85,P值均<0.01).诊断为原发性肝癌病人中,当AFP> 400 ng/ml时,AFP-L3%的阳性率为95.23%.AFP-L3%与AFP水平无相关性.结论:AFP-L3%对于肝癌的诊断有较高的灵敏性和特异性;是比AFP更好的诊断指标;可以作为诊断肝癌的独立指标.%Objective: To explore the significance of alpha - fetoprotein heterogeneity (AFP - L3) in differential diagnosis of malignant liver disease. Methods: Serum samples were collected from 240 patients, 70 cases with hep-atocellular carcinoma, 10 cases with postoperative liver cancer, 63 cases with cirrhosis, 97 cases with chronic hepatitis. AFP - L3 was isolated by microspincolumn with lens culinaris agglutinin. The levels of AFP and AFP - L3 were detected by chemiluminescence method. Results: The ratios of AFP and AFP - 13% in hepatocellular carcinoma group were significantly higher those in hepatectomy group, liver cirrhosis and chronic hepatitis group (χ2 values were 87. 30, 142.22, P <0. 01 ). For AFP - L3% , sensitivity was 86. 96% , specificity was 88. 30% , and the Youden index was 0. 75, while for AFP, sensitivity was 75. 68% , specificity was 85. 54% , Youden index was 0. 61. From the Youden index, it can be seen the AFP - L3

  11. 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.

  12. Radioimmunoassay for methotrexate

    International Nuclear Information System (INIS)

    Rabbits and a goat immunized with a methotrexate-hemocyanin conjugate produced antibodies that bind [3H]methotrexate. The goat antibodies were used for the development of a radioimmunoassay for methotrexate. The radioimmunoassay is specific: 105 to 106 times more folic acid, tetrahydrofolic acid, or folinic acid are required to compete effectively with methotrexate for the antibodies to methotrexate. As little as 100 picograms of methotrexate can be detected in the assay. The radioimmunoassay was used to follow the disappearance of methotrexate in the plasma of two patients with malignancies following its intravenous administration. (auth)

  13. Radioimmunoassay for methaqualone

    International Nuclear Information System (INIS)

    A sensitive radioimmunoassay for methaqualone and its major metabolites is described. Included within this disclosure is the preparation of novel methaqualone haptens, the preparation of antigens from such haptens by coupling to an immunogenic carrier material, the use of such antigens to elicit antibodies selective to methaqualone and its major metabolites and an improved radiolabelled derivative of a methaqualone analog used in the practice of said radioimmunoassay. 4 claims

  14. The Value of Combined Serum Alpha-fetoprotein Heteroplasmon-L3 Concentration and AFP-L3/Alpha-fetoprotein Ratio Detection in the Early Diagnosis of Primary Liver Cancer%联合检测血清甲胎蛋白异质体浓度与甲胎蛋白异质体百分含量在原发性肝癌诊断中的价值

    Institute of Scientific and Technical Information of China (English)

    黄彩云; 韩素桂; 贾红莲

    2013-01-01

    Objective To explore the value of combined serum alpha-fetoprotein inform (AFP-L3) and AFP-L3/AFP as biomarkers in the early diagnosis of hepatocellular carcinoma (HCC).Methods A total of 137 patients from the People's Hospital of Tangshan of Hebei Union University were included in our study.They were treated in hospital from March to November 201 1.All the patients were at the age of 28 to 77 years old,and 98 of them were men,and 39 were women.According to the pathological diagnosis,patients were divided into two groups.HCC group had 92 patients,and benign liver disease group had 45 patients including 37 with liver cirrhosis and 8 with chronic hepatitis.ELISA method was used to test the serum AFP-L3 level.At the same time,microcentrifugal column method was adopted to separate AFP-L3 from the patients' serum,and electrochemical luminescence method was used to detect the separated AFP-L3 and total AFP levels.Then,we calculated the ratio of AFP-L3/AFP.We also calculated the statistical indexes of AFP-L3 level and AFP-L3/AFP ratio in the diagnosis of HCC,such as sensitivity,specificity and Youden index.The above indexes of combined AFP-L3 level and AFP-L3/AFP ratio were also detected to evaluated its diagnostic value.Results HCC group AFP-L3 level (109.04 ± 62.51) ng/mL was significantly higher than that of the benign liver disease group [(25.96 ± 49.43) ng/mL; t=8.28,P < 0.001].HCC group serum AFP-L3/AFP ratio (17.35% ± 14.48%) was significantly higher than that of the benign liver disease group (5.62% ± 6.38%,t=6.545,P < 0.000 1).With serum AFP-L3 level > 38.0 ng/mL as a critical value of primary liver cancer diagnosis,the sensitivity was 83.69%,and the specificity was 88.88%.For AFP-L3/AFP ratio > 10% as a critical value in primary liver cancer diagnosis,the sensitivity was 83.69%,and the specificity was 95.55%.When serum AFP-L3 level > 38.0 ng/mL and AFP-L3/AFP ratio > 7.5% were combined in primary liver cancer diagnosis

  15. Alpha-fetoprotein triggers hepatoma cells escaping from immune surveillance through altering the expression of Fas/FasL and tumor necrosis factor related apoptosis-inducing ligand and its receptor of lymphocytes and liver cancer cells

    Institute of Scientific and Technical Information of China (English)

    Meng-Sen Li; Qiu-Ling Ma; Qian Chen; Xin-Hua Liu; Ping-Feng Li; Guo-Guang Du; Gang Li

    2005-01-01

    AIM: To investigate the mechanism of α-fetoprotein (AFP)in escaping from the host immune surveillance of hepatocellular carcinoma.METHODS: AFP purified from human umbilical blood was administrated into the cultured human lymphoma Jurkat T cell line or hepatoma cell line, Bel7402 in vitro. The expression of tumor necrosis factor related apoptosisinducing ligand (TRAIL) and its receptor (TRAILR) mRNA were analyzed by Northern blot and Western blot wasused to detect the expression of Fas and Fas ligand (FasL)protein.RESULTS: AFP (20 mg/L) could promote the expression of FasL and TRAIL, and inhibit the expression of Fas and TRAILR of Bel7402 cells. For Jurkat cell line, AFP could suppress the expression of FasL and TRAIL, and stimulate the expression of Fas and TRAILR. AFP also could synergize with Bel7402 cells to inhibit the expression of FasL protein and TRAIL mRNA in Jurkat cells. The monoclonal antibody against AFP (anti-AFP) could abolish these functions of AFP.CONCLUSION: AFP is able to promote the expression of FasL and TRAIL in hepatoma cells and enhance the expression of Fas and TRAILR in lymphocytes. These could elicit the escape of hepatocellular carcinoma cells from the host's lymphocytes immune surveillance.

  16. Methadone radioimmunoassay: two simple methods

    Energy Technology Data Exchange (ETDEWEB)

    Robinson, K.; Smith, R.N. (Metropolitan Police Forensic Science Laboratory, London (UK))

    1983-09-01

    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-/sup 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 ..cap alpha..-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/sup -1/ for blood, and 24 and 21 ng ml/sup -1/ for urine. The assay using the radioiodinated conjugate can be made more sensitive if required by increasing the specific activity of the label.

  17. Automated radioimmunoassay of nicotine

    International Nuclear Information System (INIS)

    The authors have developed an automated nonequilibrium procedure for the radioimmunoassay of nicotine. The use of a unique iodinated nicotine derivative in this procedure gave a sensitivity of 10 μg/l for nicotine with a between-run precision of 7.4% and within-run precision of 6.0%. Nicotine levels of 60 to 67 μg/l were found in subjects 15 min after smoking one standard cigarette. The technique herein reported is a very rapid, and sensitive radioimmunoassay for nicotine and facilitates the determination of nicotine in smoking subjects during the actual process of smoking. (Auth.)

  18. Radioimmunoassay the oxytocin

    International Nuclear Information System (INIS)

    The radioimmunoassay is based on a reaction antigen-antibody, whose kinetics are expressible in mathematical terms. The specificity and sensibility of the method depend fundamentally on the specificity and the likeness of the antibody. For these reasons el RIA turns out to be a simple, quick and sensitive rehearsal, specific with potential applicability to a great number of substances

  19. Radioimmunoassay of pancreatic glucagon

    International Nuclear Information System (INIS)

    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.)

  20. Radioimmunoassay of enkephalins

    International Nuclear Information System (INIS)

    Using iodine (125I) tracers of high specific radioactivity, the authors have developed the specific radioimmunoassay of methionine-enkephalin (Met-Enk) and leucine-enkephalin (Leu-Enk). The immunosera are obtained by immunizing rabbits with Met-Enk or Leu-Enk combined with ovalbumin by means of carbodiimide. The specificity and the sensitivity of these radioimmunoassays (IC50 of 0.57 nM and 0.55 nM for the assay of Met-Enk and Leu-Enk, respectively) afford a means of estimating the enkephalin levels in extracts of various regions of the rat brain. The highest levels are in the striatum and the hypothalamus and the lowest in the cerebellum and the hippocampus. (author)

  1. Radioimmunoassay of casein

    International Nuclear Information System (INIS)

    The heterogeneity of casein and its consequence on the radioimmunoassay are discussed. All protein fractions of human milk are chequed for cross reactivity. Casein K and II are labelled with 125I. The elution curves of these labelled compounds are measured by radioactivity and optical density. The sera of lactating women and of cancerous patients were chromatographed. In human milk a major peak is found in the elution region of casein. The serum of cancerous patients showed a heterogenous elution pattern

  2. Comparative study of radioimmunoassay dates

    International Nuclear Information System (INIS)

    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)

  3. Radioimmunoassay of phenytoin

    International Nuclear Information System (INIS)

    A radioimmunoassay procedure is described for the measurement of phenytoin (5,5-diphenylhydantoin) in serum samples. Antiserum to phenytoin was produced against phenytoin - valeric acid - bovine serum albumin conjugate. 125I labelled phenytoin - acetic acid - tyrosine methyl ester was used as a tracer. The assay covers a range of 10-500 ng/cm3 and has a sensitivity of 0.25 ng. The assay is validated by specificity tests, precision profile and recovery tests. (author) 8 refs.; 6 figs.; 2 tabs

  4. Radioimmunoassay of platelet proteins

    International Nuclear Information System (INIS)

    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

  5. Radioimmunoassay of serum ferritin

    International Nuclear Information System (INIS)

    Purified human spleen ferritin was labelled with 125I. On Sepharose 6-B gel filtration four species of labelled products were separated: a component with a higher molecular weight than ferritin; a component which is eluted in the same volume as unlabelled ferritin; and two labelled compounds with molecular weights lower than ferritin. When these labelled materials were used in a double antibody radioimmunoassay, the higher molecular weight fraction showed variable and high non-specific binding and was poorly displaced by unlabelled ferritin; the fraction behaving like true ferritin gave good standard curves and showed non-specific binding of less than 1%. The remaining two components showed poor binding to rabbit antiferritin. Using labelled material from the second fraction, a double antibody radioimmunoassay capable of measuring 2μg ferritin protein/litre of serum was developed. Inter-and intra-assay variation was between 3% and 8% over a concentration range of 0 to 250 μg ferritin protein/litre. Good agreement between serum ferritin levels assayed by the present method and by an immunoradiometric method was obtained. Labelled ferritin was stable for at least six weeks. The simplicity of the methodology makes it possible to assay serum ferritin in large batches. (author)

  6. Radioimmunoassay of steroid hormone

    International Nuclear Information System (INIS)

    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 3H. 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.)

  7. Retrospective study of the diagnostic value of quantitative alpha-fetoprotein in infants with terato-mas%婴儿畸胎瘤甲胎蛋白量化检测的临床意义探讨

    Institute of Scientific and Technical Information of China (English)

    何小庆; 王珊; 刘苗; 阮味维

    2014-01-01

    目的:探讨婴儿期畸胎瘤血清甲胎蛋白(alpha-fetoprotein,AFP)定量检测值与同月龄儿正常参考值上限的比值对成熟型、未成熟型及恶性畸胎瘤的辅助鉴别意义。方法回顾性分析1991年10月至2010年5月我们收治的166例畸胎瘤患儿临床资料。化学发光法定量检测血清AFP含量,将患儿检测值与正常同月龄儿参考值上限的比值作为血清AFP比值,结合成熟型、未成熟型及恶性畸胎瘤的病理诊断,分析血清AFP比值在鉴别3种不同类型畸胎瘤中的规律及其临床意义。结果①131例成熟型畸胎瘤血清AFP比值最大16.34(绝对值670 ng/mL);13例未成熟型畸胎瘤血清AFP比值差异明显,7例Ⅰ级未成熟型畸胎瘤,最小AFP比值1.25,最大39.27;5例Ⅱ级未成熟型畸胎瘤,最小AFP比值1.23,最大139.51,其中2例比值大于83.17;1例Ⅲ级未成熟型畸胎瘤,AFP超出检测值上限;22例恶性畸胎瘤最小AFP比值83.17(绝对值499 ng/mL),13例超出检测上限;②未成熟型畸胎瘤血清AFP阳性率(12例,92.31%)和恶性畸胎瘤血清AFP阳性率(22例,100%)均高于成熟型畸胎瘤(58例,44.28%)(P<0.05),未成熟型畸胎瘤与恶性畸胎瘤血清AFP阳性率无明显差异(P>0.05);<9月龄婴儿血清AFP阳性率(33例,84.62%)高于9月龄婴儿(59例,46.46%)(P<0.05);性腺内畸胎瘤血清AFP阳性率(20例,76.92%)高于性腺外(72例,51.43%)(P<0.05)。结论婴儿期畸胎瘤AFP阳性率受年龄、发病部位及肿瘤病理类型等因素的影响。血清AFP比值在不同类型畸胎瘤有相应变化规律,可用于术前辅助判断畸胎瘤良恶性及制定手术方案,对动态随访治疗效果有重要意义,结合病理检查对于术后是否化疗有决定作用。%Objetive To explored the differential diagnostic value of quantitative Serum alpha

  8. 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

  9. 血清甲胎蛋白异质体3测定在原发性肝癌早期筛查中作用%Value of serum alpha-fetoprotein detection in early screening of primary liver cancer

    Institute of Scientific and Technical Information of China (English)

    周云龙; 廖志军; 邓国孙; 丁罡

    2014-01-01

    Objective To explore the value of serum alpha-fetoprotein (AFP-L3) in early screening of primary liver cancer. Methods Lentil lectin coupled with ( LCA) micro-centrifuge column was separated to obtain AFP-L3 from 116 cases of chronic liver disease and primary liver cancer .AFP before separation and AFP-L3 after separation were detected with chemoluminescence , and their levels were then calculated .Results Statistical differences could be seen in the average value of AFP-L3 between the patients with pri-mary liver cancer and patients with chronic liver disease (P<0.01).Conclusion The level detected with micro-centrifuge column was of importance in early screening of primary liver cancer .%目的:探讨血清甲胎蛋白异质体3(AFP-L3)在原发性肝癌早期筛查中的作用。方法以装有耦联小扁豆凝集素(LCA)的微量离心柱分离慢性肝病、原发性肝癌共116例患者AFP-L3,用化学发光方法检测分离前后AFP-L3,计算AFP-L3在AFP中的含量比例。结果原发性肝癌组AFP-L3含量比例均值[(36.89±2.13)%],与慢性肝病组[(7.45±2.32)%]相比差异有统计学意义( P<0.01)。结论微量离心柱法测定AFP-L3的含量在原发性肝癌的早期筛查中具有重要意义。

  10. Digoxin radioimmunoassay kit

    International Nuclear Information System (INIS)

    The study was made with an objective of research in methodology on self-production of digoxin radioimmunoassay kit. The development had been done successfully in the second antibody liquid phase technique with 10 times cheaper in price. The assay showed good precision and reproducibility with 93.6% of the recovery result. Additionally, the anti-digoxin serum composed in this kit was very highly specific to digoxin i.e. only 0.7% cross-reactivity to digitoxin. The stability study showed that the shelf-life of the kit was up to 2 months at 40C storage. Therefore, the kit appeared suitable for routine clinical diagnosis of serum digoxin

  11. A radioimmunoassay for chicken avidin

    International Nuclear Information System (INIS)

    A double-antibody solid-phase radioimmunoassay for chicken avidin is reported. Avidin was labelled with 125I 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 [14C]biotin/bentonite method gave poor recoveries for avidin in the liver and kidney. Radioimmunoassay and the [14C]biotin/bentonite method gave similar concentrations for oviduct avidin. (author)

  12. Prognostic value of the content of lentil lectin-rcactive alpha-fetoprotein-L3 in early-stage hepatocellular carcinoma%甲胎蛋白异质体3含量对早期肝癌的预后价值

    Institute of Scientific and Technical Information of China (English)

    冯凯; 马宽生; 高峻; 刘念洲; 王曙光

    2008-01-01

    目的 探讨甲胎蛋白异质体3(lentil lectin-reactive alpha-fetoprotein-L3,AFP-L3)的含量对早期肝癌的预后价值.方法 97例早期肝癌患者根据术前AFP和AFP-13的含量分为:(1)AFP阳性、AFP-L3阴性组(29例):AFP>20 μg/L & AFP-13<15%;(2)AFP-L3、AFP均低含量组(16例):15%≤AFP-L3≤50% & 20 μg/L≤AFP≤200 μg/L;(3)AFP-L3、AFP均高含量组(13例):AFP-L3>50% & AFP>200 μg/L;(4)AFP-L3高含量、AFP低含量组(24例):AFP-13>50%& 20 μg/L≤AFP≤200 μg/L;(5)AFP-L3低含量、AFP高含量组(15例):15%≤AFP-L3≤50% & AFP>200 μg/L.对各组患者的肿瘤分化程度、术后1、2、3年生存率和无瘤生存率进行分析.结果 AFP-13阴性患者的肿瘤分化程度、术后3年生存率和无瘤牛存率明显优于AFP-L3阳性患者(χ2=21.051,10.043,4.450,6.977,25.566,P<0.05).AFP-L3高含量组患者的肿瘤分化程度、术后1、2、3年生存率和无瘤生存率明显低于低含量组(χ2=7.938,3.488,9.085,P<0.05).结论 AFP-L3含量的增高提示肿瘤恶性程度高,预后不良,尤其是AFP水平低时.手术前后检测AFP-L3含量对于患者预后的评价具有指导意义.%Objective To explore the prognostic value of the content of lentil lectin-reactive alphafetoprotein-L3(AFP-L3)in early-stage hepatocellular carcinoma(HCC).Methods According to the content of alpha.fetoprotein(AFP)and AFP-L3 before the treatment,97 patients with early-stage HCC were divided into group A(AFP>20 μg/L & AFP-L3<15%,n=29),group B(15%≤AFP-13≤50% & 20 μg/L≤AFP≤200 μg/L,n=16),group C(AFP-13>50%& AFP>200 μg/L,n=13),group D(AFP-L3>50% & 20 μg/L≤AFP≤200 μg/L,n=24)and group E(15%≤AFP-L3≤50%& AFP>200 μg/L,n=15).The degree of tumor differentiation,the 1-,2-,3-year survival rates and tumor-free survival rates of the patients were analyzed.Results The degree of tumor differentiation.3-year survival rate and tumor-free survival rate of patients in group A were significantly higher than

  13. Diagnostic value of alpha-fetoprotein for hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Hepatocellular carcinoma (HCC) variously occupies the fifth or sixth position as the most frequent neoplasia worldwide. The present work used alphafetoprotein (AFP) determinations on the ultra-micro analytical system (SUMA) as a tumoral marker in 189 cirrhotic patients evaluated at the Center for Medical and Surgical Research between January 1999 and September 2005. The principal factors associated to increases in AFP were HCC and viral cirrhosis. In all, 22 patients (11.64%) suffered from HCC, with viral cirrhosis caused mainly by hepatitis C virus infections as the most important etiological factor. AFP as a tumoral marker displayed a sensitivity of 68.18% and a specificity of 92.17%, which increased to 86.36 and 100% respectively when combined with abdominal sonography. It is concluded that AFP is valuable for the diagnosis of HCC

  14. Radioimmunoassay for serum ferritin

    International Nuclear Information System (INIS)

    We describe the development and evaluation of a serum ferritin radioimmunoassay, in which 125I-labeled ferritin and rabbit anti-ferritin antibody are used. Goat anti-rabbit gamma-globulin antibody, together with polyethylene glycol, is used as the separating reagent. The assay has a working range up to 500 μg of ferritin per litre, and a sample requirement of 75 μl of serum for assay at two dilutions. The assay requires 24 h. It has a sensitivity of 1.5 μg of ferritin per litre and a long-term precision (CV) of 13%. Reference intervals for a population of men were 18 to 330 μg/litre, with no marked age dependence, while those for a population of women older than 50 years were 18 to 200 μg/litre. Many apparently healthy women in the 20 to 50 year age group have much lower concentrations. Serum ferritin concentrations of <18 μg/litre are indicative of iron deficiency, defined as the absence of stainable iron in an aspirate of bone marrow

  15. Salivary phenytoin radioimmunoassay

    International Nuclear Information System (INIS)

    A simple, specific and rapid radioimmunoassay method for the assessment of non-protein bound ('free') phenytoin concentrations in mixed saliva using 125I-labelled phenytoin is described. Epileptic patients on maintenance phenytoin therapy have mixed saliva phenytoin concentrations similar to 'free' drug levels measured directly in serum or cerebrospinal fluid. Salivary phenytoin levels are approximately 10% of the total serum level in treated epileptic patients and in normal subjects after ingestion of a single oral dose. The half time of disappearance of phenytoin after 100 or 300 mg doses is 12.2 +- 3.0 h in serum and 12. 3 +- 3.2 h in saliva. This method of assessing the biologically active fraction of the drug may be particularly valuable in situations where serum protein binding is abnormal or in drug interactions. It is also non-invasive and requires small sample volumes (20μl) and may therefore be valuable in paediatric practice and in pharmacokinetic studies in which multiple venepunctures would otherwise be required. (orig./MG)

  16. Radioimmunoassay in the detection of insulin secretion

    International Nuclear Information System (INIS)

    Some antihypertensive drugs have been shown to cause clinically significant alteration in the endocrine function. This study was conducted to investigate the effect of a new antihypertensive drug, rilmenidine on insulin secretion, which is an important determinant for glucose metabolism. In-vitro method was used to study the direct effect of rilmenidine on glucose induced insulin secretion using isolated rat pancreas. Insulin was assayed using radioimmunoassay. Concentrations of rilmenidine used were based on the peak plasma concentration achieved with an oral standard dose of 1 mg. This study showed that rilmenidine at low concentration was able to stimulate insulin secretion whereas at higher concentration inhibited the insulin secretion. This probably was due to its effect on the imidazoline receptor and the alpha2 adrenoceptor known to induce and inhibit insulin secretion respectively. (Author)

  17. 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.

  18. Calcitonin radioimmunoassay: clinical application

    International Nuclear Information System (INIS)

    A radioimmunoassay for human Calcitonin (hCT) was established: antisera were produced by immunizing goats with synthetic hCT; 7.5 μg hCT were labelled with 1 mCi 125J; hCT of different quantities in the range between 0.1 to 20 ng/ml served as standard. Separation of free from antibody bound tracer was done using the charcoal procedure. - This RIA-system was sensitive to determine 0.1 ng/ml; the normal range lying below 0.5 ng/ml. This assay was used to study the following clinical problems: 1) in 31 patients, with a thyroid tumor, diagnosis of calcitonin producing medullary thyroid carcinoma was proven. Serum calcitonin of these patients were lying between 1.7 and 120 ng/ml. Clinical signs of this disease are nonspecific, so CT determination is of importance for early diagnosis and control of therapy. In patients with a high tumor risk pentapastrin stimulation of the C-cells reveals calcitonin secretion above normal, if a medullary thyroid carcinoma is present. 2) two patients with pheochromocytoma showed elevated levels of CT before operation; after removal of the tumor serum CT was normalized. Extracts of the adreno-medullary tumor revealed immunoreactive CT corresponding to 4 and 1 ng/ml wet weight. - 3) CT is used for therapy of Paget's disease of the bone, so control of antibody development in the patients is necessary. In 25 patients with Paget's disease no antibody production against the injected hormone was evident. (orig.)

  19. Calcitonin radioimmunoassay. Clinical application

    International Nuclear Information System (INIS)

    A radioimmunoassay for human calcitonin (hCT) was established. Antisera were produced by immunizing goats with synthetic hCT; 7.5 μg of hCT was labelled with 1 mCi of 125I; hCT served as a standard in the range between 0.1 to 20 ng/ml. Separation of free from antibody-bound tracer was done with a charcoal procedure. The RIA system had a sensitivity of 0.1 ng/ml, the normal range lying below 0.5 ng/ml. The assay was used to study the following clinical problems: (1) In 35 patients with a thyroid tumour, diagnosis of calconin-producing medullary thyroid carcinoma was proven. Serum CT of these patients lay between 1.7 and 120 ng/ml. Clinical signs of this disease are non-specific, so CT determination is important for early diagnosis and control of therapy. In patients with a high tumour risk, pentagastrin stimulation of the C-cells reveals CT secretion above normal if a medullary thyroid carcinoma is present. (2) Two patients with pheochromocytoma showed elevated levels of hCT before operation; after removal of the tumour, the serum CT levels became normal. Extracts of the adrenomedullary tumour revealed immunoreactive CT corresponding to 1 and 4 ng/mg wet weight. (3) hCT is used in the therapy of Paget's disease of the bone, so it is necessary to check the production of antibodies in the patient. In 25 patients with Paget's disease no antibody production against the injected hormone was evident. (author)

  20. Diagnostic Value of Des-gamma Carboxyprothrombin(DCP), Lectin-bound AFP(AFP-L3) and Alpha-fetoprotein (AFP) Expression in Primary Hepatocellular Carcinoma%DCP、AFP-L3和AFP在原发性肝细胞癌诊断中的价值

    Institute of Scientific and Technical Information of China (English)

    付水; 齐娟飞; 朱海燕; 祁志荣; 陈红艳

    2013-01-01

    Objective To observe the alone and joint diagnostic value of des-gamma carboxyprothrombin (DCP) , lectin-bound AFP(AFP-13) and alpha-fetoprotein(AFP) in primary hepatocellular carcinoma, and provide a novel method for diagnosis for PHC and screening for high risk population. Methods The levels of serum DCP,AFP-L3 and AFP were measured in 53 patients with hepa-tocellular carcinoma, 51 cases of cirrhosis, 60 cases of chronic hepatitis, 52 normal healthy subjects established as control group, and analyse the result statistically. Results The levels of serum DCP,AFP-L3 and AFP were significantly higher in the hepatocellular carcinoma than those in other groups. The diagnostic sensitivities of DCP,AFP-I3 and AFP to hepatocellular carcinoma were 77.36% , 69.81% and 60.38% respectively, the specificity were 89.57% , 88.34% and 77.91% , individually. The joint detection could improve sensitivity up to 94.34%. Conclusion DCP was a high sensitivity and specificity marker for diagnosis of PHC. The combined assay of serum tumor markers possess more value in the diagnosis of PHC, especially in AFP negative patients.%目的:探讨血清中脱γ一羧基凝血酶原(DCP)、甲胎蛋白异质体(AFP-L3)和甲胎蛋白(AFP)对原发性肝细胞癌(PHC)单独和联合诊断的意义,为PHC诊治提供一种新方法,以及对高危人群作筛选.方法:采集 53 例PHC、51 例肝硬化、60 例慢性肝炎和 52 例健康对照者的血清分别检测 DCP、AFP-L3 和 AFP,并对其统计分析.结果:PHC患者的 DCP、AFP-L3 和 AFP 均显著高于肝硬化、慢性肝炎和健康对照者,DCP、AFP-L3 和 AFP在PHC组中的灵敏度分别为 77.36 %、69.8 1% 和60.38 %,特异性分别为89.57 %、88.34 % 和 77.91 %.三者联检时,其诊断的敏感度可提高至 94.34 %.结论:DCP 对PHC诊断具有较好的敏感度和特异性,联检DCP、AFP-L3和AFP可有效提高PHC尤其是AFP阴性PHC的诊断效率,对PHC的早期诊治具有一定指导意义.

  1. Value of early alpha fetoprotein detection in predicting prognosis of patients with drug-induced liver injury%早期甲胎蛋白测定对药物性肝损害患者转归的预测价值

    Institute of Scientific and Technical Information of China (English)

    陈琰; 汤紫荣; 曹艳菊; 屈昌民; 梁淑文; 房慧; 胡瑾华

    2015-01-01

    Objective To investigate the clinical significance of early alpha fetoprotein (AFP) detection in predicting prognosis of patients with drug-induced liver injury (DILI).Methods Clinical data of 160 DILI patients who were admitted to 302th Hospital of Chinese People's Liberation Army from July 2011 to July 2014 were retrospectively analyzed.According to the levels of AFP within 1 week after admittion,the patients were divided into high AFP group (AFP≥20 μg/L,80 cases) and normal group (AFP <20 μg/L,80 cases).The AFP and liver function were measured.Results There were no statistical differences in sex,average age,alanine aminotransferase(ALT) and alkaline phosphatase (ALP) between high AFP group and normal group (P > 0.05).However,high AFP group showed higher level of AFP,total bilirubin and total hospitalization days compared with those in normal group[82(45,153) ng/L vs 9(3,20) ng/L,(173 ±25) μmol/L vs (70 ± 15) μmol/L,(22.3 ± 1.8) d vs (11.6 ± 1.2) d] (P < 0.01).In high AFP group the incidence of severe case was 22.5 % (18/80),statistically higher than that in normal group [1.3 % (1/80)] (P < 0.01).Conclusions High level of AFP in early stage indicates severe inflammation and poor prognosis,which may be used as an important marker for assessing the liver injury and predicting the prognosis in patients with DILI.%目的 探讨早期甲胎蛋白水平测定对药物性肝损害(DILI)转归的预测价值.方法 纳入2011年7月至2014年7月解放军第三○二医院诊断为DILI的住院患者160例,根据入院后1周内初次甲胎蛋白检测值将其分为甲胎蛋白增高组(甲胎蛋白≥20 μg/L)和甲胎蛋白正常组(甲胎蛋白<20μg/L),每组80例.对2组患者进行生存预后分析.结果 甲胎蛋白增高组和甲胎蛋白正常组年龄、性别、丙氨酸转氨酶、碱性磷酸酶比较差异无统计学意义(P>0.05);甲胎蛋白、总胆红素及住院时间比较,差异有统计学意义[82(45,153) ng/L比9

  2. Radioimmunoassay for rhesus monkey gonadotropins

    International Nuclear Information System (INIS)

    Heterologous double-antibody radioimmunoassay methods are described for the measurement of circulating levels of rhesus monkey (Macaca mulatta) FSH and LH; the latter assay is also applicable to rhesus chorionic gonadotropin (CG) estimations. The FSH assay utilizes purified rat FSH for trace, either of two anti-human FSH antisera and a semipurified rhesus pituitary standard. The LH assay utilizes purified ovine LH for trace, an anti-human CG antiserum and the same rhesus pituitary standard. The use of these systems obviates the necessity of purifying rhesus gonadotropins which are required for the development of homologous radioimmunoassay systems. (U.S.)

  3. Recent advances in steroid radioimmunoassay

    International Nuclear Information System (INIS)

    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.)

  4. Homologous radioimmunoassay of human prolactin

    International Nuclear Information System (INIS)

    Gelfiltration on Sephadex G-75 showed a heterogenity of prolactin in serum of patients with prolactinoma and in culture medium of a prolactinoma. Serum of patients with prolactinoma and culture medium of a prolactinoma were examined as possible sources of prolactin by gel filtration and ion exchange chromatography. Polyacrylamide electrophoresis revealed both preparations as contaminated by other proteins. Nevertheless prolactin isolated form culture medium of a prolactinoma is good enough as a tracer in our radioimmunoassay because contaminating proteins in this preparation do not inferfere in our system. An hPRL antiserum created in a rabbit against a crude fraction of human serum of a patient with prolactinoma was tested by titration, saturation studies, and ion exchange chromatography. In comparison with lactoperoxidase-iodinated prolactin Chloramine T iodinated prolactin showed higher loss of immunochemical properity, however higher specific activity. Specifity and precision in our radioimmunoassay system were described and the conditions of optimal sensitivity in our assay were evaluated. (orig.)

  5. A sensitive radioimmunoassay for fentanyl

    International Nuclear Information System (INIS)

    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.)

  6. Radioimmunoassay of human plasma trypsin

    International Nuclear Information System (INIS)

    A radioimmunoassay has been developed for the determination of human trypsin in plasma. It allows the measurement of trypsin concentration in spite of the presence of plasma or pancreatic inhibitors. The human trypsin used as a standard as well as for labelling was isolated from pancreatic tissue and purified by affinity chromatography. The antiserum was obtained from guinea-pigs immunized with partially purified human trypsin. In the radioimmunoassay, the values of trypsin in serial dilutions of plasma were parallel to those of the standard curves. The assay was shown to be reproducible, sensitive and specific. However, the two antisera used did not distinguish between the enzyme and its proenzyme. In normal subjects, plasma values were found to be around 400 ng/ml. They were 10-40 times higher in patients with acute pancreatitis. The method appears to be much more specific for the diagnosis of acute pancreatitis than the current determinations of amylase and lipase activity

  7. Radioimmunoassays of papain in beer

    International Nuclear Information System (INIS)

    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)

  8. Radioimmunoassay for steroid hormones, 3

    International Nuclear Information System (INIS)

    A specific, sensitive and reliable radioimmunoassay for plasma dehydroepiandrosterone (DHEA) has been developed. Anti-DHEA serum was obtained by immunizing rabbits with a DHEA-3-hemisuccinate-BSA conjugate. A useful range in the standard curve was from 10 pg to 500 pg. DHEA was separated from cross-reacting steroids by microcolumn chromatography. The coefficients of variation for within-assay and between-assay are 6.3% and 7.7%, respectively. (auth.)

  9. 甲胎蛋白对耐药基因MDR1表达及肝癌细胞化疗敏感性的影响%Influence of alpha-fetoprotein on the expression of drug-resistance gene MDR1 and chemotherapeutic sensitivity in hepatocellular carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    吴超; 杨健; 张金玲; 金涛; 何前进; 李常海

    2015-01-01

    Objective To explore the influence of alpha-fetoprotein (AFP) on the expression of drug-resistance gene MDR1 and chemotherapeutic sensitivity in hepatocellular carcinoma (HCC) cells.Methods A HCC cell line SMMC-7721/AFP, which was stably transfected with AFP gene, was established.mRNA and protein expressions of AFP and MDR1 were detected by real-time PCR and Western Blot,respectively.The sensitivity of SMMC-7721/AFP and SMMC-7721/EGFP cells with or without MDR1 silencing by siRNA to doxorubicin was tested by MTT assay.Immunohistochemistry was used to detect the expression of MDR1 genes-coded protein Pgp in 60 cases of HCC tissues, and the relationship between Pgp expression and serum AFP levels was analyzed.Results AFP mRNA and protein could be detected in SMMC-7721/AFP cells, but not in control cells, indicating that the AFP stably transfected cell line was successfully established.MDR1 mRNA and protein levels were higher in SMMC-7721/AFP cells than those in SMMC-7721/EGFP cells.MDR1 mRNA level in SMMC-7721/AFP cells was (52.7 ± 1.5) times as high as that in SMMC-7721/EGFP cells (P < 0.05).The resistance to doxorubicin was increased by (12.8 ± 1.1) times after AFP transfection (P < 0.05).The chemosensitivity to doxorubicin was increased after the expression of MDR1 was knocked down by siRNA.The expression of Pgp in HCC tissues was positively correlated with the serum AFP levels.Conclusion AFP could induce drug-resistance to doxorubicin in HCC cells by increasing the expression of MDR1.%目的 探讨甲胎蛋白(AFP)对耐药基因MDR1表达和肝癌细胞化疗敏感性的影响.方法 建立稳定表达AFP的肝癌细胞系SMMC-7721/AFP,分别通过Real-time PCR和蛋白印迹检测转染前后AFP和MDR1的表达.MTT法测定SMMC-7721/AFP和SMMC-7721/EGFP细胞对阿霉素的化疗敏感性.siRNA沉默SMMC-7721/AFP细胞中MDR1的表达,观察细胞对阿霉素化疗敏感性的变化.采用免疫组织化学染色法检测60例肝癌组织中MDR1编码蛋

  10. Radioimmunoassay of plasma corticotrophin

    International Nuclear Information System (INIS)

    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 1011 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.)

  11. 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...

  12. 甲胎蛋白与异常凝血酶原在鉴别诊断肝细胞癌中的价值%Alpha-fetoprotein and des-gamma-carboxyprothrombin in the differential diagnosis of hepatocellular carcinoma from other liver tumors

    Institute of Scientific and Technical Information of China (English)

    纪文斌; 肖年军; 罗英; 刘哲; 张宁; 孔哲; 卢实春

    2016-01-01

    目的 评估术前血清甲胎蛋白(AFP)与异常凝血酶原(DCP)在肝脏肿瘤患者中鉴别肝细胞癌的价值.方法 选取2015年1月1日至2015年5月1日解放军总医院肝胆外科收治的影像学检查证实为肝脏肿瘤的患者141例.检测患者术前血清甲胎蛋白及异常凝血酶原浓度.以病理结果或临床诊断标准(射频消融及介入栓塞治疗患者)为金标准,绘制受试者工作特征曲线并分析AFP及DCP在肝脏肿瘤患者中鉴别肝细胞癌的价值.结果 141例肝脏肿瘤患者中肝细胞癌98例,非肝细胞癌43例(包括肝内胆管细胞癌16例、肝血管瘤12例、转移性肝癌10例、局灶性结节增生3例、炎性假瘤2例).肝细胞癌组患者术前甲胎蛋白显著高于非肝细胞癌组患者[80.0(3.9~1 375.0) μg/L比2.1(1.6 ~3.2) μg/L,Z=6.98,P<0.01];肝细胞癌组患者术前异常凝血酶原显著高于非肝细胞癌组患者[141.5(24.0~978.0) AU/L比19.0(14.0 ~ 25.5) AU/L,Z=5.18,P<0.01].AFP及DCP在肝脏肿瘤患者诊断肝细胞癌的受试者工作曲线下面积(AUROC)分别为0.87(95% CI:0.81 ~0.93)及0.78(95% CI:0.69 ~0.86)(截断值分别为3.6 μg/L及35 AU/L),二者差异无统计学意义(Z=1.72,P=0.085).以20 μg/L为AFP的临界值,其诊断肝细胞癌的灵敏度为56.1%,特异度为95.4%;以40 AU/L为DCP的临界值,其灵敏度为69.4%,特异度为83.7%.在肝细胞癌患者,AFP与DCP两者检测结果相关(x2=9.12,P<0.01),相关系数r=0.292.二者联合检测(平行试验)诊断肝细胞癌的灵敏度为79.6%,特异度为81.4%.结论 DCP是肝细胞癌的有效肿瘤标志物.在肝脏肿瘤患者,DCP与AFP鉴别肝细胞癌患者能力类似;联合DCP与AFP检测有助于提高诊断灵敏度,减少漏诊.尽管肝细胞癌的诊断主要依据影像学检查,但在肿瘤影像学表现不典型时,肿瘤标志物有助于肝细胞癌的诊断.%Objective To compare the clinical utility of alpha-fetoprotein (AFP) and

  13. Optimization of AFP-radioimmunoassay using Antibody Capture Technique

    International Nuclear Information System (INIS)

    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

  14. Radioimmunoassay in children with glomerulonephritis

    International Nuclear Information System (INIS)

    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 β2-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

  15. Radioimmunoassay and other related techniques

    International Nuclear Information System (INIS)

    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

  16. The application of lentil lectin-reactive alpha-fetoprotein ratio in the differential diagnosis of primary liver cancer%甲胎蛋白异质体比率在原发性肝癌鉴别诊断中的应用

    Institute of Scientific and Technical Information of China (English)

    赵晓玲; 王晶晶; 赵巧玉; 黄梁镔

    2016-01-01

    Objective To explore the application of lentil lectin‐reactive alpha‐fetoprotein ratio (AFP‐L3% ) applied in the dif‐ferential diagnosis between hepatitis B infection of primary liver cancer and benign liver disease .Methods We included 108 cases of chronic HBV infection ,including 50 cases of primary liver cancer ,42 cases of cirrhosis ,16 cases of chronic hepatitis .Chemilumines‐cence detection was used to detect alpha‐fetoprotein (AFP) and AFP‐L3 content ,AFP‐L3 and the ratio of AFP (AFP‐L3% ) was calculated .Results AFP≥400 ng/mL as primary liver cancer diagnostic threshold ,the sensitivity and specificity were 36% ,84% , when used AFP‐L3% ≥ 10% as primary liver cancer diagnostic threshold ,the sensitivity and specificity were 62% ,83% . Conclusion AFP‐L3% is a better clinical indicator to distinguish between primary liver cancer and benign liver disease .AFP‐L3%can be used as a clinical indicator to differential diagnosis between HBV infection of primary liver cancer and benign liver disease .%目的:探讨甲胎蛋白异质体比率(A FP‐L3%)在乙型肝炎感染原发性肝癌和良性肝病鉴别诊断中的应用。方法慢性乙型肝炎病毒(HBV)感染108例,其中原发性肝癌50例、肝硬化42例、慢性肝炎16例。运用化学发光法检测甲胎蛋白(AFP)及AFP‐L3水平,计算AFP‐L3%。结果以AFP≥400 ng/mL作为原发性肝癌诊断阈值时,灵敏度及特异度分别为36%、84%;以AFP‐L3%≥10%作为原发性肝癌诊断阈值时,灵敏度及特异度分别为62%、83%。结论 AFP‐L3%更能区分原发性肝癌和良性肝病。临床上可结合A FP‐L3%作为鉴别诊断 HBV感染原发性肝癌和良性肝病的指标。

  17. 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

    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 in...... that are almost identical to naturally occurring human polypeptides, including antibody (Ab) constructs; unfortunately, all human biological agents are potentially immunogenic.An increasing number of recent studies have demonstrated the safety of influenza and pneumococcal vaccines administered to...

  18. Determination of bile acids by radioimmunoassays

    International Nuclear Information System (INIS)

    The present paper, based on the current literature, considers the practical aspects of bile acid radioimmunoassays. The problems assoziated with the raising of specific antisera and their characterization are discussed. Features of assay design for bile acids are considered. Solid-phase radioimmunoassays are described for separate determination of unconjugated cholic acid and conjugated cholic acid in serum. The clinical application of specific bile acid radioimmunoassays is shown by an 'oral cholate tolerance test' as a sensitive indicator of liver function and by an 'oral cholylglycine tolerance test' for characterization of intenstinal function in diarrheal states. (orig.)

  19. Radioimmunoassay of Atrazine in Environment

    International Nuclear Information System (INIS)

    Full text: The objective of this study was to develop a rapid and reliable solid-phase radioimmunoassay for the determination of atrazine in water. The tubes were first coated with sheep IgG anti atrazine (Guild hay,U.K.). The reaction was performed by incubating 2 hours a mixture of 100 μl of water and 200 μl of atrazine tracer in antibody coated tubes. After washing, the tubes were counted in gamma counter. The minimum detectable concentration was 0.15 ppb, which below the limits permitted by the US EPA guidelines for drinking water (3 ppb). This assay allows accurate determination of atrazine in water with good specificity, precision and accuracy, and is suitable for the rapid screening of numerous surface and subsurface water samples, as well as for a variety of other analytical application

  20. Modification of the theophylline radioimmunoassay

    International Nuclear Information System (INIS)

    The routine utilization of a commercially available radioimmunoassay (RIA) for theophylline (GammaDab), although reliable, is currently prohibited by the high cost of the reagents. In an effort to reduce these costs we have diluted the [125I]theophylline tracer and theophylline antiserum reagents by one-half, contrary to the manufacturer's recommendations. We have demonstrated that an excellent correlation exists (r . 0.968) between our modified RIA method and a conventional high-performance liquid chromatographic technique, despite reagent dilution. Accordingly, our reagent costs have been reduced by half. We conclude that the GammaDab kit reagents can be diluted twofold and still provide an accurate determination of serum theophylline. We must also emphasize that any further alteration(s) of this theophylline RIA procedure would require a thorough evaluation before its routine use could be substantiated

  1. Methadone radioimmunoassay: two simple methods

    International Nuclear Information System (INIS)

    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 [1-3H](-)-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)

  2. Measurement of antibodies to tubulin by radioimmunoassay

    International Nuclear Information System (INIS)

    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. (Auth.)

  3. Radioimmunoassay method for triiodothyronine and thyroxine

    International Nuclear Information System (INIS)

    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 125I and 131I are described

  4. Measurement of ferritin in serum by radioimmunoassay

    International Nuclear Information System (INIS)

    A method for the measurement of circulating ferritin concentrations by direct radioimmunoassay is presented. The method described is simple to operate, precise, and sensitive and is suitable for the routine diagnostic measurement of ferritin in either plasma or serum. (author)

  5. Sensitive radioimmunoassays using partially purified gamma globulins coupled to enzacryl (acrylamide polymer) solid support

    International Nuclear Information System (INIS)

    Enzacryl polythiolactone is a cross-linked acrylamide polymer with an active thiolactone group, capable of coupling to lysine, serine and tyrosine residues. Gamma globulins from antisera specific to human chorionic gonadotropin, carcinoembryonic antigen, alpha-foetoprotein or casein were isolated using Protein A Sepharose and were subsequently coupled to enzacryl. The resulting coupled antibodies were found to provide greater sensitivity and convenience in radioimmunoassay studies than conventional double antibody precipitation methods using the same antisera. (Auth.)

  6. Radioimmunoassay of triiodothyronine in urine

    International Nuclear Information System (INIS)

    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-T3 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 T3 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 T3 was normal amounting to from 34 to 87 ng/hour, although in most cases the serum T3 concentration was raised. In cases of anorexia nervosa and in obese starving subjects the excretion of T3 fell significantly, and similarly low excretion was found in some cases of debilitating diseases and myocardial infarction. (author)

  7. A radioimmunoassay for human ACTH

    International Nuclear Information System (INIS)

    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, α 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. (auth.)

  8. Specificity of radioimmunoassays for relaxin

    International Nuclear Information System (INIS)

    The specificities of two radioimmunoassays (RIA) for relaxin, based upon crude porcine relaxin (NIH-R-P1;RIA I) and a highly purified porcine relaxin (RIA II) have been studied concurrently using purified hormones and plasma samples. A labelled fraction, selected from radio-iodinated NIH-R-P1 and used in that RIA, was also bound to antiserum raised to the highly purified relaxin. Hence a third RIA was possible in which both the crude and the purified relaxins inhibited in the ng/ml range. Porcine insulin and the connecting peptide of porcine proinsulin did not inhibit any of the assay systems whereas porcine proinsulin did inhibit in each assay at the μg/ml range. Concurrent measurements by assays I and II have been made in sheep plasma obtained during both delivery of the lamb and suckling. The peak values obtained by assays I and II are 3 and 6 min out of phase during suckling and delivery respectively; the NIH-R-P1 relaxin immunoactivity appearing first. The plasma inhibition curves of both appear to be the sum of individual contributions from relaxin and relaxin-like peptides, such as prorelaxin and its fragments, as seen by different antisera. Both assays, however, give qualitatively similar indices of relaxin immunoactivity. The RIA developed for the more purified peptide would be expected to yield a better quantitative estimate of relaxin secretion but this, like specificity, cannot be shown absolutely. (author)

  9. Radioimmunoassay for C-peptide and proinsulin

    International Nuclear Information System (INIS)

    Proinsulin, the biosynthetic precursor of insulin, was discovered by Steiner et al. (1967) and shown to be converted to insulin and C-peptide in the β-cell. The first part of this paper deals with aspects of the radioimmunoassay for C-peptide with special emphasis on the development and the sources of errors encountered in our laboratory (Heding, 1975; Naithani et al., 1975). The second part deals with the many problems involved in the determination of human proinsulin and describes a direct and specific radioimmunoassay developed for measuring proinsulin in serum with a detection limit of less than 0.01 pmol/ml. (Auth.)

  10. Steroid derivatives and their use in radioimmunoassays

    International Nuclear Information System (INIS)

    Radioimmunoassay techniques have been used to determine the concentration in body fluids of various endogenous and exogenous steroids. In the development of radioimmunoassays for the various steroids, the preparation of an antigen labelled with iodine-125 is of primary concern. The chemical structure of steroids is such that it is generally not possible to radioiodinate them directly. It is necessary to utilize as a precursor of the radiolabeled antigen a derivative of the steroid to be assayed which can be readily iodinatd. The process by which steroids are chosen and structurally modified is given

  11. Radioimmunoassay of protein C system

    International Nuclear Information System (INIS)

    Protein C system is an anticoagulation pathway which consists of protein C (PC), protein S (PS), thrombomodulator (TM) and protein C inhibitor (PCI). Using the McAb SZ-57, the authors have established SZ-57-Sepharose CL-6B affinity chromatography to purify human urinary TM. A procedure for isolation and purification of PC, PS and PCI from albumin-free human plasma by rivanol precipitation was also established. The isolation steps include adsorption onto and elution from barium, PEG precipitation, ion-exchange chromatography and preparative isoelectric focusing and so on. The molecular weight, isoelectric points, amino acid contents and the functional activity of these proteins were consistent with other previous reports. Four radioimmunoassays (RIAs) of PC, PS, TM and PCI were established using the equilibrium method. 125I-PC and 125I-PS were prepared using the chloramine-T method. 125I-PCI was prepared by iodogen method and 125I-TM by Bolton-Hunter method. Their sensitivities were 3.94 μg/L, 9.87 μg/L, 6.16 μg/L and 2.58 μg/L, respectively. The recovery rates were 104.28%, 94.30%, 105.22% and 101.89. Some antiserum provided a linear response from 6.25 to 1024 μg/L for PC, 21 to 700 μg/L for PS, 8.1 to 560 μg/L for TM and 4.8 to 1024 μg/L for PCI. The intra- and inter-assay CV were 4.4% and 9.68% for PC RIA, respectively, 4.99% and 13.14% for PS RIA, 5.10% and 10.94% for TM RIA, 2.73% and 8.62% for PCI RIA. The cross reactivity with factor II, thrombin, and antithrombin III was negligible. These methods can be used as effective tools especially for diagnosis of thrombosis and basic or clinical studies of protein C system. (20 refs., 8 figs., 6 tabs.)

  12. A radioimmunoassay for the Hydra head activator

    International Nuclear Information System (INIS)

    A highly-sensitive and specific radioimmunoassay for the head activator has been developed which utilises tritiated head activator. The assay is sensitive in the range of 40-200 fmol. Immunochemical studies showed that the antiserum recognised the intact molecule better than any of the fragments or derivatives produced by enzymatic treatment or chemical synthesis. (Auth.)

  13. 南昌地区妊娠14~20周妇女血清甲胎蛋白和游离绒毛膜促性腺激素的MOM值测定及临床应用%Detection and clinical application of MOM values of serum alpha-fetoprotein and free human chorionic gonadotropin in pregnant women during 14~20 gestational weeks in Nanchang region

    Institute of Scientific and Technical Information of China (English)

    赖华; 刘艳秋; 刘淮

    2011-01-01

    目的:运用计算出的AFP及F-βHCG的MOM值与目前采用的MOM值对孕妇进行胎儿神经管缺陷畸形及唐氏综合征筛查,期待能更有效地筛查出南昌地区胎儿神经管缺陷及唐氏综合征高危孕妇.方法:依据2005~2007年到江西省妇幼保健院产前诊断门诊就诊的孕妇各孕周AFP、F-βHCG的中位数,计算出AFP、F-βHCG的MOM值,用计算出的MOM值(研究组)和目前采用的MOM值(对照组)分别对2008年来该院产前诊断门诊就诊的孕妇进行胎儿神经管缺陷畸形及唐氏综合征筛查,对筛查出的高危孕妇进行羊水检测和随访,比较两种MOM值对胎儿神经管缺陷畸形和唐氏综合征筛查的阳性率、假阴性率及假阳性率.结果:神经管缺陷畸形、唐氏综合征阳性率研究组高于对照组,具有统计学意义(P<0.01);假阴性率研究组与对照组比较,结果无统计学意义;假阳性率研究组较对照组降低,结果具有统计学意义,(神经管缺陷畸形P<0.05,唐氏综合征P<0.01).结论:应用本地区MOM值进行神经管缺陷畸形及唐氏综合征筛查,能更有效地筛查出本地区的神经管缺陷畸形及唐氏综合征胎儿.%Objective: To screen fetal neural tube defect and Down's syndrome by calculated MOM values and values currently used of serum alpha - fetoprotein (AFP) and free β human chorionic gonadotropin (F - β HCG) in pregnant women, in order to screen out the high risk pregnant women of fetal neural tube defect and Down's syndrome in Nanchang region.Methods: The MOM values of AFP and F - β HCG were calculated according to the medians of AFP and F - β HCG in pregnant women of different gostational weeks who visited the prenatal diagnosis outpatient of the hospital from 2005 to 2007, then the calculated MOM values ( study group) and the MOM values currently used ( control group) were used to screen fetal neural tube defect and Down's syndrome among the pregnant women who visited the prenatal

  14. 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.

  15. Anamnesis of the radioimmunoassay: A historical reminiscence

    International Nuclear Information System (INIS)

    Anamnesis of the Radioimmunoassay - A Historical Reminiscence: Several requirements has 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. (orig.)

  16. Radioimmunoassays of 7-hydroxymethotrexate and methotrexate

    International Nuclear Information System (INIS)

    Antisera against 7-hydroxymethotrexate were raised in rabbits by injection of 7-hydroxymethotrexate coupled to bovine serum albumin. A 125I-radiolabelled probe was synthesised by radioiodination of 7-hydroxymethotrexate conjugated to tyrosine methyl ester. We developed with these reagents a liquid phase radioimmunoassay for 7-hydroxymethotrexate in which methotrexate did not interfere significantly (cross-reactivity factor = 2.5 X 10-4). Similar techniques were used also for the development of a specific radioimmunoassay for methotrexate, which minimally cross-reacted with 7-hydroxymethotrexate. The assessment of the specificity of these assays showed that parallel monitoring of methotrexate and 7-hydroxymethotrexate in human plasma was possible. Coefficients of variation for duplicate determinations were less than 15% between 0.06 and 1.2 nmol/l for methotrexate and between 0.08 and 9 nmol/l for 7-hydroxymethotrexate. (Auth.)

  17. Development of a radioimmunoassay for formoterol

    Energy Technology Data Exchange (ETDEWEB)

    Yokoi, K.; Murase, K.; Shiobara, Y.

    1983-10-01

    The development of a radioimmunoassay (RIA) for the ..beta../sub 2/-stimulant formoterol is described. The sensitivity of the method is 0.1 ng/ml in plasma and urine, when a l-ml sample is used. The cross-reactivity of the antiserum with formoterol glucuronide was 30%. Since formoterol is metabolized extensively to formoterol glucuronide in rats, dogs and human, extraction with ethyl ether prior to the radioimmunoassay was carried out. Satisfactory agreement was obtained for levels of formoterol in plasma and urine when they were determined by RIA and gas chromatography-mass spectrometry. The concentration of formoterol was determined in dog plasma and human urine after oral administration of formoterol fumarate to dogs (61 mcg/kg) and humans (40 mcg).

  18. A radioimmunoassay for serum medroxyprogesterone acetate.

    Science.gov (United States)

    Shrimanker, K; Saxena, B N; Fotherby, K

    1978-04-01

    When injected intramuscularly in a dose of 150 mg, Depo Provera, a microcrystalline suspension of medroxyprogesterone acetate (MPA), will provide a contraceptive effect for at least 3 months. This paper describes a sensitive radioimmunoassay for MPA which has been used in the author's laboratory for the past 2 years. MPA was converted to MPA-3-CMO and the oxime was conjugated with bovine serum albumin (BSA) by the mixed anahydride method. 4 rabbits were immunized with the antiserum. A high titre of MPA antibodies was detected 6 months after immunization. Serum from the rabbit with the highest titre of antibodies to MPA was subjected to radioimmunoassay. 7 days after the intramuscular injection of 150 mg Depo-Provera, serum levels of MPA were found in the range of 1750 to 9000 pg/ml. By 75 days, the levels had decreased to 680-2600 pg/ml. The method was found to have adequate accuracy, precision and sensitivity. PMID:661315

  19. Radioimmunoassay to quantitatively measure cell surface immunoglobulins

    International Nuclear Information System (INIS)

    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/106 cells. Determination of immunoglobulins on the peripheral lymphocytes obtained from different cancer patients varied between 340 to 1040 ng/106 cells. Cultured tumor cells, on the other hand, were found to contain negligible quantities of human IgG

  20. Studies on the radioimmunoassay of thyroid hormones

    International Nuclear Information System (INIS)

    To establish radioimmunoassay (RIA) systems of 3,5,3'-triiodo-L-thyronine (T3) and thyroxine (T4), various experiments such as 125I 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)

  1. Principles and development of the radioimmunoassay

    International Nuclear Information System (INIS)

    Compared to conventional biological test methods as well as to purely physicochemical methods, the radioimmunoassay exhibits higher sensitivity, higher selectivity, and better practicability. It has thus become possible to determine many substances in the nano- and picogramme region. It should, however, be pointed out that for an efficient use of radioimmunological test methods, their indications and limitations should be studied critically, and their reliability and quality should receive constant supervision. (GSE/AK)

  2. Radioimmunoassay of calcitonin in normal human urine

    International Nuclear Information System (INIS)

    Direct radioimmunoassay of calcitonin in human urines containing 0espectively. The urine calcitonin values apparently reflect serum calcitonin concentrations (e.g., urine/serum r = 0.9873 for 40 hypercalcitonemic patients); but urine calcitonin determination has two important advantages: greater reproducibility because of decreased heterogeneity and greater differentiation of patient populations. In view of these results, the assay of urine calcitonin may prove to be a very useful clinical tool

  3. Radioimmunoassay of proinsulin and hyperproinsulinemic states

    International Nuclear Information System (INIS)

    The purpose of this paper is: (a) to present an improved radioimmunoassay for proinsulin with a lower detection limit, and discuss its sources of error, and (b) to present extended studies on serum proinsulin in patients with insulinomas and hyperthyroidism, and a thorough analysis of the percentage of immunoreactive insulin (IRI) constituted by proinsulin in controls before, during and after stimulation with oral glucose. (Auth.)

  4. Folic acid derivatives for use in radioimmunoassay

    International Nuclear Information System (INIS)

    The chemical preparation of two folic acid derivatives, labelled with 125I or 131I, 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.)

  5. Methods in radioimmunoassay, toxicology, and related areas

    International Nuclear Information System (INIS)

    The following topics were discussed at the conference: past, present, and potential of radioimmunoassay; measurement of biogenic amines at the picogram level; parathyroid hormone; in vitro radioisotope methods for clinical evaluation of vitamin B12 and folic acid metabolism; forensic toxicology; role of biotransformation reactions in prenatal and postnatal chemical toxicity; the use of electrochemical techniques in FDA analytical procedures; and atomic absorption in food analysis

  6. A radioimmunoassay for urinary and serum dexamethasone

    International Nuclear Information System (INIS)

    A radioimmunoassay suitable for measuring dexamethasone concentrations in serum and urine specimens following the low dose dexamethasone suppression test (0.25-1.0 mg dexamethasone) is reported. The assay has a sensitivity of 0.26 nmol/L, a between-assay coefficient of variation (CV) for dexamethasone concentrations between 1.15 and 15.40 nmol/L ranging from 11.7 - 5.5% and recoveries of 91 - 103%. (author)

  7. The production of antibodies for radioimmunoassay

    International Nuclear Information System (INIS)

    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

  8. Radioimmunoassay of measles virus hemagglutinin protein G

    International Nuclear Information System (INIS)

    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.)

  9. 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.

  10. Radioimmunoassay for serum 11-deoxy-17-ketosteroids

    International Nuclear Information System (INIS)

    A simplified method for evaluating serum 11-deoxy-17-ketosteroids (11-deoxy-17-KS) equivalent to dehydroepiandrosterone sulfate (DHEAS) has been developed without solvolysis and chromatography. Blood serum or plasma (5 mu l) was added to 1 ml of ethanol, mixed, and centrifuged, and 10 or 20 mu l of the supernatant was evaporated to dryness and incubated with anti-11-deoxy-17-KS antiserum obtained by immunizing a rabbit with DHEA-3.O. CO-BSA which was prepared from DHEA-3.O.COCl and containing DHEAS-7α3H, pepsin-treated human immune serum globulin and bovine serum albumin. Ammonium sulfate was used to separate free from bound DHEAS-7α3H. The accuracy, precision and sensitivity were satisfactory. The blank values could not be differentiated from zero. As the antiserum reacted not only on DHEAS but also on androsterone sulfate and etiocholanolone sulfate, serum 11-deoxy-17KS obtained by the radioimmunoassay expressed nearly the sum of 100 percent of DHEAS, 45 percent of androsterone sulfate and 35 percent of etiocholanolone sulfate in the serum. A good correlation was found between serum 11-deoxy-17-KS and DHEAS obtained by the radioimmunoassay described in a previous paper. The present radioimmunoassay is the simplest method for the evaluation of the concentrations of C19 steroids in the serum. (auth)

  11. Radioimmunoassay techniques and their application in veterinary parasitology

    International Nuclear Information System (INIS)

    In parasitology radioimmunoassay has been largely employed as a technique for detecting antibodies. Three types of this assay are outlined here and a brief comparison is made with the enzyme-linked immunosorbent assay. Examples where radioimmunoassay and enzyme-linked immunosorbent assay have been used in parasitology are given. It is concluded that in veterinary parasitology radioimmunoassay is of greater value as a tool in the study of immune mechanisms to parasites rather than as a diagnostic technique. (author)

  12. Diagnostic value of α-fetoprotein in hepatocellular carcinoma

    International Nuclear Information System (INIS)

    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)

  13. Solid phase separation technique for use in radioimmunoassays

    International Nuclear Information System (INIS)

    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

  14. Solid phase double-antibody radioimmunoassay procedure

    International Nuclear Information System (INIS)

    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

  15. Application of radioimmunoassay in food industry

    International Nuclear Information System (INIS)

    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.)

  16. Development of a radioimmunoassay for papain

    International Nuclear Information System (INIS)

    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 20C. 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.)

  17. Radioimmunoassay in the diagnosis of fascioliasis

    International Nuclear Information System (INIS)

    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 125iodine. (T.I.)

  18. Radioimmunoassay for human growth hormone (HGH)

    International Nuclear Information System (INIS)

    A radioimmunoassay for human growth hormone (HGH) has been developed, which is highly specific, sensitive, precise, and reliable. The assay is based on the competitive binding principle of radioimmunoassay and utilizes a guinea pig anti-HGH serum and 125I-HGH. The tracer can be used for 3-4 weeks without further purification. Separation of antibody-bound from free 125I-HGH is achieved by the use of an immuno-immobilized precipitating reagent in the presence of 4% polyethylenglycol. Under these conditions the unspecific binding is as low as 2% of the total radioactivity. The standard curve covers the range of 0.05 to 1.6 nmol/l. At higher HGH concentrations (0.8 to 1.6 nmol/l) the inter- and intraassay coefficients of variation are 12-14 and 5-8%, respectively. For determination of HGH concentration plasma, serum or buffer-diluted samples may be used. Recovery of HGH added to 11 individual plasma and serum samples was 103.7 +- 6.3 and 106.5+- 10.1% (x +- S.D.), respectively. Recovery of endogenous HGH in 5 acromegalic sera after dilution with buffer was 102.2 +- 4.2% (x +- S.D.). The levels of circulating HGH obtained with this assay in normal women and men were found to be in the range of 0-0.5 and 0-0.4 nmol/l, respectively. (author)

  19. Heterologous radioimmunoassay of 19-nortestosterone and trenbolone

    International Nuclear Information System (INIS)

    Using 3H-methyltrienolone (86 Ci/mmol) in a heterologous radioimmunoassay instead of 3H-19-nortestosterone (19 Ci/mmol) in the generally used homologous system the sensitivity of the test could be improved 7-fold (50% relative binding by 15 picograms of 19-nortestosterone). Due to the relatively high cross reactivity for trenbolone of about 35% (with a different antiserum 50% have been found for trenbolone-17α) it is possible to have a simultaneous control of that illegal anabolic. In case of suspicion the substances responsible for the positive radioimmunoassay can be verified with a very specific assay for trenbolone. A preliminary examination of the findings via determination of 'internal blanc values' has proved to be a good measure again. With the test using the previously described extraction and clean up procedures prosecution limits of 0.1 ng/g muscle, 1 ng/ml urine and 10 ng/ml bile might be reasonable. The blanc values depend significantly on the colour of urine samples and on the enzyme used for hydrolysis (glucuronidase from Escherichia coli gave no additional blanc in contrast to succ. helix pomatioa). (orig.)

  20. Radioimmunoassay of influenza A virus haemagglutinin. II

    International Nuclear Information System (INIS)

    Individual rabbits differed greatly in their antibody response to the ''strain-specific'' and ''cross-reactive'' antigenic determinants on the hemagglutinin (HA) subunit of influenza virus recombinant MRC11 (H3N2) and influenza virus Dunedin (H3N2), after immunization with whole virus or bromelain-released hemagglutinin (B-HA). Consequently, diverse cross-reactions between these viruses and A/Hong Kong/68 virus were found in the hemagglutination inhibition (HI) test as well as in homologous radioimmunoassay (125I-B-HA from MRC11: anti MRC11 serum, and 125I-B-HA from Dunedin: anti Dunedin serum) when sera from different animals were employed. Radioimmunoassay (RIA), over and above to the HI test, was also able to differentiate clearly the respective HAs with antisera reacting to the same HI titre with both corresponding influenza virus strains. Thus it appeared that antigenic differences could be identified with higher sensitivity by homologous RIA than by the HI test and that multiple antigenic determinants were reactive on the 125I-B-HA in the RIA procedure employed. MRC11 and A/HK/68 viruses were also compared by heterologous RIA (125I-B-HA from MRC11: anti A/HK/68 serum). It was found that preferentially antigenic determinants with a high degree of cross-reactivity could be studied in the heterologous system. (author)

  1. Effects of alpha fetoprotein on escape of Bel 7402 cells from attack of lymphocytes

    Directory of Open Access Journals (Sweden)

    Li Pingfeng

    2005-08-01

    Full Text Available Abstract Background 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. Methods 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. Results 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. Conclusions 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.

  2. Antitumor immunity induced by DNA vaccine encoding alpha-fetoprotein/heat shock protein 70

    Institute of Scientific and Technical Information of China (English)

    Xiao-Ping Wang; Guo-Zhen Liu; Ai-Li Song; Hai-Yan Li; Yu Liu

    2004-01-01

    AIM: To construct a DNA vaccine encoding human alphafetoprotein (hAFP)/heat shock protein 70 (HSP70), and to study its ability to induce specific CTL response and its protective effect against AFP-expressing tumor.METHODS: A DNA vaccine was constructed by combining hAFP gene with HSP70 gene. SP2/0 cells were stably transfected with pBBS212-hAFP and pBBS212-hAFP/HSP70eukaryotic expression vectors. Mice were primed and boosted with DNA vaccine hAFP/HSP70 by intramuscular injection, whereas plasmid with hAFP or HSP70 was used as controls. ELISPOT and ELISA were used to detect IFN-γ-producing splenocytes and the level of serum anti-AFP antibody from immunized mice respectively. In vivo tumor challenge was measured to assess the immune effect of the DNA vaccine.RESULTS: By DNA vaccine immunization, the results of ELISPOT and ELISA showed that the number of IFN-γ-producing splenocytes and the level of serum anti-AFP antibody were significantly higher in rhAFP/HSP70 group than in hAFP and empty plasmid groups (95.50±10.90IFN-γ spots/106 cells vs 23.60±11.80 IFN-γ spots/106 cells,7.17±4.24 IFN-γ spots/106 cells, P<0.01; 126.50±8.22 μg/mL vs 51.72±3.40 μg/mL, 5.83±3.79 μg/mL, P<0.01). The tumor volume in rhAFP/HSP70 group was significantly smaller than that in pBBS212-hAFP and empty plasmid groups (37.41±7.34 mm3 vs381.13±15.48 mm3, 817.51±16.25 mm3,P<0.01).CONCLUSION: Sequential immunization with a recombinant DNA vaccine encoding AFP and heat shock protein70 could generate effective AFP-specific T cell responses and induce definite antitumor effects on AFP-producing tumors, which may be suitable for some clinical testing as a vaccine for HCC.

  3. Effects of alpha fetoprotein on escape of Bel 7402 cells from attack of lymphocytes

    International Nuclear Information System (INIS)

    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

  4. Identification of pregnancy-associated glycoproteins and alpha-fetoprotein in fallow deer (Dama dama) placenta

    OpenAIRE

    Bériot, Mathilde; Tchimbou Njanjo, Aline Flora; Barbato, Olimpia; Beckers, Jean-François; Melo de Sousa, Noelita

    2014-01-01

    Background: This paper describes the isolation and characterization of pregnancy-associated glycoproteins (PAG) from fetal cotyledonary tissue (FCT) and maternal caruncular tissue (MCT) collected from fallow deer (Dama dama) pregnant females. Proteins issued from FCT and MCT were submitted to affinity chromatographies by using Vicia villosa agarose (VVA) or anti-bovine PAG-2 (R#438) coupled to Sepharose 4B gel. Finally, they were characterized by SDSPAGE and N-terminal microsequencing. Result...

  5. Maternal seric alpha-fetoprotein: determination of the medium values in Costa Rican pregnant women

    International Nuclear Information System (INIS)

    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)

  6. A competitive-inhibiton radioimmunoassay for influenza virus envelope antigens

    International Nuclear Information System (INIS)

    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. Steroid radioimmunoassay: contribution of standards to blank values, ch. 4

    International Nuclear Information System (INIS)

    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

  8. Radioimmunoassay of cholecystokinin in tissue and plasma

    International Nuclear Information System (INIS)

    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 125I 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.)

  9. A sensitive radioimmunoassay of scorpion neurotoxin

    International Nuclear Information System (INIS)

    Scorpion neurotoxins form a family of homologous proteins which are basic and have approx. mol. wt. 7000. They consist of a single peptide chain crosslinked by four disulfide bridges. The complete amino acid sequences of some of them as well as the N-terminal of others, have been determined: their comparison has led to a classification into four groups. They have been shown to affect the conduction of ions through membrane channels and are thus good tools for the study of these structures on the molecular level. Toxins I and II of Androctonus australis Hector have been labelled with 125I and specific radioactivities up to 2000 Ci/mmol have been obtained. Here the setting up of a radioimmunoassay allowing a sensitive and specific detection of toxin I of Androctonus australis Hector is reported

  10. Radioimmunoassay of influenza A virus haemagglutinin. I

    International Nuclear Information System (INIS)

    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 125I-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. Report on the national seminar on radioimmunoassays

    International Nuclear Information System (INIS)

    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 125I, 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.)

  12. Radioimmunoassay of aflatoxin M1 in milk

    International Nuclear Information System (INIS)

    The aflatoxin M1 content in samples of commercial milk as well as in milk directly from farms in Western Slovakia was determined by radioimmunoassay using a commercial Czechoslovak RIA kit. In 25 analyzed samples of commercial milk the limit of 0.1 μg kg-1 aflatoxin M1 was not exceeded. From 110 sample from milk farms, 93 samples (84.5%) contained less than 0.1 μ kg-1 aflatoxin, for 15 samples (13.6%) the content ranged between 0.1 and 0.5 μg kg-1, and 2 samples contained 0.5 to 1.0 μg kg-1 of aflatoxin M1. (author). 2 figs., 20 refs

  13. Radioimmunoassay for progesterone in bovine milk

    International Nuclear Information System (INIS)

    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)

  14. Radioimmunoassay for thyroid stimulating hormone (TSH)

    International Nuclear Information System (INIS)

    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.)

  15. Radioimmunoassay for 21-deoxycortisol: clinical applications

    International Nuclear Information System (INIS)

    A radioimmunoassay for 21-deoxycortisol is described. The immunogen, 21-deoxycortisol-3-(0-carboxymethyl) oxime-bovine serum albumin, was prepared, the antisera raised against it were studied and the reliability of the assay was checked. The antiserum selected cross-reacted with 11-deoxycortisol (0.08%), corticosterone (0.25%), cortisol (0.6%) and 17-hydroxyprogesterone (1.6%). 21-deoxycortisol was separated by celite partition chromatography and eluted in the 70/30 (v/v) isooctane/ethyl acetate fraction together with 11-deoxycortisol and corticosterone. The radioimmunoassay was used to measure 21-deoxycortisol in the plasma of normal subjects and patients with androgen excess. In normal subjects, men (0.19 ng/ml +/- 0.08) and women (0.18 ng/ml +/- 0.09) had similar basal levels (mean +/- SD). One hour after ACTH stimulation, these levels were increased by a factor of 3.5. In 7 patients treated for classical congenital adrenal hyperplasia associated with 21-hydroxylase deficiency, basal values varied between 9.1 and 39.9 ng/ml (measured at 8 a.m.). In 7 untreated women with late-onset congenital adrenal hyperplasia (with 21-hydroxylase deficiency), ACTH-stimulated levels were increased to between 9 and 25.5 ng/ml. In 14 heterozygous carriers of 21-hydroxylase deficiency, diagnosed by HLA genotyping, all ACTH-stimulated levels were well above the highest corresponding levels in normal subjects, whereas 17-hydroxyprogesterone levels remained within the normal range in 9 of the cases. (author)

  16. Radioimmunoassay for 21-deoxycortisol: clinical applications

    Energy Technology Data Exchange (ETDEWEB)

    Gueux, B.; Fiet, J.; Villette, J.M.; Brerault, J.L.; Vexiau, P.; Julien, R. (Laboratoire de Biochimie Hormonale, Hopital Saint-Louis, Paris); Pham-Huu-Trung, M.T.; Gourmelen, M. (INSERM U 142 and Laboratoire d' Explorations Fonctionnelles, Hopital Trousseau, Paris); Galons, H. (Laboratoire de Chimie Organique, Faculte de Pharmacie, Paris)

    1985-01-01

    A radioimmunoassay for 21-deoxycortisol is described. The immunogen, 21-deoxycortisol-3-(0-carboxymethyl) oxime-bovine serum albumin, was prepared, the antisera raised against it were studied and the reliability of the assay was checked. The antiserum selected cross-reacted with 11-deoxycortisol (0.08%), corticosterone (0.25%), cortisol (0.6%) and 17-hydroxyprogesterone (1.6%). 21-deoxycortisol was separated by celite partition chromatography and eluted in the 70/30 (v/v) isooctane/ethyl acetate fraction together with 11-deoxycortisol and corticosterone. The radioimmunoassay was used to measure 21-deoxycortisol in the plasma of normal subjects and patients with androgen excess. In normal subjects, men (0.19 ng/ml +/- 0.08) and women (0.18 ng/ml +/- 0.09) had similar basal levels (mean +/- SD). One hour after ACTH stimulation, these levels were increased by a factor of 3.5. In 7 patients treated for classical congenital adrenal hyperplasia associated with 21-hydroxylase deficiency, basal values varied between 9.1 and 39.9 ng/ml (measured at 8 a.m.). In 7 untreated women with late-onset congenital adrenal hyperplasia (with 21-hydroxylase deficiency), ACTH-stimulated levels were increased to between 9 and 25.5 ng/ml. In 14 heterozygous carriers of 21-hydroxylase deficiency, diagnosed by HLA genotyping, all ACTH-stimulated levels were well above the highest corresponding levels in normal subjects, whereas 17-hydroxyprogesterone levels remained within the normal range in 9 of the cases.

  17. alpha-Melanocyte-stimulating-hormone precursors in the pig pituitary

    DEFF Research Database (Denmark)

    Fenger, M

    1986-01-01

    The occurrence of intermediates from the processing of ACTH-(1-39) [adrenocorticotropic hormone-(1-39)] to alpha-melanocyte-stimulating hormone was investigated in normal pig pituitaries by the use of sensitive and specific radioimmunoassays for ACTH-(1-13), ACTH-(1-14), ACTH-(1-13)-NH2 and ACTH-...

  18. 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)

  19. Purification of antibodies anti-interleukin-1 beta for radioimmunoassay

    International Nuclear Information System (INIS)

    The sensitivity of a radioimmunoassay is dependent on the affinity of the antibody. The isolation of specific very high affinity antibodies is therefore a key requirement. In this project purified antibodies were prepared by affinity chromatography from polyclonal antisera anti-Interleukin-1 beta involving stepwise elution from pH 7.0 to 2.5 contributing to the development of a sensitive radioimmunoassay for measurement of human IL-1 beta. (author). 7 refs, 2 figs, 1 tab

  20. Radio-immunoassay of somatostatin from isolated rat pancreatic islets

    Energy Technology Data Exchange (ETDEWEB)

    Vonen, B.; Florholmen, J.; Giaever, A.K.; Burhol, P. (Tromsoe Univ. (Norway))

    1989-04-01

    Certain aspects of radio-immunoassay of somatostatin from isolated rat pancreatic islets are described. Somatostatin-14, and not somatostatin-28, is secreted from isolated rat pancreatic islets. Less somatostatin secretion is measured per islet owing to purity of tracer in the radio-immunoassay. Theophylline apparently cross-reacts with somatostatin in the assay described, and this has to be taken into consideration when studying somatostatin release induced by theophylline in isolated islets. (author).

  1. A solid-phase radioimmunoassay for detection of tetanus antibodies

    International Nuclear Information System (INIS)

    A solid-phase radioimmunoassay has been developed as a screening technique for tetanus antibodies in blood plasma. It is based on the principle of a commercial test for Hepatitis B antibody. Compared to previous screening techniques, the radioimmunoassay showed better stability with no apparent loss of sensitivity over a 2 month period. This technique has proved useful in determining tetanus immunity and in monitoring free antibody level in treated cases of clinical tetanus. (U.K.)

  2. Radio-immunoassay of somatostatin from isolated rat pancreatic islets

    International Nuclear Information System (INIS)

    Certain aspects of radio-immunoassay of somatostatin from isolated rat pancreatic islets are described. Somatostatin-14, and not somatostatin-28, is secreted from isolated rat pancreatic islets. Less somatostatin secretion is measured per islet owing to purity of tracer in the radio-immunoassay. Theophylline apparently cross-reacts with somatostatin in the assay described, and this has to be taken into consideration when studying somatostatin release induced by theophylline in isolated islets. (author)

  3. 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.

  4. Radioimmunoassay for chlorinated dibenzo-p-dioxins

    International Nuclear Information System (INIS)

    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

  5. 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.

  6. Estimation of urinary angiotensin II by radioimmunoassay

    International Nuclear Information System (INIS)

    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. (author)

  7. The radioimmunoassay of human serum inhibin

    International Nuclear Information System (INIS)

    The measurement of circulating inhibin is essential to the advancement of our understanding of its role in the control of FSH secretion. Recently, sensitive in vitro bioassay systems have been described which have allowed the measurement of inhibin bioactivity in the peripheral serum of immature female rats, sheep and women (C. Tsonis, personal communication) following ovarian hyperstimulation with exogenous gonadotrophins. Inhibin bioactivity has also been shown to rise in ovarian venous blood in the late follicular phase of women and during FSHG administration to primates. Such studies have some important limitations, in particular, the bioassay systems employed lack the sensitivity required for the study of normal physiology, and questions regarding the specificity of in vitro bioassay in application to serum samples have arisen following the reports of FSH releasing factors (transforming growth factor-beta inhibin B subunit dimers) that may confound the accurate assessment of inhibin bioactivity. This paper reports on the development of sensitive and specific radioimmunoassay (RIA) systems applicable to human serum utilizing highly purified preparations of 58kDa and 31kDa bovine follicular fluid (bFF) inhibin. The development of two RIA systems utilizing antisera raised to either or 31kDa bFF inhibin is detailed and their application to the study of inhibin physiology in the human female during ovulation inducting, the normal menstrual cycle and gestation described

  8. Radioimmunoassay of human intestinal alkaline phosphatase

    International Nuclear Information System (INIS)

    A new method of radioimmunoassay using the double antibody method for human intestinal alkaline phosphatase (ALP) was first elaborated. The following results were obtained: 1) In this system, the optimal antibody concentration is 10,000 times the dilution of the original anti-serum, and the optimal assay range is 0.5 to 25 ng. Enzymatic activity of 1 ng intestinal ALP is 4.1 King-Armstrong units. 2) In this system, the sera including intestinal ALP are divided to two groups. One group shows a dose response curve similar to that of purified intestinal ALP, and the other shows a lesser one. This reason is not clear. Hepatic ALP, osseous ALP and placental ALP in the sera show no response in this system. 3) In this system, the B/T value of 50 μg of purified human placental ALP is almost equal to 1 ng of purified human intestinal ALP. Similarly, the B/T value of 50 μg of purified human intestinal ALP is equal to almost 5 ng of purified human placental ALP. This shows that cross-reaction exists between intestinal and placental ALPs at high concentrations. (J.P.N.)

  9. Radioimmunoassay of renin in human renal tissues

    International Nuclear Information System (INIS)

    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)

  10. Radioimmunoassay study of neurophysins in human plasma

    International Nuclear Information System (INIS)

    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)

  11. Radioimmunoassay in developing countries: General principles

    International Nuclear Information System (INIS)

    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

  12. Radioimmunoassay of penicilloyl groups in biological fluids

    International Nuclear Information System (INIS)

    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

  13. Milk progesterone radioimmunoassay using radioiodinated tracers: a rapid and reliable assay system

    International Nuclear Information System (INIS)

    Both the validity and practicability of a direct progesterone radioimmunoassay based on radioiodinated progesterone tracers were studied. The results obtained show the reliability of the assay; when compared with assays based on 3H-progesterone tracers there are fewer steps for assay execution, saving time and reducing the number of reagents used. Various commercially available 125I-progesterone tracers were assayed, and only those with an 11 alpha-hemisuccinate bridge were suitably bound by antisera raised against progesterone-bovine serum albumin conjugates having identical bridge structure. The bridge effect caused no observable alteration in validity parameters. Finally, the results support the utility of this assay as a practical method of early diagnosis of pregnancy and as a reliable experimental technique to monitor cow ovarian function

  14. Radioimmunoassay of human epidermal growth factor (urogastrone)

    International Nuclear Information System (INIS)

    Epidermal growth factor (EGF) is a polypeptide hormone that stimulates growth of a variety of tissues. Although it was originally discovered in male mouse submaxillary glands, EGF has recently been isolated from human urine. A heterologous radioimmunoassay for human EGF (hEGF) has been developed, using purified hEGF as reference standard and radioiodinated tracer and antibodies raised against mouse EGF. Purified hEGF specifically displaced radioiodinated hEGF from the antibodies; no other human peptide hormone tested demonstrated any cross-reaction. Twenty-four hour urinary excretion of RIA-hEGF in normal adult males and females was 97.8 +- 10.7 and 72.0 +- 4.5 (mean +- SE) μg/total volume, or 51.7 +- 4.5 and 57.0 +- 4.9 μg/g of creatinine, respectively. Urinary excretion in normal children increased with age, from less than 40 μg/24 h at 4 years of age to adult levels at about the age of puberty. The concentration of RIA-hEGF in human saliva ranged from 5.6 to 16.8 ng/ml, was about 80 ng/ml in human milk and was undetectable in human amniotic fluid (<1.4 ng/ml). It was recently been suggested that human EGF is identical with human urogastrone. However, the tissue secreting this ''new'' human hormone and the role of hEGF in health and disease have yet to be determined

  15. Radioimmunoassay of thyrotropin concentrated from serum

    Energy Technology Data Exchange (ETDEWEB)

    Nisula, B.C.; Louvet, J.P.

    1978-05-01

    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 ..mu..U/ml. The mean of detectable serum hTSH levels in normal adult women (n = 11) was 1.54 +- 1.03 ..mu..U/ml (mean +- SD) and in normal men (n = 9) was 2.02 +- 1.15 ..mu..U/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.

  16. Radioimmunoassay of ADH in human urine

    International Nuclear Information System (INIS)

    A simple efficient procedure for extracting and concentrating arginine-8-vasopressin (AVP) from urine has been coupled with a specific and sensitive radioimmunoassay in order to measure antidiuretic hormone (ADH) excretion in normal human under various physiological stimuli. Antisera have been raised in rabbits injected with lysine-vasopressin (LVP) or AVP coupled with bovine serum albumin. The antiserum selected for the assay which inhibits the antidiuresis induced in the rat by AVP is used at a final dilution of 1 : 50,000 and possesses a high association constant of 1x1011 l.mol-1. The limit of detection of the RIA system is 0.5 μUI/ml of urine (1.25 pg). Urinary ADH has been extracted from urine by Miller and Moses method. Mean recovery of added vasopressin averaged 90.2%+-11 (SD) and assay of serial dilutions of such extracts showed that they behave in the assay system in the same way as synthetic AVP standards. Moreover comparison of the results obtained by the RIA to those given by the biological method using the ethanol anesthetized rat showed excellent correlation (r=0.9p<0.001). Under ad libitum fluid and food intake, mean daily urinary excretion of AVP (uncorrected for recovery) determined in 22 subjects was found to be 30.58+-11.64mU/h with no significant difference between men and women. In response to an oral waterload ADH became undetectable at the peak of diuresis. Following a 16 hr fluid deprivation, ADH rose moderately. A significant correlation has been found between urine osmolality and AVP excretion rate

  17. Radioimmunoassay of drugs: Phenytoin, clonidine and theophylline

    International Nuclear Information System (INIS)

    The development of radioimmunoassay procedures using 125I labelled tracers, for the drugs phenytoin, clonidine and theophylline is described. These drugs are conjugated to tyrosine methyl ester (TME) and bovine serum albumin (BSA) after suitable chemical modification, for preparation of radioiodinated drugs and antisera respectively. Phenytoin was modified to phenytoin valeric acid (P-VA) and phenytoin acetic acid (p-AA). In the case of clonidine, 4 carboxy clonidine (4-CLD) was used as the starting material, while for theophylline, 7-propionic acid (7-PAT) and 8-butyric acid (8-BAT) derivatives were made. Phenytoin and clonidine derivatives were coupled to BSA by the carbodiimide method. Theophylline derivatives were coupled to BSA by the mixed anhydride method. The drug-BSA conjugates were purified by dialysis and emulsified in complete Freund's adjuvant for immunization in rabbits. The different drug derivatives were coupled to TME using carbodiimide. The conjugates were purified by thin layer chromatography (TLC). Iodine-125 labelled tracers were prepared from the drug-TME derivatives using the chloramine-T method and purified by TLC or gel filtration. In phenytoin and theophylline good separation was obtained with TLC, while in clonidine, gel filtration was found to be better. Dextran coated charcoal was used as the separation system for clonidine, while polyethylene glycol was used for phenytoin and theophylline. Various factors such as time, temperature, concentration of reagents, serum interference and separation system were optimized to arrive at a suitable assay system for each of the drugs. The optimized assay systems were validated by precision profiles, interassay and intra-assay variations and specificity studies. These optimized systems have adequate sensitivity and can be used for the measurement of the drugs in serum without prior extraction of the samples. (author). 18 refs, 3 figs, 4 tabs

  18. Radioimmunoassays for protein A of Staphylococcus aureus

    International Nuclear Information System (INIS)

    Radioimmunoassays have been developed that can detect nanogram amounts of protein A (SpA), a product generated by Staphylococcus aureus that binds selectively to the Fc region of IgG from most mammalian species. Competition assays for fluid phase SpA utilize antibodies produced in chickens, 125I-labeled SpA as the tracer molecule, and either F(ab')2 fragments of rabbit IgG anti-chicken IgG or 40% ammonium sulfate as the precipitating agent to separate antigen-antibody complexes from free antigen. The double antibody assay could be carried out in serum from species that form only soluble complexes with SpA (e.g., rabbit), that react poorly with SpA (e.g., rat) or under appropriate conditions in serum from species (e.g., dog) that show high reactivity with SpA and form precipitating complexes. Chicken antibodies prepared by affinity chromatography on SpA-Sepharose and labeled with 125I were used in a direct binding assay for SpA present either on the cell wall of Cowan strain I or Wood 46 bacteria, in insoluble complexes prepared from SpA and whole serum or purified IgG, or in C1q binding complexes that were formed by passage of serum from normal or tumor bearing humans or dogs over SpA-collodion charcoal. Since both types of assays could detect SpA even in the presence of serum or IgG, they offer advantages over other techniques in which the SpA-Fc interaction may interfere. (Auth.)

  19. Estimation of antibodies to human cytomegalovirus by immunofluorescence and radioimmunoassay

    International Nuclear Information System (INIS)

    The 125I 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)

  20. Immunodiagnosis of opportunistic mycoses: detection of fungal antigenemia by radioimmunoassays in systemic candidiasis and aspergillosis

    International Nuclear Information System (INIS)

    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.)

  1. Detection of sputum eicosanoids in cystic fibrosis and in normal saliva by bioassay and radioimmunoassay.

    Science.gov (United States)

    Zakrzewski, J T; Barnes, N C; Piper, P J; Costello, J F

    1987-01-01

    We have measured arachidonic acid (AA) metabolites, leukotrienes (LTs) and prostanoids (Ps), in sputum of patients with cystic fibrosis (CF) and in normal saliva using bioassay and radioimmunoassay (RIA). Almost three times as much LTB4 is present in CF extracts compared with slow reacting substances (SRSs). Leukotrienes were not detected in normal saliva. In CF sputum there is a three-fold increase in the level of the vasodilator prostanoid prostaglandin E2 (PGE2) and the stable metabolite of prostacyclin, 6-oxo PGF1 alpha compared with the vasoconstrictor prostaglandin F2 alpha (PGF2 alpha) and thromboxane B2 (TxB2), a hydrolysis product of thromboxane A2. Experiments with BW755c (25 micrograms ml-1, n = 3) indicated that the majority of this activity was not produced during the extraction procedure. The detection of LTs and Ps in sputum of CF patients shows that these substances are present at biologically active concentrations and may contribute to the pathophysiology of this disease. PMID:3028454

  2. Radioimmunoassay for mono-(2-ethylhexyl) phthalate in unextracted plasma

    International Nuclear Information System (INIS)

    We report a radioimmunoassay for mono-(2-ethylhexyl) phthalate that has been coupled to a protein carrier as a radioligand. Competitive interference tests with a variety of related compounds indicated the assay to be highly specific. Quantitative comparison of mono-(2-ethylhexyl) phthalate values in plasma and serum samples between the radioimmunoassay and gas chromatographic procedures indicated a high reliability. Because this potentially toxic compound can leach into plasma from polyvinyl plastics, this assay should be particularly useful for those involved in the manufacture or use of medical devices made of them

  3. A new radioimmunoassay of thyrotropin-releasing hormone

    International Nuclear Information System (INIS)

    An approach entirly different to previous radioimmunoassays for the measurement of thyrotropin-releasing hormone has been the introduction of an amide bond between pGlu-His-Pro-OH and NH2-groups of a protein carrier resulting in the formation of a TRH-like structure, i.e., pGlu-His-Pro-NH-protein. The attachment of TRH to hemocyanin (HC) using the bifunctional reagent 1,5-difluoro-2,4-dinitrobenzene (DFDNB) is reported here. Administration of the conjugate to rabbits elicited the production of antisera, which were used in the radioimmunoassay of the hypothalamic hormone

  4. Radioimmunoassay of methotrexate: use of 75Se-labelled methotrexate

    International Nuclear Information System (INIS)

    75Se labelled methotrexate has been used as the radiolabel in a radioimmunoassay for methotrexate and its use compared with that of 3H- methotrexate. The gamma-labelled drug has proved to be extremely reliable and easier to use than 3H- methotrexate. Results obtained with it compare well with those obtained with tritiated drug. There are practical advantages in using a gamma-emitter, and it is suggested that the modified radioimmunoassay is invaluable for routine monitoring of methotrexate in patients undergoing chemotherapy with the drug. (author)

  5. A double antibody radioimmunoassay specific for placental alkaline phosphatase

    International Nuclear Information System (INIS)

    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.)

  6. A sensitive radioimmunoassay for a component of mouse casein

    International Nuclear Information System (INIS)

    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 [125I]-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

  7. Logistics of national radioimmunoassay services and radioimmunoassay trouble shooting in developing countries

    International Nuclear Information System (INIS)

    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)

  8. Development of T3 radioimmunoassay kit

    International Nuclear Information System (INIS)

    Thyroxine T4 radioimmunoassay (RIA) kit has already been developed in this laboratory. For an efficient diagnosis of thyroid disease, however, it is well known that the T3 RIA should also be carried out in addition to the T4 RIA. Accordingly, the development of T3 RIA kit was urgently desired to match the T4 kit and to hold a sound domestic supply systems. The high specific activity T3125I about 3,000 μCi/μg T3) could be obtained by radioiodinating T2 with chloramine-T, and the labelled product could be stahilized. In the preparation of T3 free serum, charcoal was eliminated easily from serum by high speed centrifugation, and the resulting T3 free serum was used for the preparation of T3 standard serum solutions. RIA buffer system could be improved with the use of 0.025M barbital buffer, pH 8.2, containing 0.1% BSA, 0.5% bovine aamma globulin and 0.02% merthiolate. Antibody titer was increased threefold by using the 0.025M barbital buffer; the titer was 8,000: 1 in the 0.078M borbital buffer, pH 8.6, containing 0.1% BSA and 0.1% NaN3 while it was 26,000 : 1 in the above described 0.025M barbital buffer. The modified buffer system was also efficient for the use in T4 RIA since it increased the T4 antibody titer twofold. When the same buffer system was used in T3 RIA, no significant difference was observed between the use of HSA and of BSA in so far as 0.5% bovine gamma globulin was added to the buffer, contradicting those in the reference. The resalts indicated that the cost for the preparation of both kits can be saved. Quality guaranteed kits could be prepared by careful control of the assay values in comparing with those of the reference control sera. In consequence, there is not any technical difficulty in routine production. (Author)

  9. Clinical application of radioimmunoassay of dehydroepiandrosterone sulfate (DHA-S)

    International Nuclear Information System (INIS)

    A radioimmunoassay was developed for the measurement of dehydroepiandrosterone Sulfate (DHA-S) in serum. Normal values for the different age groups were determined in 146 normal individuals. Patients with different adrenal and hypophyseal diseases were also investigated and the data on DHA-S levels are presented. (Author)

  10. Specific and direct radioimmunoassay for human proinsulin in serum

    International Nuclear Information System (INIS)

    A routine radioimmunoassay for human proinsulin in serum has been developed. The reagents used were: antibodies against the C-peptide part of the proinsulin molecule, human proinsulin as the standard and 125I-labelled sythetic human Tyr-C-peptide as the tracer. (orig.)

  11. Development of a radioimmunoassay of tetracycline and its derivatives

    International Nuclear Information System (INIS)

    A radioimmunoassay for tetracycline was developed in the context of the present work. The determination of tetracycline content in biological samples is to be made valid using tetracycline RIA. As well as the carbodiimide method, a by radioimmunologists extremely seldomly used way involving a condensation reaction between protein and haptene via the Mannich reaction was successfully applied. Antibodies were produced using a conventional immunisation method after Abraham, the other applied alternative method after Vaitukaitis et al. The general working methods had to be adapted to the tetracycline RIA. All variable parameters of the antigen-antibody bond were tested to optimize the incubation conditions of the system as well as to control the tracer for a degradation. The detection limit of RIA is 10-12, the measuring range from 10-12 to 10-10 mol for tetracycline hydrochloride and rolitetracycline respectively and up to 10-9 mol for its derivates. The investigations for cross reactions showed a high specificity for tetracycline (100%) and its intravenously appliable pyrrolidino-methyl derivative rolitetracycline (88%). Standard curves could be drawn up using either of the two compounds tetracycline and rolitetracycline as standard. The pharmaco-kinetic behaviour of the parenteral administrable tetracycline was analyzed as example for the possible applications of the tetracycline radioimmunoassay. Parallel animal tests administring 3H tetracycline hydrochloride were performed for radioimmunoassay reference. The possible application of tetracycline radioimmunoassay in food analysis is discussed. (orig./MG)

  12. Direct salivary cortisol radio-immunoassay determination. Clinical applications

    International Nuclear Information System (INIS)

    Salivary cortisol levels reflect the biologically active free fraction of blood cortisol. The authors describe the results obtained with the aim of a radio-immunoassay commercial serum cortisol kit, without prealable extraction in different physiological and pathological situations. Salivary cortisol determination appears performant both in nycthemeral studies and in stimulation or freination tests

  13. Effect of proteases on the β-thromboglobulin radioimmunoassay

    International Nuclear Information System (INIS)

    Rat peritoneal mast cells and mast cell granules were evaluated by radioimmunoassay for the presence of β-thromboglobulin and platelet factor 4. The initial assays indicated that a β-thromboglobulin cross reacting material was released from mast cells by compound 48/80 in a similar dose-dependent manner as histamine release. The material was also found to be associated with purified granules. However, the use of protease inhibitors in the buffers completely abolished the positive assays. Further evaluation of the effects of various proteases on the β-thromboglobulin assay indicated that elastase would also generate a false positive assay which could then be neutralized by the use of α1-antitrypsin as a protease inhibitor. There was no protease effect on the platelet factor 4 radioimmunoassay which always showed no detectable amounts with mast cells, granules or proteases. These results clearly indicate the artifactual positive assays which can arise when using certain radioimmunoassay tests in the presence of cell proteases. The use of protease inhibitors is a necessary control when applying a radioimmunoassay to a system with potentially active proteases. 24 references, 2 figures, 4 tables

  14. Double-antibody radioimmunoassay for staphylococcal enterotoxin C2

    International Nuclear Information System (INIS)

    A sensitive double-antibody radioimmunoassay for staphylococcal enterotoxin C2 is described. The assay procedure employs anti-rabbit gamma globulin, prepared in goats, to precipitate the antigen-antibody complex of enterotoxin C2 and anti-enterotoxin C2. The test is sensitive to 100 pg of enterotoxin

  15. Development of a specific radioimmunoassay for the detection of clenbuterol residues in treated cattle

    Energy Technology Data Exchange (ETDEWEB)

    Delahaut, Ph.; Dubois, M. (Lab. d' hormonologie, Marloie (Belgium)); Pri-Bar, I.; Buchman, O. (Israel Atomic Energy Commission, Beersheba (Israel). Nuclear Research Center-Negev); Degand, G.; Ectors, F. (Liege Univ. (Belgium))

    A radioimmunoassay for clenbuterol detection in cattle has been validated and used to monitor treated cattle. The tracer used was 4-amino-3,5-dichloro-{alpha}(tert-butylaminomethyl) benzyl alcohol (benzyl-{sup 3}H)(clenbuterol) prepared by catalytic tritiation with tritium gas of 4-amino-3,5-dibromo-{alpha}-(tert-butylamino)-acetophenone, followed by chlorination at positions 3 and 5 in the aromatic ring. The rabbit antiserum was raised against a diazotized clenbuterol/human serum albumin conjugate. The assay described was sensitive (7.8 pg/tube) and reproducible. The intra- and inter-assay variability, which was assessed by measuring known quantities of clenbuterol in plasma, urine and faeces, was satisfactory for RIA. When this assay was used to monitor treated cattle the concentrations of clenbuterol in plasma, urine and faeces were directly related to the administered dose. The absorption and elimination of clenbuterol in cattle was rapid. Data obtained were consistent with results obtained in other species where a rapid clearance rate was also demonstrated. (author).

  16. Radioimmunoassay of steroids in homogenates and subcellular fractions of testicular tissue

    International Nuclear Information System (INIS)

    Radioimmunoassays for testosterone (T), dihydrotestosterone (DHT) and 5alpha-androstan-3alpha, 17beta-diol (DIOL) in homogenates of whole testis, interstitial tissue and seminiferous tubules as well as subcellular fractions of the latter were developed. Steroids were extracted with acetone, submitted to several solvent partitions and isolated by a celite: propylene glycol: ethylene glycol column chromatography. Anit-T serum was used for the assay of T and DTH, and a specific anti-Diol serum for DIOL. Subcellular fractions were separated by differential centrifugation. The nuclear fraction was purified by centrifugation in a dense sucrose buffer followed by several washings. Losses were corrected according to recovery of DNA. Optimal conditions for purification of acetone extracts at minimal losses were established. Validation of the method was studied testing linear regression of logit-log transformations of standard curves and parallelism with unknowns. T was the steroid present in higher concentrations in all samples studied. It is concluded that the present method for determination of endogenous androgen concentrations in testicular tissue is valid and might be useful in studing testicular function. (orig.)

  17. Gene Cloning of Murine α-Fetoprotein Gene and Construction of Its Eukaryotic Expression Vector and Expression in CHO Cells

    Institute of Scientific and Technical Information of China (English)

    易继林; 田耕

    2003-01-01

    To clone the murine α-fetoprotein (AFP) gene, construct the eukaryotic expression vector of AFP and express in CHO cells, total RNA were extracted from Hepa 1-6 cells, and then the murine α-fetoprotein gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant of vector was identified by restriction enzyme analysis and sequencing. A fter transient transfection of CHO cells with the vector, Western blotting was used to detect the expression of AFP. It is concluded that the 1.8kb murine α-fetoprotein gene was successfully cloned and its eukaryotic expression vector was successfully constructed.

  18. Is inconsistency of α-fetoprotein level a good prognosticator for hepatocellular carcinoma recurrence?

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    AIM: To identify the clinical outcomes of hepato-cellular carcinoma (HCC) patients with inconsistent α-fetoprotein (AFP) levels which were initially high and then low at recurrence.METHODS: We retrospectively included 178 patients who underwent liver resection with high preoperative AFP levels (≥ 200 ng/dL). Sixty-nine HCC patients had recurrence during follow-up and were grouped by their AFP levels at recurrence: group Ⅰ, AFP ≤ 20 ng/dL (n = 16); group Ⅱ, AFP 20-200 ng/dL (n = 24); and group Ⅲ, AFP ≥ 200 n...

  19. Coefficient Alpha

    OpenAIRE

    Panayiotis Panayides

    2013-01-01

    Heavy reliance on Cronbach’s alpha has been standard practice in many validation studies. However, there seem to be two misconceptions about the interpretation of alpha. First, alpha is mistakenly considered as an indication of unidimensionality and second, that the higher the value of alpha the better. The aim of this study is to clarify these misconceptions with the use of real data from the educational setting. Results showed that high alpha values can be obtained in multidimensional scale...

  20. Overcoming the radiological and legislative obstacles in radioimmunoassay

    International Nuclear Information System (INIS)

    The safety of radioimmunoassays especially the safety of protein iodination with 125I is discussed. Two types of users, namely the research worker and the routine user in the clinical laboratory, should be regarded. The possible hazards of the so-called 'do-it-yourself' iodinations are described and compared with those of routine use of commercial kits and reagents. As there is no significant hazard involved in radioimmunoassay in clinical chemistry laboratories using commercially supplied reagents, there should be, to the author's opinion, no significant legislative difficulties. The interpretation of the ICRP recommendations and the practice of legislation in the USA and Europe, especially the United Kingdom and teh Federal Republic of Germany is described. (MG)

  1. Homologous radioimmunoassay for guinea pig corticosteroid-binding globulin

    International Nuclear Information System (INIS)

    A rapid, specific, and sensitive (requiring only 20 fmole of antigen equivalent to 0.007) μl of serum) radioimmunoassay (RIA) was developed for the measurement of guinea pig corticosteroid-binding globulin (CBG). CBG was purified to homogeneity from guinea pig serum by affinity chromatography and used for immunization, as the standard and as the radiolabeled trace in the RIA. The antiserum to CBG was raised in rabbits. It was judged specific by immunoelectrophoresis and by comparison of RIA values with steroid-binding assay profiles obtained on serum separated on the basis of size and ion-exchange properties. The results of the radioimmunoassays agree with those of a steroid-binding assay run on identical samples. The sensitivity of the assay allows detection of CBG in serial serum samples, other biologic fluids such as milk, and cell culture supernatants

  2. Measurements of urinary kinins by HPLC-radioimmunoassay

    International Nuclear Information System (INIS)

    In this paper the authors describe a method for the individual determination of urinary kinins. Extraction from the urine is performed on an Amberlite CG-50 column and kinins are eluted with formic acid. The samples are further purified and kinins are separated by reversed phase HPLC. Bradykinin and lysylbradykinin are quantified by a sensitive radioimmunoassay capable of detecting 0.1 fmol of either peptide. Procedural losses are monitored by measuring the recovery of [3H]bradykinin and [3H]lysylbradykinin. Simple methods for labeling of bradykinin and lysylbradykinin with tritium are also presented. Recoveries of [3H]bradykinin and [3H]lysylbradykinin from biological material ranged between 77 and 91%. The combination of HPLC with radioimmunoassay makes it possible to determine kinin concentrations of biological samples with a higher sensitivity and greater specificity than previous methods. (Auth.)

  3. Detection of gonococcal antigens in urine by radioimmunoassay

    International Nuclear Information System (INIS)

    A method of detecting gonococcal antigens by solid-phase radioimmunoassay with radioactively labelled antibody is described. A specificity test has been developed that enables this method to be used to detect gonococcal antigens in urine sediments. When sediments from samples of urine from male patients with gonorrhoea were tested, 31 (74%) of 42 gave positive results, clearly distinguishing them from sediments from urine samples from men with non-specific urethritis, none of which was positive. Ten of 14 urine sediments from urine samples from women with gonorrhoea gave positive results, as did 3 of 18 sediments from urine samples from women patients without gonorrhoea.These experiments demonstrate that gonococcal antigens can be detected in urine by radioimmunoassay; the method could be useful in diagnosis if, after refinement, its sensitivity and specificity were to be increased. (author)

  4. Radioimmunoassay for nortriptyline (and other tricyclic antidepressants) in plasma

    International Nuclear Information System (INIS)

    The radioimmunoassay for nortriptyline described here can detect as little 1 μg/liter of plasma. Within-day precision and day-to-day precision (CV) were +-6 and +-11%, respectively, over the concentration range 100 to 200 μg/liter. The major metabolite hydroxy-nortriptyline does not cross react with the antiserum. Results so obtained correlate closely with results by a double-isotope derivative dilution technique. The major advantages of this technique over currently available methods are its sensitivity, convenience (many samples can be processed in one day), simplicity, and cost. Further, prior extraction of plasma samples is not required. Cross-reactivity studies have been carried out with all other available tricyclic antidepressants. The antiserum has the ability to bind these drugs, thus radioimmunoassay for all the tricyclic antidepressant drugs can be set up because concurrent use of more than one of these drugs is rare

  5. Radioimmunoassay of steroids in Czechoslovakia - current state and prospects

    International Nuclear Information System (INIS)

    The experience is summarized with radioimmunoassay of steroids during the last 15 years, and the expected trends in this field are dealt with. The following problems connected with the radioimmunoassay of steroids are discussed: 1) ways leading to improved antisera specificity (choice of immunogen, use of the principle of immunological tolerance, use of antisera mixtures, preparation of monoclonal antibodies); 2) methods of ligand labelling (steroids labelled in the ring, radioiodine labelled derivative); 3) separation techniques of bound and free ligands; 4) sample preparation with regard to automation and 5) importance of and approaches to the determination of free, plasma protein non-bound steroids. The survey is complemented with a list of antisera to steroids, prepared in the laboratories of the authors. (author)

  6. Development of a radioimmunoassay for pig pancreatic kallikrein

    International Nuclear Information System (INIS)

    A radioimmunoassay for the determination of pig pancreatic kallikrein was developed. The chloramine-T method was employed for the labelling of the antigen with 125I. The assay allows the determination of kallikrein in concentrations as low as 0.4 μg/l. Pig urinary and pig submandibular kallikreins are indistinguishable from pig pancreatic kallikrein by the assay. No cross reactivity was observed for bovine trypsin and chymotrypsin, porcine trypsin and kallikreins of guinea pig submandibular glands and guinea pig coagulation glands. Because of the high specificity of the assay, which is not attainable with conventional assays based on the enzymatic activity, the radioimmunoassay is highly suited for investigations into the physiological role and the pharmacological mechanism of action of pig glandular kallikreins. (orig.)

  7. Development and applications of a radioimmunoassay to human plasma kallikrein

    International Nuclear Information System (INIS)

    A radioimmunoassay to human plasma kallikrein (Hu PK) was developed using specific antibodies against the purified activates enzyme. The antiserum showed identity in the precipitation, double immuno-diffusion, immobility, immuno-electrophoresis, using as normal plasma as purified kallikrein. The protein 125 I-kallikrein was prepared by chloramine-t method and presented homogeneity, stability, immunoactivity and high specific activity, so that a sensitive assay was assured. 193 refs., 19 figs., 4 tabs

  8. Development and validation of a radioimmunoassay for thyrotropin in cattle

    OpenAIRE

    Guyot, Hugues; Sulon, Joseph; Beckers, Jean-François; Closset, Jean; Lebreton, Pascal; Alves de Oliveira, Laurent; Rollin, Frédéric

    2007-01-01

    In mammals, thyrotropin, or thyroid-stimulating hormone (TSH), assay is used for the diagnosis of primary hypothyroidism. Hypothyroidism is the most common type of thyroid disorder in cattle. The aim of this study was to develop and validate, under physiologic and pathologic conditions, a radioimmunoassay (RIA) for bovine TSH (bTSH). Double RIA was performed with purified bTSH and specific bovine antiserum. Laboratory validation included research of minimal detection limit, accuracy, and repr...

  9. Enkephalin: radioimmunoassay and radioreceptor assay in morphine dependent rats

    International Nuclear Information System (INIS)

    A sensitive and specific radioimmunoassay for enkephalins is described which discriminates between leucine-enkephalin and methionine-enkephalin. Total opioid peptide activity was assayed by the ability of brain extracts to compete for opiate receptor binding. In rats treated with morphine and/or naloxone, opiates were separated from opioid peptides by Dowex AG-50W column chromatography prior to radioreceptor assay. Chronic administration of morphine failed to alter immunoreactive enkephalin levels or total opioid activity in radioreceptor assays

  10. Typing of murine cell-surface antigens by cellular radioimmunoassay

    International Nuclear Information System (INIS)

    A cellular radioimmunoassay utilizing 125I-labelled Protein A was used for detecting antigen-antibody complexes on gultaraldehyde fixed cells attached to microtiter plates. This method is rapid, sensitive and specific for revealing H-2 private and public specificities as well as Ia and Lyt antigens. As plates may be kept for months, several reactivities can be tested in one step on a large panel rendering a regular supply of animals unnecessary. (Auth.)

  11. Semiautomated radioimmunoassay for mass screening of drugs of abuse

    International Nuclear Information System (INIS)

    A rapid, semiautomated radioimmunoassay system for detection of morphine, barbiturates, and amphetamines is described. The assays are applicable to large drug abuse screening programs. The heart of the system is the automatic pipetting station which can accomplish 600 pipetting operations per hour. The method uses 15 to 30 μl for the amphetamine and combined morphine/barbiturate assays. A number of other drugs were tested for interference with the assays and the results are discussed

  12. Radioimmunoassay for human placental alkaline phosphatase and clinical significance

    International Nuclear Information System (INIS)

    A radioimmunoassay specific for placental alkaline phosphatase (PALP) has been performed. Sera from blood donnors contain less than 15 μg of PALP per liter. The amounts of PALP found in sera of pregnant women are higher, as soon as the first trimester of the pregnancy, increasing untill delivery (50-600 μg of PALP/l). Only 3,5% of the patients with various cancer diseases have amounts higher than 25 μg PALP/l

  13. Measurement of trypsin in duodenal juice by radioimmunoassay.

    OpenAIRE

    Lake-Bakaar, G; McKavanagh, S; Rubio, C E; Epstein, O; Summerfield, J A

    1980-01-01

    Trypsin in duodenal aspirate and pure pancreatic juice samples has been measured by both radioimmunoassay and enzymatic mathods. The radioimmunossay has been shown to be specific and to detect trypsin in the presence of aprotinin (Trasylol). In duodenal juice samples from control subjects and from patients with primary biliary cirrhosis a good correlation was obtained between both immunoreactive trypsin concentration and trypsin activity. The immunoreactive trypsinogen concentration in pure p...

  14. The cooperation of radioimmunoassays - a survey of experience

    International Nuclear Information System (INIS)

    Exceptional methods of determinations of hormones, tumor promoters and other parameters of laboratories with the radioimmunoassay lead to increasing costs. For this reason in a test model through cooperation of 3 nuclear medicine institutes rarely performed methods of examinations were made. It is a question of the determination of calcitonin, peptide hormones, ferritin, immunoglobulines. The examination shows the necessity of cooperation in the field of the laboratory diagnostics of more rare methods of examination. (eva)

  15. Antibody coated tubes in T3 - solid phase radioimmunoassay

    International Nuclear Information System (INIS)

    The aim of this study was to develop a simple and inexpensive form of solid phase radioimmunoassay of T3 (3,5,3'-L- triiodothyronine); for the preparation of solid phase, the adsorption of anti-T3 antibodies to polystyrene tubes has been used. The polystyrene tubes were used without washing or other treatment; each tube was coated by addition of an uniform volume (175 μl) of diluted antiserum of moderately high titer. Antiserum dilution was 1:3000 and the optimal pH of buffer solution was 8.4 - 8.6. The best results were achieved with an exposure time to antiserum of 40 h at 4 deg C. The antibody - coated tubes prepared in this way were verified by using them to the radioimmunoassay of T3. The results obtained with the above mentioned solid phase in T3-RIA of three level control serums were found to be successful for setting up T3- solid phase radioimmunoassay of high precision. (authors)

  16. Simultaneous detection of morphine and barbiturates in urine by radioimmunoassay

    International Nuclear Information System (INIS)

    This report describes a radioimmunoassay for the simultaneous detection of morphine and barbiturates. Morphine and barbiturate antibodies, obtained from goats, were mixed with 125I-labeled antigens. By adjusting concentrations of the morphine and barbiturate antibodies and radiolabeled antigens, closely superimposed standard curves for the two drugs would be obtained. As a consequence, similar response curves were obtained for urine specimens containing morphine or barbiturates. Although concentrations as low as 25 μg/liter could be measured, to ensure against false positive reactions the test should be performed at the 100 μg/liter concentration. Unknown samples positive by the dual assay were confirmed by separately testing the specimens with the individual radioimmunoassay specific for morphine or barbiturate. Equivalency tests of urines positive for morphine, positive for barbiturates, or negative for both demonstrated complete correlation between the single and dual assays. The mixed reagent retained its sensitivity and specificity for at least three months when stored at 4 or 250C. The dual radioimmunoassay is a rapid, simple procedure that can be adapted to automated processes and that is suitable for large- and small-scale screening

  17. Diagnostic value of prostatic acid phosphatase as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Serum concentrations of prostatic acid phosphatase (PAP) were determined with 4 different radioimmunoassays and with the standard enzymatic method (p-nitrophenylphosphate) in 35 patients with prostatic carcinoma. Staging of localized tumors was based on histopathological evaluation after radial prostatectomy and pelvic lymphnode dissection (pTsub(1-3), pN0). In tumor lesions Tsub(1-2) N0 M0 elevated PAP-serum concentrations were found by RIA-determination in only one patient. Increased PAP serum levels were observed in 43-78% of carcinomas stage T3 N0 M0 and in 54-83% in stage Tsub(2-4) Nsub(x) M1 tumors, depending on the test kit used for the PAP determination. Concentrations for PAP obtained with the 4 different RIA-kits used, varied significantly and thus are not comparable. No false positive results were observed in sera of 9 patients with benign prostatic hyperplasia. Elevated PAP serum levels were found in a significantly higher frequency when determined by radioimmunoassay than by the enzymatic method. The results clearly indicate, that PAP is of no value for early recognition of carcinoma of the prostate even when measured by radioimmunoassay. However, the RIA-method seems to be of clinical importance in estimating the course of advanced local and metastasizing carcinoma of the prostate. (orig.)

  18. Thrittene radioimmunoassay. Description and application of a novel method

    International Nuclear Information System (INIS)

    In the present paper the development and application of a novel thrittene radioimmunoassay (RIA) are described. 125I-labeling of Tyr(0)-thrittene was performed by the iodogen-method and the mono-iodinated peptide, as RIA tracer, was separated by reversed-phase high performance liquid chromatography (HPLC). The RIA results show that the antiserum used in the radioimmunoassay turned to be C-terminal specific, without significant affinity to other members of the somatostatin peptide hormone family. Detection limit of the assay was 0.2 fmol/ml. This highly specific and sensitive thrittene RIA was used to investigate the distribution of thrittene in the rat gastrointestinal tract and other tissue samples. Different areas of the gastrointestinal tract and other tissues were removed from rats and after extraction the samples were processed for thrittene radioimmunoassay. Highest concentrations were found in the duodenum samples followed by jejunum and ileum, however, all the examined tissues contained highly enough thrittene for the measurement. (author)

  19. The Predictive Power of Serum α-Fetoprotein and Des-γ-Carboxy Prothrombin for Survival Varies by Tumor Size in Hepatocellular Carcinoma.

    Science.gov (United States)

    Tsugawa, Daisuke; Fukumoto, Takumi; Kido, Masahiro; Takebe, Atsushi; Tanaka, Motofumi; Kuramitsu, Kaori; Matsumoto, Ippei; Ajiki, Tetsuo; Koyama, Tatsuki; Ku, Yonson

    2016-01-01

    Alpha-fetoprotein (AFP) and des-γ-carboxy prothrombin (DCP) are frequently used as tumor markers in hepatocellular carcinoma (HCC). The authors hypothesized different patient populations with varying tumor sizes would influence the predictive power of tumor markers for survival in HCC patients. The authors investigated the influence of tumor size on predictive powers of AFP and DCP. 181 patients underwent hepatectomy for HCC from 2003 to 2008 at Kobe University Hospital. Tumor markers were measured before and at 1 month post-hepatectomy. The Cox proportional-hazards model revealed that preoperative serum AFP was associated with survival; its effects depended on tumor size. Hazard ratios (HRs) for preoperative AFP were maximum for medium-sized HCC, and for DCP, HRs were maximum in small-sized tumors. Post-hepatectomy, both tumor markers were associated with survival, revealing significant interactions with tumor size. HRs for postoperative AFP were greater than 1 for relatively wide range tumors (3-11 cm). HRs for postoperative DCP increased with tumor size, with a strong prognostic predictive power for tumors >5 cm. The predictive power of serum tumor markers varied by tumor size in HCC patients. By selecting the appropriate tumor marker, its predictive power can be improved. PMID:27363395

  20. Alpha-amidated peptides derived from pro-opiomelanocortin in normal human pituitary

    DEFF Research Database (Denmark)

    Fenger, M; Johnsen, A H

    1988-01-01

    Normal human pituitaries were extracted in boiling water and acetic acid, and the alpha-amidated peptide products of pro-opiomelanocortin (POMC), alpha-melanocyte-stimulating hormone (alpha MSH), gamma-melanocyte-stimulating hormone (gamma 1MSH), and amidated hinge peptide (HP-N), as well as their...... glycine-extended precursors, were characterized by sequence-specific radioimmunoassays, gel-chromatography, h.p.l.c. and amino acid sequencing. alpha MSH and gamma 1MSH constituted 0.27-1.32% and 0.10-5.10%, respectively, of the POMC-derived products [calculated as the sum of adrenocorticotropic hormone...

  1. Detection and purification of rat and goat immunoglobulin G antibodies using protein G-based solid phase radioimmunoassays

    International Nuclear Information System (INIS)

    Using the newly described streptococcal surface protein, protein G, which has powerful immunoglobulin G binding properties, solid-phase radioimmunoassays were developed for the quantitation of goat and rat immunoglobulin G bound to the plastic surface of microtiter plates. The binding of goat immunoglobulin G to the surface via a specific antigen (guinea pig alpha1-microglobulin) permitted the determination of antigen-specific antibodies with a detection limit of 50-100 ng. Optimum assay conditions were established and the whole assay procedure could be brought to completion at room temperature in less than a working day. The value of the assays was illustrated by monitoring rat and goat immunoglobulin G antibodies during their purification from whole sera by classical chromatographic procedures. (Auth.)

  2. Association of liver inflammation with alpha-fetoprotein and treatment response in hepatitis C virus genotype 4 patients

    OpenAIRE

    Samar Samir Youssef; Sameh Mohamed Seif

    2014-01-01

    Objectives: Chronic hepatitis C (CHC) or liver inflammation resulting from infection with hepatitis C virus (HCV) is the cause of chronic liver disease and leads to cirrhosis and hepatocellular carcinoma. The burden of chronic HCV-related liver disease in Egypt continues to rise and the interaction of liver inflammation with biomarkers and response to therapy is scarcely discussed. Moreover, serum alanine transaminase (ALT) is considered as a moderately accurate test for indicating liver infl...

  3. Gastric choriocarcinoma admixed with an α-fetoprotein-producing adenocarcinoma and separated adenocarcinoma

    Institute of Scientific and Technical Information of China (English)

    Bang Wool Eom; So-Youn Jung; Hongman Yoon; Myeong-Cherl Kook; Keun Won Ryu; Jun Ho Lee; Young-Woo Kim

    2009-01-01

    We report a case of gastric choriocarcinoma admixed with an α-fetoprotein (AFP)-producing adenocarcinoma .A 70-year-old man was hospitalized for gastric cancer that was detected during screening by esophagogastroduodenoscopy (EGD). Initial laboratory data showed the increased serum level of AFP and EGD revealed a 5-cm ulcerofungating mass in the greater curvature of the gastric antrum.The patient underwent radical subtotal gastrectomy with D2 lymph node dissection and Billroth Ⅱ gastrojejunostomy. Histopathological evaluation confirmed double primary gastric cancer: gastric choriocarcinoma admixed with an AFP-producing adenocarcinoma and separated adenocarcinoma. At 2 wk postoperatively, his human chorionic gonadotropin and AFP levels had reduced and six cycles of adjuvant chemotherapy were initiated. No recurrence or distant metastasis was observed at 4 years postoperatively.

  4. 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 with...... ASD and 243 control subjects were included in a case–control study using a historic birth cohort maintained at Statens Serum Institute. Measurements of MS-AFP were obtained from a multicentre screening program, whereas clinical data were obtained from nationwide registers. Association between MS-AFP...... and ASD status was analyzed using a logistic regression model and nonparametric tests. Results: Crude, but not adjusted estimates, showed that MS-AFP levels were slightly, but significantly, higher in mothers of children with ASD, compared with their control subject counterparts. People with ASD had...

  5. Misdiagnosis of an α-fetoprotein-producing esophageal carcinoma: A case report and literature review

    Science.gov (United States)

    SUN, NINGBO; YIN, XUNLU; ZHONG, YUREN; ZHANG, XIAOTIAN; XIE, YAN; MENG, XIANGFANG; ZANG, QI

    2016-01-01

    α-fetoprotein (AFP)-producing esophageal carcinoma is a rare type of esophageal cancer, with its characteristics not yet fully clarified. In the present study, a case of esophageal carcinoma was misdiagnosed as an AFP-producing esophageal carcinoma. The patient was a 50-year-old woman who was referred to Qianfoshan Hospital Affiliated to Shandong University in November 2014 with a 3-month history of progressive dysphagia. A chest computed tomography (CT) scan showed thickening of the wall of the esophagus, corresponding regions of luminal stenosis and massive lymph node swelling around the lesser curvature of the esophagus. A laboratory investigation showed that the serum AFP levels of the patient were elevated to 18.97 ng/ml (normal range <12 ng/ml). These laboratory investigation findings combined with the aforementioned pathological diagnosis supported a diagnosis of AFP-producing esophageal carcinoma. An abdominal ultrasound was performed and a cystic low-density measuring 5×4 mm was identified. No metastases were revealed in the liver. The boundary of the focal low density was clear, which indicated a clinical diagnosis of liver cyst. A radical esophagectomy was performed on December 5, 2014. Microscopically, the tumor was a moderately differentiated squamous cell carcinoma invading the serous layer, with no hepatoid features. Immunohistochemistry showed that the cells were diffusely negative for AFP expression. Histopathological examination revealed the absence of hepatoid features. According to these findings, the tumor was diagnosed as a moderately differentiated squamous cell carcinoma. In the present study, the case of a patient with squamous cell carcinoma that was misdiagnosed as an α-fetoprotein-producing esophageal carcinoma was reported, with a review of the literature.

  6. Radioimmunoassay in the evaluation of the hormonal activity of hypophyseal adenoma

    Energy Technology Data Exchange (ETDEWEB)

    Snigireva, R.Ya.; Kasumova, S.Yu.; Barabanov, V.M. (Akademiya Meditsinskikh Nauk SSSR, Moscow. Inst. Nejrokhirurgii; Akademiya Meditsinskikh Nauk SSSR, Moscow. Inst. Morfologii Cheloveka)

    1983-08-01

    Direct correlation was found between the results of a radioimmunoassay electron microscopy and immunohistochemistry in cases of STH-secreting and hormonally inactive hypophyseal adenomas. In hyperprolactinemia and a simultaneous increase in the levels of 2 or more hormones the results of the radioimmunoassay coincided with those of electron microscopy and immunohistochemistry in 50-70% of the cases.

  7. Prediction of ovulation in women using a rapid progesterone radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Fleming, R.; Coults, J.R.T. (Glasgow Univ. (UK))

    1982-02-01

    A rapid (3-h) radioimmunoassay of plasma progesterone has been developed and used successfully to predict the time of ovulation in women undergoing artificial insemination. The results obtained using progesterone levels to date the stage of the cycle were analysed retrospectively by (1) estimation of the length of the ensuing luteal phases and comparison of these with luteal phase lengths of a control group (2) comparison of the dating using progesterone levels with retrospective determination of LH values and (3) by analysis of the dating in cycles in which conception occurred.

  8. Radioimmunoassay for determining the digoxin content of a sample

    International Nuclear Information System (INIS)

    A competitive radioimmunoassay for the measurement of digoxin in a serum sample in which the nonradioactive digoxin competes with a constant amount of a radioactively labeled digoxin tyramine analog comprising 3-O-(4-Hydroxyphenethylcarbamoyl) digoxigenin for binding cites on a limited amount of digoxin antibody. The percentage of the radioactively labeled digoxin which is bound to the antibody is inversely proportional to the concentration of digoxin in the serum sample. The concentration of digoxin in the serum is determined by the comparison with standards containing measured amounts of unlabeled digoxin. 7 claims, 1 drawing figure

  9. Investigations of serum HPL during pregnancy using two different radioimmunoassays

    International Nuclear Information System (INIS)

    The interassay investigations showed that it is absolutely necessary to standardize the HPL antisera as well as the standard sera, as it is otherwise impossible to compare and interpret the findings of different HPL radioimmunoassays. The investigations have shown that in addition to conventional clinical examinations and laboratory test methods (urine estriol determination, DHEAS-dehydroepiandrosterone sulphate test-, urine pregnandiol determination, and determination of heat-resisting alkaline serum phosphatase), HPL concentration determination is a parameter of the nutritive function of the placenta. (orig.)

  10. Radioimmunoassay of CRF-like material in rat hypothalamus

    International Nuclear Information System (INIS)

    Corticotropin releasing factor (CRF) was recently isolated from ovine hypothalami by its ability to stimulate adrenocorticotropin (ACTH) and beta-endorphin release from dispersed rat pituitary cells. In order to study the physiology of this peptide, we have developed a sensitive and specific radioimmunoassay (RIA) for CRF. Synthetic CRF was conjugated to bovine thyroglobulin and emulsified with complete Freund's adjuvant. A suitable antiserum was obtained which showed no crossreactivity with eight naturally occurring peptides. N-Tyr-CRF was iodinated and used as tracer. With this assay, CRF-like immunoreactivity which coeluted with ovine CRF on Sephadex G50 was detected in rat hypothalami

  11. Determination of chloramphenicol (CAP) residues in trout by radioimmunoassay

    International Nuclear Information System (INIS)

    Chloramphenicol (CAP) concentrations in the muscle of rainbow trout from natural lakes, culture pools and markets were determined by radioimmunoassay technique. Chloramphenicol concentrations in the fish muscle samples from natural lakes were found in the ranges of 0-0.8 ng/g. In the fish muscle samples from markets and aqua cultured Chloramphenicol was not observed. The results from natural lakes will provide a basis for determination of the detection limit of CAP for other laboratories to be established in the future where analyses will be carried out regularly. (author)

  12. Development of a radioimmunoassay for human deoxyribonuclease I

    International Nuclear Information System (INIS)

    A reliable radioimmunoassay (RIA) for human deoxyribonuclease I (DNase I) is described. Using delayed addition of tracer antigen, the method is sensitive, reproducible and specific. A good parallel relationship was observed between the standard curve and dilution curves for human urine and human pancreatic juice. G-actin, a naturally occurring DNase I inhibitor, caused no change in the immunoreactivity of DNase I. In healthy individuals, aged 11-90 yr, the mean serum DNase I was 18.4 ng/ml. Increased serum DNase I occurred in patients with acute pancreatitis, renal failure, and in about one-third of patients with various malignant tumors. (Auth.)

  13. Quality control in radioimmunoassay for Argentina, Chile, Paraguay and Uruguay

    International Nuclear Information System (INIS)

    The purpose of this project was the development of an External Quality Control Programme on Radioimmunoassay among laboratories from Argentina and other Latinoamerican countries. The purpose of this programme was to improve the performance of the participating laboratories in order to increase the diagnostic value of patient results. The number of returns of this programme (Buenos Aires I) were compared against those of the previous one (Programme I) (Contract 2737/RB) to determine if the objectives were fulfilled. The preparation of the samples and the evaluation of the results of the first six samples were analyzed in the progress report sent on December 30th, 1983 and were reanalysed including late results

  14. Isotopic methods or immuno diagnosis: The Radioimmunoassay and immunoradiometric assay

    International Nuclear Information System (INIS)

    This work offers an explanation about the more used isotopic techniques for immuno diagnosis: the radioimmunoassay (RIA) and immunoradiometric assay (IRMA). It describes the basic principles of these assays, the antigen-antibody reaction, the radioiodination methods with I-125 for antigens and antibodies, the purification and characterization of labelled compounds. On the order hand they present work gives a review of the methods for separate the bound and free fractions. At the end it offers the principles of the quality control of immunoassay and the future lines of research in the field of RIA and IRMA

  15. Intrinsic factor in human amniotic fluid as determined by radioimmunoassay

    International Nuclear Information System (INIS)

    The intrinsic factor (IF) concentration in 55 human amniotic fluid specimens was determined by radioimmunoassay (RIA). The antiserum was produced by immunizing rabbits with the cobalamin-IF complex isolated from human gastric juice. The median concentration of IF was 0.17 nmol/l and the extreme values <0.07-2.51 nmol/l. Three specimens with a clearly elevated level (0.96, 1.11 and 2.51 nmol/l) were observed. The highest value was associated with a fetal malformation, viz. obstruction of the proximal gut. There was no evident correlation between the concentration of IF in amniotic fluid and gestational age. (author)

  16. Preliminary evaluation of a radioimmunoassay for platelet derived growth factor

    International Nuclear Information System (INIS)

    Together with insulin and epidermal growth factor, platelet derived growth factor (PDGF) is one of the most important and powerful mitogens. The authors developed a radioimmunoassay (RIA) with a double incubation solid phase for human PDGF, in some biological fluids. Immunoglobulins were immobilized via protein A. using purified recombinant protein A at 1 mg/L with monoclonal antibody against human recombinant PDGF (BB) at 1000 fmol/tube, a binding capacity of approx. 17% was obtained. Non-specific binding (NSB%) was 50 ng/mL in 9 of 10 serum samples tested. (author). 18 refs, 8 figs

  17. Development of high throughput radioimmunoassay systems for steroid hormone analysis

    International Nuclear Information System (INIS)

    Improvements have been developed in the radioimmunoassays (RIAs) for the naturally occurring steroids oestriol, cortisol and testosterone and for the synthetic progestagen, norethisterone. Throughputs for these assays have been increased by up to four-fold compared to conventional assay procedures. Except for oestriol, running costs were also considerably lower in the improved RIAs. The assays were validated by accepted criteria and shown to be acurate and precise. These improved RIAs have been shown to be most useful in providing rapid, accurate results to clinicians in time to influence patient treatment. (U.K.)

  18. Value of myoglobin radio-immuno-assay in myocardial infarction

    International Nuclear Information System (INIS)

    Serum myoglobin radio-immuno-assay was performed in 137 patients with myocardial infarction. Two groups, of 102 and 35 patients respectively, were seen in two different hospitals. Serum myoglobin was increased in 72 patients from the first group, and in 30 patients from the second group. In 10 patients from the first group, during the first 24 hours, the increase in serum myoglobin was isolated, without increases in CPK, LDH or SGOT. In the second group, a high serum myoglobin lebel with normal CPK was found in five patients. Increased serum myoglobin is not a specific indicator of myocardial infarction

  19. Radioimmunoassay for determination of thyroglobulin in human serum

    International Nuclear Information System (INIS)

    We described the development and analytical evaluation of a radioimmunoassay with double antibody in liquid phase for human serum thyroglobulin determination using a set of reagents produced in the National Institute of Endocrinology. The reference interval for normal population was 5.7 - 44 ng/ml (X± 2DS; n=170). The intra-assay precipision was 8.1 % (control serum A) and 7.0 (control serum B) and the inter-assay precision was 9.1 % (control serum A) and 9.2 % (control serum B). The sensibility was 4.7 ng/ml and the recovery was 95 %

  20. Radioimmunoassay of serum β2-microglobulin in donor's blood

    International Nuclear Information System (INIS)

    Serum β2-microglobulin (β2-MG) was tested by radioimmunoassay in 149 donors' and 54 healthy volunteers' blood. The results were 203 +- 33.0 nmol/l and 176 +- 26.2 nmol/l, respectively. There was significant difference statistically between them (P2-MG content. In order to increase the quality of donated blood and to keep the health of blood donor, it is suggested that the high content of serum β2-MG is the indicator of too frequent blood donating. The results also showed that the content of β2-MG in donor's blood is not a normal reference value

  1. Radioimmunoassay for free and bound forms of abscisic acid

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) for the quanitation of abscisic acid (ABA) has been developed. The assay is extremely sensitive and measuring ranges extend from 10 pg to 10 ng. Although the assay was free of contaminant interference when applied to avocado material, crude extract analysis yielded a composite of free and bound forms of ABA. Equivalents of 20 mg of plant material were spotted onto silica gel plates (GF245 solvent:toluene:ethyl acetate : acetic acid 25:15:3), developed and the relative Rf zones removed and subjected to RIA. The technique was tested on avocados

  2. Improvements in the automated radioimmunoassay for cAMP or cGMP

    International Nuclear Information System (INIS)

    The work others in developing antibodies and the original radioimmunoassay for cyclic nucleotides provides the basis for these sensitive assays. The acetylation radioimmunoassay for cyclic nucleotides has enabled the measurement of cyclic AMP and cyclic GMP in very small biological samples. This is because accurate determinations can be made in samples containing less than 1 fmol of cyclic AMP or cyclic GMP. The Gamma-Flo automated radioimmunoassay system has been adapted to these assays such that cyclic nucleotides can be automatically measured at a rate of about 60 samples/hr. The Gamma-Flo instrument provides high-precision assays and eliminates human intervention in all steps of the radioimmunoassay. The automated assay has been in continuous operation in our laboratory over the last 10 years and this chapter summarizes the methodology and delineates improvements which have occurred over that time frame. Details for the preparation of the radioligands apply also to the manual acetylated radioimmunoassay for cyclic nucleotides

  3. The development of a general solid-phase radioimmunoassay method. Application to follicle stimulating hormone and to luteinizing hormone radioimmunoassays

    International Nuclear Information System (INIS)

    A solid phase method of radioimmunoassay utilizing a second antibody adsorbed onto tubes was developed. Polyethylene tubes were selected for their adsorption capacity. The following topics were emphasized: the rate of labelled antigen uptake on the second antibody adsorbed on the tubes through the medium of the first antibody; the influence of the second antibody on the antigen-first antibody reaction and comparison with the immunoprecipiting technique; the various factors able to influence the calibration curve and applications to assay optimization; the performances of hFSH AND hLH assays

  4. Radioimmunoassay of human cardiac tropomyosin in acute myocardial infarction

    International Nuclear Information System (INIS)

    Tropomyosin was prepared from fresh human myocardium and antisera raised in rabbits. A sensitive radioimmunoassay was developed for the detection of human cardiac 125I-labelled tropomyosin in human sera down to levels of 1 ng/ml. Values for human cardiac tropomyosin in normal patients ranged from less than 1 to 3 ng/ml. In 18 patients with acute myocardial infarction all had elevated tropomyosin levels ranging from 41 to above 200 ng/ml with a mean peak level of 101 ng/ml. In this study there were no false positive or false negative results. In the initial stages of infarction the time course of appearance and peak levels of cardiac tropomyosin, total creatine kinase and creatine kinase MB isoenzyme were similar. Although total creatine kinase and creatine kinase MB isoenzyme levels were normal after 72 h in patients with single, uncomplicated infarction, cardiac tropomyosin levels were still significantly elevated above normal after this time, being 30-60% of peak values. Radioimmunoassay of human cardiac tropomyosin may prove useful in the diagnosis and in the management of patients with acute myocardial infarction, particularly in the long-term postinfarction period. (author)

  5. Design and optimization of radioimmunoassay for T4

    International Nuclear Information System (INIS)

    Design and optimization of radioimmunoassay for T4. The results of a radioimmunoassay are influenced by the equipment, the laboratory personnel, the quality of the reagents used and especially by the assay design. A good essay design should yield analytical results with high precision and sensitivity within the desired concentration range and can be performed rapidly and easily at low cost. In order to optimize the assay design for T4 the following parameters were studied: amount and dilution of reagents, incubation temperature and length of time prior to separation, reagent volume and sequence of reagent addition. Optimal results for T4 assay were achieved using 25 ul of T4 standard (or samples), 100 ul of 125I labelled T4 (30,000 cpm) and 100 ul of T4 antiserum (1:200 dilution), incubated at room temperature for 1 hour or at 370C for 45 minutes. The following results were obtained under these conditions: a non specific binding value of (6,8+-0,7)%, a maximum binding (B/T) of (67,8+-2,2)%, a value of (5,0+-0,7)ug/dl for ED50. Results for low medium, and high quality control samples were (1,8+-0,1)ug/dl, (5,6+-0,3)ug/dl, and (14,9+-0,6)ug/dl respectively. (author). 12 figs, 6 refs

  6. Radioimmunoassay of TSH subunits in thyroid diseases and endocrine opthalmopahty

    International Nuclear Information System (INIS)

    Highly sensitive radioimmunoassays of hTSH sub-units were developed. The hormone preparations were labelled with 125-iodine according to a modified chloramine -T method, and purified by chromatography using biogel P6 and P60. Rabbit antisera were used as antibodies. Separation of the antibody-bound and of the free antigens was carried out via the double antibody method. The antiserum required for this purpose was obtained from a goat. The sensitivity of the assay was influenced by changing the protein content of the buffer, the incubation volume, the tracer amounts, the incubation time and the incubation temperature. For hTSH-α, the lowest detectable limit was found to be 50 pg/ml, for hTSH-#betta# 20 pg/ml. Thus, the sub-units could be determined for 98% of the patients under review. The #betta#-TSH radioimmunoassay is largely specific, TSH cross-reacts to a degree of 5%. The computerized evoluation was carried out by means of Spline approximation using the Siemens 4004 computer. Precision and accurateness are in compliance with generally accpted criteria. The serum levels of α and #betta# sub-units showed no discordancy with regard to TSH. In all groups of patients examined, the levels of the hormone-specific #betta#-chain were found to be exclusively dependent upon the actual thyroid activity. (orig.)

  7. Radioimmunoassay of prostaglandin F in plasma of pregnant women

    International Nuclear Information System (INIS)

    The aim of this dissertation was to determine quantitatively prostaglandin-F in the plasma of pregnant women in order to obtain further knowledge on changes in PG-F during pregnancy, in particular during the last three months. The plasma of women with clinically normal pregnancies was taken. Prior to radioimmunoassay the plasma was extracted (separation of PG from other plasma components) and chromatography carried out (group separation of PG). The efficiency of this process, as measured by the recovery rate of 3H-PGF, lies between 60.99% and 93.01% for extraction and between 80.58% and 92.16% for chromatography. The plasma was extracted and analysed chromatographically for the assay. The radioimmunoassay was carried out according to the procedure recommended by the manufacturer. A calibration curve was produced without difficulty. The results of the examination of plasma samples were unsatisfactory because of the low sensitivity of the assay; PG-F values of the same order were obtained for all weeks of pregnancy. (orig./MG)

  8. Radioimmunoassay for mammalian type C viral reverse transcriptase

    International Nuclear Information System (INIS)

    Radioimmunological techniques were applied to the analysis of reverse transcriptases of mammalian type C RNA viruses. The polymerase of Rauscher mouse leukemia virus was purified by ion exchange and sequential affinity chromatography. Radioimmunoassays that utilized the viral enzyme as a probe detected as little as 1 ng of purified polymerase. No cross-reactivity could be demonstrated between the reverse transcriptase and other known virus-coded proteins. By comparing the immunological reactivity of the purified enzyme with the reactivity of detergent-disrupted virions, Rauscher mouse leukemia virus was shown to contain the antigenic equivalent of 40 molecules of reverse transcriptase. In a homologous competition immunoassay, the Rauscher viral enzyme demonstrated type-specific antigenic determinants, which distinguish it from other mouse type C viral polymerases. In a broadly reactive interspecies immunoassay, the reverse transcriptases of a number of mammalian type C viruses were cross-reactive, indicating their shared antigenic determinants. Various treatments that inhibited or inactivated DNA polymerase activity had little or no effect on the immunological properties of the enzyme. Thus, radioimmunoassays should be useful in the search for type C viral reverse transcriptase as a marker of subviral expression

  9. A radioimmunoassay for equilin in equine pregnancy plasma

    International Nuclear Information System (INIS)

    It is well known that the pregnant mare produces large quantities of the ring B unsaturated steroid equilin in addition to classical oestrogens. However, the precise biogenesis of this unusual steroid remains a mystery. To facilitate a study of the interrelationship of the steroids present during equine pregnancy, a radioimmunoassay for the measurement of equilin in peripheral plasma was developed. As equilin is thought to be a product of the foeto-placental unit, such an assay may also be of use as an index of foetal well-being. Oestrone and equilin are present in similar concentrations in equine pregnancy plasma so it was important that an antiserum was produced which could differentiate between these two steroids. An antigen was synthesised in which equilin is linked to a protein carrier through the 17-position in order that the C7-C8 double bond might be fully exposed for immune recognition. This stratagem proved successful as the antiserum obtained gave a cross-reaction of only 7.3% for oestrone. The use of a radioimmunoassay incorporating this antiserum is demonstrated by measuring equilin concentrations in plasma samples taken from a mare at weekly intervals from day 60 of pregnancy through to parturition. The corresponding oestrone concentrations are also recorded and demonstrate the validity of the equilin assay in this situation

  10. Double-antibody radioimmunoassay for factor VIII-related antigen

    International Nuclear Information System (INIS)

    A plasma protein required for the support of ristocetin-induced platelet aggregation was isolated from antihemophilic factor concentrate and radiolabeled with 125I. A double-antibody radioimmunoassay was developed, with use of specific rabbit anti-VIII related antigen serum and goat anti-rabbit globulin. The assay is sensitive, reproducible, and technically simple to perform. Values obtained in normal subjects ranged from 0.65 to 1.53 units, similar to our normal range for VIII coagulant activity (0.67 to 1.43 units). However, normal or increased values of VIII-related antigen were observed in VIII coagulant-deficient hemophiliacs. Also, concentrations of VIII-related antigen significantly exceeded coagulant concentrations in several patients with liver disease or disseminated intravascular coagulation, or both. Of a broad selection of congenital coagulation disorders examined, only patients with von Willebrand's disease had decreased VIII-related antigen concentrations, and these corresponded to the lowered concentration of ristocetin cofactor in the patients. In three transfused patients, VIII-related antigen values correlated with the concentration of the cofactor. Our results suggest that the radioimmunoassay of VIII-related antigen is a simple and valuable adjunct in the study of patients with clotting abnormalities

  11. Development of radioimmunoassay method for 16alpha-OH-DHEA using DHEA-17-oxime-BSA

    International Nuclear Information System (INIS)

    Microdetermination of 16α-OH-DHEA (known as a main precursor of placental estriol which is significantly increased in pregnancy) using RIA was developed and offered for clinical application. 1) Microbiological biosynthesis of 3H-labeled 16α-OH-DHEA with high specific activity was performed to correct the recovery factor in the extraction-purification process which is necessary for RIA of hormones of the steroid group. The purity was 97.4%, and the specific activity (S.a.) was 13.9 Ci/mM. 2) Applying the fact that the anti-serum against DHEA-17-oxime-BSA conjugate also presents a cross reaction of 40% with 16α-OH-DHEA, a special assay system using this anti-serum was created. Isolation of other steroids such as Δ5 Androstendiol, Pregnenolone and 17α-OH-Pregnenolone, which present cross reactions with the antiserum, was performed using thin layer chromatography. Therefore, these steroids of 0 to 300 pg could be determined with satisfactory accuracy and precision. 3) 16α-OH-DHEA was determined with the steroids, DHEA and estrogen in embryo and maternal blood. The value of 16α-OH-DHEA, which was the highest in the two umbilical arteries, was about twice as high as that of DHEA. These facts indicate that the role of the steroids as precursors of placental estrogen is important. (Ichikawa, K.)

  12. A new approach to the use of α-fetoprotein as surveillance test for hepatocellular carcinoma in patients with cirrhosis

    OpenAIRE

    Biselli, M; Conti, F.; Gramenzi, A; M. Frigerio; Cucchetti, A; Fatti, G; D'Angelo, M; Dall'Agata, M; Giannini, E G; Farinati, F; Ciccarese, F; Andreone, P; Bernardi, M.; Trevisani, F

    2014-01-01

    Background: Surveillance for hepatocellular carcinoma (HCC) is recommended in patients with cirrhosis. As α-fetoprotein (AFP) is considered a poor surveillance test, we tested the performance of its changes over time. Methods: Eighty patients were diagnosed with HCC (cases) during semiannual surveillance with ultrasonography and AFP measurement were recruited and matched for age, gender, etiology and Child-Pugh class with 160 contemporary cancer-free controls undergoing the same surveillance ...

  13. Electrochemical Enzyme-Linked Immunosorbent Assay (ELISA) for α-Fetoprotein Based on Glucose Detection with Multienzyme-Nanoparticle Amplification

    OpenAIRE

    Ning Gan; Bo Li; Li Lin; Bo Situ; Han-Kun Zhou; Xiao-Mao Yin; Xiao-Hui Yan; Qin-Lan Liu; Lei Zheng

    2013-01-01

    Since glucose biosensors are one of the most popular and widely used point-of-care testing devices, a novel electrochemical enzyme-linked immunosorbent assay (ELISA) for protein biomarkers has been developed based on a glucose detection strategy. In this study, α-fetoprotein (AFP) was used as the target protein. An electrochemical ELISA system was constructed using anti-AFP antibodies immobilized on microwell plates as the capture antibody (Ab1) and multi-label bioconjugates as signal tracer....

  14. Increased concentration of. cap alpha. - and. gamma. -endorphin in post mortem hypothalamic tissue of schizophrenic patients

    Energy Technology Data Exchange (ETDEWEB)

    Wiegant, V.M.; Verhoef, C.J.; Burbach, J.P.H.; de Wied, D.

    1988-01-01

    The concentrations of ..cap alpha..-, ..beta..- and ..gamma..-endorphin were determined by radioimmunoassay in HPLC fractionated extracts of post mortem hypothalamic tissue obtained from schizophrenic patients and controls. The hypothalamic concentration of ..cap alpha..- and ..gamma..-endorphin was significantly higher in patients than in controls. No difference was found in the concentration of ..beta..-endorphin, the putative precursor of ..cap alpha..- and ..gamma..-endorphins. These results suggest a deviant metabolism of ..beta..-endorphin in the brain of schizophrenic patients. Whether this phenomenon is related to the psychopathology, or is a consequence of ante mortem farmacotherapy, remains to be established.

  15. Early α-fetoprotein response predicts survival in patients with advanced hepatocellular carcinoma treated with sorafenib

    Directory of Open Access Journals (Sweden)

    Lee SH

    2015-04-01

    Full Text Available Sangheun Lee,1,* Beom Kyung Kim,2–5,* Seung Up Kim,2–5 Jun Yong Park,2–5 Do Young Kim,2–5 Sang Hoon Ahn,2–6 Kwang-Hyub Han2–6 1Department of Internal Medicine, International St Mary’s Hospital, Catholic Kwandong University, Incheon Metropolitan City, Republic of Korea; 2Department of Internal Medicine, 3Institute of Gastroenterology, 4Liver Cancer Special Clinic, Yonsei University College of Medicine, Seoul, Republic of Korea; 5Liver Cirrhosis Clinical Research Center, Seoul, Republic of Korea; 6Brain Korea 21 Project for Medical Science, Seoul, Republic of Korea.   *These authors contributed equally to this work Background: It is not clear whether tumor marker responses can predict survival during sorafenib treatment in hepatocellular carcinoma (HCC. We investigated whether the α-fetoprotein (AFP response is associated with survival in patients with advanced HCC treated with sorafenib. Methods: We retrospectively reviewed the records of 126 patients with advanced HCC treated with sorafenib between 2007 and 2012. An AFP response was defined as >20% decrease from baseline. At 6–8 weeks after commencing sorafenib, AFP and radiological responses were assessed by modified Response Evaluation Criteria in Solid Tumors. Results: The median overall survival (OS and progression-free survival (PFS were 6.2 and 3.5 months, respectively. Of the study population, a partial response (PR was identified in 5 patients (4.0%, stable disease (SD in 65 patients (51.6%, and progressive disease (PD in 57 patients (44.4%, respectively. AFP non-response was an independent prognostic factor for poor OS (median 10.9 months for AFP response vs 5.2 months for AFP non-response, together with Child-Pugh B, tumor diameter ≥10 cm, and portal vein invasion (all P<0.05, and PFS (median 5.3 months for AFP response vs 2.9 months for AFP non-response, together with tumor diameter ≥10 cm and portal vein invasion (all P<0.05. SD or PR was more frequently found

  16. Development of a new radioimmunoassay for erythropoietin using recombinant erythropoietin

    Energy Technology Data Exchange (ETDEWEB)

    Mason-Garcia, M.; Beckman, B.S.; Brookins, J.W.; Powell, J.S.; Lanham, W.; Blaisdell, S.; Keay, L.; Li, S.C.; Fisher, J.W. (Tulane Univ. School of Medicine, New Orleans, LA (USA))

    1990-11-01

    The development of a 24 hour radioimmunoassay for erythropoietin (EPO) using EPO derived from recombinant DNA as both immunogen and ligand is described in the present paper. Mixed breed rabbits immunized with 10 micrograms/kg of EPO derived from a stably transfected cell line (MD) produced antibodies to EPO with high titer (up to 1:896,000 final dilution in the tube), high affinity (8.4 x 10(11) liter/M), and good specificity. Purified EPO from the above source or from AmGen Biologicals (AG) were successfully radioiodinated with the chloramine-T method and used as ligand in the radioimmunoassay. Standard dose-response curves prepared with EPO from both commercial sources were not significantly different and showed a sensitivity of 0.75 to 0.96 mU/tube. The dose-response curves in both systems also showed parallelism with serially diluted serum from a patient with aplastic anemia. Within-assay and between-assay precision were determined by assaying multiple replicates of a serum pool. Recovery of exogenous EPO added to a serum pool averaged 97% for both systems. The range of normal human serum EPO was determined by assaying the sera of 153 hematologically-normal adult subjects and was found to be 1.1 to 27.3 mU/ml for MD EPO and 0.5 to 16.7 mU/ml for AG EPO. Sera from several patients with hematologic abnormalities were also assayed, including those of 36 patients with anemia of end-stage renal disease (mean +/- SEM, 29.5 +/- 4.0 mU/ml; P less than 0.01). In conclusion, this new, more rapid and sensitive radioimmunoassay system can be used to measure EPO levels in sera from normal human subjects and patients with several types of anemia, and should also be very useful in therapeutic drug monitoring of patients receiving EPO from various commercial sources.

  17. Prognostic value of α-fetoprotein and des-γ-carboxy prothrombin responses in patients with hepatocellular carcinoma treated with transarterial chemoembolization

    International Nuclear Information System (INIS)

    Alpha-fetoprotein (AFP) and des-gamma-carboxy prothrombin (DCP) have been used as diagnostic tools for hepatocellular carcinoma (HCC). However, prediction of outcome using AFP and DCP has not been elucidated. We investigated the clinical role of AFP and DCP as predictors of treatment outcome in patients with HCC undergoing trans-arterial chemoembolization (TACE). Between January 2003 and December 2005, we enrolled 115 treatment-naïve patients who received TACE as an initial treatment modality. An AFP or DCP response was defined as a reduction of more than 50% from the baseline level 1 month after TACE. Patients with AFP < 20 ng/mL or DCP < 20 mAU/mL were excluded. The median age was 59 years and the male gender predominated (n = 81, 70.4%). AFP and DCP response was identified in 91 (79.1%) and 77 (66.9%) patients after TACE. Although progression-free survival (PFS) did not differ according to AFP response (P = 0.150), AFP responders showed significantly better overall survival (OS) than non-responders (34.9 vs. 13.2 months; P = 0.002). In contrast, DCP response did not influence either PFS or OS (all P > 0.05). Multivariate analyses showed that gamma-glutamyltranspeptidase and baseline AFP were predictors of PFS (all P < 0.05) and that male gender, the presence of liver cirrhosis, baseline DCP, number of measurable tumors and AFP response were independent predictors of OS (all P < 0.05). AFP response and higher baseline DCP level are significant predictors of OS in treatment-naïve patients with HCC receiving TACE who showed pretreatment elevation of both AFP and DCP

  18. Radioimmunoassay of 17-OH progesterone and its clinical implications

    International Nuclear Information System (INIS)

    Using highly sensitive radioimmunoassay method the serum level of 17-OH progesterone is estimated, at 2- or 4-day intervals, in forty-one healthy women with a normal menstrual and ovulatory cycle. The alteration in its concentration is compared with the one of progesterone. It is found that the curve of 17-OH progesterone secretion closely follows the curve of progesterone secretion which may be used as a sign of ovulatory cycle occurrence. The level of 17-OH progesterone is assayed also in three children presenting congenital adrenal hyperplasia and fourteen women with ovarial hyperandrogenism of nontumor origin. It is established that 17-OH progesterone level is considerably raised among the patients with congenital adrenal hyperplasia, and may therefore serve as a reliable diagnostic criterion. 17-OH progesterone level in women presenting ovarial hyperandrogenism cannot be used as a diagnostic indicator because its increase is statistically insignificant

  19. Local development of TSH radioimmunoassay and it's evaluation

    International Nuclear Information System (INIS)

    Pakistan is an iodine deficient country. Thyroid diseases are very common and their investigation requires heavy foreign exchange to import relevant commercial kits, AEMC, Multan therefore tried to develop adequate and economical protocol for TSH radioimmunoassay. TSH-RIA protocol supplied by the commercial source did not provide the desired quality and evaluation of thyroid diseases. Our method was therefore optimized using precision profile as the criteria of performance. A total of six assay systems were studied, out of which the most precise one was selected. The new system gives results within expected limits. The sensitivity of the system, 0.3 m IU/ml is suitable for low dose measurements. The working range, 0.6-25.1 m IU/ml covers hypo, hyper and euthyroid levels of the hormone. Comparison with other laboratories shows an agreement of values. A low cost, locally developed good quality TSH RIA is therefore in our hand. (author)

  20. Engineering of radioimmunoassay (RIA) IP10.1

    International Nuclear Information System (INIS)

    Engineering of Radioimmunoassay (RIA) IP10.1 is an innovative by PRPN - BATAN in 2010. Innovations made to the device IP10.1 RIA is the sample changer system, sample changer system on the device RIA IP10.1 applied 2 linear axis system (X, Z) with AC servo motor. In the RIA IP10.1 also use 5 pieces of the detector, so the enumeration process 50 (Fifty) sample is expected to be faster. Like its predecessor, The whole enumeration, data collection procedures and mechanisms operating within this system is entirely controlled by a PC via an electronic module. Electronics module consists of a high voltage module, amplifier and signal processor module, the module enumerators, low-voltage module, the module driver motor controller and a USB interface. The data acquisition and communication system using a USB port with the computer. (author)

  1. Radioimmunoassay of hair for determining opiate-abuse histories

    International Nuclear Information System (INIS)

    Heroin and morphine metabolites can be detected in hair with the use of commerically available radioimmunoassay reagents and with minor sample preparation. Hair samples obtained from morphine-treated mice and heroin users contained nanogram levels of the drug per milligram of hair (single human hair). The results of the hair analyses for all subjects admitting the use of heroin were positive, whereas the results of only 30% of thin-layer chromatographic urinanalyses of these same subjects were positive. In addition, differences in drug concentration for sections of hair near the scalp and near the distal end correlated with the length of time the drug had been used. These results exemplify the potential advantages of the use of hair analysis over urine and serum analyses in terms of accessibility, sample stability, and long-term retention of information

  2. Importance of radioimmunoassay of insulin secretion disorder as atherogenic factor

    International Nuclear Information System (INIS)

    Using a radioimmunoassay a C-peptide levei was revealed in children, pregnant and lying-in women as well as in patients with insulin-dependent diabetes mellitus. After breakfast and insulin administration wich curative purposes the IRI concentration in children increased whereas the C-peptide level changed insignificantly. Changes of the insulin secretion were more noticeable in severe diabetes mejlitus with vascular complications and in disease decompensation. The atherogenic nature of the lipid metaboiism (an increase in the cholesterol, triglyceride and β-lipoprotein levels), changes in the liver and tendency to vascular involvement are results of insulin effect inadequacy. Such metabolic derangements in pregnant women create unfavorable conditions for the development of fetus and may lead to early atherogenic processes

  3. The radioimmunoassay for type III procollagen peptide. A technical note

    International Nuclear Information System (INIS)

    We have studied the immunoreactivity of human sera and synovial fluids in a radioimmunoassay which utilizes an anti-bovine procollagen type III antiserum and the bovine type III procollagen peptide as standard and tracer. We have noticed a complete cross section reaction between bovine col 1-3 and human synovial fluid, which thus contains substance(s) having the same antigenic determinant(s) as col 1-3. On the contrary, we have demonstrated non parallelism between col 1-3 and human serum, whether the serum was obtained from normal subjects or patients suffering from various diseases. These results suggest that immunoreactive substances related to col 1-3 and detected in sera are at least partially different from those detected in synovial fluids

  4. High sensitivity solid phase exchange radioimmunoassay for morphine

    International Nuclear Information System (INIS)

    An immobilized antibody technique is described which has several advantages over other opiate measuring methods. It is exceptionally useful in determining morphine concentrations in biological fluids and tissues. Antiserum was prepared by injection of a morphine-6-succinyl bovine serum albumen suspension into rabbits. Strips of water swollen unsintered polyvinylidene fluoride film were submerged in antiserum solution for 16 hr. The impregnated strips were washed, air-dried and stored. The activity of the immobilized antibody did not change significantly over 3 months. This very firm binding is believed to comprise physical absorption aided by entanglement of the protein in the strands of the polymer. Results of tests of radioimmunoassay capability are given. (author)

  5. Radioimmunoassay for the determination of free and conjugated abscisic acid

    International Nuclear Information System (INIS)

    The characterization and application of a radioimmunoassay specific for free and conjugated abscisic acid (ABA) is reported, The antibodies produced against a bovine serum albumin-(+-)-ABA conjugate have a high affinity for ABA (Ka= 1.3 x 109 l mol-1). Trans, trans-ABA and related compounds, such as xanthoxin, phaseic acid, dihydrophaseic acid, vomifoliol or violaxanthin do not interfere with the assay. The detection limit of this method is 0.25 x 10-12 mol ABA, the measuring range extends to 20 x 10-12 mol, and average recoveries are 103%. Because of the high specificity of this immunoassay, no extract purification steps are required prior to analysis. Several hundred plants can be analyzed per day in a semi-automatic assay performance. ABA has been detected in all higher plant families examined, but was absent in the blue-green alga, Spirulina platensis, the liverwort Marchantia polymorpha, and two species of fungi. (orig.)

  6. Effect of radioimmunoassay procedures on therapeutic drug monitoring

    International Nuclear Information System (INIS)

    Methods for the measurement of therapeutic drugs have covered every aspect of analysis from extraction to derivatization. In general, published methods were modified to shorten drug extractions and overall analysis time. The use of different standards, as well as the frequent omission of internal standards, often produced large and clinically unacceptable analytical variations. As a result, physicians would adjust drug dosages according to the physiological response to a standard dose. The introduction of radioimmunoassay techniques for the quantitation of therapeutic drugs have made a significant impact on the clinical chemistry laboratory. The similarities of the various assay methods and the technologists' familiarity with the assay protocols have produced clinically relevant results. Clinical laboratories are now able to frequently analyze a large number of samples with acceptable accuracy and precision. The esoteric test once performed infrequently is today a routine analytical assay often performed STAT. Therapeutic drug monitoring has become a major activity in many clinical laboratories

  7. Provision of radioimmunoassays for reproductive hormones in developing countries

    International Nuclear Information System (INIS)

    The Special Programme of Research, Development and Research Training in Human Reproduction of the World Health Organization (WHO) has, for the last eight years, distributed sets of matched assay reagents for the radioimmunoassay of hormones to approximately 75 laboratories in 45 countries. The Matched Reagent Programme has enabled these centres to carry out research that would otherwise have been difficult, if not impossible, for them to undertake. Between-centre comparability, as judged by WHO External Quality Assurance Scheme data, has improved and at least five hundred scientific papers have been published as a consequence of the availability of WHO reagents. National and regional reagent programmes and associated schemes are being encouraged. These activities should help to improve the availability of research and diagnostic facilities for patient care in developing countries, which is consistent with the WHO policy of technology transfer. (author)

  8. Standardisation of radioimmunoassay for human insulin employing magnetizable cellulose particles

    International Nuclear Information System (INIS)

    We describe a convenient and flexible solid phase radioimmunoassay for human insulin employing magnetizable cellulose particles. Anti-porcine insulin antibody was covalently linked to magnetizable cellulose particles to form a stable and economical solid phase immunosorbent system. The tracer was prepared by radioiodinating insulin with 125I using Chloramine-T oxidation method. The analytical sensitivity of assay observed was 5.5 μIU/mL. Intra-assay and inter-assay variations were found to be <12 % along with analytical recovery of 93-109 %. The developed assay can be used for the routine analysis of clinical samples. In addition, concentration of the solid phase magnetizable immunosorbent can be easily varied as per the specific requirement for research purposes. (author)

  9. Radioimmunoassay data processing program for IBM PC computers

    International Nuclear Information System (INIS)

    The Medical Applications Section of the International Atomic Energy Agency (IAEA) has previously developed several programs for use on the Hewlett-Packard HP-41C programmable calculator to facilitate better quality control in radioimmunoassay through improved data processing. The program described in this document is designed for off-line analysis using an IBM PC (or compatible) for counting data from standards and unknown specimens (i.e. for analysis of counting data previously recorded by a counter), together with internal quality control (IQC) data both within and between batch. The greater computing power of the IBM PC has enabled the imprecision profile and IQC control curves which were unavailable on the HP-41C version. It is intended that the program would make available good data processing capability to laboratories having limited financial resources and serious problems of quality control. 3 refs

  10. Production of biological reagents for radioimmunoassay second antibody

    International Nuclear Information System (INIS)

    The experimental production of second antibody to be used in hormonal assays, in which the first antibody is raised in rabbits, is described. Four sheep were immunized with the rabbit immunoglobulin prepared at IPEN-CNEN laboratory. Their antisera were evaluated by the human thyrotropin radioimmunoassay employing materials provided by the National Hormone and Pituitary Program (USA), in comparison with a reference antiserum of known quality, produced in goat by the Radioassay Systems Laboratories - RSL (USA). From the fourth booster injection the animals developed antiserum with titer similar to that exhibited by the commercial product, even presenting higher values. These antisera are now being examinated for the optimal conditions of precipitation before be packed for future use and distribution. (author)

  11. The radioimmunoassay of human placental protein 14 (PP14)

    International Nuclear Information System (INIS)

    The development and validation of a radioimmunoassay for the measurement of human placental protein 14 in maternal serum is described. The mean concentration of this protein in serum from 22 normal pregnant women showed a decline during the third trimester from 120 μg/l at 27 weeks gestation to 65 μg/l at term. Serum samples from 16 patients with intra-uterine growth retardation tended to contain lower concentrations of placental protein 14, these results reaching significance at weeks 36-38 of gestation. Of seven patients with pre-eclampsia from whom two or more blood samples were taken, four showed increases in concentration of this protein as pregnancy proceeded, compared with the normal pattern of decreasing values. (Auth.)

  12. Measurement of plasma canine C peptide by radioimmunoassay

    International Nuclear Information System (INIS)

    A sensitive radioimmunoassay for canine C peptide (CCP) was established using synthetic CCP, a specific antiserum, and rabbit anti-guinea pig serum. Radioiodination was performed according to a modified chloramine T method. Tracer preparations have been used for 6 weeks after iodination. The standard curve ranges from 0.028 to 3.0 nmol/l. The intra-assay coefficient of variation (CV) was 3-5% and the inter-assay CV was 6-9% in the optimal range between 0.3 and 0.8 nmol/l. The average recovery of CCP added to plasma samples was 100.6% (n = 9). Canine insulin, porcine proinsulin, bovine proinsulin, and human C peptide exhibited no cross-reactivity. The mean fasting plasma CCP concentration was 0.089 +- 0.021 nmol/l in normal dogs and -0.005 +- 0.007 nmol/l (mean +- SEM) in diabetic dogs, respectively. (author)

  13. Production of antisera for radioimmunoassay of human proinsulin: preliminary studies

    International Nuclear Information System (INIS)

    In attempt to the production of anti proinsulin antisera for the radioimmunoassay, five guinea pigs received 50 μg of biosynthetic human proinsulin (Eli Lilly and Company, US) injected subcutaneously. Booster injections were administered at 3 weeks intervals and blood was taken 14 days after the injection. Just after the tenth week, three guinea pigs developed antisera with titers of 1:15.000, 1:1.500 and 1:300 and one of them presented titer lower than 1:300. The evaluation of the specificity of those three antisera showed cross-reactivity only with insulin, which antibodies were not sufficiently sensitive for the assay of endogenous proinsulin from healthy subjects, being only suitable for the measurement of circulating proinsulin in patients with hyper proinsulinemia. Alternative schedules of immunization in order to obtain more sensitive antisera are discussed. (author)

  14. Mechanical design sample changer in device radioimmunoassay (RIA)

    International Nuclear Information System (INIS)

    The mechanical system of sample changer is to change the sample in the Radioimmunoassay (RIA) for accounting the sample. To obtain the optimum result it is expected that the sample can reach inside the collimator and detector at a certain. This mechanical system is also expected to change in amount of samples simultaneously into detector hole. It is necessary to design a mechanical system which can change the sample at the optimal depth and can change a 25 sample continuously. Carousel is design to change 25 sample into detector hole correctly for one process continuously with diameter caro sel cap is 52 cm. Meanwhile eccentric is design with 9 cm diameter to maximize the Up and Down motion of caro sel and to change the sample with the rotating sample. To cycle the eccentric used motor DC, 60 watt. Here, it is expected that this design can solve the problem to optimize the counter of amount of sample in a detector. (author)

  15. Scintillation proximity radioimmunoassay utilizing 125I-labeled ligands

    International Nuclear Information System (INIS)

    A unique type of radioimmunoassay is described that does not require centrifugation or separation. Microbeads containing a fluorophor are covalently linked to antibody. When an 125I-labeled antigen is added it binds to the beads and, by its proximity, the emitted short-range electrons of the 125I excite the fluorophor in the beads. The light emitted can be measured in a standard scintillation counter. Addition of unlabeled antigen from tissue extracts displaces the labeled ligand and diminishes the fluorescent signal. Application of scintillation proximity immunoassay to tissue enkephalins, serum thyroxin, and urinary morphine is described. Applications of the principle to study the kinetics of interaction between receptors and ligands are discussed

  16. The application of radioimmunoassay to diagnosis of liver diseases

    International Nuclear Information System (INIS)

    The concentrations of serum hyaluronic acid and serum prolactin were determined by radioimmunoassay in patients with liver diseases. The results show that the level of serum hyaluronic acid in patients with CAH or liver cirrhosis is significantly higher than those of normal control subjects. The difference is remarkable (P<0.01). The mean value of patient group gradually increases with the increasing of liver damage. The serum hyaluronic acid can be used as a predictive indicator to diagnose CAH and early liver cirrhosis. In addition, the level of serum prolactin in patients with liver cirrhosis is also much higher than those of normal control subjects. Thus the serum prolactin may be used as an associate biochemical marker for detecting liver cirrhosis. The mechanism of the change of serum hyaluronic acid and prolactin in patients with liver diseases is presented

  17. Development of radioimmunoassay. Preparation of radiolabeled tracers theophylline

    International Nuclear Information System (INIS)

    Therapeutic monitoring of theophylline can be accurately performed by radioimmunoassay (RIA). It is radioactive tracer as an essential reagent for the development of very sensitive RIA. Direct radiolabeling of theophylline with 125I is very difficult due to the absence of appropriate functional groups. Hence carboxylic acid of theophylline was tagged to tyrosine methyl ester and then radiolabeled. The derivatives of theophylline, bearing a propionic acid and butyric acid side chains at seventh and eight position of theophylline, were synthesised and coupled to tyrosine methyl ester. Theophylline-tyrosine methyl ester conjugates were labeled with 125I using chlora mine-T. Radiolabeled theophylline was purified by solvent extraction followed by thin layer chromatography. The purified radiolabeled compound were assessed for their radiochemical purity, specific activity and immunoreactivity. Stability studies of radiolabeled compounds were performed with different solvents at different temperatures. Theophylline serum samples analysed using developed and commercial kits showed the correlation coefficient of 0.961 (n=9). (author)

  18. Problems associated with the radioimmunoassay of serum trypsin

    International Nuclear Information System (INIS)

    A commercial trypsin radioimmunoassay (RIA) kit was tested for its ability to measure trypsin bound to the serum protease inhibitors, α2macroglobulin (α2M) and α1 anti-trypsin (α1AT). Only 20% of trypsin bound to α2M and 70% bound to α1AT was detected by the assay system. Recovery of trypsin added to human serum varied from 0 to 20%. Standard curves prepared from purified human cationic trypsin did not exhibit parallelism with the kit standard curves. Inclusion of horse serum in the standard solutions improved the parallelism observed. Immunoreactive trypsin (IRT) levels obtained for serum samples were found to vary considerably depending on the standard curve used to calculate the assay results. Lower IRT levels were observed when trypsin standards prepared in the absence of horse serum were used as reference. (Auth.)

  19. Cerebrospinal fluid beta-endorphin radioimmunoassay: Methodological problems

    International Nuclear Information System (INIS)

    This note describes the technical details of a radioimmunoassay for #betta#-endorphin (#betta#-EP). An antiserum raised in rabbits against synthetic #betta#-EP measures a ''#betta#-EP-like'' immunoreactivity (#betta#-EP and #betta#-LPH). Thus, filtration chromatography is used to separate both peptides. #betta#-EP is the 31 amino-acid C-terminal end of #betta#-LPH, with potent opiate-like activity. The measurement of CSF #betta#-EP in man may advance the understanding of numerous brain functions especially in pain problems. This determination is difficult since #betta#-LPH contains the immunological determinants of both #betta#-LPH and #betta#-EP. The measurement of #betta#-EP in human CSF has been approached by using gel filtration of CSF extracts followed by a RIA employing an anti-#betta#-EP antibody which cross reacts with #betta#-LPH. (author)

  20. Purification, characterization and development of radioimmunoassay of human liver ribonuclease

    International Nuclear Information System (INIS)

    Human liver ribonuclease (RNase) was purified 3600-fold into an electrophoretically homogeneous state by column chromatography on phosphocellulose, gel filtration, poly(G) affinity chromatography, and heparin affinity chromatography. The molecular weight of the RNase estimated by SDS disc electrophoresis was 19500. RNase was a heat- and pH-stable protein, and optimum activity was obtained at pH 7.0. The radioimmunoassay (RIA) for human liver RNase has been developed and the assay was shown to be sensitive (20 ng/ml), reproducible and specific. A good parallel relationship was observed between the standard curve and the dilution curves for serum and urine. No cross-reactivity was demonstrated between human liver and pancreatic RNase (less than 1%). In 44 normal subjects, the mean serum concentration of liver RNase determined by the RIA was found to be 99.4 ng/ml (SD+-66.3). (Auth.)

  1. Production of Anti-triiodothyronine sulfate antibody for radioimmunoassay applications

    International Nuclear Information System (INIS)

    Triiodothyronine sulfate (T3S) may be an obligatory intermediate metabolic of the metabolism of thyroid gland hormones invertebrates in peripheral during the process of deiodination of the inactive form of the thyroid gland hormones, thyroxine(T4), into the active form triiodothyronine (1,2). Construction of a reliable procedure for the estimation of T3S accurately in blood serum will be of great importance for medical, biochemical and physiological investigations. In this work we developed a robust method for the production of anti-triiodothyronine sulfate polyclonal antiserum with good specifications using a derivatized immuno gen and a modified immunization process and a sensitive radioimmunoassay system was designed and developed

  2. Radioimmunoassay for pregnancy-associated plasma protein A

    International Nuclear Information System (INIS)

    A specific and highly sensitive radioimmunoassay for determination of pregnancy-associated plasma protein A in human serum is described. The minimum detection limit for this protein was 2.9 μg/L. The within- and between-assay coefficients of variation were 4.0 and 4.5%, respectively. The circulating protein was detected within 32 days of conception in eight normal pregnancies and within 21 days in a twin pregnancy. Circulating concentrations in the mother at term were consistently higher (10-fold) than in matched amniotic fluid; none was detected in the umbilical circulation. This protein was also detected in the circulation of patients with hydatidiform mole. This assay will permit investigations into the clinical evaluation of measurements of the protein during early pregnancy and trophoblastic disease

  3. Farm application of radioimmunoassay technology in dairy cattle management

    International Nuclear Information System (INIS)

    Monitoring of progesterone concentrations in milk or blood plasma of farm animals, using radioimmunoassay technology is presented in this report. This was instituted among 103 dairy cows managed by dairy cooperatives under smallholder level in Sta. Cruz-Pagsanjan, Laguna and Sariaya, Quezon (n=103), and under communal level Pontevedra, Capiz (n=48). The authors observed that the measurement of progesterone in milk/plasma was proven useful as a diagnotic aid in dairy cattle production studies such as: (a) early pregnancy diagnosis; (b) identification of fertile and abnormally cycling/subestrus or anestrous cows, and (c) appropriate timing for breeding services especially at post-partum stage. This information is relevant where appropriate management intervention measures are indicated to improve dairy cattle production in the country. (author)

  4. Radioimmunoassay of hair for determining opiate-abuse histories

    Energy Technology Data Exchange (ETDEWEB)

    Baumgartner, A.M.; Jones, P.F.; Baumgartner, W.A.; Black, C.T.

    1979-07-01

    Heroin and morphine metabolites can be detected in hair with the use of commerically available radioimmunoassay reagents and with minor sample preparation. Hair samples obtained from morphine-treated mice and heroin users contained nanogram levels of the drug per milligram of hair (single human hair). The results of the hair analyses for all subjects admitting the use of heroin were positive, whereas the results of only 30% of thin-layer chromatographic urinanalyses of these same subjects were positive. In addition, differences in drug concentration for sections of hair near the scalp and near the distal end correlated with the length of time the drug had been used. These results exemplify the potential advantages of the use of hair analysis over urine and serum analyses in terms of accessibility, sample stability, and long-term retention of information.

  5. The development of determining human prostatic acid phosphatase by radioimmunoassay

    International Nuclear Information System (INIS)

    We purified human prostatic acid phosphatase (hPAP) from prostatic tissues by affinity chromatography, DEAE cellulose and gel filtration and also examined physicochemical properties of highly purified PAP. We developed a double-antibody radioimmunoassay for hPAP in serum, with use of antiserum raised in rabbit against highly purified PAP. The antiserum did not cross react with acid phosphatase from platelets and red blood cells. Experimental detail are outlined to assess the reproducibility and reliability of the method under various conditions. The upper limit of the serum PAP levels in the present assay was set at 3.0 ng/ml by 162 determinations of samples. The serum PAP levels of 2 untreated patients with prostatic carcinoma were higher than 3.0 ng/ml and 39 patients with benign prostatic hyperplasia were an average value of 1.9 ng/ml. (author)

  6. Development and characterization of a homologous radioimmunoassay for equine prolactin

    International Nuclear Information System (INIS)

    A specific and sensitive homologous radioimmunoassay has been developed for equine prolactin, suitable for measuring prolactin concentrations in serum of horses. The sensitivity of the assay ranged from 0.4 to 0.6 ng/ml and the intra- and inter-assay coefficients of variation averaged 6.9 and 15.4%, respectively, for five doses of hormone. Cross-reactivity with other mammalian and nonmammalian prolactins and growth hormones was less than 20 and 0.3%, respectively. Cross-reactivity with equine growth hormone was less than 0.07%. Equine serum and pituitary extracts showed parallel dilution-response curves with equine prolactin. The percentage recovery of exogenous equine prolactin in serum was 89%. Preliminary analysis of several physiological samples (stallions, pregnant, and nonpregnant mares) yielded values from 0.6 to 12.0 ng/ml

  7. Radioimmunoassay techniques and reproductive management of livestock in North Africa

    International Nuclear Information System (INIS)

    The report summarizes the principal applications of progesterone radioimmunoassay (P4-RIA) in Morocco for evaluating and improving fertility in cattle, sheep and camels. In cattle, P4-RIA of blood or milk helped to determine the time for onset of sexual functions after parturition and the incidence of silent oestrus. It is especially important to assess the first factor in semi-extensively managed herds, while the second factor occurs mainly in intensively managed herds. P4-RIA is an important tool in fertility improvement programmes involving induction and synchronization of oestrus and testing for early pregnancy. In sheep, P4-RIA helped to define the optimum age at first mating of ewe lambs and the optimum mating season of the year for adult ewes. In camels, analysis of the profile of plasma progesterone before and after mating suggests that P4-RIA could be used for early pregnancy testing. (author)

  8. Development of a radioimmunoassay for triamcinolone acetonide in horse plasma

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) was developed for the detection of triamcinolone acetonide (TAAc) in equine blood plasma. These antibodies exhibited cross reactions of 0,015% with cortisol and of 0,1% with other endogenous glucocorticoids. Four different synthetic corticosteroids interfered in a range of 0,21 to 0,93%. In vitro 86 of TAAc could be recovered. This method proved sufficient reproducibility down to a limit of 131,7 fmol/ml = 57,2 pg/ml. The TAAc RIA is suitable for the detection of a TAAc application particularly during the 1st day p.i. The results may obtain a higher limit of confidence by the simultaneous demonstration of cortisol suppression. By the use of this test more detailed conclusions may be drawn about presence and duration of a pharmacodynamic action originating from the TAAc depot. (orig./TRV)

  9. Simultaneous radioimmunoassay of thyrotropin and thyroxine in human serum

    International Nuclear Information System (INIS)

    The importance of early diagnosis and treatment of congenital hypothyroidism has been well established, and several screening programs have been undertaken to detect neonates with this disorder by measurement of concentrations of thyrotropin or thyroxine in the serum. However, measurement of either hormone alone may fail to identify all affected patients. Accordingly, we have established a simulatneous double-antibody, dual-isotope radioimmunoassay for both. Sensitivity, slope, analytical recovery, and precision characteristics of the simultaneous assay do not differ from those of each assay performed separately. Values for the two analyses in the single and simultaneous assays correlate well (r = 0.951 for thyroxine, 0.983 for thyrotropin). This assay system permits determination of both hormones within 72 h after specimen collection and thus should allow more rapid evaluation, diagnosis, and treatment of infants with congenital hypothyroidism

  10. A different approach to the radioimmunoassay of thyrotropin releasing hormone

    International Nuclear Information System (INIS)

    Thyrotropin releasing hormone (TRH) was linked to hemocyanin by means of a dinitrophenylene moiety. TRH (pGlu-His-Pro-NH2) was made to react with a large excess of 1,5-difluoro-2,4-dinitrobenzene to yield Nsup(im)-[5-fluoro-2,4-dinitrophenyl]TRH. After removal of excess reagent the derivative was coupled to hemocyanin with a minimum of side-reactions. From two rabbits out of four immunized with this material valuable antisera were obtained, which were used in the radioimmunoassay of the hypothalamic hormone at a final dilution of 1:7,500 and 1:15,000, respectively. The properties, especially with regard to specificity, of these antisera were studied and compared with another antiserum, which was obtained using a conjugate having TRH linked to thyroglobulin via a p-azophenyl-acetyl moiety. Despite the difference between the derivatives, i.e. the nature and the point of attachment of the side chains, the specificities of the assays were very similar. Deamidation of TRH, deletion of either one of the terminal residues, hydrolysis of the lactam of the pyroglutamyl residue, and replacing Pro-NH2 by Pro-Gly-NH2 or by an octapeptide chain yield peptides with strongly diminished cross-reactivities. However, Nsup(im)-benzyl-TRH and pGlu-Phe-Pro-NH2 were 5-10 times as active as TRH probably due to a closer physico-chemical similarity to the arrangement of the haptens in the conjugates. This suggests that the sensitivity of the radioimmunoassay may be increased markedly by conversion of TRH into the Nsup(im)-dinitrophenyl derivative and by using a related compound for radioiodination. (orig.)

  11. Cholangiolocellular carcinoma with rapid progression initially showing abnormally elevated serum alfa-fetoprotein.

    Science.gov (United States)

    Yoh, Tomoaki; Kato, Tatsushi; Hirohata, Yoshiaki; Nakamura, Yuya; Nakayama, Hiroyuki; Okamura, Ryuji

    2016-08-01

    Cholangiolocellular carcinoma (CoCC) is a rare malignant liver tumor derived from hepatic progenitor cells, which exist in the canals of Hering. We encountered a case of CoCC with an extremely poor clinical course, initially showing abnormally elevated serum alfa-fetoprotein (AFP). A 72-year-old male presented with a liver tumor and abnormally elevated serum AFP levels (16,399 ng/ml). We preoperatively diagnosed hepatocellular carcinoma and performed extended right hepatectomy, after which the serum AFP levels remarkably decreased to 97 ng/ml. Postoperatively, the disease was pathologically diagnosed as CoCC. Furthermore, immunohistochemical pathological findings were alcian blue negative, cytokeratin (CK) 7 partially positive, CK19 positive, hepatocyte paraffin-1 negative, membranous negative for epithelial membrane antigen, and AFP negative. Fifty-five days later, intra- and extrahepatic recurrence developed, and the patient died 65 days after surgery. Although CoCCs show favorable outcomes, these characteristics of our case were not previously reported. It is necessary to accumulate more information on CoCC. PMID:27363839

  12. α-Fetoprotein-producing ovarian tumor in a postmenopausal woman with germ cell differentiation.

    Science.gov (United States)

    Meguro, Shiori; Yasuda, Masanori

    2013-02-01

    α-Fetoprotein (AFP)-producing ovarian tumors (APOTs) are rarely encountered in postmenopausal women, irrespective of whether they are of the germ cell or non-germ cell type. The APOTs that do occur in postmenopausal women are characterized by variable histologies such as hepatoid carcinoma, yolk sac tumor, and epithelial malignancies, most of which are combined. We herein present a case with APOT, which arose in a 58-year-old, gravida 2, para 2, postmenopausal woman. Preoperatively, the tumor, which was in the right ovary, was found to produce AFP (102768.0 ng/mL). The tumor was evenly composed of glands mimicking secretory endometrial gland or fetal gut accompanied by abundant stroma. Immunohistochemically, these glands were positive for SALL4, glypican-3, and hepatocyte nuclear factor 1β. We considered the present case as an AFP-producing adenocarcinoma with adenofibroma showing germ cell differentiation, but it seemed controversial that this tumor should be designated as a yolk sac tumor of the glandular type. The expression profiles of SALL4, OCT4, glypican-3, and hepatocyte nuclear factor 1β were thought to provide interesting implications to characterize the present case. PMID:22056036

  13. 亲和层析微柱法测定肝癌特异性AFP及其在肝癌诊断中的临床价值%Quantitative analysis of hepatoma-specific α-fetoprotein (HS-AFP) by a new mini-column affinity chromatography and its clinical value in diagnosis of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Wei Wu; Dengfu Yao; Liwei Qiu; Xiaoxiao Gu; Xinhua Wu

    2008-01-01

    Objective: To establish a convenient and economic method to determine hepatoma-specific α-fetoprotein (HS-AFP) for diagnosis of hepatocellular carcinoma (HCC). Methods: HS-AFP from serum of HCC patients was separated by a mini-column Lens culinaris agglutinin (LCA)-affinity chromatography. The levels of serum total AFP and separated HS-AFP were detected by radioimmunoassay (RIA). Results: Circulating AFP was separated into three peaks (AFP-1, AFP-2, and AFP-3) by LCA-affinity chromatography. During the elution course, the AFP-1 and AFP-2 could be eluted with TE buffer. HS-AFP (AFP-3) from sera of HCC patients was eluted dearly on the LCA-sepharose gel mini-column with a solution containing a-methyl-D-mannoside. It was a part of total AFP and only found in sera of HCC patients. A ratio of more than 15% for HS-AFP to total AFP in serum was considered as a specific marker for HCC diagnosis with higher sensitivity (92.7%) and specificity(88.2%). Conclusion: The new assay for circulating HS-AFP analysis is more sensitive, repeatable, and convenient. Its clinical application would be useful to early diagnosis of HCC.

  14. Radioimmunoassay for human pancreatic amylase: comparison of human serum amylase by measurement of enzymatic activity and by radioimmunoassay

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) for human pancreatic amylase has been developed for the determination of human serum amylase content. The assay was shown to be sensitive (7 ng/mI), reproducible and specific, but human pancreatic amylase and salivary amylase could not be distinguished by the antiserum used. In normal subjects, the mean concentration of amylase determined by the RIA was found to be 122.1 ng/ml (range: 55-250 ng/ml). A good correlation was observed between the concentration of amylase and its enzymatic activity in normal subjects. In some instances with high amylase activity, however, the rise in enzymatic activity was not accompanied by increasing amount of amylase content. (Auth.)

  15. Preparation and characterization of 125I-labelled proinsulin for radioimmunoassay (Preprint No. CT-62)

    International Nuclear Information System (INIS)

    Bovine proinsulin is iodinated with 125I. The labelled proinsulin was purified over sephadex. The preparation was characterized for stability and immunoreactivity. A radioimmunoassay was standardized using this labelled preparation. (author)

  16. Antibodies to cytoskeletal proteins as evidenced by immunofluorescence microscopy and radioimmunoassay

    International Nuclear Information System (INIS)

    In patients suffering from chronic hepatitis, collagenosis and infectious mononucleosis, resp., as well as in blood donors antibodies against cytoskeletal antigens such as actin, myosin, actinin, desmin, keratin, and tubulin were determined by radioimmunoassay

  17. Development of a radioimmunoassay for pig pancreatic kallikrein and its application in physiological studies

    International Nuclear Information System (INIS)

    A RIA for pig pancreatic kallikrein has been developed. Purified kallikrein was labelled with 125I. The use of radioimmunoassays in investigating the intestinal absorption of pig pancreas kallikrein is dealt with. (VJ)

  18. Control of routine radioimmunoassays: a computer program for calculation of control charts for precision and accuracy

    International Nuclear Information System (INIS)

    A computer program is proposed allowing the automatic calculation of control charts for accuracy and precision. The calculated charts enable the analyst to control easily the daily results for a determined radioimmunoassay. (Auth.)

  19. Radioimmunoassay for detection of VP1 specific neutralizing antibodies of foot and mouse disease virus

    International Nuclear Information System (INIS)

    A solid-phase radioimmunoassay was developed for the detection of antibodies against a specific region of the VP1 protein of the A24 and O1 serotypes of foot and mouth disease virus. The antibody titers from the radioimmunoassay showed a positive correlation with neutralizing antibody titers determined by a mouse protection assay. The specificity of the assay resides in the peptide used as antigen. The assay is rapid, reproducible and does not require the use of whole virions. (orig.)

  20. $\\alpha_s$ review (2016)

    CERN Document Server

    d'Enterria, David

    2016-01-01

    The current world-average of the strong coupling at the Z pole mass, $\\alpha_s(m^2_{Z}) = 0.1181 \\pm 0.0013$, is obtained from a comparison of perturbative QCD calculations computed, at least, at next-to-next-to-leading-order accuracy, to a set of 6 groups of experimental observables: (i) lattice QCD "data", (ii) $\\tau$ hadronic decays, (iii) proton structure functions, (iv) event shapes and jet rates in $e^+e^-$ collisions, (v) Z boson hadronic decays, and (vi) top-quark cross sections in p-p collisions. In addition, at least 8 other $\\alpha_s$ extractions, usually with a lower level of theoretical and/or experimental precision today, have been proposed: pion, $\\Upsilon$, W hadronic decays; soft and hard fragmentation functions; jets cross sections in pp, e-p and $\\gamma$-p collisions; and photon F$_2$ structure function in $\\gamma\\,\\gamma$ collisions. These 14 $\\alpha_s$ determinations are reviewed, and the perspectives of reduction of their present uncertainties are discussed.

  1. Dual-signal amplification strategy for ultrasensitive photoelectrochemical immunosensing of α-fetoprotein.

    Science.gov (United States)

    Li, Yong-Jie; Ma, Meng-Jie; Zhu, Jun-Jie

    2012-12-01

    An ultrasensitive photoelectrochemical immunoassay of cancer biomarker α-fetoprotein (AFP) is proposed that uses titanium dioxide (TiO(2)) coupled with AFP-CdTe-GOx bioconjugate, which featured AFP antigen and glucose oxidase (GOx) labels linked to CdTe quantum dots (QDs) for signal amplification. The synthesized CdTe QDs yielded a homogeneous and narrow size distribution, which allowed the binding of AFP and GOx on CdTe QDs. Greatly enhanced sensitivity for AFP came from a dual signal amplification strategy. First, an effective matching of energy levels between the conduction bands of CdTe QDs and TiO(2) allowed for fast electron injection from excited CdTe QDs to TiO(2) upon irradiation, which reduced the recombination process of electron-hole pairs and prompted photoelectrochemical performance. Second, GOx enzyme could catalyze glucose to produce H(2)O(2), which acted as a sacrificial electron donor to scavenge the photogenerated holes in the valence band of CdTe QDs, further causing an enhanced photocurrent. Thus, on the basis of the dual signal amplification strategy, the competitive immunosensor based on the specific binding of anti-AFP antibodies to AFP and AFP-CdTe-GOx bioconjugates was achieved. This proposed biosensor for AFP possessed largely increased linear detection range from 0.5 pg/mL to 10 μg/mL with a detection limit of 0.13 pg/mL. The proposed amplification strategy shows high sensitivity, stability, and reproducibility and can become a promising platform for other protein detection. PMID:23140135

  2. Expression and location of α-fetoprotein during rat colon development

    Institute of Scientific and Technical Information of China (English)

    Xiao-Yan Liu; Dan Dong; Peng Sun; Jun Du; Luo Gu; Ying-Bin Ge

    2009-01-01

    AIM: To investigate the expression of a-fetoprotein (AFP), a cancer-associated fetal glycoprotein, and its involvement during rat colon development.METHODS: Colons from Sprague-Dawley rat fetuses, young and adult (8 wk old) animals were used in this study. Expression levels of AFP in colons of different development stage were detected by reversetranscriptase PCR (RT-PCR) and Western blotting. To identify the cell location of AFP in the developing rat colons, double-immunofluorescent staining was performed using antibodies to specific cell markers and AFP, respectively. RESULTS: The highest levels of AFP mRNA were detected in colons of rats at embryonic day 18.5 (e18.5). Compared to e18.5 d, the AFP expression was significantly decreased during rat development [85% for e20.5, P < 0.05, 58% for postnatal day 0.5 (P0.5), P < 0.05, 37% for P7, P < 0.05, 24% for P14, P < 0.05, and 11% for P21, P < 0.05] and undetected in adult rats. Only the 72-kDa isoform of AFP was detected by Western blotting, the expression pattern was similar to AFP mRNA and conformed to the results of mRNA expression. The AFP positive staining was identical to different distribution patterns in fetuses, young and adult animals and positive staining for both AFP and vimentin was overlapped in mesenchymal cells at each stage tested. CONCLUSION: This study has for the first time This study has for the first time demonstrated that AFP is localized in the mesenchyme of rat colon from the embryo to the weaning stage by immunofluorescence and presents 72-kDa isoform in the developing rat colons by Western blotting. The dynamic expression of AFP in the various developmental stages of the colon indicates that AFP might be involved in many aspects of colon development.

  3. Prognostic value of serum α-fetoprotein in ovarian yolk sac tumors: A systematic review and meta-analysis

    OpenAIRE

    GUO, YANG-LONG; Zhang, Ying-Li; Zhu, Jian-Qing

    2014-01-01

    This study was conducted to determine the prognostic value of serum α-fetoprotein (AFP) levels in patients with ovarian yolk sac tumor (OYST). We performed a systematic review and meta-analysis to assess the associations between serum AFP level and prognosis in OYST. A total of 12 quantitative studies met the inclusion criteria. Preoperative AFP was not found to be associated with overall survival (OS) [odds ratio (OR)=0.84, 95% confidence interval (CI): 0.43–1.62] in OYST. However, a high po...

  4. Serum Aminoglycoside Assay by Enzyme-Mediated Immunoassay (EMIT): Correlation with Radioimmunoassay, Fluoroimmunoassay, and Acetyltransferase and Microbiological Assays

    OpenAIRE

    White, L O; Scammell, L. M.; Reeves, D S

    1981-01-01

    Enzyme-mediated immunoassay (EMIT) serum aminoglycoside assay results were accurate and precise and correlated well with radioimmunoassay, fluoroimmunoassay, and acetyltransferase and microbiological assay determinations.

  5. A radioimmunoassay for determination of anti-actin antibodies

    International Nuclear Information System (INIS)

    The reaction of spontaneously occurring human anti-actin antibodies and experimentally produced rabbit anti-actin antibodies was investigated in a solid-phase radioimmunoassay (RIA). Three structurally different in vitro forms of actin, monomeric G-actin, filamentous F-actin and aggregated denatured actin were used as antigens. Human anti-actin antibodies reacted with F- and G-actin but not with aggregated actin, while rabbit anti-actin antibodies gave a strong reaction with all 3 forms of actin indicating differences in antibody specificities. The results of the anti-actin RIA were compared with those obtained by indirect immunofluorescence (IFL) on cryostat sections of rat stomach. The anti-actin RIA discriminated between patients' sera and control sera in most cases, although the indirect IFL test gave more conclusive results. The seemingly low sensitivity of the anti-actin RIA compared with that of indirect IFL test for detection of human anti-actin antibodies is probably due to favourable antigen distribution in tissue, not available in the solid phase. The anti-actin RIA was able to detect anti-actin antibodies in 8 out of 8 immunized rabbits although only two produced antibodies detectable by indirect IFL. The differences in reactivity between the two methods may depend on the presence of aggregated denatured actin in the antigen preparation used for immunization and exposure of the corresponding antigenic determinants of actin on the solid phase. (Auth.)

  6. Radioimmunoassay of pepsinogens I and II in human serum

    International Nuclear Information System (INIS)

    Radioimmunoassay (RIA) for pepsinogens I and II (PG I and II) in serum is developed by using the purified PG I and II from human gastric mucosa. The assay range was 8∼256 μg/L for PG I and 2∼64 μg/L for PG II, the sensitivity was 1.3 and 0.6 μg/L, respectively. The reproducibilities of PG I -RIA (106.9%) and PG II-RIA (106.7%) are quite well. The within- and between-assay coefficient of variation (CV)of each RIA was 5.1% and 6.3% for PG I, 7.2% and 8.9% for PG II. The serum PG I level in healthy volunteer was 60.41 +- 14.98 μg/L (mean +- SD), significantly higher than the serum PGII level which was 20.70 +- 9.64 μg/L. Therefore, the PG I-RIA and PG II-RIA are useful tools in studying the relation between serum pepsinogen levels and various peptic disorders

  7. The development of a radioimmunoassay for 18-hydroxy-corticosterone

    International Nuclear Information System (INIS)

    It has been suggested that 18-hydroxy-corticosterone (18-OH-B) is a precursor for aldosterone in the biosynthetic pathway. This steroid had not been measured in peripheral plasma and therefore to evaluate its clinical role a sensitive and specific radioimmunoassay was developed. The assay is described and was applied to measure the levels of 18-hydroxy-corticosterone in normal human subjects on (a) an ad libitum sodium intake whilst lying or standing, (b) on a high or low sodium intake whilst recumbent, (c) before and after treatment with either ACTH, metyrapone or dexamethasone on a high or low sodium intake whilst recumbent (d) after insulin induced hypoglycaemia and (e) after infusions of 18-hydroxy-corticosterone. The levels of 18-hydroxy-corticosterone were also measured in patients with (a) 17α-hydroxylase deficiency, (b) primary or secondary aldosteronism, (c) Bartter's syndrome, (d) Cushing's syndrome, (e) adrenalectomised patients or patient's with Addison's disease and (f) patients with essential hypertension. (author)

  8. Radioimmunoassay for the simultaneous determination of morphine and codeine

    International Nuclear Information System (INIS)

    Antiserum against morphine was produced in rabbits immunized with morphine hapten conjugated to bovine serum albumin. The carrier protein was conjugated to the nitrogen atom of the opiate alkaloid in order to make the phenolic hydroxy group on C3 and the alcoholic group on C6 as determinant groups. The antibody does not recognize codeine or the major metabolite of morphine, 3-O-monoglucuronide. This antibody was used in conjunction with an antobody prepared against 3-O-carboxymethylmorphine to develop a radioimmunoassay which can measure codeine in the presence of morphine. The assay was used to follow both the plasma and brain levels of codeine and its biotransformation to morphine. Codeine when administered at a dose of 5 mg/kg i.v. showed a biphasic plasma decay curve the first phase of which had a tsub(1/2) of 26 min. Peak concentrations of morphine were detected in the plasma following that dose of codeine at 0.5 h 30 min after the injection of 20 mg/kg i.p. codeine, the brain levels of morphine were only 2% that of codeine. Thereafter, the brain levels of morphine slowly declined

  9. Sensitive radioimmunoassay method for urinary kinins in man

    International Nuclear Information System (INIS)

    A sensitive and specific radioimmunoassay method for urinary kinins was developed, which uses antiserum against synthetic bradykinin in combination with labeled tyr8-bradykinin. The assay detects as little as 6 pg per tube of bradykinin. The dose-response curves of bradykinin, lysyl-bradykinin (kallidin), and methionyl-lysyl-bradykinin were almost identical. This suggests that the values estimated with bradykinin as the standard exhibit the total urinary kinins which contain these three peptides. The assay was performed without extraction of urinary samples, since a chromatographic study demonstrated that no interfering substances in urine samples remain in our assay system. The urinary levels of total kinins in 10 normal male subjects, three male patients with chronic renal failure, and 12 male patients with essential hypertension were 37.9 +- 3.9 μg/day (mean +- S.E.M.), 9.0 +- 5.1 μg/day, and 24.2 +- 5.2 μg/day, respectively

  10. Radioiodination and quality control of human thyrotropin for radioimmunoassay

    International Nuclear Information System (INIS)

    This work reports the radioiodination of human thyrotropin (hTSH) in our laboratory (IPEN) and evaluates its quality in comparison with a commercial product. The radioiodination yield obtained in 20 experiments ranged from 18.5 to 56.3%, while the purification recovery ranged from 75.5 to 124.0% and the specific activity ranged from 1.01 to 3.10 MBq μg-1. The values for the distribution coefficient revealed in the purification of radioiodinated hTSH ranged from 0.232 to 0.371. When tested concomitantly in the same radioimmunoassay system, the IPEN and the commercial tracer presented parallel standard curves. A highly significant correlation ion was observed between the quality control samples estimated through both curves (p < 0.001). These results confirm the quality of the hTSH radioiodinated at IPEN and suggest the acquirement of self-sufficiency in this 'in vitro' nuclear technology. (author) 26 refs.; 5 figs.; 2 tabs

  11. Specific and direct proinsulin radioimmunoassay for the evaluation of insulinomas

    International Nuclear Information System (INIS)

    The paper describes a highly specific human proinsulin (hPI) radioimmunoassay (RIA) developed by using biosynthetic hPI as immunogen, standard and tracer. Proinsulin was radioiodinated by the iodogen method and purified on QAE-Sephadex A-25 to a specific activity of 6.5 MBq/μg. The antiserum was raised in a guinea pig and then adsorbed against insulin and C-peptide conjugate to Sepharose to improve its specificity. The range of the standard curve extended from 0.004 to 16 pmol/mL, with a 50% displacement between 0.19-0.26 pmol/mL and the minimal detectable concentration between 0.01-0.04 pmol/mL. This sensitive and specific RIA proved suitable for measurements of serum hPI concentrations in patients with insulinomas, which levels ranged from 0.11 to 1.80 pmol/mL (n = 8). (author). 18 refs, 6 figs, 1 tab

  12. Radioimmunoassay for C-peptide in diabetic children

    International Nuclear Information System (INIS)

    Direct insulin radioimmunoassay (RIA) studies in a diabetic are no longer meaningful once insulin therapy has been instituted. For this reason, use is made of RIA for blood C-peptide, a proinsulin component reflecting endogenous insulin secretion independently of insulin therapy. The paper reports experience with C-peptide RIA studies carried out on blood from 273 diabetic children of normal body weight and 11.3 years average age, as well as 31 healthy children (control group). Diabetes duration ranged from 7 days to 14 years. The basic level of C-peptide in diabetic children is lower than that of healthy ones. Glucose stimulation produces C-peptide elevation in healthy but not in diabetic children. Glucagon stimulation produced a further rise of blood C-peptide in the healthy children. Diabetics showed very modest response to glucagon stimulation. C-peptide secretion in diabetic children proved to be inversely proportional to the duration of the diabetes. These findings in children with diabetes mellitus indicated their insulin secretion by beta cells of the pancreatic islets of Langerhans to be substantially decreased and unresponsive to glucose and glucagon stimulation. 3 figs, 1 tab

  13. Radioimmunoassay for plasma corticotropin in frogs (Rana esculenta L.)

    International Nuclear Information System (INIS)

    A radioimmunoassay technique has been developed for measuring frog plasma corticotropin (ACTH) without prior extraction. Using synthetic porcine ACTH as a reference standard, 131I-labeled synthetic human ACTH (sp act greater than 500 mCi/mg) as tracer and rabbit anti-porcine ACTH serum, the lower measurable value was estimated at about 4 pg ACTH. Only human and porcine ACTH, ACTH, and frog pituitary ACTH reacted with the rabbit anti-porcine ACTH serum. No cross-reactivity has been found with synthetic ACTH, αMSH, and bovine βMSH. Appearance of damaged 131I-h ACTH components after storage in plasma solutions was followed for 7 days. The conditions making it possible to reduce ACTH damage have been ascertained. The average plasma corticotropin level (+- CI) was found to be 38.8 +- 7.8 pg/ml without any significant difference between males and females. These results suggest that frog ACTH secretion has much in common with mammalian secretions

  14. An Interlaboratory study of lipid effects on steroid radioimmunoassay

    International Nuclear Information System (INIS)

    The lipid effects on the performances of routine steroid radioimmunoassay (RIA) have been assessed using the scheme of the CNR interlaboratory quality control program. Cortisol, estradiol, progesterone and testosterone assays have been considered. In the study, ca 80 laboratories were supplied with two sets of control plasma samples with different triglyceride contents (pool N, ca 120 mg/dl; pool L, ca 310 mg/dl), each corresponding to three steroid levels (level 0: charcoaldeprived samples; levels 1 and 2: the same added with increasing steroid amounts). A comparison of the neat results obtained by partecipants for both levels 1 and 2 of N and P panels - after subtraction of the concentrations estimated for level 0 - gave a direct information on lipid effects (triglycerides being assumed as an index of lipemia). In no case the abnormality high triglyceride content proved effective in practical terms, though significant differences were observed for testosterone and progesterone (ca 10% underestimation in pool L) and for estradiol (ca 10% overestimation in pool L)

  15. A radioimmunoassay for lignin in plant cell walls

    International Nuclear Information System (INIS)

    Lignin detection and determination in herbaceous tissue requires selective, specific assays which are not currently available. A radioimmunoassay (RIA) was developed to study lignin metabolism in these tissues. A β-aryl ether lignin model compound was synthesized, linked to keyhole limpet hemocyanin using a water-soluble carbodiimide, and injected into rabbits. The highest titer of the antiserum obtained was 34 ηg/mL of model derivatized BSA. An in vitro system was developed to characterize the RIA. The model compound was linked to amino activated polyacrylamide beads to mimic lignin in the cell walls. 125I Radiolabelled protein A was used to detect IgG antibody binding. The RIA was shown in the in vitro system to exhibit saturable binding. The amount of antibody bound decreased when the serum was diluted. Immunoelectrophoresis and competitive binding experiments confirmed that both aromatic rings of the lignin model compound had been antigenic. Chlorogenic acid, a phenolic known to be present in plant cells, did not compete for antibody binding. The RIA was used to measure lignin in milled plant samples and barley seedlings. Antiserum binding to wheat cell walls and stressed barley segments was higher than preimmune serum binding. Antibody binding to stressed barley tissue decreased following NaClO2 delignification. The RIA was found to be less sensitive than expected, so several avenues for improving the method are discussed

  16. The radioimmunoassay of clomipramine (Anafranil-Geigy). A tricyclic antidepressant

    International Nuclear Information System (INIS)

    A radioimmunoassay has been developed for the tricyclic antidepressant, clomipramine (Anafranil-Geigy) which allows accurate determination of plasma levels without a pre-assay purification step. This is achieved by generation of specific antisera using an antigen produced by conjugation of clomipramine to bovine serum albumin via the 10, 11 bridge positions. As expected, cross-reaction of the pharmacologically active major metabolite, desmethylclomipramine was <5% and that of didesmethylclomipramine <1%. Specificity was confirmed by comparing titres achieved in the routine assay with those observed in an assay incorporating a pre-assay thin-layer chromatographic purification step. Pharmacokinetic data were in agreement with double radioisotope derivative assays and also with previously reported assays using GC or GC/MS techniques. The sensitivity is superior to any previous assay known to us for this class of compound. The specificity and precision, coupled with the high sample turnover (greater than 300 samples a week per technician), make the assay ideal for supervision of patient compliance and routine assay of samples generated in large clinical trials. (author)

  17. Development and clinical application of human gastrin radioimmunoassay

    International Nuclear Information System (INIS)

    The determination of human gastrin levels in the blood is very important for diagnosis of gastrointestinal disorders. This work describes the radioimmunoassay of gastrin developed according to Russell et al. and its clinical application measuring fasting levels of this hormone in normal subjects, gastrectomized, chagasics, patients with chronic renal failure (CRF), pernicious anemia (PA) and Zollinger-Ellison syndrome (ZES). Synthetic human gastrin was used for radioiodination and as standard, while the specific antibody was raised in rabbits. Gastrin was radioiodinated by a modification of the chloramine T technique and purified by anion exchange chromatography in QAE-Sephadex A-25 to a specific activity around 200 uCi/ug. The assays were performed by incubation of 125I-gastrin, standard gastrin (zero to 500 pmol/l) or unknown samples with the antiserum for 4 days at 40C. The antibody bound and free 125 I-gastrin was separated by adsorption of the latter to the charcoal. The basal gastrin values of normal subjects ranged from 2 to 74 pmol/l, being these levels higher in the chagasics (from 6 to 261 pmol/l). Higher levels of gastrin were determined in patients with CRF (from 12 to 350 pmol/l), PA (from 160 to 680 pmol/l) and with ZES(1010 pmol/l), while very low levels were confirmed in gastrectomized (from 1 to 8 pmol/l). (author)

  18. Multiple steroid radioimmunoassays and automation. Versatile techniques for reproductive endocrinology

    International Nuclear Information System (INIS)

    The combination of the efficient steroid-separating properties of a lipophilic Sephadex derivative Lipidex-5000sup(TM) and the use of antibodies with carefully selected specificity allows the quantitative determination of pregnenolone, progesterone, 17α-hydroxyprogesterone, androstenedione, testosterone, 5α-dihydrotestosterone, 5α-androstanedione, androsterone and 5α-androstane-3α, 17β-diol in 1- to 2-ml samples of both blood serum and amniotic fluid as well as in 300- to 600-mg pieces of prostatic tissue. The adaptation of the pipetting unit and incubator of a discrete clinical chemical analyser, System Olli 3000, for the automation of the radioimmunoassays has resulted in a greatly increased through-put and has decreased the experimental error of the procedure. In studies on reproductive endocrinology, the methodology developed has allowed the detection of a sex difference in androgen composition of the amniotic fluid early in pregnancy. Further, it is very likely that the decline in steroid production by the testis seen during the first year of life and then in senescence is affected by basically different mechanisms. There are also important differences in the steroid content of normal, hyperplastic and carcinomatous prostate. (author)

  19. An Evaluation by TSH Radioimmunoassay on Familial Thyroid Disorders

    International Nuclear Information System (INIS)

    The occurrence of thyroid disorders is connected with iodine deficiency, defective synthesis or releasing of thyroid hormone and endemicity. Genetic factors are known as a single gene defects, interaction of multiple genes with environmental factors, as well as chromosomal aberrations. Diofnosis thyroid disorders is enforced by 13I uptake test, thyroid scanning with 131I or 99mTc and serum radioimmunoassays of T3, T4, free T4 and TSH. They were largely classified as hypothyroidism, hyperthyroidism, simple goiter and normal. The pedigree of 58 families was drawn by propositus, and then the correlation between thyroid disorders and TSH levels was analyzed. The results are as follows: 1) The offsprings and their mothers of 15 families were hypothyroidism, THS level was 5 folds for offsprings and 4 folds for mothers in comparison with control group. 2) 13 families were hyperthyroidism in siblings but their mothers were normal in thyroid function, TSH level of the siblings was lower than control group. 3) Though the offsprings and their mothers of 10 families were similar to TSH level of control group, they are all simple goiter, familial thyroid disorders, in other thyroid function test. The familial thyroid disorders suggested that these transmitted from mothers to offsprings with X-linked dominant or autosomal dominant inheritance.

  20. Multiple steroid radioimmunoassays and automation: versatile techniques for reproductive endocrinology

    International Nuclear Information System (INIS)

    The combination of the efficient steroid separating properties of a lipophilic Sephadex derivative Lipidex-5000sup(TM), with the use of antibodies with carefully selected specificity allows the quantitative determination of pregnenolone, progesterone, 17α-hydroxyprogesterone, androstenedione, testosterone, 5α-dihydrotestosterone, 5α-androstanedione, androsterone and 5α-androstane-3α, 17β-diol from 1-2 ml samples of blood serum, amniotic fluid or 300-600 mg pieces of prostatic tissue. The adaptation of the pipetting unit and incubator of a discrete clinical chemical analyzer, System Olli 3000, for the automation of the radioimmunoassays has resulted in a greatly increased through-put and decreased experimental error of the procedure. In studies on reproductive endocrinology, the methodology developed has allowed the detection of a sex difference in androgen composition of the amniotic fluid early in pregnancy. Further, it is very likely that the decline in steroid production by the testis seen during the first year of life and then in senescence is affected by basically different mechanisms. There are also important differences in the steroid content of normal, hyperplastic and carcinomatous prostate. (orig.)

  1. Measurement of guinea pig eosinophil major basic protein by radioimmunoassay

    International Nuclear Information System (INIS)

    Guinea pig eosinophil major basic protein (MBP) was measured by radioimmunoassay (RIA) using 131I-MBP. Two critical features of the assay were: (1) alkylation of the MBP with iodoacetamide prior to radioiodination and (2) inclusion of another basic protein, either protamine or histone, in the phosphate buffer. Freshly isolated non-alkylated MBP was immunologically deficient when compared to alkylated or reduced MBP, but its reactivity could be redtores by reduction with dithiothreitol and alkylation. Reduction and alkylation also restored the immunoreactivity of polymerized MBP. MBP levels were not elevated in sera from guinea pigs parasitized with Trichinella spiralis and having peripheral blood eosinophilia. Muscle extracts from Trichinella infected animals showed significantly higher levels of MBP activity than normal controls. MBP was measurable in extracts of untreated eosinophils, but reduction and alkylation of these extracts increased MBP activity several fold. The RIA permits detection of MBP in body fluids and tissues at levels as low as 2 ng./ml. The RIA is useful in assessing increased or decreased levels of MBP activity in samples from experimental animals when compared to samples from controls. (author)

  2. Laboratory training manual on radioimmunoassay in animal reproduction

    International Nuclear Information System (INIS)

    Reproduction must always be regarded as one of the major limiting factors in animal production and many of the modern methods for improving reproduction rely heavily on the ability to measure hormone levels in blood and milk. This has produced a world-wide demand for laboratory facilities to carry out hormone assays and the need for specialist training to allow these assays to be undertaken. The need to measure nanogram and picogram quantities and the use of radionuclides require a good deal of skill and care and this Manual has been prepared to aid training and provide the sort of information that rarely appears in scientific papers. It represents a further step in the Joint FAO/IAEA Division's series of Laboratory Training Manuals, and has been designed to aid training programmes of the type carried out during the Joint FAO/IAEA Interregional Training Course on Radioimmunoassay and its Application in Research on Animal Reproduction at Cornell University in July 1982. Many of the laboratory exercises described in this Manual are based on those conducted during the course

  3. Specific radioimmunoassay of human β-endorphin in unextracted plasma

    International Nuclear Information System (INIS)

    With an antiserum against human β-endorphin (β-EP) crossreacting <2% with human β-lipotropin (β-LPH) by weight we have developed a radioimmunoassay that can detect 1 pg β-EP in diluted raw plasma. In a.m. fasting plasma of 14 normal subjects β-EP ranged from <5 to 45 pg/ml. β-EP was elevated in untreated, but normal in successfully treated Cushing's disease; undetectable in a patient with adrenal adenoma; extremely high in Nelson's syndrome; and elevated in a patient with bronchogenic carcinoma before, but undetectable after tumor resection. In subjects with intact hypothalamic-pituitary-adrenal axis, β-EP was undectectable after dexamethasone and increased after metyrapone administration and insulin-induced hypoglycemia. β-EP concentration was considerably lower in serum than in simultaneously collected plasma, but increased in serum left unfrozen for several hours after clot removal. Thus, β-EP behaves like a hormone responding to the same stimuli as ACTH and β-LPH and blood appears to contain enzymes both generating and destroying immunoreactive β-EP

  4. Radioimmunoassay of fibrinopeptide A: Modifications of technique and clinical trials

    International Nuclear Information System (INIS)

    In 1971 Nossel and coworkers developed a highly sensitive radioimmunoassay for the determination of plasma concentration of fibrinopeptide A. Our own experience gained using this test is reported here as well as a number of improvements in methodology which have not only led to a shortening of the whole test procedure to one day but also to greater ease of handling. The modifications concern a gel adsorption procedure for the extraction of FPA from defibrinated plasma samples, the double antibody method for separating free and antibody-bound tracer-phase in assay and automatic evaluation of test results with the aid of a suitable computer programme. Considerations concerning the specificity of conclusions drawn from the test, in particular the necessity of keepin gout interfering plasmin-induced fibrinogen break-down products, play an important role in all changes of methodology. An attempt was made to define a normal range for FPA-plasma levels and a mean value of 1.51 ng/ml at a standard deviation of 0.69 ng/ml was obtained from 42 clinically healthy test patients. Pathologically evaluated FPA-levels were regularly found in patients with septic or thrombotic illness. (orig./MG)

  5. The $\\alpha-\\alpha$ fishbone potential revisited

    CERN Document Server

    Day, J P; Elhanafy, M; Smith, E; Woodhouse, R; Papp, Z

    2011-01-01

    The fishbone potential of composite particles simulates the Pauli effect by nonlocal terms. We determine the $\\alpha-\\alpha$ fishbone potential by simultaneously fitting to two-$\\alpha$ resonance energies, experimental phase shifts and three-$\\alpha$ binding energies. We found that essentially a simple gaussian can provide a good description of two-$\\alpha$ and three-$\\alpha$ experimental data without invoking three-body potentials.

  6. Alpha One Foundation

    Science.gov (United States)

    ... Tested Find Support Find Doctor What Is Alpha-1? Alpha-1 Antitrypsin Deficiency (Alpha-1) is a ... results for inhaled augmentation More News Our Number One Goal: Find a cure for Alpha-1. Website ...

  7. Alpha-1 Antitrypsin Test

    Science.gov (United States)

    ... helpful? Also known as: Alpha 1 -antitrypsin; A1AT; AAT Formal name: Alpha 1 Antitrypsin; α1-antitrypsin Related ... know? How is it used? Alpha-1 antitrypsin (AAT) testing is used to help diagnose alpha-1 ...

  8. Alpha spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Krueger, Felix; Wilsenach, Heinrich; Zuber, Kai [IKTP TU-Dresden, Dresden (Germany)

    2014-07-01

    Alpha decays from long living isotopes are one of the limiting backgrounds for experiments searching for rare decays with stringent background constrains, such as neutrinoless double beta decay experiments. It is thus very important to accurately measure the half-lives of these decays, in order to properly model their background contribution. Therefore, it is important to be able to measure half-lives from alpha decays of the order of 1 x 10{sup 15} yr. A measurement of such a long lived decay imposes, however, a series of challenges, where the correct discrimination between background and true signal is critical. There is also a more general interest in such long living half-life measurements, as their value depends crucially on the underlying nuclear model. This work proposes a setup to measure long lived alpha decays, based on the design of the Frisch-Grid ionisation chamber. It is shown that the proposed design provides a good separation of signal and background events. It is also demonstrated that, with pulse shape analysis, it is possible to constrain the source position of the decay, further improving the quality of the data. A discussion of the characterisation of the detector is also presented as well as some results obtained with calibration sources.

  9. Alpha spectroscopy

    International Nuclear Information System (INIS)

    Alpha decays from long living isotopes are one of the limiting backgrounds for experiments searching for rare decays with stringent background constrains, such as neutrinoless double beta decay experiments. It is thus very important to accurately measure the half-lives of these decays, in order to properly model their background contribution. Therefore, it is important to be able to measure half-lives from alpha decays of the order of 1 x 1015 yr. A measurement of such a long lived decay imposes, however, a series of challenges, where the correct discrimination between background and true signal is critical. There is also a more general interest in such long living half-life measurements, as their value depends crucially on the underlying nuclear model. This work proposes a setup to measure long lived alpha decays, based on the design of the Frisch-Grid ionisation chamber. It is shown that the proposed design provides a good separation of signal and background events. It is also demonstrated that, with pulse shape analysis, it is possible to constrain the source position of the decay, further improving the quality of the data. A discussion of the characterisation of the detector is also presented as well as some results obtained with calibration sources.

  10. A Rat α-Fetoprotein Binding Activity Prediction Model to Facilitate Assessment of the Endocrine Disruption Potential of Environmental Chemicals

    Science.gov (United States)

    Hong, Huixiao; Shen, Jie; Ng, Hui Wen; Sakkiah, Sugunadevi; Ye, Hao; Ge, Weigong; Gong, Ping; Xiao, Wenming; Tong, Weida

    2016-01-01

    Endocrine disruptors such as polychlorinated biphenyls (PCBs), diethylstilbestrol (DES) and dichlorodiphenyltrichloroethane (DDT) are agents that interfere with the endocrine system and cause adverse health effects. Huge public health concern about endocrine disruptors has arisen. One of the mechanisms of endocrine disruption is through binding of endocrine disruptors with the hormone receptors in the target cells. Entrance of endocrine disruptors into target cells is the precondition of endocrine disruption. The binding capability of a chemical with proteins in the blood affects its entrance into the target cells and, thus, is very informative for the assessment of potential endocrine disruption of chemicals. α-fetoprotein is one of the major serum proteins that binds to a variety of chemicals such as estrogens. To better facilitate assessment of endocrine disruption of environmental chemicals, we developed a model for α-fetoprotein binding activity prediction using the novel pattern recognition method (Decision Forest) and the molecular descriptors calculated from two-dimensional structures by Mold2 software. The predictive capability of the model has been evaluated through internal validation using 125 training chemicals (average balanced accuracy of 69%) and external validations using 22 chemicals (balanced accuracy of 71%). Prediction confidence analysis revealed the model performed much better at high prediction confidence. Our results indicate that the model is useful (when predictions are in high confidence) in endocrine disruption risk assessment of environmental chemicals though improvement by increasing number of training chemicals is needed. PMID:27023588

  11. α-fetoprotein involvement during glucocorticoid-induced precocious maturation in rat colon

    Institute of Scientific and Technical Information of China (English)

    Min Chen; Peng Sun; Xiao-Yan Liu; Dan Dong; Jun Du; Luo Gu; Ying-Bin Ge

    2011-01-01

    AIM: To investigate the role of α-fetoprotein (AFP), a cancer-associated fetal glycoprotein, in glucocorticoidinduced precocious maturation in rat colon. METHODS: Colons from suckling Sprague-Dawley rats were used in this study. Corticosterone acetate at a dose of 100 μg/g body weight was given to normal pups on days 7, 9 and 11 after birth to induce hypercorticoidism. Control animals were injected with identical volumes of normal saline. Some rats receiving corticosterone 7 d after birth were also treated with mifepristone (RU38486), a glucocorticoid cytoplasm receptor antagonist to investigate the effects of glucocorticoids (GCs). The morphological changes of the crypt depth and villous height of the villous zone in colon were observed as indices of colon maturation. Expression levels of AFP in colons were detected by reverse transcriptase polymerase chain reaction and Western blotting. To identify the cellular localization of AFP in developing rat colons, double-immunofluorescent staining was performed using antibodies to specific mesenchymal cell marker and AFP. RESULTS: Corticosterone increased the crypt depth and villous height in the colon of 8- and 10-d-old rats with hypercorticoidism compared with that in the control animals (120% in 8-d-old rats and 118% in 10-d-old rats in villous height, P = 0.021; 145% in 8-d-old rats and 124% in 10-d-old rats in crypt depth, P = 0.017). These increases were accompanied by an increase of AFP expression in both mRNA and protein (2.5-folds in 8-dold and 2.5-folds in 10-d-old rats higher than in control animals, P = 0.035; 1.8-folds in 8-d-old and 1.3-folds in 10-d-old rats higher than in control animals, P = 0.023). Increased crypt depth and villous height and increased expression of AFP in the colon of rats with hypercorticoidism were blocked by mifepristone. Both had positive staining for AFP or vimentin, and overlapped in mesenchymal cells at each tested colon.

  12. Solid-phase radioimmunoassay for measurement of circulating antibody titres to wheat gliadin and its subfractions in patients with adult coeliac disease

    Energy Technology Data Exchange (ETDEWEB)

    Ciclitira, P.J.; Ellis, H.J. (Guy' s Hospital, London (UK)); Evans, D.J. (Hammersmith Hospital, London (UK))

    1983-08-26

    A solid-phase radioimmunoassay for the measurement of circulating antibody titres to wheat gliadin is described. Using this assay, the authors have measured antibody titres to unfractionated gliadin in normal healthy controls, in coeliac patients on a gluten-free or a normal diet, and in patients with ulcerative colitis and Crohn's disease. High titres of antibodies to unfractionated gliadin were observed only in the patients with untreated coeliac disease. Antibody titres to ..cap alpha.., ..beta.., ..gamma.. and ..omega.. gliadin subfractions were measured in patients with untreated coeliac disease and compared with titres in normal controls. Patients with untreated coeliac disease had higher antibody titres to the gliadin subfractions. No specific pattern of circulating antibody titres to gliadin subfractions was observed in the untreated coeliac patients which would provide a diagnostic profile. These results suggest shared antigenicity between the gliadin subfractions.

  13. Radioimmunoassay principles and techniques. January, 1975-September, 1981 (citations from the International Information Service for the Physics and Engineering Communities data base). Report for Jan 75-Sep 81

    International Nuclear Information System (INIS)

    This bibliography contains citations concerning the general principles, methodology and the evaluation of radioimmunoassay. Many assays (including T3, T4,TSH, HPAP) are discussed and computer assisted radioimmunoassay analysis is considered

  14. Radioimmunoassay of human growth hormone: characterization of a monoclonal antibody

    International Nuclear Information System (INIS)

    The introduction of cell-hybridization techniques by Koehler and Milstein in 1975 to produce monoclonal antibodies (MA) was a definite improvement in methodological tools of radioimmunoassay, quite apart from all other applications of this technique in immunohistochemistry, affinity chromatography, target-directed drug delivery systems etc. MAs would ideally be suited in RIAs, when the specificity is the crucial aspect of the determination. However, for reasons which are not completely understood, assays with MAs very often lack the attribute of being highly sensitive. Despite several reports in the literature on MAs against human growth hormone (HGH), none of these seem sensitive enough to be of use in clinical chemistry, where a too strongly marked specificity may even be unwanted. However, from the scientific point of view, MAs against polypeptide hormones are of great interest. An MA to HGH was developed with a sensitivity limit of 0.2 ng. The titre of the ascites fluid is higher than 1:106 and the specificity against human placental lactogen, human prolactin and rat growth hormone is nearly complete. A critical step of the RIA procedure is the separation of bound and free hormone. A combination of human immunoglobulin (Sandoglobulin) with polyethylene glycol gives optimal results. A Scatchard plot reveals an affinity constant of 4x10-11M-bar and a maximal binding capacity of 2x108cpm/μL. In conclusion, our monoclonal antibody represents an excellent investigational tool for endocrine research and it seems to be the most sensitive and specific MA to HGH described to date. However, for practical clinical applications, there seems to be little advantage of an MA over a conventional polyclonal antiserum. (author)

  15. Development of a Radioimmunoassay for Bovine Chymosin B

    Directory of Open Access Journals (Sweden)

    Borceux, JP.

    2007-01-01

    Full Text Available The present study was conducted to develop and validate a specific radioimmunoassay system for measurement of bovine chymosin B (bChyB concentrations in plasma samples. Bovine ChyB was used for immunization of rabbits and as standard and tracer. Chymosin B concentrations were measured in plasma samples from two groups of calves (Group 1: calves sampled from birth to 24 hours; Group 2: calves sampled from Day 1 to 21 after birth and from one cow during the peri-partum period. Detection limit of the assay was 9.0 ng/ml. Recovery was higher than 89.3%. Repeatability and reproducibility ranged from 1.52% to 5.23% and from 1.52% to 12.57% respectively. No cross-reaction was found with pepsinogen A from bovine, porcine or human origins. In Group 1, bChyB concentrations increased from 47.3 ± 45.1 ng/ml (5 min after birth to 325.5 ± 161.2 ng/ml (12 hours after birth, then no significant change was observed till 24 hours after birth (293.0 ± 161.5 ng/ml. In Group 2, concentrations decreased from Day 1 (455.3 ± 191.1 ng/ml to Day 21 (117.9 ± 85.1 ng/ml. In adult cow, mean concentration was 136.0 ± 32.3 ng/ml. In conclusion, bChyB is able to cross the stomach basal membrane and to reach the blood circulation at detectable levels in both young calves and adult cows.

  16. Radioimmunoassay methods for the study of communicable diseases

    International Nuclear Information System (INIS)

    Radioimmunoassay (RIA) and related methods should prove particularly suitable for the diagnosis and study of common communicable diseases. While hepatitis caused by the B virus is currently the only major communicable disease for which such methods are routinely used, in the near future RIA methods will be introduced for a variety of bacterial, fungal, viral, protozoan and helminthic pathogens. These methods will be most helpful for highly prevalent disease states in which rapid, low-cost diagnostic methods are not now available. The methods should be directed towards the detection of organism-specific antigens. The selection of appropriate antigens is of primary importance in the development of new methods. In the area of communicable diseases special consideration should be given to the immunochemical specificity of antigens, standardization of antigen preparations, validation of assay methods, and the safety of assay constituents and samples. Tuberculosis is an ideal disease for the introduction of RIA methods. The development of an assay which detects specific heat-altered tuberculoprotein may offer a rapid low-cost method for use in the diagnosis and management of active tuberculosis. A method which detects heat-altered antigen is dependent upon the antigen retaining its immunochemical species-specificity in the heat-altered state and allows for the decontamination of samples, the dissociation of antigen-antibody complexes, and the denaturation of endogenous antibody. In addition to viral hepatitis and tuberculosis, a number of communicable diseases are now being approached by RIA methodology. In general, these methods have not as yet been well validated or subjected to extensive clinical trial. In the near future it may be possible to review the results obtained with these assays. (author)

  17. Development of reagents for radioimmunoassay of: triiodothyronine, thyroxine and thyrotrophin

    International Nuclear Information System (INIS)

    The radioimmunoassay (RIA) of thyroid hormones it is the but it frequents of all the studies carried out by RIA in the laboratories of Nuclear Medicine, these essays are carried out with imported reagents. In the ININ the reagents and the necessary methodology have been developed for the triiodothyronine (T3), thyroxine (T4) and thyrotrophin (TSH). The good titles of the antibodies (Ac) primary for each hormone were of 1:4,000; 1:750 and 1:1,500. The used separation system was of double Ac with PEG to 10%, with titles of 1:10 for the second Ac of lamb. The specific activity for 125-I-T3 and 125-I-T4 oscillate between 850 at 900 μCi / μ g: being this of 90 μ Ci /μg for TSH. To the first two hormones they were added 1-8 aniline naftalen sulfonic acid (ANS) to concentrations of 3 and 2 mg/ml respectively. As buffer for T3 and T4 it was used Tris-HCl pH 8.6 and PBS with normal serum of rabbit (SNC) for TSH. The standards got ready in buffer or free serum of thyroid hormones. The slope of the standard curves varied between -2.3 to -2.7 and the variation intra and inter assay among 4 to 10%. It is had at the moment in the ININ with standardized reagents for the RIA of T3, T4 and TSH, it is hoped to carry out tests in other laboratories and to establish the conditions of stability more appropriate to begin the preparation of pilot reagents. (Author)

  18. Quantification of the adrenal cortex hormones with radioimmunoassay

    International Nuclear Information System (INIS)

    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 α-OH-PRG and of the DHEA-SO4 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)

  19. The reproductive performance in female Damascus goats using progesterone radioimmunoassay

    International Nuclear Information System (INIS)

    An experiment was conducted on Damascus goats' female kids and does to study the reproductive performance by identifying some reproductive parameters such as age and weight at puberty, start of the breeding season, duration of pregnancy and other related parameters. The above objectives were met by means of the measurement of progesterone levels in blood sera using radioimmunoassay. The results could be summarized as follows: 1- Average weight of female kids at age of 7-8 months was 30.2 kg, at puberty was 35.6 ky and at mating was 41.6 kg. 2- There were individual differences in the age at puberty ranging from 266 to 653 days. 3- At age of 7-8 months, progesterone level was 0 n mol/1 in 8, and 0.1 and 0.2 n mol/1 in the 2 remaining female kids. This level increased to an average of 6.81 n mol/1 at puberty. 4- Average duration of pregnancy was 149.0 days, overall kids' birth weight was 3.73 kg, for singles was 4.43 kg, whereas for twins was 3.21 kg, for the kids produced by the female kids. Average kids' birth weight produced by the does was: overall 4.63 kg, singles 4.02 kg and for twins was. 4.18 kg. 5- Average progesterone e level on day 21 post-mating was 23.02 n mol/1. Therefore, the accuracy of early pregnancy diagnosis was 100%. 6- The breeding season for the experimental does started in September. (author)

  20. Process for preparation of a solid-phase radioimmunoassay support and use thereof

    International Nuclear Information System (INIS)

    A process is described for the preparation of a support useful in radioimmunoassay chromatographic columns. The process involves the preparation of a chromatographic gel capable of selectively retaining one or more components contained in an antigen-antibody-containing solution. The gel is bound to the appropriate antiserum, then freeze-dried, pulverized and compressed into a tablet. The tablet support swells upon contact with an antigen-antibody-containing solution to conform to the shape of the columns. An example of the application of this support in the radioimmunoassay of thyroid-stimulating hormone is described. This type of support is also particularly useful in second antibody solid phase radioimmunoassays since there is no limit to the size of the antigen to which this technology may be applied. (U.K.)

  1. Direct radioimmunoassay of rat urinary kallikrein and comparison with other measures of urniary kallikrein activity

    International Nuclear Information System (INIS)

    A highly sensitive radioimmunoassay for rat urinary kalikrein (minimal detectable amount, 40 pg/tube) has been developed. The assay uses a sheep antibody (Keq = 3.25 x 1010M-1) against purified Sprague-Dawley rat urinary kallikrein in a final dilution of 1:1,200,000. The assay incorporates a convenient and inexpensive PEG technique for separation of free from bound antigen. Parallel standard curves with rat urine or kidney homogenates were obtained. No cross-reactivity with human or dog urine samples or purified human urinary kallikrein was seen. Correlations among this assay, an esterolytic method, and a kininogenase radioimmunoassay for kallikrein were highly significant, with only the esterolytic assay demonstrating any significant nonspecificity. The radioimmunoassay can detect changes in urinary kallikrein levels produced by a maneuver known to alter urinary kallikrein excretion

  2. Luteinizing hormone-releasing hormone: radioimmunoassay and extraction from human serum

    International Nuclear Information System (INIS)

    Luteinizing hormone-releasing hormone (LRH) is a decapeptide which releases both LH and FSH in man. Its chemical synthesis allowed sufficient amounts to become available for clinical studies and for the generation of specific antibodies. Radioimmunoassay techniques have advantages over bioassays in relation to sensitivity and specificity. Radioimmunoassays have been applied to quantify exogenously administered LRH but data on the endogenous LRH levels of unextracted serum samples are still conflicting. The low concentrations of various polypeptide hormones make extraction procedures almost essential for their assay, in particular for improvement of specificity and for removal of interfering substances. The present paper concerns the extraction of LRH from biological fluids and the development of radioimmunoassay for testing the extracted samples

  3. Evaluation of a commercially available radioimmunoassay kit for measurement of doxorubicin in plasma

    International Nuclear Information System (INIS)

    We evaluated a commercially available (Diagnostic Biochemistry Inc.) doxorubicin 125I radioimmunoassay kit. This kit gave a high apparent doxorubicin concentration (> 12 μg/L), which was not linearly related to dilution, for two pools of normal human serum and plasma and also for samples collected from patients before they received the drug. In contrast, a doxorubicin 3H radioimmunoassay developed by us gave a low blank (2 μg/L), which was linearly related to dilution, for the same pools and patients' samples. Doxorubicin concentrations in the plasma of patients receiving the drug were compared by the two methods; the kit gave results five- to 10-fold those obtained with our assay. High nonspecific interference by serum and plasma as measured by the 125I radioimmunoassay must therefore be borne in mind by users of the kit, and we suggest that results should be corrected for these nonspecific effects

  4. α-Fetoprotein promoter-driven Cre/LoxP-switched RNA interference for hepatocellular carcinoma tissue-specific target therapy.

    Directory of Open Access Journals (Sweden)

    Yuan-Fei Peng

    Full Text Available BACKGROUND: RNA interference (RNAi has recently emerged as a potential treatment modality for hepatocellular carcinoma (HCC therapy, but the lack of cellular targets and sustained efficacy limits its application. The purpose of this study is to develop an HCC tissue-specific RNAi system and investigate its possibility for HCC treatment. METHODS: Two different HCC-specific RNAi systems in which therapeutic miRNA or shRNA against target gene (Beclin 1 was directly or indirectly driven by alpha-fetoprotein promoter (AFP-miRNA and AFP-Cre/LoxP-shRNA were constructed. Human HCC cell lines (HepG2, Hep3B and HCCLM3 and non-HCC cell lines (L-02, Hela and SW1116 were infected with the systems. The effectiveness and tissue-specificity of the systems were examined by Q-PCR and western blot analysis. The efficacy of the systems was further tested in mouse model of HCC by intravenous or intratumoral administration. The feasibility of the system for HCC treatment was evaluated by applying the system as adjuvant therapy to enhance sorafenib treatment. An AFP-Cre/LoxP-shRNA system targeting Atg5 gene (AFP-Cre/LoxP-shRNA-Atg5 was constructed and its efficacy in sensitizing HCC cells (MHCC97L/PLC to sorafenib treatment was examined by apoptosis assay in vitro and tumorigenesis assay in vivo. RESULTS: The AFP-miRNA system could silence target gene (Beclin 1 but required a high titer which was lethal to target cells. The AFP-Cre/LoxP-shRNA system could efficiently knockdown target gene while maintain high HCC specificity. Intratumoral injection of the AFP-Cre/LoxP-shRNA system could efficiently silence target gene (Beclin 1 in vivo while intravenous administration could not. The AFP-Cre/LoxP-shRNA system target Atg5 gene could significantly sensitize MHCC97L/PLC cells to sorafenib-induced apoptosis in vitro and tumor growth suppression in vivo. CONCLUSIONS: An efficient HCC tissue-specific RNAi system (AFP-Cre/LoxP-shRNA was successfully established. The system

  5. Examinations of hens' eggs on residues of Chloramphenicol using a radioimmunoassay

    International Nuclear Information System (INIS)

    In the Federal Republic of Germany the application of Chloramphenicol to animals used for human food supply is restricted by law. Milk and dairy products as well as hen's eggs and egg products are not allowed to contain more than 1 ppb Chloramphenicol. 18 hens were fed with water treated with Chloramphenicol. The eggs of the treated animals were then analysed by means of radioimmunoassay. The applied radioimmunoassay is suitable for routine analysis to a minimum detection limit of 1 ppb. 378 eggs from the Weser-Ems district were tested for Chloramphenicol. No sample contained Chloramphenicol. (orig.)

  6. Specific antiserum to Leu-enkephalin and its use in a radioimmunoassay

    International Nuclear Information System (INIS)

    This paper describes the preparation of an immunogenic protein-conjugate of Leu-enkephalin and the specificity of an antiserum generated to this antigen in rabbits. Such an antiserum, when used in a radioimmunoassay as described herein, should provide a useful tool for studying the role of both Leu- and Met-enkephalin in the 'Pain Pathway'. According to a recent hypothesis, enkephalin levels in the central nervous system should determine not only the pain threshold but phenomena such as tolerance, physical dependence and the withdrawl syndrome as well. From this respect, the importance of a radioimmunoassay is obvious

  7. What type of statistical model to choose for the analysis of radioimmunoassays

    International Nuclear Information System (INIS)

    The current techniques used for statistical analysis of radioimmunoassays are not very satisfactory for either the statistician or the biologist. They are based on an attempt to make the response curve linear to avoid complicated computations. The present article shows that this practice has considerable effects (often neglected) on the statistical assumptions which must be formulated. A more strict analysis is proposed by applying the four-parameter logistic model. The advantages of this method are: the statistical assumptions formulated are based on observed data, and the model can be applied to almost all radioimmunoassays

  8. IN VITRO CELL CULTURE AND HORMONE RADIOIMMUNOASSAY OF HUAMAN PITUITARY ADENOMAS

    Institute of Scientific and Technical Information of China (English)

    陆汉魁; 林祥通; 等

    1994-01-01

    Tissues from 30 human pituitary adenomas are monolayer-cell-cultured in vitro.Hormone secretion of GH,PRL,TSH,LH and FSH by cells into medium is detected by radioimmunoassay .The pattern and amount of hormone(s0 in the medium are used to determine the nature of the cells and thus to establish functional classification of pituitary adenomas.The results show that cell culture technique provides and easy and suitable mode for investigating the nature of pituitary adenomas.Hormone radioimmunoassay of culture medium is precise and reliable and represents the whole adenoma tissue.Further studies can lead to clearer understandngs of the pathology of pituitary adenomas.

  9. Differing results of direct and indirect solid phase radioimmunoassay for HBsAg in acute hepatitis

    International Nuclear Information System (INIS)

    In 54 patients suffering from active viral hepatitis the indirect solid phase radioimmunoassay (ind-SPRIA) for HBsAg was positive in 9 cases the direct solid phase radioimmunoassay (d-SPRIA) being negative. In 2 further cases ind-SPRIA was positive during several weeks but d-SPRIA only once. AntiHBc could be detected in 9 of these patients. In 7 patients the usual decrease of the transaminase activity was followed by a second elavation with prolongation of the disease. The unknown factor detected by ind-SPRIA suggests a special of acute hepatitis. (orig.)

  10. The use of the radioimmunoassay for detection of diethylstilbestrol in fecal samples of fattened calves

    International Nuclear Information System (INIS)

    A radioimmunoassay for diethylstilbestrol (DES) in fecal samples of fattened calves is described. The antibody used in this test showed no cross reactions with natural estrogens or other steroids. Blank values (fecal samples without DES) varied between 0 an 4,9 ng DES-equivalent/g sample. The distribution of blank values shows the significant proof of existence of DES in concentrations above 10 ng/g sample. The method has been applied for the control of living calves at the producer. Examples of illegal DES applications are given. The described radioimmunoassay is suitable for routine analysis as a control for illegal estrogen treatment. (orig.)

  11. Labelling of human follicle stimulant hormone with 125I, for radioimmunoassay

    International Nuclear Information System (INIS)

    An efficient labeling of human Follicle Stimulant Harmone is essential to development of sensitive radioimmunoassays. Iodination by Chloramine T method frequently is subject to severe iodination damage and some preparations are unaccetable for radioimmunoassays. Modifications to the Hunter method, changing incubation time, reaction temperature and reducing Chloramine T amount used in the reaction, were performed in obtaining a more effective labeling. FSH-125 I fraction obtained from Sephadex G-75 column purification presented excellent immunoreactivity and quality control of the steps of the reaction demonstrated a high percentage (90%) of intact Follicle Stimulant Hormone

  12. Assessment of a method for measuring serum thyroxine by radioimmunoassay, with use of polyethylene glycol precipitation

    International Nuclear Information System (INIS)

    We assessed the efficacy of a new thyroxine radioimmunoassay kit (Abbott) in which polyethylene glycol is used to separate bound from free hormone. Mean serum thyroxine was 88 +- 15 (+-SD) μg/liter for 96 normal persons. Results for hypothyroid and hyperthyroid persons were clearly separated from those for normal individuals. Women taking oral contraceptive preparations showed variable increases in their serum thyroxine values. The coefficient of variation ranged from 1 to 3% within assay and from 5.4 to 11% among different assays. Excellent parallelism was demonstrated between thyroxine values estimated by this method and those obtained either by competitive protein binding or by a separate radioimmunoassay for the hormone

  13. Heterophilic antibodies interfering with radioimmunoassay. A false-positive pregnancy test

    International Nuclear Information System (INIS)

    A young woman with amenorrhea had a consistently positive pregnancy test result (serum radioimmunoassay measurement of #betta#-human chorionic gonadotropin hormone). No fetal or placental tissue was found after uterine curettage and exploratory laparotomy. The false-positive pregnancy test result was due to heterophilic antibovine and antigoat antibodies in the patient's serum. These antibodies interfered with radioimmunoassays using goat antibodies. This case shows that serum heterophilic antibodies can interfere with immunoassays and result in unnecessary diagnostic procedures and/or unnecessary treatment

  14. Development of radioimmunoassay system for osteocalcin and applications in clinics

    International Nuclear Information System (INIS)

    The authors have developed an in-house, heterologous radioimmunoassay (RIA) system for human osteocalcin (Oc) using bovine OC as antigen, reference preparation and tracer and separation with polyethylene glycol (PEG). This RIA system is based on a satisfactory cross-reaction of antibovine serum with human Oc. The standard curve doses ranged between 1.5 and 6.0 ng/mL. The intra- and interassay coefficients of variation (%CV), from 20 batches, were 5-7% and 10-15%, respectively. This Oc-RIA system was applied to estimate the normality range in human subjects of both sexes, aged 1 to 70 years. In children and adolescents high Oc values and rather large variations between individuals were found. The OC serum levels decrease in young and adult subjects but increase significantly through decades seven and eight (p < 0.001) possibly due to the osteoporotic processes in the aged. Children and adolescents of both sexes with longitudinal growth deficiency (n = 318) were submitted to the insulin stimulation test (IST). According to the human growth hormone (hGH) increase in their sera they were grouped as responders or non-responders. The Oc measured in basal conditions showed non-significant differences between these two groups except at the age of 10 years (p < 0.01): the hGH responder group presented the peak increase earlier than the hGH non-responder group, with the peak values at the age of 13 years. Further, the serum Oc variations were followed in the sera of three patients diagnosed as having parathyroid adenoma. The results showed that high pre-operatory Oc levels increased a short time after surgery (patients No. 1) and decreased slowly thereafter (patient No. 2) or increased (patient No. 3). The serum Oc is a sensitive marker of bone growth in normal children and in children with growth deficiency. It may be used as an indicator of osteoporosis in aged subjects and may occasionally replace parathormone measurement. However, in interpreting an Oc-RIA result

  15. Production and use of thyroxine antisera in radioimmunoassay technique

    International Nuclear Information System (INIS)

    This study describes the production of antisera from sheep and its use in the determination of thyroxine hormone (T4) level in serum using radioimmunoassay (RIA) technique. In this study two local sheep (Ovis aris) were subjected to immunization against human T4 immunogen, sera obtained from both sheep after each injection were subjected to evaluation through titration in a purified and non purified form. The produced antibodies were used to assemble a kit for the determination of total human serum thyroxine. Different separation techniques were tried, (second antibody polyethylene glycol (PEG) assisted precipitation, polystyrene beads and magnetisable particles solid phases). For the PEG assisted precipitation, local antiserum and that produced by the North East Thamus Region Immunoassay (NETRIA donkey anti-sheep serum (DASS) as second antibodies) were tried. The final dilutions of the anti-T4 antibody used were 1/4000 in a liquid phase using second antibody PEG assisted separation, 1/3000 using magnetizable particles and a dilution of 1/10,000 using polystyrene beads solid phase for separation. Optimization of T4 assay conditions including incubation temperature and reaction time were done. Tests for T4 assay validation (linearity, recovery and responsibility) were carried out. For linearity and recovery tests, the regression coefficient ranges were found to be from (0.8 to 0.9) and (0.88 to 0.98) respectively. The assay was found to be reproducible where the coefficients of variation within and between assays were less than 10%. The locally developed assay was found to be comparable with NETRIA assay as a reference method with a correlation coefficient of 0.88, 0.93 and 0.87 for PEG assisted separation, magnetizable particles and polystyrene beads techniques respectively. The clinical validation tests showed a reliable sensitivity, specificity and efficiency with values of 97%, 94% and 96% respectively. When the T4 concentrations measured using the locally

  16. Quality control scheme for thyroid related hormones measured by radioimmunoassay

    International Nuclear Information System (INIS)

    A regional quality control scheme for thyroid related hormones measured by radioimmunoassay is being established in the Middle East. The scheme started in January 1985, with eight laboratories which were all from Iraq. At the present nineteen laboratories from Iraq, Jordan, Kuwait, Saudi Arabia and United Arab Emirates (Dubai) are now participating in the scheme. The scheme was supported by the International Atomic Energy Agency. All participants received monthly three freeze dried quality control samples for assay. Results for T3, T4 and TSH received from participants are analysed statistically batch by batch and returned to the participants. Laboratories reporting quite marked bias results were contacted to check the assay performance for that particular batch and to define the weak points. Clinical interpretation for certain well defined samples were reported. A regular case study report is recently introduced to the scheme and will be distributed regularly as one of the guidelines in establishing a trouble shooting programme throughout the scheme. The overall mean between the laboratory performance showed a good result for the T4, moderate but acceptable for T3 and poor for TSH. The statistical analysis of the results based on the concept of a ''target'' value is derived from the believed correct value the ''Median''. The overall mean bias values (ignoring signs) for respectively low, normal and high concentration samples were for T4 18.0 ± 12.5, 11.2 ± 6.4 and 11.2 ± 6.4, for T3 28.8 ± 23.5, 11.2 ± 8.4 and 13.4 ± 9.0 and for TSH 46.3 ± 50.1, 37.2 ± 28.5 and 19.1 ± 12.1. The scheme proved to be effective not only in improving the overall performance but also it helped to develop awareness of the need for internal quality control programmes and gave confidence in the results of the participants. The scheme will continue and will be expanded to involve more laboratories in the region. Refs, fig and tabs

  17. Chemotherapy by Intravenous Administration of Conjugates of Daunomycin with Monoclonal and Conventional Anti-Rat α -fetoprotein Antibodies

    Science.gov (United States)

    Tsukada, Yutaka; Hurwitz, Esther; Kashi, Rina; Sela, Michael; Hibi, Nozomu; Hara, Akihiko; Hirai, Hidematsu

    1982-12-01

    Monoclonal antibodies to rat α -fetoprotein (AFP) were produced by hybridization of mouse myeloma cells with spleen cells from mice immunized with rat AFP. The monoclonal antibodies as well as horse anti-rat AFP were coupled via a dextran bridge to daunomycin. Both types of conjugates were tested in vitro and in vivo for their anti-tumor activity. They were equally cytotoxic to rat AH66 hepatoma cell line in culture. Rats challenged with hepatoma cells were treated with the conjugates either by intraperitoneal or intravenous injections. Daunomycin conjugates with horse anti-AFP and monoclonal mouse anti-AFP were capable of delaying the tumor development more efficiently than the controls of antibodies or free drug, mixtures of drug with antibodies, and a conjugate of drug and normal immunoglobulin. The specific conjugates were considerably more effective when the treatments were given intravenously. The specific conjugates produced 60% long-term survival, whereas the controls delayed only slightly tumor development.

  18. Radioimmunoassay for bupropion in human plasma: comparison of tritiated and iodinated radioligands

    International Nuclear Information System (INIS)

    We evaluated the potential usefulness of 125I-labeled p-hydroxybupropion in a direct radioimmunoassay for bupropion in human plasma as compared with a currently used [3H]bupropion dextran-coated charcoal method. In both radioimmunoassay methods succinoylpropylbupropion antiserum was used that was highly specific for unchanged drug, cross reactivities with known bupropion metabolites being less than 0.3%. However, the use of 125I-labeled p-hydroxybupropion afforded greater sensitivity (0.3 microgram/L vs 0.6 microgram/L with [3H]bupropion) and was readily adaptable to the more convenient polyethylene glycol separation method. Between-assay CVs were 3.8 to 12.2% (mean 7.6%) with the 125I-based radioimmunoassay and 5.1 to 11.5% (mean 7.5%) with the 3H-based assay. Agreement between the two radioimmunoassay determinations of buproprion in human plasma samples collected over a 60-h period after oral drug administration was excellent (slope . 1.086, r . 0.989). We find the 125I-based assay a convenient and suitable alternative to the [3H]bupropion assay in pharmacokinetic studies in humans

  19. Radioimmunoassay for bupropion in human plasma: comparison of tritiated and iodinated radioligands

    Energy Technology Data Exchange (ETDEWEB)

    Butz, R.F.; Smith, P.G.; Schroeder, D.H.; Findlay, J.W.

    1983-03-01

    We evaluated the potential usefulness of /sup 125/I-labeled p-hydroxybupropion in a direct radioimmunoassay for bupropion in human plasma as compared with a currently used (/sup 3/H)bupropion dextran-coated charcoal method. In both radioimmunoassay methods succinoylpropylbupropion antiserum was used that was highly specific for unchanged drug, cross reactivities with known bupropion metabolites being less than 0.3%. However, the use of /sup 125/I-labeled p-hydroxybupropion afforded greater sensitivity (0.3 microgram/L vs 0.6 microgram/L with (/sup 3/H)bupropion) and was readily adaptable to the more convenient polyethylene glycol separation method. Between-assay CVs were 3.8 to 12.2% (mean 7.6%) with the /sup 125/I-based radioimmunoassay and 5.1 to 11.5% (mean 7.5%) with the /sup 3/H-based assay. Agreement between the two radioimmunoassay determinations of buproprion in human plasma samples collected over a 60-h period after oral drug administration was excellent (slope . 1.086, r . 0.989). We find the /sup 125/I-based assay a convenient and suitable alternative to the (/sup 3/H)bupropion assay in pharmacokinetic studies in humans.

  20. Determination of progesterone for reproduction control in cows using a 3H radioimmunoassay. 1

    International Nuclear Information System (INIS)

    For verification of cow fertility a 3H radioimmunoassay of progesterone in milk and blood plasma was developed. It is of high specificity and accuracy as well. Extraction of progesterone from milk was facilitated by application of alcohol. Suggested differences in milk and plasma progesterone levels between pregnant and nonpregnant cows could be revealed

  1. A comparative study on bioassay and radioimmunoassay of vasopressin in human urine

    International Nuclear Information System (INIS)

    The excretion of vasopressin in urine from healthy human subjects under different stages of hydration was estimated in urine extracts by bioassay (rat antidiuresis) and radioimmunoassay. In normally hydrated subjects the excretion was 490 plus minus 164 μU/h and 430 plus minus 133μU/h for bioassay and radioimmunoassay respectively (mean plus minus SEM, n=5). After total fluid restriction for 10 to 12 h the excretion increased to 1370 plus minus 329 μU/h for bioassay and 1163 plus minus 279 μU/h for radioimmunoassay (mean plus minus SEM, n=6). An oral water load (25 ml/kg) reduced the value to 169 μU/h (bioassay) and 118 μU/h (radioimmunoassay) (mean, n=2). In general the biological estimations were 20 % higher than the immunological estimations (P<0.01). The urinary vasopressin excretion was positively correlated to urine osmolality. Synthetic arginine vasopressin (AVP) and urine extracts were both heterogeneous in ion exchange chromatography. The immunoreactive material was always eluted in 2 peaks, situated at the same places in the elution diagram. The material in the second peak was biologically active. The first peak from the AVP standard was biologically inactive, whereas the same peak from urine extract was active. This peak was shown to be an artefact formed during the extraction procedure. (author)

  2. Automated approach to radioimmunoassays of somatotropin (human growth hormone) and insulin

    International Nuclear Information System (INIS)

    The adaptation of human somatotropin and insulin assays to the automated Centria radioimmunoassay system [Clin. Chem. 21, 1305 (1975)] is reported. For the somatotropin assay, reaction conditions include a borate/bovine serum albumin buffer (pH 8.4) and a 20-h incubation at 40C. Assay results for clinical samples compared favorably (correlation coefficient = 0.930) with values obtained from a reference laboratory. The means determined for 92 patients' samples were 4.3 μg/liter (reference laboratory) and 5.1 μg/liter (Centria). Intra- and inter-run precision ranged from 3.2 to 15.9%. For the insulin assay, a phosphate/bovine serum albumin buffer (pH 7.4) is used, with a 20-h incubation at 40C. Previously analyzed insulin samples from a reference laboratory were determined by the Centria analyzer with excellent correlation (r = 0.965). Means for patients' samples were 43.0 milli (USP) units of insulin per liter (reference laboratory) and 47.5 milliunits of insulin per liter (Centria). In both assays an anionicexchange gel is used in the separation step. The criterion of parallelism, an indication of the validity of a radioimmunoassay, was satisfied in both assays. The Centria radioimmunoassay system offers the advantage of automating all the critical steps of these radioimmunoassays

  3. Preparation of iodine - 125 - labeled insulin for radioimmunoassay: comparison of chloramine T and iodogen iodination

    International Nuclear Information System (INIS)

    Stoichiometric iodination of porcine insulin was performed to the general method of Hunter and Greenwood with modifications recommended by Roth. These method was compared with radioidination using Iodogen. Films of Iodogen react rapidly in the solid phase with aqueous mixtures of I -and proteins. For two methods satisfactory activity of the labeled porcine insulin was obtained and characteristics of the radioimmunoassay were studied. (author)

  4. Detection of viral hepatitis B markers by radioimmunoassay in medical personnel

    International Nuclear Information System (INIS)

    Results of revealing B markers of viral hepatitis (VHB) in medical personnel in the city of Zaporozhe by radioimmunoassay method are presented. By the frequency of revealing two markers (HB sAg and anti-HBs) risk groups are indicated depending on the profession, age (length of service) of medical personnel

  5. Improvements in the labelling technique of human proinsulin for radioimmunoassay use

    International Nuclear Information System (INIS)

    Changes in the labelling technique of human proinsulin were done viewing the attainment of a tracer with a higher specific activity (SA) and consequently a increasing in the radioimmunoassay sensitivity: employment of polypropylene vials recovered with Iodogen dissolved in dichloromethane and evaporates used up to 15 months and the tracers showed high SA and purity, with a shelf-live of a months. (author)

  6. Radioimmunoassay of plasma corticotropin in the edible Frog Rana esculenta L

    International Nuclear Information System (INIS)

    In the green Frog (Rana esculenta) the plasma contains a polypeptide immunologically related to human and porcine corticotropins. A radioimmunoassay capable of detecting 4.10-12 g hog ACTH has been used for a direct plasma ACTH assay in the Frog. Using this method the ACTH rate was determined both in untreated frogs and in animals under various experimental conditions

  7. Radioimmunoassay in situ used to detect products of cloning mycoplasma pneumoniae DNA and clinical micro samples

    International Nuclear Information System (INIS)

    From the recombinants, which constructed in the laboratory, using plasmids as vector, one strain which express antigen of Mycoplasma pneumoniae by detecting in situ radioimmunoassay was obtained. The authors detected 7 children samples which were throat washing liquids of children suspected of atypical pneumoniae, 4 of them positive by using above method. This data indicated that it was suitable for clinical diagnosis directly

  8. Radioimmunoassay in the evaluation of the hormonal activity of hypophyseal adenoma

    International Nuclear Information System (INIS)

    Direct correlation was found between the results of a radioimmunoassaly electron microscopy and immunohistochemistry in cases of STH-secreting and hormonally inactive hypophyseal adenomas. In hyperprolactinemia and a simultaneous increase in the levels of 2 or more hormones the results of the radioimmunoassay coincided with those of electron microscopy and immunohistochemistry in 50-70% of the cases

  9. Comparative determination of phenytoin by spectrophotometry, gas chromatography, liquid chromatography, enzyme immunoassay, and radioimmunoassay

    International Nuclear Information System (INIS)

    Sera from patients being treated with phenytoin were analyzed for the drug by spectrophotometry, gas chromatography, radioimmunoasay, enzyme immunoassay, and liquid chromatography. The assay values obtained were intercompared statistically. Enzyme immunoassay and liquid chromatography appear to be attractive alternatives to the more traditional methods of spectrophotometry and gas chromatography. Our radioimmunoassay data correlated poorly with results by the four other methods

  10. Radioimmunoassay measurement of plasma oxytocin and vasopressin in cows during machine milking

    Energy Technology Data Exchange (ETDEWEB)

    Landgraf, R.; Wehowsky, G.; Schulz, J.; Schulze, H.; Bothur, D. (Forschungsinstitut fuer Koerperkultur und Sport, Leipzig (German Democratic Republic); Karl-Marx-Universitaet, Leipzig (German Democratic Republic). Sektion Tierproduktion und Veterinaermedizin)

    1982-07-01

    The response of plasma oxytocin and vasopressin to machine milking in cows was studied by radioimmunoassay. Depending on the method of machine milking used, plasma oxytocin increased to a greater or lesser degree after teat cup application. Plasma vasopressin was not affected by the milking procedures.

  11. Measurement of Steroids in Rats after Exposure to an Endocrine Disruptor: Mass Spectrometry and Radioimmunoassay Demonstrate Similar Results

    Science.gov (United States)

    Commercially available radioimmunoassays (RIAs) are frequently used in toxicological studies to evaluate effects of endocrine disrupting chemicals (EDCs) on steroidogenesis in rats. Currently there are limited data comparing steroid concentrations in rats as measured by RIAs to t...

  12. A fluorescent turn-on detection scheme for α-fetoprotein using quantum dots placed in a boronate-modified molecularly imprinted polymer with high affinity for glycoproteins

    International Nuclear Information System (INIS)

    This article describes a fluorescent molecularly imprinted polymer (MIP) capable of selective fluorescent turn-on recognition of the tumor biomarker α-fetoprotein. The technique is making use of amino-modified Mn-doped ZnS quantum dots (QDs) as solid supports, 4-vinylphenylboronic acid and methyl methacrylate as the functional monomers, γ-methacryloxypropyl trimethoxysilane as the grafting agent, and α-fetoprotein as a template. A graft imprint is created on the surface of the QDs. The functional monomers are shown to play an important role in the formation of the binding sites and in preventing nonspecific protein binding. The resulting MIP-QDs display a good linear response to α-fetoprotein in the 50 ng · L−1 to 10 μg · L−1 concentration range, and the limit of detection is 48 ng · L−1. In our perception, the method has a wide scope in that it may be adapted to various other glycoproteins. (author)

  13. Application of a sepharose bead immunofluorescence assay and a solid-phase radioimmunoassay to the bovine leukemia virus system

    International Nuclear Information System (INIS)

    Several fluorescence assays with bovine leukemia virus (BLV) conjugated to activated Sepharose 4B were used for the detection of BLV and anti-BLV antibodies. These tests were compared with a solid-phase radioimmunoassay and found to be in the same sensitivity range. Sepharose bead immunofluorescence assay and solid-phase radioimmunoassay can be applied to the diagnosis of BLV infection in cattle. (author)

  14. Application of a monoclonal antibody to a comparative study of alpha-lactalbumins from various species

    International Nuclear Information System (INIS)

    A monoclonal antibody to bovine alpha-lactalbumin was prepared and purified. The binding ability of alpha-lactalbumin from different species (cow, goat, giraffe, horse, pig, human, monkey, and guinea pig) was examined by a competitive radioimmunoassay. The order in strength of the binding affinity was cow goat, giraffe, horse, cynomolgus monkey and human, pig, and guinea pig. The order of evolutional divergence calculated from the amino acid composition was cow, goat, giraffe, horse, pig, guinea pig and human, and monkey. The orders in both cases were similar. Hence, it is suggested that immunological divergence as deduced by a monoclonal antibody is likely to be close to the evolutional divergence of alpha-lactalbumin

  15. Radioimmunoassay of diagoxin with the aid of the solid phase - microtitre plating technique

    International Nuclear Information System (INIS)

    Preliminary results are reported here on the development of a digoxin-radioimmunoassay with an anti-digoxin antibody (goat) in a solid phase technique (mictrotitre plate). The advantages compared to conventional RIAs are: Cross reactions towards digoxin is minimal, both in vitro and in vivo. The calibraton range extends from 0.25 to 8 ng/ml. The radioactive load could be reduced significantly by use of smaller amounts of tracer (0.004 μCi/single determination) and by reduction of waste volume (solid), waste weight (solid) and liquid waste. The DIGOXIN RIA BIOTEST MTP is, in addition, the only digoxin radioimmunoassay where radioactive waste is produced in a sealed form. The test is a simple one and can be carried out without the need for complicated apparatus and techniques. (orig./MG)

  16. Development of radioimmunoassay for gamma-glutamyl transferase using pancreatic enzyme

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) for the determination of gamma-glutamyl transferase (GGT) was developed using human pancreatic enzyme as antigen. The assay allows the determination of GGT in concentrations as low as 80 ng/ml, and it is reproducible and specific. A good parallel relation was demonstrated between the standard curve and dilution curves for serum, urine, bile, and partially purified kidney GGT. In normal individuals, the mean serum concentration of GGT determined by RIA was found to be 3.43 μg/ml (SD+-1.20). Enzyme activity calculated from the GGT concentration measured by the radioimmunoassay using a regression equation was approximately twice as great as that determined by conventional enzyme assay. (author)

  17. A solid phase radioimmunoassay for free triiodothyronine in serum:assay development and validation

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    A solid phase radioimmunoassay for free triiodothyroninein serum was developed based on double-antibody coated tubes.The method was turned out to be reliable with good reproducibility,higher sensitivity and easy performance.The measurable range of FT3 in serum was 1.2 to 38pmol/L.The mean coefficients of variationwithin and between assays were 1.79%~3.18% and 4.72%~9.31%,respectively.The FT3 concentrations in euthyroid serum as determined by this methodwere 2.8 to 7.8pmol/L.The FT3 values determined by this new methodcorrelated well with those measured by a commercial radioimmunoassay(r=0.853).

  18. Characterization of a radioimmunoassay for ovine FSH utilizing an anti-bovine FSH serum

    International Nuclear Information System (INIS)

    In the radioimmunoassay for ovine FSH, utilizing a bovine FSH antiserum and 125I-labelled bovine FSH tracer, the sensitivity was shown to be 5 ng NIH-FSH-S6/ml and cross-reactivities of other ovine pituitary glyco-protein and protein hormones (LH,TSH, growth hormone and prolactin) were less than 1.0%. Both pituitary and serum FSH of the sheep cross-reacted in a manner parallel with that of standard NIH-FSH-S6. Serum FSH concentrations in 4 ewes at various times after the onset of oestrous peaked more or less coincidently with the preovulatory increase of serum LH. The serum levels of FSH and LH did not correlate proportionally but fluctuated considerably in individual animals. The radioimmunoassay system appeared to be sensitive and specific for measuring serum concentrations of FSH in rams and ewes in different physiological states. (author)

  19. Model for validation of radioimmunoassay kit reagents: measurement of follitropin and lutropin in blood and urine

    International Nuclear Information System (INIS)

    We measured lutropin and follitropin in blood and urine with radioimmunoassay kits from Diagnostic Products Corporation and compared the results with those obtained by use of re agents from the National Institutes of health (NIH) and the World Health Organization (WHO). The urine standard (second IRP-HMG) from WHO, the blood standard (LER-907) from NIH, and the commercial standards all effected similar displacement of trace material when the commercial gonadotropin kit reagents were used. Highly significant correlations were achieved for these hormones in blood or urine on comparing commercial and NIH/WHO reagents. Serial dilutions of urine samples produced similar relative potencies with the commercial reagents. Conversion factors are presented to relate results for LER-907, second IRP, or commercial standards. Commercially available reagents can provide a practical and reliable means of gonadotropin radioimmunoassay in blood or urine

  20. Importance of radioimmunoassays in the diagnosis evaluation of severity and prognosis of myocardial infarction

    International Nuclear Information System (INIS)

    The authors presented the results of a radioimmunoassay of myoglobin (MG), MV-fraction of creatine phosphokinase (CPK), hypophyseal thyrotropic hormone, peripheral thyroid hormones and testosterone in 150 patients with acute myocardial infarction. A high informative value of the radioimmunoassay was shown. The MG level and MV-CPK activity as well as the functioning of the hypophysealthyroid system were shown to be directly correlated with the severity of disease and were of prognostic value. A considerable suppression of hypophyseal and thyroid function coinciding with the maximum values of myoglobinemia in the most severe cases was noted. Three variants of the testosterone content were singled out in patients with acute myocardial infraction. Paroxysmal rhythm disorders in the 1 st variant (up to 300 ng/100 ml) and A-V blockades (over 1000 ng/100 ml) in the 3 rd variant were frequently noted

  1. Determination of degree of oncologic disease spreading with the use of radioimmunoassay

    International Nuclear Information System (INIS)

    An effort to evaluate the possibility of applying tumor morkers to determine the degree of oncologic desease propagation is made. Radioimmunoassay of 431 patients with tumors with different localization is performed. In patients with lung, esophagus, stomach, rectum cancer the level of carcinoembryonic antigen (CEA) in the blood serum is studied, with mammary gland cancer - the CEA, ferritin and prolactin levels. An increase of CEA level is detected under wide-spread forms of lung, esophagus, stomach and rectum cancer, and an increase of CEA, ferritin and prolactin levels-under mammary gland cancer. It is shown that radioimmunoassay of tumor marker level is quite information content as to the evaluation of tumor process propagation under digestive tract, lung and mammary gland cancer. 12 refs.; 5 tabs

  2. Ultrasensitive radioimmunoassay for direct determination of free triiodothyronine concentration in serum

    International Nuclear Information System (INIS)

    Free triiodothyronine (T3) in serum has been measured directly in dialysates of serum, using a wick chromatographic radioimmunoassay. Adequate sensitivity was attained by the use of [125I]T3 with a very high specific activity (2,000 to 3,000μCi/μg). Sera were dialysed against a Krebs-Ringer bicarbonate buffer modified so as to be similar to plasma water. Dialysis took place under carefully controlled circumstances. The influence on the equilibrium of total to free T3 of temperature, serum dilution and dialysis time was studied. By the present method, free T3 in serum from groups of subjects including 20 men, 10 women taking oral contraceptives and 20 women with normal menstrual cycles were identical, averaging 5.2 pg/ml. A chromatographic radioimmunoassay of total T3 using high specific activity [125I]T3 and a very small test sample is also described. (auth.)

  3. Quality characteristics of chemicals for the radioimmunoassay of thyroxine and thyrotropin

    International Nuclear Information System (INIS)

    Radioimmunoassay is a form of saturation analysis in which the test material competes with labelled antigen for a limited amount of antibody, the amount of label displaced being a measure of the antigen in the test sample. In this country, the kits for Radioimmunoassay (RIA) are imported, and this increase the cost of it. Because this lack of production, the National Institute of Nuclear Research (ININ) has developed RIA's kits for the thyroxine (T4), Thyrotropin (TSH) and Triyodotironine (T3) hormones. This work presents the conclusions of the test recommended by the WHO. The quality test were: recuperation, cross reactions, basic parameters, intra and inter assay variations, sensibility and others. The results show that the RIA's kits of the ININ have a good behavior and can be use in the clinical laboratory. (Author)

  4. Interferences in radioimmunoassay of aflatoxins in food and fodder samples of plant origin

    International Nuclear Information System (INIS)

    Cross-reactions and resulting nonspecific binding of substances with structures resembling aflatoxins (derivatives of coumarin, and cinnamonic and benzoic acids, etc.) were investigated. The concentrations of these substances causing erroneously high or false positive values in radioimmunoassay were determined. One μg aflatoxin B1/kg sample may be simulated by the occurrence of 5 g coumarin, 10 g caffeic acid, 16 g chlorogenic acid, or 15 g vanillin/kg fodder or food sample

  5. External quality assessment scheme for the radioimmunoassay of thyroid related hormones

    International Nuclear Information System (INIS)

    An external quality assessment programme to evaluate performance of laboratories estimating thyroid related hormones by radioimmunoassay techniques was undertaken under the auspices of International Atomic Energy Agency, Vienna and Bhabha Atomic research Centre, Bombay. 35 laboratories in different parts of the country participated in the programme. The report summarises the findings. The programme helped to identify the steps that required trouble shooting and to improve the performance of some laboratories. (author). 3 refs., 4 figs., 12 tabs

  6. Immunoradiometric assay for prolactin in serum and tissue; Comparison with radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Ohnami, Shumpei; Nakata, Hajime; Eto, Sumiya (University of Occupational and Environmental Health Hospital, Kitakyushu, Fukuoka (Japan))

    1990-09-01

    Prolactin (PRL) concentrations in sera and tumors of patients with various pituitary tumors were measured by both immunoradiometric assay (IRMA) and radioimmunoassay (RIA). PRL concentrations in sera and tumor tissues measured by IRMA were well correlated with those measured by RIA. PRL concentrations in sera reflected those of tumors removed. This IRMA is a simple and useful method for PRL determination in serum and tissue. (author).

  7. Radioimmunoassay for determination of the hormonal and immune states in patients aczema; psoriasis and neurodermatitis

    International Nuclear Information System (INIS)

    The hormonal and immune status was investigated by a radioimmunoassay in 105 patiets with dermatosis (55 female and 50 male patients aged 15 to 80): 51 suffered frm ecsema, 41 -from psoriasis, and 13 - from neurodermatitis. Serum concentrations of T3, T4, TSH, insulin, trypsin, C-peptide, cortisol, and IgE were investigated. Disorders of the hormonal and immune status were noted in the examinees with relation to sez, type of disease, season, time-period and extent of disease

  8. The insulin radioimmunoassay kit prepared by IPEN-CNEN/SP - Brazil

    International Nuclear Information System (INIS)

    The specification and methodological aspects of the insulin radioimmunoassay kit produced by IPEN-CNEN/SP - Brazil are described. The limitations taking care and the following quality control parameters or procedures are discussed: specific radioactivity, comparison between two insulin - 125I purification procedures, affinity constant 'K' of the antigen - antibody reaction, minimal detectable dose (MDD), kinetics degradation of the radioinsulin, radioassay imprecision profile, radioassay performance temperature dependence and normal values histogram. (Author)

  9. Development of a radioimmunoassay for pig pancreatic kallikrein and its application in physiological studies

    International Nuclear Information System (INIS)

    The radioimmunoassay was employed to investigate whether pig pancreatic kallikrein is absorbed by the intestine. The ductus thoracicus of albino rats was canulated with a U-like polyehtylene tube and the lymphatic fluid collected into tubes which were changed at regular intervals. Kallikrein was found in concentrations up to 200 ng/ml. If pig serum was subjected to gel filtration two peaks of kallikrein were detected in elution positions as described above for rat lymphatic fluid. (orig./AJ)

  10. Bepaling van Diethylstilbestrol in urine m.b.v. Radio-Immuno-Assay RIA

    OpenAIRE

    Berghmans, M.C.J.; Driessen-Lankveld, W.D.M.; Polman, Th.H.G.; Ruig, de, W.G.

    1981-01-01

    Om inzicht te krijgen in de opname en uitscheiding van het groeihormoon dethylstilbestrol (DES) bij mannelijke runderen en in het verschil t.o.v. onbehandelde dieren werd een stier ingespoten met 100 mg DES dipropionaat in olie (intramusculaire toediening). Van beide dieren werd gedurende enige weken urine opgevangen, om deze te onderzoeken op de aanwezigheid van DES d.m.v. Radio-Immuno-Assay.

  11. Quantification of adsorbed human serum albumin : a comparison between radioimmunoassay and simple null ellipsometry

    OpenAIRE

    Benesch, Johan; Askendal, A.; Tengvall, P.

    2000-01-01

    Radioimmunoassay (RIA) and null ellipsometry are two common methods to quantify adsorbed proteins. However, the accuracy of null ellipsometry with a constant protein refractive index (n 1.465, k 0) at l 632.8 nm has this far not been explored. The present study compared the methods, and the degree of agreement between the simplified single wavelength null ellipsometry and RIA to quantify adsorbed proteins was explored on different surfaces. The quantification methods agreed well w...

  12. Radioimmunoassay of haloperidol in human serum: correlation of serum haloperidol with serum prolactin

    International Nuclear Information System (INIS)

    A radioimmunoassay (RIA) for measurement of serum haloperidol is described. Compared to gaschromatography (GC), RIA vaues average 40% higher. However, a simple organic extraction of serum yields statistically equivalent RIA and GC haloperidol determinations. For both men and women combined, there was a positive correlation between dose (mg/kg/day) and steady-state serum haloperidol level (r = +0.86) and between steady-state serum haloperidol and serum prolactin (PRL) concentration

  13. Comparison of radioimmunoassay and fluorescent polarization immunoassay for quantitative determination of vancomycin concentrations in serum.

    OpenAIRE

    Ackerman, B H; Berg, H G; Strate, R G; Rotschafer, J C

    1983-01-01

    A new fluorescent polarization immunoassay (Abbott Laboratories, Diagnostics Division, North Chicago, Ill.) was compared with a standard radioimmunoassay (American Diagnostics Corp., Newport Beach, Calif.) in 34 patients being treated with vancomycin. A total of 123 serum samples were divided and quantitatively analyzed for vancomycin by both assay methods. The results obtained indicated that the two assay methods are comparable (y = 1.01x - 0.81; r = 0.99).

  14. Pregnancy diagnosis in mares by estrogen determination in faeces: radioimmunoassay and enzymatic method

    International Nuclear Information System (INIS)

    As estrogens are also excreted in faeces, this fact could be used for pregnancy confirmation. Sampling of faeces offers the advantage of a non-invasive test and an easy sample collection which can be done by the farmer. In this study two extraction procedures for the radioimmunoassay of estrogens in faeces and an enzymatic method for the determination of estrogens are compared as regards their practicability. (Auth.)

  15. Radioimmunoassay detection of levels of triiodothyronine and thyroxine in Mangalarga Marchador equine

    International Nuclear Information System (INIS)

    Serum levels of triiodothyronine (T3) and thyroxine (T4) were determined in equine of Mangalarga Marchador breed through radioimmunoassay. Forty-two animals (17 males and 25 females), with age ranging from two to eighteen years, were utilized. The values recorded for males and females were, respectively: 101.68 ± 23.44 and 71.14 ± 18.82 ng/d l of T3 (P4 (P<0.05). (author). 10 refs, 1 tab

  16. Measurement of dihydro testosterone by radioimmunoassay after celite column chromatography

    International Nuclear Information System (INIS)

    A method for measuring dihydro testosterone after celite column chromatography is developed. One milliliter of serum containing 1000 cpm of tritiated dihydro testosterone was extracted with hexane: ethyl acetate (2:3): dried, diluted with non saturated iso octane and injected in the column previously washed with 3.5 ml of pure iso octane. The serum was eluted from the column with pure iso octane (3.5 ml) followed by 5% ethyl acetate in iso octane. The quantity of tritiated dihydro testosterone which was recovered ranged from 50% to 80% in all assays. The sensitivity of the method was 4 ng/d l. The intra-assay variation was less than 9% and the inter-assay variation was less than 9,7%. It was measured dihydro testosterone, testosterone and testosterone/dihydro testosterone ratio in the following groups: Group 1- forty-one normal adult subjects in basal conditions, Group 2 - six normal adult subjects, evaluated in basal conditions and after stimulus with 6000 International Unity of human Chorionic Gonadotropin; Group 3- six pre-puberal children with unilateral cryptochidism. Group 4- eight patients with male pseudo hermaphroditism due to 5-alpha-reductase deficiency in basal conditions and after HCG. (author)

  17. A solid-phase radioimmunoassay of serum dehydroepiandrosterone sulphate using a monoclonal antibody

    International Nuclear Information System (INIS)

    A monoclonal antibody directed against dehydroepiandrosterone, but with high affinity for dehydroepiandrosterone sulphate (DHA-S), has been used to develop a solid phase radioimmunoassay for measuring serum DHA-S. The antibody was covalently linked to polyacrylamide microbeads with no change in binding characteristics. The procedure requires only the chromatography of serum on anion-exchange cellulose before assaying the equivalent of 0.25 μl serum. The method is precise, accurate and specific and can detect 19.5 pg of DHA-S. Serum DHA-S levels measured by this method were in good agreement with those found in a validated radioimmunoassay method involving hydrolysis. The method is quick and one operator could assay 50 blood specimens per day. DHA-S levels in serum from 50 men and 86 women were in agreement with those in the literature. With the availability of theoretically limitless quantities of consistently high quality monoclonal antibodies the advantages of developing solid phase radioimmunoassays for steroids is discussed. (author)

  18. Influence of various desinfectants on the radioimmuno-assay for Australia antigen

    International Nuclear Information System (INIS)

    At normal room temperature dilution series were produced out of pooled serum, serum previously treated with UV irradiation and beta propiolacton, and serum of patients with hepatitis type B and various desinfectants. After differing incubation times the Australia antigen titre was measured in the radioimmunoassay. Electronmicroscopic examinations should detect possible morphologic changes of the Dane particles. The counts/min. measured for Australia antigen after an incubation with aldehyde-containing preparations, are below the limit value with serum treated previously with UV light and beta propiolacton; in negative Australia-antigen-positive serum the counts/min. are close to the limit value. The antigenity is clearly reduced. The comparison with an insulin containing serum showed that also the radioimmunoassay was influenced by the aldehyde. A direct influence of the aldehydes on the protein seems to be possible. From this results that the radioimmunoassay for Australia antigen cannot be used as the exclusive method for measuring the efficacy of desinfectants compared to Dane particles. The morphologic changes of the Dane particles observed in the electronic microscope confirm the supposition that the desinfectants influence the hepatitis viruses. (orig./MG)

  19. Simultaneous detection of morphine and barbiturates in urine by radioimmunoassay. [/sup 125/I

    Energy Technology Data Exchange (ETDEWEB)

    Usategui-Gomez, M.; Heveran, J.E.; Cleeland, R.; McGhee, B.; Telischak, Z.; Awdziej, T.; Grunberg, E.

    1975-09-01

    This report describes a radioimmunoassay for the simultaneous detection of morphine and barbiturates. Morphine and barbiturate antibodies, obtained from goats, were mixed with /sup 125/I-labeled antigens. By adjusting concentrations of the morphine and barbiturate antibodies and radiolabeled antigens, closely superimposed standard curves for the two drugs would be obtained. As a consequence, similar response curves were obtained for urine specimens containing morphine or barbiturates. Although concentrations as low as 25 ..mu..g/liter could be measured, to ensure against false positive reactions the test should be performed at the 100 ..mu..g/liter concentration. Unknown samples positive by the dual assay were confirmed by separately testing the specimens with the individual radioimmunoassay specific for morphine or barbiturate. Equivalency tests of urines positive for morphine, positive for barbiturates, or negative for both demonstrated complete correlation between the single and dual assays. The mixed reagent retained its sensitivity and specificity for at least three months when stored at 4 or 25/sup 0/C. The dual radioimmunoassay is a rapid, simple procedure that can be adapted to automated processes and that is suitable for large- and small-scale screening. (auth)

  20. Development and characterization of a radioimmunoassay to measure human tissue kallikrein in biological fluids

    International Nuclear Information System (INIS)

    A direct radioimmunoassay has been developed to measure tissue kallikrein in human biological fluids, including serum, plasma, urine, pancreatic juice and saliva. Purified kallikreins from human urine and human saliva were used to raise rabbit antibody and each labelled with Na125I for use in the radioimmunoassay. Comparison of the different antigen-antibody systems was then made. Bound and free enzyme were separated by a double-antibody technique. The usable range of the standard curve was from 2.5 to 100 μg kallikrein/1. The intra-assay coefficient of variation was 4.7%, the interassay coefficient of variation 8.9% and the recoveries of purified kallikrein added to the samples were 99.3, 96.0, 110.8 and 81.2% for urine, saliva, serum and plasma respectively. Parallel dilution curves were obtained for serum and plasma, as well as urine, saliva and pancreatic juice. Plasma anticoagulated with EDTA or heparin gave consistently lower values than serum, when measured in the radioimmunoassay. From eight different subjects plasma (EDTA) values were on average 50% lower than those of serum, and subsequent experiments revealed that treatment of blood with some anticoagulants, in particular heparin and EDTA, resulted in a marked reduction in measurable tissue kallikrein. (author)

  1. A sensitive and selective radioimmunoassay for serum 24, 25-dihydroxycholecalciferol in man

    International Nuclear Information System (INIS)

    A sensitive and selective radioimmunoassay for 24,25-dihyroxycholecalciferol (24,25(OH)2D3) has been developed. Antisera with a high titre and affinity to 24,25(OH)2D3 were raised in rabbits immunized with a protein conjugate of the 3-hemisuccinate derivative of 24,25(OH)2D3. Serum samples were extracted by methanol/dichloromethane and the lipid extracts purified on Sephadex LH 20 and chromatographed by single step HPLC on a straight phase silica column. The radioimmunoassay is capable of measuring 24,25(OH)2D3 in the '24,25(OH)2D complex' isolated by HPLC and containing the comigrating metabolites: 24,25(OH)2D3, 24,25(OH)2D2, 25(OH)D3-26,23 lactone and 25,26(OH)2D2. The detection limit of 2 pg/assay tube is marked improvement compared to the competitive protein binding assay using rachitic rat serum. Measured by the radioimmunoassay 24,25(OH)2D3 ranged from 0.05 to 1.96 ng/ml with a mean of 0.85 ng/ml in 34 healthy adults. For comparison 24,25(OH)2D, measured simultaneously by competitive protein binding with rachitic rat serum, ranged from 0.1 to 4.0 ng/ml with a mean of 1.76 ng/ml. (author)

  2. Electrochemical enzyme-linked immunosorbent assay (ELISA) for α-fetoprotein based on glucose detection with multienzyme-nanoparticle amplification.

    Science.gov (United States)

    Liu, Qin-Lan; Yan, Xiao-Hui; Yin, Xiao-Mao; Situ, Bo; Zhou, Han-Kun; Lin, Li; Li, Bo; Gan, Ning; Zheng, Lei

    2013-01-01

    Since glucose biosensors are one of the most popular and widely used point-of-care testing devices, a novel electrochemical enzyme-linked immunosorbent assay (ELISA) for protein biomarkers has been developed based on a glucose detection strategy. In this study, α-fetoprotein (AFP) was used as the target protein. An electrochemical ELISA system was constructed using anti-AFP antibodies immobilized on microwell plates as the capture antibody (Ab1) and multi-label bioconjugates as signal tracer. The bioconjugates were synthesized by attaching glucoamylase and the secondary anti-AFP antibodies (Ab2) to gold nanoparticles (AuNPs). After formation of the sandwich complex, the Ab2-glucoamylase-AuNPs conjugates converted starch into glucose in the presence of AFP. The concentration of AFP can be calculated based on the linear relation between AFP and glucose, the concentration of which can be detected by the glucose biosensor. When the AFP concentration ranged from 0.05 to 100 ng/mL, a linear calibration plot (i (µA) = 13.62033 - 2.86252 logCAFP (ng/mL), r = 0.99886) with a detection limit of 0.02 ng/mL was obtained under optimal conditions. The electrochemical ELISA developed in this work shows acceptable stability and reproducibility, and the assay for AFP spiked in human serum also shows good recovery (97.0%-104%). This new method could be applied for detecting any protein biomarker with the corresponding antibodies. PMID:24129276

  3. Electrodeposition of CdSe quantum dots and its application to an electrochemiluminescence immunoassay for α-fetoprotein

    International Nuclear Information System (INIS)

    We report on the first label-free electrochemiluminescence (ECL) immunosensor for α-fetoprotein (AFP). It is based on the use of CdSe quantum dots that were electrodeposited directly on a gold electrode from an electrolyte (containing cadmium sulfate, EDTA and selenium dioxide) by cycling the potential between 0 and -1.2 V (vs. SCE) for 60 s. The electrodeposited dots were characterized by scanning electron microscopy and energy dispersive spectroscopy. Under optimal conditions, the specific immunoreaction between AFP and anti-AFP resulted in a decrease of the ECL signal because of the steric hindrance and the transfer inhibition by peroxodisulfate. The quenching effect of the immunoreaction on the intensity of the ECL was used to establish a calibration plot which is linear in the range from 0.05 to 200 ng mL-1. The detection limit is 2 pg mL-1. The assay is highly sensitive and satisfactorily reproducible. In our opinion it opens new avenues to apply ECL in label-free biological assays. (author)

  4. The Immunogenicity of the Tumor-Associated Antigen α-Fetoprotein Is Enhanced by a Fusion with a Transmembrane Domain

    Directory of Open Access Journals (Sweden)

    Lucile Tran

    2012-01-01

    Full Text Available Aim. To investigate the ability of recombinant modified vaccinia virus Ankara (rMVA vector to induce an immune response against a well-tolerated self-antigen. Methods. rMVA vectors expressing different form of α-fetoprotein (AFP were produced and characterized. Naïve mice were vaccinated with MVA vectors expressing the AFP antigen in either a secreted, or a membrane-bound, or an intracellular form. The immune response was monitored by an IFNΓ ELISpot assay and antibody detection. Results. Vaccination with the membrane-associated form of AFP induced a stronger CD8+ T-cell response compared to the ones obtained with the MVA encoding the secreted or the intracellular forms of AFP. Moreover, the vaccination with the membrane-bound AFP elicited the production of AFP-specific antibodies. Conclusions. The AFP transmembrane form is more immunogenic. Expressing a membrane-bound form in the context of an MVA vaccination could enhance the immunogenicity of a self-antigen.

  5. Simultaneous determination of carcinoembryonic antigen and α-fetoprotein using an ITO immunoelectrode modified with gold nanoparticles and mesoporous silica

    International Nuclear Information System (INIS)

    We report on an electrochemical immunoassay for simultaneous detection of the tumor markers carcinoembryonic antigen (CEA) and α-fetoprotein (AFP). Gold nanoparticles (AuNPs) were synthesized inside the internal pore walls of mesoporous silica (MPS). Then, Methylene blue (MB) and 6-ferrocenylhexanethiol (FeC), which act as electrochemical substrates, were incorporated into the channels via the formation of Au-S bonds. Finally, monoclonal antibodies against CEA (anti-CEA) and AFP (anti-AFP) were immobilized in the channels of MB-AuNPs-MPS and FeC-AuNPs-MPS, respectively. Suspensions of anti-CEA/MB-AuNPs-MPS and of anti-AFP/FeC-AuNPs-MPS were deposited on two different sites of the ITO electrode. When incubated with samples containing the two analytes, the formation of electrically nonconductive immunoconjugates blocks electron transfer. Simultaneous detection of CEA and AFP was accomplished by monitoring the current change before and after an immunoreaction has occurred. The AuNPs confined inside the channels are promoting the electron transport and thus improve detection limits. The immunoelectrode responds to CEA in the 0.5–50 ng mL−1 concentration range, and to AFP in concentrations between 0.5 and 100 ng mL−1. The detection limits (at an S/N of 3) are 0.1 ng mL−1 in both cases. (author)

  6. Prognostic value of serum α-fetoprotein in ovarian yolk sac tumors: A systematic review and meta-analysis.

    Science.gov (United States)

    Guo, Yang-Long; Zhang, Ying-Li; Zhu, Jian-Qing

    2015-01-01

    This study was conducted to determine the prognostic value of serum α-fetoprotein (AFP) levels in patients with ovarian yolk sac tumor (OYST). We performed a systematic review and meta-analysis to assess the associations between serum AFP level and prognosis in OYST. A total of 12 quantitative studies met the inclusion criteria. Preoperative AFP was not found to be associated with overall survival (OS) [odds ratio (OR)=0.84, 95% confidence interval (CI): 0.43-1.62] in OYST. However, a high postoperative AFP level was associated with worse OS (OR=0.16, 95% CI: 0.05-0.48) and relapse-free survival (RFS) (OR=0.18, 95% CI: 0.08-0.43) compared to a low postoperative AFP level in patients with OYST. In addition, a postoperative AFP level of >1,000 ng/ml was associated with a decrease in OS (OR=0.16, 95% CI: 0.05-0.50) and RFS (OR=0.21, 95% CI: 0.08-0.57). In conclusion, the postoperative, but not the preoperative, AFP level was found to be a prognostic factor in patients with OYST. In particular, a postoperative AFP level of >1,000 ng/ml was an indicator of poor prognosis in patients with OYST. PMID:25469282

  7. Ab initio alpha-alpha scattering.

    Science.gov (United States)

    Elhatisari, Serdar; Lee, Dean; Rupak, Gautam; Epelbaum, Evgeny; Krebs, Hermann; Lähde, Timo A; Luu, Thomas; Meißner, Ulf-G

    2015-12-01

    Processes such as the scattering of alpha particles ((4)He), the triple-alpha reaction, and alpha capture play a major role in stellar nucleosynthesis. In particular, alpha capture on carbon determines the ratio of carbon to oxygen during helium burning, and affects subsequent carbon, neon, oxygen, and silicon burning stages. It also substantially affects models of thermonuclear type Ia supernovae, owing to carbon detonation in accreting carbon-oxygen white-dwarf stars. In these reactions, the accurate calculation of the elastic scattering of alpha particles and alpha-like nuclei--nuclei with even and equal numbers of protons and neutrons--is important for understanding background and resonant scattering contributions. First-principles calculations of processes involving alpha particles and alpha-like nuclei have so far been impractical, owing to the exponential growth of the number of computational operations with the number of particles. Here we describe an ab initio calculation of alpha-alpha scattering that uses lattice Monte Carlo simulations. We use lattice effective field theory to describe the low-energy interactions of protons and neutrons, and apply a technique called the 'adiabatic projection method' to reduce the eight-body system to a two-cluster system. We take advantage of the computational efficiency and the more favourable scaling with system size of auxiliary-field Monte Carlo simulations to compute an ab initio effective Hamiltonian for the two clusters. We find promising agreement between lattice results and experimental phase shifts for s-wave and d-wave scattering. The approximately quadratic scaling of computational operations with particle number suggests that it should be possible to compute alpha scattering and capture on carbon and oxygen in the near future. The methods described here can be applied to ultracold atomic few-body systems as well as to hadronic systems using lattice quantum chromodynamics to describe the interactions of

  8. Ab initio alpha-alpha scattering

    Science.gov (United States)

    Elhatisari, Serdar; Lee, Dean; Rupak, Gautam; Epelbaum, Evgeny; Krebs, Hermann; Lähde, Timo A.; Luu, Thomas; Meißner, Ulf-G.

    2015-12-01

    Processes such as the scattering of alpha particles (4He), the triple-alpha reaction, and alpha capture play a major role in stellar nucleosynthesis. In particular, alpha capture on carbon determines the ratio of carbon to oxygen during helium burning, and affects subsequent carbon, neon, oxygen, and silicon burning stages. It also substantially affects models of thermonuclear type Ia supernovae, owing to carbon detonation in accreting carbon-oxygen white-dwarf stars. In these reactions, the accurate calculation of the elastic scattering of alpha particles and alpha-like nuclei—nuclei with even and equal numbers of protons and neutrons—is important for understanding background and resonant scattering contributions. First-principles calculations of processes involving alpha particles and alpha-like nuclei have so far been impractical, owing to the exponential growth of the number of computational operations with the number of particles. Here we describe an ab initio calculation of alpha-alpha scattering that uses lattice Monte Carlo simulations. We use lattice effective field theory to describe the low-energy interactions of protons and neutrons, and apply a technique called the ‘adiabatic projection method’ to reduce the eight-body system to a two-cluster system. We take advantage of the computational efficiency and the more favourable scaling with system size of auxiliary-field Monte Carlo simulations to compute an ab initio effective Hamiltonian for the two clusters. We find promising agreement between lattice results and experimental phase shifts for s-wave and d-wave scattering. The approximately quadratic scaling of computational operations with particle number suggests that it should be possible to compute alpha scattering and capture on carbon and oxygen in the near future. The methods described here can be applied to ultracold atomic few-body systems as well as to hadronic systems using lattice quantum chromodynamics to describe the interactions of

  9. Connexin-43 can delay early recurrence and metastasis in patients with hepatitis B-related hepatocellular carcinoma and low serum alpha-fetoprotein after radical hepatectomy

    International Nuclear Information System (INIS)

    We studied the relationships among Cx43, CD105, and VEGF in specimens of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) with different serum AFP levels with respect to recurrence and metastasis. Expressions of Cx43, CD105, and VEGF in 234 HCC tissue specimens were examined using tissue microarray and immunohistochemistry. Cx43 mRNA expression was examined in 38 frozen HCC specimens using RT-PCR. Correlations between these expressions and tumor recurrence, metastasis, and prognosis were analyzed using Kaplan–Meier and Cox regression analyses. Cx43 expression correlated with early tumor recurrence (P = 0.001), disease-free survival (P = 0.026), and overall survival (P = 0.000) in patients with serum AFP < 400 ng/ml, but not in those with serum AFP ≥ 400 μg/L. Cx43 expression is an independent predictor of later recurrence and longer overall survival and is inversely correlated with expression of CD105 and VEGF (P = 0.018 and 0.023, respectively), histological differentiation (P = 0.002), vessel tumor embolism (P = 0.029), and focal number (P = 0.017). Immunohistochemistry showed that Cx43 expression in patients with low AFP was lower in patients with distant metastases than in those with no metastasis or those with liver metastasis. Patients with early recurrence expressed less Cx43 mRNA than did those with no recurrence (χ2 = 9.827, P = 0.002). Cx43 expression can delay early HCC recurrence, metastasis, and poor prognosis after radical hepatectomy in patients with HBV-related HCC and low AFP

  10. AFP-L3在肝细胞癌诊断中的临床价值%Diagnostic role of alpha-fetoprotein variants in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    韩鹏; 褚瑞海; 张彩彩; 郭欣

    2012-01-01

    目的 探讨血清中AFP-L3含量对肝细胞癌诊断的临床意义.方法 利用微量离心柱法分离得到AFP-L3,采用化学发光法分别检测总AFP与AFP-L3含量,进而计算AFP-L3与总AFP的比值.结果 肝细胞癌患者血清中AFP及AFP-L3的水平均显著高于其他各组,而不同AFP浓度的肝细胞癌患者组AFP-L3的阳性率无明显差异.结论 AFP-L3作为肿瘤标志物对肝细胞癌的早期诊断及良、恶性肝病的鉴别有一定的临床价值.

  11. Faddeev calculation of 3 alpha and alpha alpha Lambda systems using alpha alpha resonating-group method kernel

    CERN Document Server

    Fujiwara, Y; Kohno, M; Suzuki, Y; Baye, D; Sparenberg, J M

    2004-01-01

    We carry out Faddeev calculations of three-alpha (3 alpha) and two-alpha plus Lambda (alpha alpha Lambda) systems, using two-cluster resonating-group method kernels. The input includes an effective two-nucleon force for the alpha alpha resonating-group method and a new effective Lambda N force for the Lambda alpha interaction. The latter force is a simple two-range Gaussian potential for each spin-singlet and triplet state, generated from the phase-shift behavior of the quark-model hyperon-nucleon interaction, fss2, by using an inversion method based on supersymmetric quantum mechanics. Owing to the exact treatment of the Pauli-forbidden states between the clusters, the present three-cluster Faddeev formalism can describe the mutually related, alpha alpha, 3 alpha and alpha alpha Lambda systems, in terms of a unique set of the baryon-baryon interactions. For the three-range Minnesota force which describes the alpha alpha phase shifts quite accurately, the ground-state and excitation energies of 9Be Lambda are...

  12. Review of alpha_s determinations

    CERN Document Server

    Pich, Antonio

    2013-01-01

    The present knowledge on the strong coupling is briefly summarized. The most precise determinations of alpha_s, at different energies, are reviewed and compared at the Z mass scale, using the predicted QCD running. The impressive agreement achieved between experimental measurements and theoretical predictions constitutes a beautiful and very significant test of Asymptotic Freedom, establishing QCD as the fundamental theory of the strong interaction. The world average value of the strong coupling is found to be alpha_s(M_Z^2)= 0.1186 \\pm 0.0007.

  13. Electrochemical Enzyme-Linked Immunosorbent Assay (ELISA for α-Fetoprotein Based on Glucose Detection with Multienzyme-Nanoparticle Amplification

    Directory of Open Access Journals (Sweden)

    Ning Gan

    2013-10-01

    Full Text Available Since glucose biosensors are one of the most popular and widely used point-of-care testing devices, a novel electrochemical enzyme-linked immunosorbent assay (ELISA for protein biomarkers has been developed based on a glucose detection strategy. In this study, α-fetoprotein (AFP was used as the target protein. An electrochemical ELISA system was constructed using anti-AFP antibodies immobilized on microwell plates as the capture antibody (Ab1 and multi-label bioconjugates as signal tracer. The bioconjugates were synthesized by attaching glucoamylase and the secondary anti-AFP antibodies (Ab2 to gold nanoparticles (AuNPs. After formation of the sandwich complex, the Ab2-glucoamylase-AuNPs conjugates converted starch into glucose in the presence of AFP. The concentration of AFP can be calculated based on the linear relation between AFP and glucose, the concentration of which can be detected by the glucose biosensor. When the AFP concentration ranged from 0.05 to 100 ng/mL, a linear calibration plot (i (µA = 13.62033 − 2.86252 logCAFP (ng/mL, r = 0.99886 with a detection limit of 0.02 ng/mL was obtained under optimal conditions. The electrochemical ELISA developed in this work shows acceptable stability and reproducibility, and the assay for AFP spiked in human serum also shows good recovery (97.0%–104%. This new method could be applied for detecting any protein biomarker with the corresponding antibodies.

  14. Using serum α-fetoprotein for prognostic prediction in patients with hepatocellular carcinoma: what is the most optimal cutoff?

    Directory of Open Access Journals (Sweden)

    Chia-Yang Hsu

    Full Text Available The prognostic ability of α-fetoprotein (AFP for patients with hepatocellular carcinoma (HCC was examined by using different cutoff values. The optimal AFP cutoff level is still unclear.A total of 2579 HCC patients were consecutively enrolled in Taiwan, where hepatitis B is the major etiology of chronic liver disease. Four frequently used AFP cutoff levels, 20, 200, 400, 1000 ng/mL, were investigated. One-to-one matched pairs between patients having AFP higher and lower than the cutoffs were selected by using the propensity model. The adjusted hazard ratios of survival difference were calculated with Cox proportional hazards model.Patients with a higher AFP level were associated with more severe cirrhosis, more frequent vascular invasion, higher tumor burden and poorer performance status (all p0.05. Patients with AFP <20 ng/mL had significantly better long-term survival than patients with AFP ≧20 ng/mL (p<0.0001, and patients with AFP <400 ng/mL had significantly better overall outcome than patients with AFP ≧400 ng/mL (p = 0.0186. There was no difference of long-term survival between patients divided by AFP levels of 200 and 1000 ng/mL. The adjusted hazard ratios of AFP ≧20 ng/mL and AFP ≧400 ng/mL were 1.545 and 1.471 (95% confidence interval: 1.3-1.838 and 1.178-1.837, respectively.This study shows the independently predictive ability of baseline serum AFP level in HCC patients. AFP levels of 20 and 400 ng/mL are considered feasible cutoffs to predict long-term outcome in unselected HCC patients.

  15. TGF-β Signaling Cooperates with AT Motif-Binding Factor-1 for Repression of the α-Fetoprotein Promoter

    Directory of Open Access Journals (Sweden)

    Nobuo Sakata

    2014-01-01

    Full Text Available α-Fetoprotein (AFP is known to be highly produced in fetal liver despite its barely detectable level in normal adult liver. On the other hand, hepatocellular carcinoma often shows high expression of AFP. Thus, AFP seems to be an oncogenic marker. In our present study, we investigated how TGF-β signaling cooperates with AT motif-binding factor-1 (ATBF1 to inhibit AFP transcription. Indeed, the expression of AFP mRNA in HuH-7 cells was negatively regulated by TGF-β signaling. To further understand how TGF-β suppresses the transcription of the AFP gene, we analyzed the activity of the AFP promoter in the presence of TGF-β. We found that the TGF-β signaling and ATBF1 suppressed AFP transcription through two ATBF1 binding elements (AT-motifs. Using a heterologous reporter system, both AT-motifs were required for transcriptional repression upon TGF-β stimulation. Furthermore, Smads were found to interact with ATBF1 at both its N-terminal and C-terminal regions. Since the N-terminal (ATBF1N and C-terminal regions of ATBF1 (ATBF1C lack the ability of DNA binding, both truncated mutants rescued the cooperative inhibitory action by the TGF-β signaling and ATBF1 in a dose-dependent manner. Taken together, these findings indicate that TGF-β signaling can act in concert with ATBF1 to suppress the activity of the AFP promoter through direct interaction of ATBF1 with Smads.

  16. Review of alpha_s determinations

    OpenAIRE

    Pich, Antonio

    2013-01-01

    The present knowledge on the strong coupling is briefly summarized. The most precise determinations of alpha_s, at different energies, are reviewed and compared at the Z mass scale, using the predicted QCD running. The impressive agreement achieved between experimental measurements and theoretical predictions constitutes a beautiful and very significant test of Asymptotic Freedom, establishing QCD as the fundamental theory of the strong interaction. The world average value of the strong coupl...

  17. World Summary of $\\alpha_s$ (2015)

    CERN Document Server

    Bethke, Siegfried; Salam, Gavin P

    2015-01-01

    This is a preliminary update of the measurements of α s and the determination of the world average value of α s (M Z 2 ) presented in the 2013/2014 edition of the Review of Particle Properties [1]. A number of studies which became available since late 2013 provide new results for each of the (previously 5, now) 6 subclasses of measurements for which pre-average values of $\\alpha_s (M_Z^2)$ are determined.

  18. A peptide derived from α-fetoprotein prevents the growth of estrogen-dependent human breast cancers sensitive and resistant to tamoxifen

    OpenAIRE

    James A. Bennett; Mesfin, Fassil B.; Andersen, Thomas T.; Gierthy, John F.; Jacobson, Herbert I.

    2002-01-01

    An 8-mer peptide (EMTOVNOG) derived from α-fetoprotein was compared with tamoxifen for activity against growth of human breast cancer xenografts implanted in immune-deficient mice. Both peptide and tamoxifen prevented growth of estrogen-receptor-positive MCF-7 and T47D human breast cancer xenografts. A subline of MCF-7, made resistant to tamoxifen by a 6-month exposure to this drug in culture, was found to be resistant to tamoxifen in vivo. Peptide completely prevented the xenograft growth of...

  19. Standardization of the radioimmunoassay technique for the determination of human gastrin and its clinical application

    International Nuclear Information System (INIS)

    It was developed and standardized a system of radioimmunoassay for the determination of gastrin, employing synthetic human gastrin for radioiodination and preparation of standard as well as specific antibody raised rabbits. The hormone was labeled with 125I by the Cloramine T techique and purified by anion exchange chromatography in QAE-Sephadex A-25, being determined its specific activity. The tracer thus obtained was submitted to analysis of purity by poliacrilamide gel eletrophoresis and precipitation of proteins by trichloroacetic acid. Its stability evaluated according to the time of storage, being its purity and adequation for the use in radioimmunoassay also compared to a tracer obtained from a commercial diagnosis kit. The assays were performed by incubation of radioiodinated gastrin, standard gastrin prepared in plasma free from this hormone (from zero to 500 pmol/l) or samples to be assayed with the antiserum for 4 days at 40C. The separation between the free gastrin and the gastrin bound to the antibody was carried out by adsorption of the free hormone to the charcoal, whose ideal concentration was previously determined. Plasma free from gastrin was obtained from time-expired blood bank plasma submitted to extraction with charcoal. When performed the quality control, this radioimmunoassay was shown specific, accurate, precise and sensitive, allowing the performance of valid assays. Its validation was even confirmed by clear discrimination not only of the gastrin concentration in subjects with very low levels (gastrectomized) and extremely high levels (Zollinger-Ellison syndrome) as well as gastrin concentrations in subjects with other diseases, such as Chagas disease, pernicious anemia and chronic renal failure. (author)

  20. Studies on quantitative determination of urokinase by radioimmunoassay and its clinical application, 1

    International Nuclear Information System (INIS)

    A new radioimmunoassay method by which UK was capable of being measured quantitatively was developed. With the method of White et al., purified UK was obtained which had an activity of 20,000 Ploug units (PKU) per mg protein and was used as an antigen in this assay system. Anti-UK antisera (antibodies) were obtained from rabbits and guinea pigs immunized with a mixture of UK and Freund's complete adjuvant. The most suitable antibody among them was used for the radioimmunoassay. Labelling of UK with 125I-Na was carried out according to the Hunter and Greenwood method. Separation of the antibody-bound 125I-UK from free 125I-UK was done with a solid phase method using disposable microtiter plates. A standard curve was obtained using 125I-UK incubated with serial additions of unlabelled UK and a certain amount of diluted antibody (x 8,000). It showed an exponential linear line with a step slope ranging from 0.2 to 32 PKU, and it was capable of determing UK concentrations as low as 0.2 PKU per ml of urine and specifically without any interference from other substances in the urine. The amount of UK determined by the radioimmunoassay correlated well with the biological activities of the fibrinolytic test tube method with the same urine samples. Using this method, the UK concentration and the daily excretion in the urine of healthy adults was investigated. The urinary concentration of UK ranged from 0.4 to 7.0 PKU per ml, and the UK excretion in 24 hours was 2.230 +- 910 PKU. (JPN)