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Sample records for alpha fetoprotein radioimmunoassay

  1. The radioimmunoassay of serum alpha-fetoprotein levels

    International Nuclear Information System (INIS)

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

    1982-01-01

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

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

    International Nuclear Information System (INIS)

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

    1981-01-01

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

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

    International Nuclear Information System (INIS)

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

    1982-01-01

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

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

    International Nuclear Information System (INIS)

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

    1980-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1980-01-01

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

  6. 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)

    Mann, K.; Lamerz, R.; Hellmann, Th.; Kuemper, H.J.; Staehler, G.; Karl, H.J.

    1980-01-01

    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 125 I-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 125 I-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.)

  7. Subcloning and expression of human alpha-fetoprotein gene in ...

    African Journals Online (AJOL)

    Subcloning and expression of human alpha-fetoprotein gene in Pichia pastoris. ... in inducing protein production in auxotrophic media lacking histidine. This protein could be useful in monoclonal antibody production and in diagnostic kit preparations. Keywords: Alpha-fetoprotein, Pichia pastoris, cloning, expression ...

  8. [Increase of alpha-fetoprotein in pancreatic endocrine tumors with hepatic metastases. Apropos of 2 cases].

    Science.gov (United States)

    Lesur, G; Bergemer, A M; Turner, L; Parlier, H; Bernades, P; Dupuy, P

    1996-03-01

    We report two cases of metastatic non-functioning pancreatic endocrine tumour with very elevated plasma levels of alpha-fetoprotein. In these two cases, serial plasma levels of alpha-fetoprotein, initially normal, correlated well with hepatic tumour progression and were associated with fatal outcome. These results suggest that elevated plasma concentration of alpha-fetoprotein may be caused by metastatic pancreatic endocrine tumour and than alpha-fetoprotein serial measurement may be useful in prognostic evaluation.

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

    African Journals Online (AJOL)

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

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

    African Journals Online (AJOL)

    EB

    2013-09-03

    Sep 3, 2013 ... Background: The prognosis of patients with hepatocellular carcinoma(HCC) is generally very poor with a 5-year survival rate of less than 15% since most of ... antigen (CEA), CD34, alpha fetoprotein (AFP) were usually used to .... and a number of immunomarkers have been widely utilized to facilitate the ...

  11. Serum alpha-fetoprotein level is higher in hepatitis C than hepatitis ...

    African Journals Online (AJOL)

    Background: The frequency of raised serum alpha-fetoprotein may vary in relation to hepatitis B or C infection in chronic liver disease (CLD). The study evaluated the frequency of hepatitis B and C in patients with chronic liver disease and correlated the levels of serum alpha-fetoprotein with hepatitis B and C infection in the ...

  12. Cellular immunolocalization of alpha-fetoprotein in rat liver.

    Science.gov (United States)

    Woods, J A

    1983-10-01

    Increased synthesis of alpha-fetoprotein (AFP) was induced in rat liver by the administration of 3'-methyl-4-dimethyl-aminoazobenzene. The indirect immunoperoxidase technique was used to detect AFP. Cellular localization of AFP was studied using a number of different fixation procedures. Serial sections stained with immunoglobulin served to determine the extent of diffusion of serum proteins into liver cells during fixation. Background staining was minimized when Lillie's neutral buffered formalin plus acetic acid was used as the fixative. After 3'-methyl-4-dimethylaminoazobenzene ingestion, bile duct cell proliferation occurred. The serum AFP was positive in all rats after 17 days on the diet. In rats with AFP-positive sera the immunohistochemical reaction in mature hepatocytes was positive while bile duct cells and small hepatocytes were negative for AFP.

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

    Science.gov (United States)

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

    2001-08-01

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

  14. Comparison of double-antibody radioimmunoassay with Farr-technique radioimmunoassay and double-antibody enzyme immunoassay for α-fetoprotein

    International Nuclear Information System (INIS)

    Kirkpatrick, A.; Wepsic, H.T.; Nakamura, R.M.

    1977-01-01

    We describe double-antibody procedures for determining α-fetoprotein in human serum. An equilibrium procedure can be done in 24 h with a sensitivity of at least 4 μg/liter and coefficient of variation of 5.5%. There are no interferences from normal human sera or sera with certain commonly seen chemical abnormalities. We also describe and discuss sequential procedures that range in sensitivity from 250 ng to 1 μg/liter and require 24 to 48 h incubation. The precise (mid-range) portion of the dose/response curve for sequential procedures can be shifted to higher or lower values by an adjustment of the time of preliminary incubation of antibody with unlabeled antigen. With a 37 0 C incubation, a sequential procedure can be completed in 7 h. Sensitivity is 1 μg/liter, and coefficient of variation 8.0%. The relative merits of the above assay procedures are discussed. The double-antibody radioimmunoassay is twice as sensitive as the Farr procedure and it is free of the large and variable nonspecific precipitation that accompanies the precipitation of bound antigen with sodium sulfate solution. Double-antibody radioimmunoassay is superior to enzyme immunoassay in both sensitivity and precision

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

    OpenAIRE

    Taketa, Kazuhisa; Ichikawa, Eriko; Taga, Hiroko; Hirai, Hidematsu

    1984-01-01

    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 ...

  16. Maternal serum alpha fetoprotein among pregnant females in Alexandria.

    Science.gov (United States)

    Baghagho, Ehssan E; Kharboush, Ibrahim F; El-Kaffash, Dalal M; KarKour, Tarek A; Ismail, Suzan R; Mortada, Mona M

    2004-01-01

    Maternal serum alpha fetoprotein (MSAFP) was introduced as a screening test for congenital malformations especially neural tube defects (NTDs) two decades ago. However, many factors were known to affect its level. From these are racial differences and maternal weight. The aim of the present work is to illustrate the normal distribution of MSAFP among working pregnant women in Alexandria in gestational age 16-18 weeks, to identify some of its determinants, and to determine the specificity and sensitivity of MSAFP for the detection of congenital anomalies and adverse pregnancy outcome. A sample of 608 pregnant working women who were 16-18 week gestation was recruited for the study from the antenatal clinic affiliated to Gamal Abdel Nasser Health Insurance Hospital in Alexandria. The enrolled women were interviewed using a structured questionnaire and a blood sample was collected from each of them to measure the level of MSAFP. At the expected time of delivery, Gamal Abd el Nasser Health Insurance Hospital was visited to collect data about the outcome of pregnancy of the enrolled women. The median of MSAFP level for deliveries with no congenital anomalies were 25.5, 33.5, and 53.2 IU/ml, at gestational weeks 16, 17 and 18 respectively. The significant variables related positively to MSAFP level included abortion or stillbirth, congenital anomalies in the index pregnancy, gestational age, bleeding during pregnancy, gestational diabetes, twin pregnancy, consanguinity between maternal parents, history of congenital or genetic diseases in maternal family, and caesarian section deliveries. Fatigue score was negatively correlated to MSAFP level. Using MSAFP multiples of median (MOM), 42.9 % of abortions and stillbirths, 57.1 % of twin pregnancies, 31.25 % of preterm deliveries and 27.3 % of low birth weight had levels of 3 MOM or more. One fourth of the congenital anomalies were below 0.5 MOM and 41.7 % were at or above 3 MOM. The sensitivity of MSAFP test for the detection

  17. 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...

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

    International Nuclear Information System (INIS)

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

    1979-01-01

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

  19. 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

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

    International Nuclear Information System (INIS)

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

    2007-01-01

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

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

    DEFF Research Database (Denmark)

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

    2011-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Rino A Gani

    2016-05-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-01-06

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

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

    Directory of Open Access Journals (Sweden)

    Sirasa Yodmongkol

    2016-03-01

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

  5. Alpha fetoprotein

    Science.gov (United States)

    ... Creasy and Resnik's Maternal-Fetal Medicine: Principles and Practice . 7th ed. Philadelphia, PA: Elsevier Saunders; 2014:chap 30. Review Date 8/26/2017 Updated by: Peter J Chen, MD, FACOG, Associate Professor of OBGYN at Cooper Medical School at Rowan ...

  6. 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

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

    NARCIS (Netherlands)

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

    2012-01-01

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

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

    Science.gov (United States)

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

    2018-02-20

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

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

    DEFF Research Database (Denmark)

    Schmidt, Lars E; Dalhoff, Kim

    2005-01-01

    An increase in alpha-fetoprotein (AFP) following hepatic necrosis is considered indicative of hepatic regeneration. This study evaluated the prognostic value of serial AFP measurements in patients with severe acetaminophen-induced liver injury. Prospectively, serial measurements of AFP were...... performed in 239 patients with acetaminophen intoxication and a peak alanine aminotransferase (ALT) level above 1000 U/L. AFP was measured using an enzyme-linked immunoassay (EIA) with a detection limit below 0.4 microg/L. The optimum threshold of AFP to discriminate nonsurvivors was identified. An increase......%. In conclusion, an increase in AFP was strongly associated with a favorable outcome in patients with acetaminophen-induced liver injury. AFP may be useful as a supplement to existing prognostic criteria. We suggest that the introduction of highly sensitive EIAs for the detection of AFP will require...

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

    Directory of Open Access Journals (Sweden)

    Ji He Kim

    2018-01-01

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

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

    Science.gov (United States)

    Wang, Xiaoping; Lin, Huanping; Wang, Qiaoxia

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

  12. 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

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

    International Nuclear Information System (INIS)

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

    2009-01-01

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

  14. Routine ultrasonography compared with maternal serum alpha-fetoprotein for neural tube defect screening.

    Science.gov (United States)

    Norem, Carol T; Schoen, Edgar J; Walton, David L; Krieger, Robyn C; O'Keefe, Jennifer; To, Trinh T; Ray, G Thomas

    2005-10-01

    This study was done to estimate the value of prenatal maternal serum alpha-fetoprotein (MSAFP) screening compared with that of routine ultrasonography in the diagnosis of neural tube defects (NTDs). An integrated database was used retrospectively to identify cases of NTDs among 219,000 consecutive pregnancy outcomes observed during a 7-year period at 40 Kaiser Permanente facilities in Northern California. We specifically examined types of NTD and the tests used to diagnose cases. We identified 189 NTD cases, 102 of which had received MSAFP screening. Results of MSAFP testing were negative in 25 (25%) of these 102 cases. Without other testing, these 25 NTD diagnoses would have been missed. These included 15 (38%) of the 40 spina bifida cases screened, 6 (67%) of the 9 encephalocele cases screened, and 4 (8%) of the 53 anencephaly cases screened. Of the 186 NTD cases diagnosed prenatally, 115 (62%) were initially detected by routine ultrasonography administered during the second trimester without knowledge of MSAFP values; 69 (37%) were diagnosed by targeted ultrasonography after MSAFP screening indicated a higher risk for NTD; and 2 (1%) were diagnosed by pathology examination after miscarriage. Compared with MSAFP performed alone for screening, routine second-trimester ultrasonography was more likely to discover an NTD.

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

    Science.gov (United States)

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

    2016-12-12

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

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

    Science.gov (United States)

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

    2017-08-01

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

  17. First trimester maternal serum alpha-fetoprotein is not raised in pregnancies with open spina bifida.

    Science.gov (United States)

    Spencer, Kevin; Khalil, Asma; Brown, Louise; Mills, Ian; Horne, Hannah

    2014-02-01

    Two recent studies have suggested that maternal serum alpha fetoprotein (AFP) levels are increased in the first trimester of pregnancies in which the fetus has an open spina bifida. This is contrary to previously published studies. This study assesses further whether maternal serum AFP is elevated in the first trimester in cases with open spina bifida. Cases with open spina bifida were identified from our fetal database, and corresponding first trimester screening samples were retrieved and analysed for maternal serum AFP. A control group was selected by taking three samples matched for gestational age (exact day), ethnicity and smoking status and received in the laboratory on the same day. AFP was measured with the Kryptor platform and free β-hCG and pregnancy-associated plasma protein A results were available from the fetal database. Thirty-nine open spina bifida cases were identified with a control group of 126 cases. The median multiple of the median AFP in the cases were not significantly different from the controls (0.92 vs 1.06 p = 0.3511) as was the case for free β-hCG (0.87 vs 0.95 p = 0.7146) and pregnancy-associated plasma protein A (1.04 vs 1.04 p = 0.261). Our results confirm that maternal serum biochemical markers in the first trimester are unable to distinguish cases in which the fetus has open spina bifida. © 2013 John Wiley & Sons, Ltd.

  18. Role of Maternal Serum Alpha-Fetoprotein and Ultrasonography in Contemporary Detection of Spina Bifida.

    Science.gov (United States)

    Racusin, Diana A; Villarreal, Sarah; Antony, Kathleen M; Harris, R Alan; Mastrobattista, Joan; Lee, Wesley; Shamshirsaz, Alireza A; Belfort, Michael; Aagaard, Kjersti M

    2015-12-01

    Midtrimester maternal serum alpha-fetoprotein (MSAFP) and sonographic evaluation have been used to screen for spina bifida. With the increased uptake of cell-free DNA (cfDNA) and first trimester screening, MSAFP levels may no longer be obtained routinely. Our aim was to evaluate a pediatric neurosurgical referral center database of spina bifida cases to determine the antenatal detection rate and means of diagnosis. Nested case series of all spina bifida cases referred postnatally from 2007 to 2013. Data were abstracted from the maternal record and rates of antenatal detection with MSAFP and sonographic screening were determined. Of the 105 postnatally referred cases, 11.4% (12/105) were not identified until delivery. Overall, 39% of the cases had MSAFP screening. The odds ratio for sonogram-based detection of spina bifida was 4.9 (95% confidence interval, 2-11.9). Of the neonatally detected cases, 100% had prenatal care and 91.6% (11 of the 12 cases) had documented sonography. We have found that 11.4% of the spina bifida cases were not detected before delivery. Nine out of the 12 cases of antenatally missed spina bifida were not screened using MSAFP. Our findings support the approach of midtrimester MSAFP screening combined with sonographic evaluation. We speculate that prenatal screening with MSAFP is underutilized. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

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

    Science.gov (United States)

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

    2016-01-15

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

  20. Fast detection of alpha-fetoprotein-L3 using lens culinaris agglutinin immobilized gold nanoparticles.

    Science.gov (United States)

    Sun, Ying; Qin, Liubin; Liu, Dejun; Liu, Changhong; Sun, Yongwei; Duan, Yourong

    2014-06-01

    Colloidal gold is extensively used for molecular sensing because that the surface plasmon resonance (SPR) bands are affected by changes in the dielectric properties in the close vicinity of these structures due to the binding of ligands to the corresponding receptor molecules immobilized onto the nanostructures through chemi- or physisorption. We describe a simple method for the detection of Alpha-Fetoprotein-L3 which is a new generation of tumor marker for hepatocellular carcinoma (HCC) based on the aggregation of Lens culinaris agglutinin (LCA) Immobilized Gold Nanoparticles. The LCA conjugated GNPs obtained were 15-20 nm in diameter. The visible color change of the gold nanoparticles from purple to blue on interaction with 100 ng/mL of AFP-L3 is the principle applied here for the sensing of AFP-L3 level. UV/Vis spectroscopy also allows assay monitoring by quantifying the red shift of the plasmon resonance wavelength. With this method, the protein AFP-L3 can be rapidly detected as demanded for clinical diagnosis.

  1. Quantum dot-based immunochromatography test strip for rapid, quantitative and sensitive detection of alpha fetoprotein.

    Science.gov (United States)

    Yang, Qiuhua; Gong, Xiaoqun; Song, Tao; Yang, Jiumin; Zhu, Shengjiang; Li, Yunhong; Cui, Ye; Li, Yingxin; Zhang, Bingbo; Chang, Jin

    2011-12-15

    Rapid, quantitative detection of tumor markers with high sensitivity and specificity is critical to clinical diagnosis and treatment of cancer. We describe here a novel portable fluorescent biosensor that integrates quantum dot (QD) with an immunochromatography test strip (ICTS) and a home-made test strip reader for detection of tumor markers in human serum. Alpha fetoprotein (AFP), which is valuable for diagnosis of primary hepatic carcinoma, is used as a model tumor marker to demonstrate the performance of the proposed immunosensor. The principle of this sensor is on the basis of a sandwich immunoreaction that was performed on an ICTS. The fluorescence intensity of captured QD labels on the test line and control line served as signals was determined by the home-made test strip reader. The strong luminescence and robust photostability of QDs combined with the promising advantages of an ICTS and sensitive detection with the test strip reader result in good performance. Under optimal conditions, this biosensor is capable of detecting as low as 1 ng/mL AFP standard analyte in 10 min with only 50 μL sample volume. Furthermore, 1000 clinical human serum samples were tested by both the QD-based ICTS and a commercial electrochemiluminescence immunoassay AFP kit simultaneously to estimate the sensitivity, specificity and concordance of the assays. Results showed high consistency except for 24 false positive cases (false positive rate 3.92%) and 17 false negative cases (false negative rate 4.38%); the error rate was 4.10% in all. This demonstrates that the QD-based ICTS is capable of rapid, sensitive, and quantitative detection of AFP and shows a great promise for point-of-care testing of other tumor markers. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. 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.

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

    Science.gov (United States)

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

    2018-05-01

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

  4. 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

  5. The association between hypertensive disorders of pregnancy and abnormal second-trimester maternal serum levels of hCG and alpha-fetoprotein

    NARCIS (Netherlands)

    Morssink, LP; Heringa, MP; Beekhuis, [No Value; DeWolf, BTHM; Mantingh, A

    Objective: To examine the association between hypertensive disorders of pregnancy and second-trimester maternal serum alpha-fetoprotein (MSAFP) and hCG levels. Methods: The proportions of abnormal second-trimester MSAFP and hCG levels in the serum samples from 65 women with true pregnancy-induced

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

    NARCIS (Netherlands)

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

    1995-01-01

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

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

    NARCIS (Netherlands)

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

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

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

    Directory of Open Access Journals (Sweden)

    G. Martínez-Mier

    2017-10-01

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

  9. Carcinosarcoma of the Uterine Corpus with Alpha-Fetoprotein-Producing Hepatoid Adenocarcinoma: A Report of Two Cases

    Directory of Open Access Journals (Sweden)

    Ryuji Kawaguchi

    2011-07-01

    Full Text Available We report two cases of uterine carcinosarcoma associated with alpha-fetoprotein (AFP-producing hepatoid adenocarcinoma. Samples were obtained from two women aged 63 and 82 years. Serum AFP levels of the two samples were 10,131 and 401 ng/ml, respectively. Histologically, in both cases the tumor cells were composed of hepatoid adenocarcinoma component and sarcoma component including rhabdomyosarcoma. Immunohistochemical analyses revealed that AFP was expressed in the cytoplasm of the carcinomatous component. After surgery, the patients received six courses of carboplatin/paclitaxel chemotherapy, and the serum levels of AFP decreased to normal range. The first patient is alive and well at the 2-year follow-up, while the second patient died of disease 1 year after initial operative treatment. This is, to our knowledge, the second report of carcinosarcoma of the uterine corpus with AFP-producing hepatoid adenocarcinoma, as proven by immunohistochemical analyses.

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

    Science.gov (United States)

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

    1990-01-01

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

  11. Blood Serum Alpha Fetoprotein Enhancer Binding Protein, a Tumor Suppressor, Decreases in Chronic HBV Hepatitis Patients as Hepatocellular Cancer Appears

    Directory of Open Access Journals (Sweden)

    James N. Riggins

    2010-01-01

    Full Text Available Chronic hepatitis increases the risk of hepatocellular carcinoma (HCC. To test whether circulating proteins reflect hepatic carcinogenesis, sera from patients and controls were albumin depleted, enriched for glycoproteins, digested with trypsin, and subjected to reverse phase chromatography and tandem mass spectrometry. Alpha-fetoprotein enhancer binding protein (AFPebp, a tumor suppressor, was repeatedly identified in sera from chronic HBV hepatitis patients. We independently identified and quantified AFPebp with a deuterated, phenylisocyanate-labeled synthetic peptide standard. Elevated AFPebp levels in sera from chronic HBV hepatitis patients decreased as cancer developed. These data suggest that rising AFPebp levels in chronic HBV hepatitis may be protective, while falling levels may contribute to HCC development.

  12. 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.

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

    Science.gov (United States)

    Huang, Ting-Shuo; Shyu, Yu-Chiau; Turner, Robin; Chen, Huang-Yang; Chen, Pei-Jer

    2013-06-06

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

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

    International Nuclear Information System (INIS)

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

    1985-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Joon Seong Ahn

    2013-06-01

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

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

    Science.gov (United States)

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

    2015-01-01

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

  17. Post-synthetic modification of human alpha-fetoprotein revealed by isoelectric focusing controls its immunosuppressive potency

    Energy Technology Data Exchange (ETDEWEB)

    Lester, E. P.; Miller, J. B.; Yachnin, S.

    1977-01-01

    We have demonstrated 3 variants of human alpha-fetoprotein (HAFP) by crossed immunoelectrophoresis, and have correlated the capacity of HAFP isolates to suppress human lymphocyte transformation in vitro with the relative proportion of the electronegative variant, HAFP-3, present in each isolate. We have now isolated HAFP from the serum, ascitic fluid, and saline extract of tumor from a single hepatoma patient, and from an homogenate of fetal livers. When tested for their capacity to inhibit human lymphocyte transformation in vitro, tumor and fetal liver HAFP were found to be extremely potent; serum HAFP had intermediate potency, and ascitic fluid HAFP was the least potent. Analysis of these HAFP isolates by crossed immunoelectrophoresis confirmed the correlation between the proportion of HAFP-3 and the immunosuppressive potency of each isolate. In addition, analysis of these HAFP isolates by isoelectric focusing in polyacrylamide gels containing 8 M urea revealed further evidence of microheterogeneity; at least 6 molecular variants were apparent. The proportion of one of these variants, termed HAFP-3a, in each isolate was correlated with the immunosuppressive potency of the isolate. The sialic acid content of the various HAFP isolates did not vary significantly. Our data suggest that a post-synthetic modification of HAFP occurs, which modulates its immunosuppressive potency.

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

    International Nuclear Information System (INIS)

    Dao, Mouhamadou.

    1976-03-01

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

  19. Enzyme-catalyzed silver deposition on irregular-shaped gold nanoparticles for electrochemical immunoassay of alpha-fetoprotein.

    Science.gov (United States)

    Lai, Wenqiang; Tang, Dianping; Que, Xiaohua; Zhuang, Junyang; Fu, Libing; Chen, Guonan

    2012-11-28

    A new and disposable electrochemical immunosensor was designed for detection of alpha-fetoprotein (AFP), as a model analyte, with sensitivity enhancement based on enzyme-catalyzed silver deposition onto irregular-shaped gold nanoparticles (ISGNPs). The assay was carried out with a sandwich-type immunoassay protocol by using ISGNP-labeled anti-AFP antibodies conjugated with alkaline phosphatase (ALP-Ab(2)) as detection antibodies. The enzymatically catalytic deposition of silver on the electrode could be measured by stripping analysis in KCl solution due to the Ag/AgCl solid-state voltammetric process. Several labeling protocols including spherical gold nanoparticle-labeled ALP-Ab(2) and ISGNP-labeled ALP-Ab(2) were investigated for determination of AFP, and improved analytical properties were achieved with the ISGNP labeling. With the ISGNP labeling method, the effects of incubation time and incubation temperature for antigen-antibody reaction, and deposition time of silver on the current responses of the electrochemical immunosensors were also monitored. Under optimal conditions, the electrochemical immunosensor exhibited a wide dynamic range from 0.01 ng mL(-1) to 200 ng mL(-1) with a detection limit of 5.0 pg mL(-1) AFP. The immunosensor displayed a good stability and acceptable reproducibility and accuracy. No significant differences at the 95% confidence level were encountered in the analysis of 10 clinical serum samples between the developed immunoassay and the commercially available electrochemiluminescent method for determination of AFP. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Oral antiviral therapy improves the diagnostic accuracy of alpha-fetoprotein levels in patients with chronic hepatitis B.

    Science.gov (United States)

    Shim, Jae-Jun; Kim, Jung Wook; Lee, Chang Kyun; Jang, Jae Young; Kim, Byung-Ho

    2014-09-01

    Analysis of alpha-fetoprotein (AFP) levels affords limited diagnostic accuracy because of the high false-positive rates, especially in those with active chronic hepatitis B (CHB). We measured AFP levels before and after commencement of oral antiviral therapy and explored the utility of these data in terms of early detection of hepatocellular carcinoma (HCC) in patients with CHB. A total of 207 patients with CHB who were treated with an oral antiviral agent were consecutively included. Dynamic changes in AFP levels and the diagnostic utility of such changes for HCC detection during the therapy were explored. The proportions of patients showing elevated AFP levels (≥ 20 ng/mL) were 22.2%, 5.5%, and 1.3% at baseline; and at 6 and 12 months after commencement of antiviral therapy, respectively. All patients who did not suffer from HCC exhibited normalization of AFP levels at 12 months. The cumulative incidence of HCC was 9.5% during 36 months of follow-up. If AFP levels were over 20 ng/mL after 12 months of antiviral treatment, the probability of HCC development approached certainty. The positive predictive value for HCC development remained at 100% in patients prescribed long-term (≥ 12 months) antiviral therapy, if AFP levels persistently or abruptly elevated more than 12 ng/mL. In the era of oral antiviral agents, AFP might be a useful biomarker for HCC surveillance in patients with CHB. © 2014 Journal of Gastroenterology and Hepatology Foundation and Wiley Publishing Asia Pty Ltd.

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

    International Nuclear Information System (INIS)

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

    2014-01-01

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

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

    Science.gov (United States)

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

    2014-09-01

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

  3. 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.

  4. p53, Ki-67, and serum alpha feto-protein as predictors of hepatocellular carcinoma recurrence in liver transplant patients.

    Science.gov (United States)

    Guzman, Grace; Alagiozian-Angelova, Victoria; Layden-Almer, Jennifer E; Layden, Thomas J; Testa, Guiliano; Benedetti, Enrico; Kajdacsy-Balla, André; Cotler, Scott J

    2005-11-01

    Patients with hepatocellular carcinoma who undergo orthotopic liver transplantation (OLT) are at risk for post-transplant tumor recurrence. The aim of this study was to evaluate whether expression of p53 and Ki-67 in hepatocellular carcinoma lesions present in explanted liver tissue was associated with time to tumor recurrence after OLT. Subjects consisted of 20 consecutive patients who underwent OLT and were found to have hepatocellular carcinoma in the liver explant. Immunostaining for p53 and Ki-67 was performed by standard methods. The presence of nuclear immunostaining in >10% of the tumor tissue was considered positive. Time to recurrence of hepatocellular carcinoma after OLT was compared between patients with positive and negative immunostaining by the log rank test. Multivariate analysis was performed using a Cox regression model to control for potentially confounding clinical factors. Time to post-transplant hepatocellular carcinoma recurrence was significantly more rapid in p53+ (P=0.0007) and Ki-67+ cases (P=0.001). These associations remained significant in multivariate analysis. Furthermore, time to recurrent hepatocellular carcinoma was significantly shorter in patients with a serum alpha feto-protein (AFP) level >or=100 ng/ml at time of diagnosis, compared to those with an AFP level or=100 ng/ml were associated with more rapid recurrence of hepatocellular carcinoma after OLT. Identification of patients at risk for early post-transplant recurrence could be used to guide surveillance and adjuvant treatment strategies. Modern Pathology (2005) 18, 1498-1503. doi:10.1038/modpathol.3800458; published online 8 July 2005.

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

    Science.gov (United States)

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

    2017-10-26

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

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

    Science.gov (United States)

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

    2015-07-01

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

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

    International Nuclear Information System (INIS)

    Hudig, D.; Sell, S.

    1979-01-01

    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 125 I 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 E 280 sup(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)

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-07-30

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

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

    International Nuclear Information System (INIS)

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

    1979-01-01

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

  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)

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

    1997-01-01

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

  11. 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

    Directory of Open Access Journals (Sweden)

    S.J. Guo

    2013-05-01

    Full Text Available 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.

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

    Science.gov (United States)

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

    2012-01-01

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

  13. 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.

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

    International Nuclear Information System (INIS)

    Dambuyant, C.; Sizaret, Ph.

    1975-01-01

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

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

    Science.gov (United States)

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

    2013-11-01

    Based on the reaction of the active -OH group in fullerol (F) with the dissociated -COOH group in fluorescein isothiocyanate (FITC) to form an F-FITC and the enhanced effect of N, N-dimethylaniline (DMA) on phosphorescence signal of F-FITC, a new phosphorescent labeling reagent (DMA-F-FITC) was developed. What's more, a phosphorescent sensor for the determination of alpha-fetoprotein variant (AFP-V) has been designed via the coupling technique of the high sensitivity for affinity adsorption-solid substrate-room temperature phosphorimetry (AA-SS-RTP) with the strong specificity reaction between DMA-F-FITC-Con A and AFP-V. The DMA-F-FITC increased the number of luminescent molecules in the biological target which improved the sensitivity of phosphorescent sensor. The proposed sensor was responsive, simple, selective and sensitive, and it has been applied to the determination of trace AFP-V in human serum and the forecast of human diseases using phosphorescence emission wavelength of F or FITC, with the results agreed well with those obtained by enzyme-linked immunoassay (ELISA). Meanwhile, the mechanisms for the labeling reaction and the sensing detection of AFP-V were discussed. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2015-04-01

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

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

    Science.gov (United States)

    Sando, Shota; Zhang, Bo; Cui, Tianhong

    2017-12-01

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

  18. Combining des-gamma-carboxyprothrombin and alpha-fetoprotein for hepatocellular carcinoma diagnosing: an update meta-analysis and validation study.

    Science.gov (United States)

    Chen, Huaping; Chen, Siyuan; Li, Shan; Chen, Zhijian; Zhu, Xuan; Dai, Meiyu; Kong, Lingxi; Lv, Xiaodan; Huang, Zhili; Qin, Xue

    2017-10-27

    Controversies about the combination of des-gamma-carboxyprothrombin (DCP) and alpha-fetoprotein (AFP) for hepatocellular carcinoma diagnosing still exist. Hence, we performed this updated meta-analysis to estimate the diagnostic value of DCP , AFP and DCP + AFP in HCC. In addition, we conducted a validation study to analyze the performance of the candidate makers. After a systematic literature review, 27 studies from 20 articles were identified from four major databases. The pooled sensitivity and specificity were 69% and 89%, respectively, for DCP; for AFP, they were 65% and 88%, respectively; and they were 82% and 85%, respectively, for DCP + AFP. The values of the area under the curve (AUC) for DCP, AFP, DCP + AFP, respectively, were 0.88, 0.75, and 0.90. The validation study confirmed that the performance of DCP + AFP (sensitivity = 84%, specificity = 86%; AUC = 0.887) was higher than that of DCP (sensitivity = 76%, specificity = 92%; AUC = 0.843) or AFP (sensitivity = 73%, specificity = 92%; AUC = 0.837) alone.

  19. Second trimester serum alpha-fetoprotein level is a significant positive predictor for intrauterine growth restriction in pregnant women with hyperemesis gravidarum

    Science.gov (United States)

    Özkaya, Enis; Çakır, Evrim; Çınar, Mehmet; Altay, Metin; Gelişen, Orhan; Kara, Fadıl

    2011-01-01

    Objective The aim of this study was to determine the association between three parameters of second trimester serum secreening and preterm labor and intrauterine growth restriction (IUGR) in patients with hyperemesis gravidarum (HG). Material and Methods A prospective study on 429 pregnancies with HG was designed to determine the association between alpha-fetoprotein (AFP), unconjugated estriol (uE3), human chorionic gonadotropin (HCG) and pregnancy prognosis in terms of preterm labor, IUGR and birth weight. Results In our study group the mean age of patients was 25.4±3.8 years. Mean birth weight was 3180±555 g. Mean AFP, uE3, hCG levels in the study group were 1.44±0.65 MoM, 0.91±0.38 MoM, 1.09±0.64 MoM, respectively. Twenty nine (6.8%) patients delivered before 37 weeks of gestation and 52 (12.1%) patients developed IUGR. Mean MoM values of AFP among patients with preterm labor, IUGR and normal delivery were 1.35±0.45, 1.97±0.81, 1.34±0.58 MoM, respectively (p1.55 was 3.73 (95% CI, 1.99–6.98, phyperemesis gravidarum. The second trimester screening test can predict poor outcome in HG. PMID:24591998

  20. Development of an automated wax-printed paper-based lateral flow device for alpha-fetoprotein enzyme-linked immunosorbent assay.

    Science.gov (United States)

    Preechakasedkit, Pattarachaya; Siangproh, Weena; Khongchareonporn, Nanthika; Ngamrojanavanich, Nattaya; Chailapakul, Orawon

    2018-04-15

    In this study, a novel wax-printed paper-based lateral flow device has been developed as an alternative approach for an automated and one-step enzyme-linked immunosorbent assay (ELISA). The design pattern consisted of a non-delayed channel, a wax-delayed channel, a test zone and a control zone. This system was easily fabricated on a nitrocellulose membrane using a wax-printing method and then baked in an oven at 100°C for 1min. The four barriers of the wax-delayed channel could delay the flow time for 11s compared to the flow time of the non-delayed channel. To use the device under optimal conditions, alpha-fetoprotein (AFP) was detected at a limit of detection of 1ngmL -1 and assessed with the naked eye within 10min. A colorimetric intensity was also measured using a smart phone and computer software at a linear range of 0.1-100ngmL -1 with a good correlation. Furthermore, the proposed device was successfully applied to detect AFP in human serum. Therefore, the wax-printing demonstrates a user-friendly, easy and quick method for the fabrication of the device, which could be used as a one-step, portable, disposable, low-cost, simple, instrument-free and point-of-care device for the automated ELISA. Copyright © 2017. Published by Elsevier B.V.

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    Franziska Fettke

    2016-12-01

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

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

    Science.gov (United States)

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

    2015-10-22

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

  4. 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.

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

    Science.gov (United States)

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

    2015-12-01

    The objective of this study was to determine the ability of biochemical analytes to identify adverse outcomes in pregnancies with Turner syndrome. Maternal serum and amniotic fluid (AF) marker concentrations were measured in 73 singleton pregnancies with Turner syndrome (10-22 weeks of gestation). Fetal Turner syndrome was definitively established by cytogenetic analysis. Two subgroups, fetuses with hydrops fetalis versus fetuses with cystic hygroma, were compared. Receiver operating characteristic curves and relative risk were established for a cut-off multiples of the median ≥3.5 for β-subunit of human chorionic gonadotropin (hCG) or AF alpha-fetoprotein (AFP). Forty-nine (67%) of 73 pregnant women had an abnormal maternal serum. While levels of pregnancy-associated plasma protein-A and free β-subunit (fβ)-hCG were not different to those of the control group, AFP, unconjugated estriol and β-hCG concentrations were significantly different in the study group (P < 0.05), when compared to those of unaffected pregnancies. Levels of β-hCG in pregnancies with hydrops fetalis were significantly higher than in those with cystic hygroma (P <0.0001), as were AF-AFP concentrations (P <0.0015). In addition, abnormalities in both maternal serum β-hCG and AF-AFP predicted fetal death. The relative risk of adverse obstetric outcome was 10.667 (P = 0.0004; 95% confidence interval [CI]: 1.554-73.203) for β-hCG and 2.19 (P = 0.0256; 95% CI: 1.001 to 4.779), for AF-AFP. Maternal serum β-hCG and AF-AFP levels may preferentially identify those Turner syndrome pregnancies with the highest risk of fetal death. © 2015 Japan Society of Obstetrics and Gynecology.

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

    Science.gov (United States)

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

    2018-01-01

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

  7. 8-Quinolineboronic acid as a potential phosphorescent molecular switch for the determination of alpha-fetoprotein variant for the prediction of primary hepatocellular carcinoma

    International Nuclear Information System (INIS)

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

    2010-01-01

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

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

    Science.gov (United States)

    Liang, Rong-Liang; Xu, Xu-Ping; Liu, Tian-Cai; Zhou, Jian-Wei; Wang, Xian-Guo; Ren, Zhi-Qi; Hao, Fen; Wu, Ying-Song

    2015-09-03

    Alpha-fetoprotein (AFP), a primary marker for many diseases including various cancers, is important in clinical tumor diagnosis and antenatal screening. Most immunoassays provide high sensitivity and accuracy for determining AFP, but they are expensive, often complex, time-consuming procedures. A simple and rapid point-of-care system that integrates Eu (III) chelate microparticles with lateral flow immunoassay (LFIA) has been developed to determine AFP in serum with an assay time of 15 min. The approach is based on a sandwich immunoassay performed on lateral flow test strips. A fluorescence strip reader was used to measure the fluorescence peak heights of the test line (HT) and the control line (HC); the HT/HC ratio was used for quantitation. The Eu (III) chelate microparticles-based LFIA assay exhibited a wide linear range (1.0-1000 IU mL(-1)) for AFP with a low limit of detection (0.1 IU mL(-1)) based on 5ul of serum. Satisfactory specificity and accuracy were demonstrated and the intra- and inter-assay coefficients of variation (CV) for AFP were both <10%. Furthermore, in the analysis of human serum samples, excellent correlation (n = 284, r = 0.9860, p < 0.0001) was obtained between the proposed method and a commercially available CLIA kit. Results indicated that the Eu (III) chelate microparticles-based LFIA system provided a rapid, sensitive and reliable method for determining AFP in serum, indicating that it would be suitable for development in point-of-care testing. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. 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.

  10. 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.

  11. Análisis de las malformaciones congénitas detectadas por el programa alfafetoproteína-ultrasonido genético Analysis of the congenital malformations detected by the alpha-fetoprotein-genetic ultrasound program

    Directory of Open Access Journals (Sweden)

    Aicha Julia Llamos Paneque

    2007-03-01

    Full Text Available La alfafetoproteína es una glicoproteína específica del plasma fetal, cuya determinación en suero materno se realiza entre las 15 y 19 semanas de gestación. Para conocer el comportamiento del programa alfafetoproteína-ultrasonido genético en el municipio 10 de Octubre se realizó esta investigación. En ella se encontró que 862 gestantes presentaron alfafetoproteína elevada en suero materno en el período analizado, y las principales causas encontradas dependientes de la madre fueron: el error en la fecha de última menstruación, seguida de la amenaza de aborto; y las malformaciones congénitas más frecuentemente encontradas fueron los defectos de cierre del tubo neural, seguidos de las malformaciones cardiovasculares.The alpha-fetoprotein is a specific glycoprotein of the fetal plasma, whose determination in maternal serum is performed from the 15th to the 19th week of gestation. This research was conducted to know the behavior of the alpha-fetoprotein-genetic ultrasound program in “10 de Octubre” municipality. It was found that 108 pregnant women presented elevated alpha-fetoprotein in maternal serum during the analyzed period. The main causes depending on the mother were: error in the date of the last menstruation and threatened abortion. The most frequent congenital malformations were the defects of the neural tube closure, and the cardiovascular malformations.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2015-09-03

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

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

    Directory of Open Access Journals (Sweden)

    Prabhu P. Gounder

    2016-05-01

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

  14. Conductive carbon nanoparticles-based electrochemical immunosensor with enhanced sensitivity for alpha-fetoprotein using irregular-shaped gold nanoparticles-labeled enzyme-linked antibodies as signal improvement.

    Science.gov (United States)

    Tang, Juan; Su, Biling; Tang, Dianping; Chen, Guonan

    2010-08-15

    A new electrochemical immunoassay protocol for sensitive detection of alpha-fetoprotein (AFP, as a model) is designed using carbon nanoparticles (CNPs)-functionalized biomimetic interface as immunosensing probe and irregular-shaped gold nanoparticles (ISNGs)-labeled horseradish peroxidase-anti-AFP conjugates (HRP-anti-AFP-ISNG) as trace label. The low-toxic and high-conductive CNPs provided a high capacity nanoparticulate immobilization surface and a facile pathway for electron transfer. In comparison with conventional label methods, i.e. spherical gold nanoparticles-labeled HRP-anti-AFP and HRP-labeled anti-AFP, the electrochemical immunosensor using HRP-anti-AFP-ISNGs as trace labels exhibited high bioelectrocatalytic response toward enzyme substrate and a wide dynamic range from 0.02 to 4.0 ng/mL with a low detection limit of 10 pg/mL toward AFP (at 3sigma). The developed immunoassay method showed good selectivity and acceptable reproducibility. Clinical serum samples with various AFP concentrations were evaluated by using the electrochemical immunosensor and the referenced enzyme-linked immunosorbent assay (ELISA), respectively, and received in good accordance with results obtained from these two methods. Copyright 2010 Elsevier B.V. All rights reserved.

  15. 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

    2015-12-01

    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.

  16. Prognostic Value of Preoperative Serum Levels of Five Tumor Markers (Carcinoembryonic Antigen, CA19-9, Alpha-fetoprotein, CA72-4, and CA125) in Gastric Cancer.

    Science.gov (United States)

    Kim, Ji-Hyun; Jun, Kyong-Hwa; Jung, Hun; Park, In-Soo; Chin, Hyung-Min

    2014-05-01

    There is no known specific tumor marker for gastric cancer, although several tumor markers have been used. The aim of this study was to investigate the prognostic significance of preoperative carcinoembryonic antigen (CEA), CA 19-9, alpha-fetoprotein (AFP), CA 72-4, and CA 125 levels in patients with gastric cancer. Medical records of 1,253 patients who were diagnosed with gastric adenocarcinoma were retrospectively reviewed. The clinicopathologic characteristics and disease-free survival rate of the patients were compared between positive and negative CEA, CA 19-9, AFP, CA 72-4, and CA 125 groups of patients. Additionally, the prognostic significance of each tumor marker was assessed by multivariate analysis. CEA, CA19-9, and CA72-4 were more frequently positive in patients with lymphatic and venous invasion, serosal involvement, and lymph node metastasis. The 5-year overall survival and disease free survival rates were significantly associated with elevated serum levels of CEA, CA 19-9, and CA 72-4. The depth of invasion and CA 19-9 were independent prognostic factors. Patients with elevated serum levels of CA 19-9 showed a 3.35-fold higher risk of death than patients with low levels of the marker. CA 19-9 has prognostic significance in gastric cancer, and a high preoperative serum level of CA 19-9 can be useful for estimating worse prognosis and a higher recurrence of gastric cancer.

  17. 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)

    Liu, Keke; Zhang, Jianrong; Liu, Qing; Huang, Haiping

    2013-01-01

    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][BF 4 ]). 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

  18. Simultaneous electrochemical immunosensing of alpha-fetoprotein and prostate specific antigen using a glassy carbon electrode modified with gold nanoparticle-coated silica nanospheres and decorated with Azure A or ferrocene carboxylic acid

    International Nuclear Information System (INIS)

    Zhao, Junqing; Guo, Zilin; Guo, Jinjin; Wang, Junchun; Zhang, Yuzhong; Feng, Dexiang

    2015-01-01

    We describe an electrochemical immunosensor for the simultaneous determination of alpha-fetoprotein (AFP) and prostate specific antigen (PSA) via a modified glassy carbon electrode. Silica nanoparticles (200–300 nm i.d.) with good monodispersity and uniform shape were synthesized, and the following species were then consecutively immobilized on their surface: gold nanoparticles (AuNPs; 5–15 nm i.d.), secondary antibody (Ab 2 ) and the redox-probes Azure A or ferrocenecarboxy acid (Fc). In parallel, two types of primary antibodies (Ab 1 ) were co-immobilized on the surface of the dissolved reduced graphene oxide sheets (rGO) that were also decorated with AuNPs. In the presence of antigens (AFP or PSA), the Ab 2 /Si-AuNPs carrying Azure A and Fc are attached to the AuNP/rGO conjugate via a sandwich type immunoreaction. Differential pulse voltammetry (DPV) was employed to measure the resulting changes in the signal of Fc or Azure A. Two well-resolved oxidation peaks, one at −0.48 V (corresponding to Azure A) and other at + 0.12 V (corresponding to Fc; both vs. SCE) can be observed in the DPV curves. Under optimal conditions, AFP and PSA can be simultaneously determined in the range from 0.01 to 25 ng mL -1 for AFP, and from 0.012 to 25 ng mL -1 for PSA. The detection limits are 3.3 pg mL -1 for AFP and 4.0 pg mL -1 for PSA (at a signal-to-noise ratio of 3). The method was applied to (spiked) real sample analysis, and the recoveries are within 96.0 and 107.2 % for PSA, and within 100.9 and 105.8 % for AFP, indicating that this dual immunosensor matches the requirements of clinical analysis. (author)

  19. Radioimmunoassays for tumor diagnosis

    International Nuclear Information System (INIS)

    Dressler, J.

    1983-01-01

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

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

    International Nuclear Information System (INIS)

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

    2013-01-01

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

  1. Radioimmunoassay apparatus

    International Nuclear Information System (INIS)

    1981-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Rejane Mattar

    2002-06-01

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

  3. Progesterone radioimmunoassay

    International Nuclear Information System (INIS)

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

    1981-01-01

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

  4. 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

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

    International Nuclear Information System (INIS)

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

    1991-01-01

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

  6. 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

  7. Radioimmunoassay of thymosin alpha 1

    International Nuclear Information System (INIS)

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

    1983-01-01

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

  8. 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.

  9. Solid phase radioimmunoassays

    International Nuclear Information System (INIS)

    Wide, L.

    1977-01-01

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

  10. Radioimmunoassay for biopterin

    International Nuclear Information System (INIS)

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

    1979-01-01

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

  11. Radioimmunoassay of pancreatic glucagon

    International Nuclear Information System (INIS)

    Nooijen, W.J.

    1979-01-01

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

  12. Radioimmunoassay of norandrosterone

    International Nuclear Information System (INIS)

    Hampl, R.; Starka, L.; Putz, Z.; Protiva, J.

    1983-01-01

    A simple radioimmunoassay of the main nortestosterone metabolite, 3α-hydroxy-5α-estran-17-one (norandrosterone) is described and evaluated. The method employs 3 H-labelled ligand prepared from nortestosterone by three-step synthesis and rabbit antiserum against norandrosterone-17-carboxymethyloxime bovine serumalbumin. (author)

  13. Automation systems for radioimmunoassay

    International Nuclear Information System (INIS)

    Yamasaki, Paul

    1974-01-01

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

  14. Radioimmunoassay in endocpinology

    International Nuclear Information System (INIS)

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

    1983-01-01

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

  15. Comparative study of radioimmunoassay dates

    International Nuclear Information System (INIS)

    Venegas Sanchez, Ruth.

    1986-01-01

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

  16. Radioimmunoassay of buprenorphine

    Energy Technology Data Exchange (ETDEWEB)

    Bartlett, A.J.; Lloyd-Jones, J.G.; Rance, M.J.; Flockhart, I.R.; Dockray, G.J.; Bennett, M.R.D.

    1980-01-01

    Antisera to buprenorphine were obtained in rabbits immunised with 3-0-carboxymethylbuprenorphine and N-hemisuccinyl-norbuprenorphine conjugated to bovine serum albumin. Using the latter antiserum and tritium labelled buprenorphine a radioimmunoassay having good accuracy and precision was developed for concentrations as low as 50 picograms in 1 ml of plasma. The N-hemisuccinyl antiserum crossreacted with norbuprenorphine, and the 3-0-glucuronide conjugate with the 3-0-carboxy-methyl antiserum. Cross-reactivity of both antisera to other pharmacologically related compounds was negligible. The assay was employed to determine plasma buprenorphine concentration following its parenteral administration to dog and man.

  17. Radioimmunoassay for Melatonin

    International Nuclear Information System (INIS)

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

    1980-01-01

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

  18. 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

  19. Fundamental and clinical evaluation of 'AMERWELL AFP RIA-kit' for radioimmunoassay of α-fetoprotein

    International Nuclear Information System (INIS)

    Kihara, Kazuhiro; Iino, Yuuko; Sakata, Hiroyuki; Sugimura, Hidekazu; Sato, Kunio; Demura, Hiroshi

    1988-01-01

    A commercial ''AMERWELL AFP RIA-kit'' was fundamentally and clinically evaluated. The present assay kit was simple to treat for measurement. Laboratory findings were encouraging enough to use it in routine practice. There were extremely small quantities of radioactive waste. A good correlation was observed between the present assay kit and ''AFP EIKEN RIA kit''. The lower detection limit for AFP in serum was 1.5 IU/ml. The upper limit for serum AFP was 3 IU/ml in normal controls (n = 112). Serum AFP concentrations were 1.5 - 34.8 IU/ml for acute hepatitis, 1.5 - 191.4 IU/ml for chronic hepatitis, 1.5 - 398.1 IU/ml for liver cirrhosis, and 1.5 - 268,350 IU/ml for hepatocellular carcinoma. The positive rates of AFP for these diseases were 46 %, 47 %, 79 %, and 93 %, respectively. Falling concentrations of serum AFP were associated with favorable management. The results indicate the usefulness of the present assay kit in the diagnosis of various liver diseases, understanding of pathophysiology, and postoperative management for hepatocellular carcinoma. (Namekawa, K.)

  20. Radioimmunoassay of platelet proteins

    International Nuclear Information System (INIS)

    Pepper, D.S.

    1987-01-01

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

  1. Radioimmunoassay of steroid hormone

    International Nuclear Information System (INIS)

    Murakami, Tadashi

    1975-01-01

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

  2. Diagnostic Utility of Alpha-fetoprotein and Des-gamma ...

    African Journals Online (AJOL)

    2017-12-05

    Dec 5, 2017 ... (DCP) have been extensively studied as biomarkers for the diagnosis of and prognostication in hepatocellular carcinoma (HCC). However there are only few reports on the clinical characteristics of hepatocellular carcinoma in relation to .... measured by ultrasound and/or computed tomography scan.

  3. Diagnostic Utility of Alpha-fetoprotein and Des-gamma ...

    African Journals Online (AJOL)

    2018-02-07

    ] High DCP levels also reflect .... using the multiple logistic regression models. P < 0.05 was considered statistically significant. .... that C-reactive protein has significant diagnostic power in AFP-negative, HBV-related HCC.[23].

  4. Radioimmunoassay of erythropoietin

    International Nuclear Information System (INIS)

    Goldwasser, E.; Sherwood, J.B.

    1981-01-01

    A brief review of the historical development of the radioimmunoassay (RIA) for serum erythropoietin is given. It has been shown that there is reasonable agreement between the results obtained by RIA and those obtained by the previously used bioassay. By RIA, a mean normal serum titre of 18 mu/ml erythropoietin has been determined in a study of 445 individuals. Serum erythropoietin results for patients with polycythaemia vera have not been shown to be significantly different from normal values but in patients with secondary polycythaemia, serum titres averaging 94 mu/ml have been found. The predicted physiological changes in erythropoietin titre have also been demonstrated in humans using the RIA; increasing after bleeding and decreasing after red cell administration. Studies of erythropoietin levels in experimental animals have shown that, with the particular antiserum used, the sensitivity of the RIA is markedly reduced. (U.K.)

  5. Radioimmunoassay of anabolic steroids

    International Nuclear Information System (INIS)

    Hampl, R.; Stranska, I.; Starka, L.; Picha, J.; Chundela, B.

    1978-01-01

    Alternative antisera against 17 α-methyltestosterone and 19-nortestosterone were raised and used for radioimmunoassay of anabolic steroids. Tritiated compounds were used as radioligands. The RIA method suitable for doping control is proposed for 17 α-alkylated anabolic steroids in both plasma and urine, using qoat antiserum against methyltestosterone-3-carboxymethyloxime-BSA. Sensitivity of the method was expressed as least amount of nonradioactive methandienone which, when added to normal urine or plasma, caused statistically significant decrease of measured supernatant radioactivity at 99% level. The amounts from 50 to 500 pg were tested, each in eight parallel determinations. The amounts of 100 pg for plasma and 200 pg for urine met these criteria. The respective coefficients of variation did not depend on the amount of steroid added in this range. They averaged 4.60% for plasma and 4.95% for urine, respectively. (T.I.)

  6. Radioimmunoassay for serum paraquat

    International Nuclear Information System (INIS)

    Fatori, D.; Hunter, W.M.

    1980-01-01

    Two variants of a radioimmunoassay for the bipyridylium herbicide Paraquat are described. Both employ antiserum raised to Paraquat-BSA which has been covalently linked to particulate solid-phase support media. The rapid assay for clinical use employs a [ 3 H] Paraquat tracer, requires no agitation and yields results in the range 10-2500 ng/ml serum in 20 min from receipt of sample. The more sensitive assay, designed for research purposes, employs a 125 iodinated tracer, requires 2 h continuous agitation but can detect Paraquat at 0.1 ng/ml in simple aqueous solution or 0.25 ng/ml serum. Results from rapid clinical assay agree well with the existing colorimetric method. (Auth.)

  7. A radioimmunoassay for chicken avidin

    International Nuclear Information System (INIS)

    Kulomaa, M.S.; Elo, H.A.; Tuohimaa, P.J.

    1978-01-01

    A double-antibody solid-phase radioimmunoassay for chicken avidin is reported. Avidin was labelled with 125 I by the chloramine-T method. The bound and free avidin were separated with a second antibody bound to a solid matrix. In the logit-log scale the standard curve was linear from 1-2 to 100-200ng of avidin/ml. Cross-reaction of ovalbumin was less than 0.015%. Saturation of biotin-binding sites of avidin with an excess of biotin decreased radioimmunoassay values by about 15%. Recovery studies indicated that avidin can be assayed from all chicken tissues studied with radioimmunoassay, whereas the [ 14 C]biotin/bentonite method gave poor recoveries for avidin in the liver and kidney. Radioimmunoassay and the [ 14 C]biotin/bentonite method gave similar concentrations for oviduct avidin. (author)

  8. Automatic computation of radioimmunoassay data

    International Nuclear Information System (INIS)

    Toyota, Takayoshi; Kudo, Mikihiko; Abe, Kanji; Kawamata, Fumiaki; Uehata, Shigeru.

    1975-01-01

    Radioimmunoassay provided dose response curves which showed linearity by the use of logistic transformation (Rodbard). This transformation which was applicable to radioimmunoassay should be useful for the computer processing of insulin and C-peptide assay. In the present studies, standard curves were analysed by testing the fit of analytic functions to radioimmunoassay of insulin and C-peptides. A program for use in combination with the double antibody technique was made by Dr. Kawamata. This approach was evidenced to be useful in order to allow automatic computation of data derived from the double antibody assays of insulin and C-peptides. Automatic corrected calculations of radioimmunoassay data of insulin was found to be satisfactory. (auth.)

  9. Clinical indications of prolactin radioimmunoassay

    International Nuclear Information System (INIS)

    Lengyel, A.M.J.; Vieira, J.G.H.; Zanella, M.R.; Zampieri, M.; Chacra, A.R.

    1980-01-01

    Is a review is presented of the main clinical uses of prolactin measurements, including the galactorrhea-amenorrhea syndrome, an experiment employing the prolactin radioimmunoassay is related. (Author) [pt

  10. Radioimmunoassay of polypeptide hormones and enzymes

    International Nuclear Information System (INIS)

    Felber, J.P.

    1974-01-01

    General principles of radioimmunoassay are reviewed. Detailed procedures are reviewed for the following hormones: insulin, pituitary hormones, gonadotropins, parathyroid hormone, ACTH, glucagon, gastrin, and peptide hormones. Radioimmunoassay of enzymes is also discussed. (U.S.)

  11. Calcitonin radioimmunoassay: clinical application

    International Nuclear Information System (INIS)

    Raue, F.; Minne, H.; Streibl, W.; Ziegler, R.; Ulm Univ.

    1977-01-01

    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 125 J; 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.) [de

  12. Radioimmunoassay for rhesus monkey gonadotropins

    International Nuclear Information System (INIS)

    Faiman, C.; Stearns, E.L.; Winter, J.S.D.; Reyes, F.I.; Hobson, W.C.

    1975-01-01

    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.)

  13. Radioimmunoassay of Human Chorionic Gonodotropin

    International Nuclear Information System (INIS)

    Majali, M.A.; Shah, K.B.; Mani, R.S.

    1981-01-01

    A rapid and sensitive radioimmunoassay has been developed for the measurement of Human Chorionic Gonadotropin (HCG) in serum. Human chorionic gonadotropin has been labelled with 125 I to attain a specific activity between 80-120 μCi/μg. Aqueous dioxane (74 vol.%) has been employed to separate the bound and freehormone in the radioimmunoassay. The sensitivity of this technique has been found to be approximately 1.5 mIU/ml of HCG. The intra assay variation has been found to be less than 5% and inter assay variation has been found to be less than 12%. (author)

  14. Peptide radioimmunoassays in clinical medicine

    International Nuclear Information System (INIS)

    Geokas, M.C.; Yalow, R.S.; Straus, E.W.; Gold, E.M.

    1982-01-01

    The radioimmunoassay technique, first developed for the determination of hormones, has been applied to many substances of biologic interest by clinical and research laboratories around the world. It has had an enormous effect in medicine and biology as a diagnostic tool, a guide to therapy, and a probe for the fine structure of biologic systems. For instance, the assays of insulin, gastrin, secretin, prolactin, and certain tissue-specific enzymes have been invaluable in patient care. Further refinements of current methods, as well as the emergence of new immunoassay techniques, are expected to enhance precision, specificity, reliability, and convenience of the radioimmunoassay in both clinical and research laboratories

  15. Simultaneous use of two prostaglandin radioimmunoassays employing two antisera of differing specificity. II. Relative stability of prostaglandins E1, E2, and F1alpha in cell cultures of BALB/c 3T3 and SV3T3 mouse fibroblasts

    Energy Technology Data Exchange (ETDEWEB)

    Ritzi, E.M.; Stylos, W.A.

    1976-11-01

    The relative stability of Prostaglandins (PGs) E1, E2 and F1..cap alpha.. in cultures of BALB/c 3T3 and SV3T3 cells has been evaluated using 3 different approaches. First, total recovery of tritium in the ethyl acetate phase following incubation and extraction of PGF1..cap alpha.. and PGE1 demonstrated greater stability for PGF1..cap alpha.. (88.8 percent) than PGE1 (65.9 percent). Second, analysis of incubated, extracted, tritiated PGs by thin layer chromatography revealed decreases of up to 23 percent in the PGE zone following incubation of 3H-PGE1. With increasing time of incubation, decreases in the PGE zone were accompanied by increase in PGA-like compounds. 3H-PGF1..cap alpha.. demonstrated greater stability, having greater than 90 percent recovery of the tritium in the PGF zone. A third approach to the assessment of PG stability in culture was the comparison of the production of individual PGs by radioimmunoassay (RIA). The data obtained by RIA indicated a lag in the increase of PGA and PGB, until an initial rise in PGE was noted, suggesting that PGA and PGB may be secondary products arising from PGE which exhibits only partial stability in culture. By employing two RIAs, one for total PGE and one for PGA and PGB, the composite determination PG (E + (A + B)) can be used to provide a more meaningful determination of PG production because of the instability of the PGs. On the other hand, individual determinations are helpful in assessing the stability of PGEs in cell cultures.

  16. Recent advances in steroid radioimmunoassay

    International Nuclear Information System (INIS)

    Jeffcoate, S.L.

    1977-01-01

    The advances since 1974 in the techniques of measuring steroid molecules by radioimmunoassay are reviewed in this paper. They are considered under the following headings: preparation and use of antisera; preparation and use of tracers; preparation of biological samples before assay; dispensing of the reagents in the assay; separation of free and bound radioactivity; counting and data processing; quality control and standardization. (orig.) [de

  17. A sensitive radioimmunoassay for fentanyl

    International Nuclear Information System (INIS)

    Michiels, M.; Hendriks, R.; Heykants, J.

    1977-01-01

    Antiserum to fentanyl was obtained in rabbits repeatedly injected with carboxyfentanyl conjugated to bovine serum albumin. Using the antiserum, a highly sensitive radioimmunoassay has been developed, based on the dextran-coated charcoal method. It proved possible to assay the drug directly in plasma, in amounts as small as 30 picogram in 0.5 ml. The antibody was highly specific for fentanyl and no cross-reaction was observed with its major metabolites. This sensitive and specific radioimmunoassay method was employed to determine fentanyl in plasma from six volunteers after an intravenous bolus of 0.2 mg, and in plasma from dogs treated both intravenously and subcutaneously with 0.02 mg/kg. The plasma level of fentanyl could be followed for up to 6 h after a therapeutic dose in dogs and man. (orig.) [de

  18. Homologous radioimmunoassay of human prolactin

    International Nuclear Information System (INIS)

    Felixberger, F.X.

    1980-01-01

    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.) [de

  19. Homologous radioimmunoassay for human prolactin

    International Nuclear Information System (INIS)

    Reuter, A.M.; Kennes, F.; Gevaert, Y.; Franchimont, P.

    1976-01-01

    Although thee are descriptions of a range of radioimmunoassays for human prolactin in various biological fluids, only one of these is an homologous assay using human prolactin as the reference standard and tracer as well and an anti-human prolactin antiserum (Sinha, Y.N., Selby, F.W.; Lewis, U.; and Vanderlaan, W.P., 1973, J. Clin. Endocr., Vol. 36, 509). A homologous radioimmunoassay using human putuitary prolactin has been developed. The separation method is based on the double antibody solid phase system. Cross reactivity with human growth hormone (GH), placental lactogen (HPL), the pituitary protein hormones and prolactins of various species were studied as were values found in normal subjects in basal conditions and after a TRH injection. (author)

  20. In vitro inhibition of enterobacteria-reactive CD4+Tumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    Mangano, K.; Sardesai, N.; D'Alcamo, M.

    2008-01-01

    VGX-1027 is an isozaxoline compound that has recently been found to primarily target the function of murine macrophages but not of T cells, inhibiting secretion of tumor necrosis factor (TNF)-alpha in response to different Toll-like receptor agonists in vitro and in vivo. The well-defined role of...

  1. Radioimmunoassay of plasma corticotrophin

    International Nuclear Information System (INIS)

    Hummer, L.

    1977-01-01

    An assay has been established, based on antibodies against the N-terminal part of the ACTH molecule with a high affinity, a detection limit of 2 pg ACTH when assaying 200 ul unextracted plasma, and implying a total incubation time of three days. The antibody has been obtained by immunizing guinea-pigs with synthetic human 1-24 ACTH coupled to bovine serum albumin. The selected antibody has an equilibrium constant of 4 x 10 11 litres/mole in a final dilution of 1/320.000. The antiserum reacts with synthetic human 1-39 ACTH as well as with synthetic human 1-24 ACTH, and the hormonally inactive synthetic human 11-24 ACTH fragment as well as alpha- and beta-melanocyte-stimulating hormones do not cross-react in the assay. The interassay coefficient of variation of replicate estimates was 11-13%. The reproducibility of the standard curve have been evaluated by calculating the amount of ACTH corresponding to 5% of the (B/T)o value, 1.2+-0.4 pg ACTH/tube+-SD and 50% of (B/T)o value, 15.7+-2.6 pg ACTH/tube+-SD. Validation of the assay has been obtained by assaying samples from patients with verified adrenal disorders, and the accuracy is supported by ACTH determinations in test, where metyrapon had been administered intravenously. A stimulation of ACTH production by insulin-induced hypoglycaemia has been found as well. Special attention is always paid as to the conditions for the blood sampling. A reference interval of 10-76 ng/l has been found (115 normal subjects). (orig.) [de

  2. Prognostic value of α-fetoprotein response in treatment of hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    JIA Jia

    2013-08-01

    Full Text Available Primary hepatocellular carcinoma (HCC is the most common malignant tumor of the liver. Accurate assessment of treatment outcome and prognosis in HCC patients is very important due to its high malignancy and poor prognosis. Currently, radiological imaging, though a standard method for prognostic evaluation of HCC, has a lot of limitations. As an important serological marker of HCC, alpha-fetoprotein (AFP has been widely used in the screening, diagnosis, and prognostic evaluation of HCC. The studies regarding the prognostic value of AFP response in HCC patients were summarized. Generally, AFP has good prognostic value in HCC patients treated with radiofrequency ablation, transarterial chemoembolization, yttrium-90 radioembolization, molecular targeted agents like sorafenib, systemic chemotherapy, hepatic artery infusion chemotherapy, or concurrent chemoradiotherapy.

  3. 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...... discuss the use of these and other biological agents targeting B lymphocytes, T-lymphocyte interaction with antigen-presenting cells, or cytokines in the future treatment of GO Udgivelsesdato: 2008/6/9...

  4. Radioimmunoassay method for detection of gonorrhea antibodies

    International Nuclear Information System (INIS)

    1975-01-01

    A novel radioimmunoassay for the detection of gonorrhea antibodies in serum is described. A radionuclide is bound to gonorrhea antigens produced by a growth culture. In the presence of gonorrhea antibodies in the serum, an antigen-antibody conjugate is formed, the concentration of which can be measured with conventional radiometric methods. The radioimmunoassay is highly specific

  5. Radioimmunoassay and other related techniques

    International Nuclear Information System (INIS)

    Zarkawi, Moutaz

    1993-04-01

    The article reviews principles, requirements and reliability criteria of radioimmunoassay (RIA). Since basic reactions involved in RIA and related techniques are derived from reactions which take place in the immune system (IS) of humans and animals, the IS and the way it works will be described. In addition to RIA which involves the use of isotopes as tracers (labels), other non-radioisotopic and recent immunoassay techniques i.e. enzyme-linked immunosorbent assay (ELISA), chemiluminescence immunoassay (CLIA) and fluoroimmunoassay (FIA) will be dealt with. Some important and related terms will be defined and explained. (author). 59 refs., 4 figs

  6. Radioimmunoassay in children with glomerulonephritis

    International Nuclear Information System (INIS)

    Ametov, A.S.; Gavryushova, L.P.; Shashinka, M.; Mumladze, Eh.B.; Tvorogova, T.M.; Dakhuk, B.; Dronova, V.I.; Toritsina, D.K.; Petrova, T.V.

    1985-01-01

    Proceeding from a radioimmunoassay of various biologically active substances in the blood and urine (ACTH, cortisol, FSH, LH, prolatin, progesterone, estradiol, plasma renin activity and β 2 -microglobulin) of 220 children with glomerulonephritis change of all indices with relation to the type and gravity of glomerulonephritis as well as renal function was revealed. The nature of influence on corticosteroid and immunosuppressive therapy was shown. The authors consider it appropriate to use the determination of biologically active substances in the blood and urine for a more profound estimation of a child's status

  7. 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

    , and for the shortest possible time should therefore greatly reduce the risk of infections. Infection is a major co-morbidity in rheumatoid arthritis (RA), and conventional disease-modifying anti-rheumatic drugs (DMARDs) can increase the risk of their occurrence, including tuberculosis. TNF-alpha plays a key role....../inflammatory conditions, and current therapies have the aim of providing adequate (low) compensatory doses, the timing of GC administration, such as during the nocturnal turning-on phase of tumour necrosis factor (TNF) secretion, can be extremely important. The use of the lowest possible GC dose, at night...

  8. Purification of subunits from human chorionic gonadotropin and radioimmunoassay

    International Nuclear Information System (INIS)

    Okumura, Hajime; Sudo, Tadamitsu; Fujisawa, Munetoshi; Namba, Shuichi; Matsushima, Sanae

    1976-01-01

    A crude hCG with an activity of about 3,000 IU per mg was purified to 10,000-15,000 IU per mg of dry weight using Amberlite CG-50 chromatography combined with DEAE-Sephadex A-50 and Sephadex G-75. The alpha and beta subunits of hCG were prepared by urea-treatment of the hormone and were isolated by DEAE-Sephadex A-50 chromatography. Further purification of the subunits was achieved by gel filtration on a Sephadex G-57 column. For radioimmunoassay, hCG was iodinated by the DMSO-chloramine T method. Iodination of hCG with non-radioactive iodine revealed that the addition of DMSO to the iodination mixture seemed to reduce the iodination damage to the antigenic activity of the hormone. The hCG iodinated with non-radioactive iodine (accomplished by the DMSO-chloramine T method) was 1.5 times more immunoreactive in the hCG radioimmunoassay then the hCG iodinated by the usual chloramine T method. The radioimmunoassay of the hCG-beta subunit developed in our laboratory was satisfactory with respect to specificity. The hLH, hFSH, hTSH and hCG-alpha subunits tested cross-reacted very poorly in our assay system. Desialylated-hCG and subunits, in which the biologic potency was almost zero, also exhibited decreased immunoreactivity, about 30% of the native hormone with grossly unimpaired parallelism in their respective homologous radioimmunoassays. The concentrations of hCG and the subunits were determined on human sera from pregnant patients. The hCG levels reached to a peak at the first trimester of the pregnancy, but the hCG-beta subunits varied poorly in their concentrations throughout the periods of pregnancy. The hCG-alpha levels, on the other hand, showed two distinct peaks, in the early period and at the term of pregnancy. (J.P.N.)

  9. Apparatus for use in radioimmunoassays

    International Nuclear Information System (INIS)

    Chen, C-H.; Tsay, H-M.; Heyer, R.E.

    1979-01-01

    Apparatus for solid-phase antibody separation techniques used in radioimmunoassays is described in this invention. It consists of a rectangular prism tray with multiple wells protruding into its interior from one side. Near the base of the tray is an orifice used for creating evacuated condition within the structure. At the base of each well there is an orifice of such size and shape as to retain an aqueous liquid under given pressure conditions but permit the evacuation of this liquid at reduced pressure. The outlet of these orifices is in the shape of an inverted conical frustrum. Each of the wells contains an antibody coated disc of porous cellulose paper surrounded by a plastic support. The porous nature of the cellulose paper ensures contact between the antibody coating and the antigen. The use of antibody coated porous cellulose paper in combination with the vacuum operated apparatus simplifies the manipulative steps whilst still maintaining the sensitivity of the radioimmunoassay. It also obviates the need for aspiration and thus lessens the risk of contamination from one sample to another. (UK)

  10. Pancreatic elastase in human serum. Determination by radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Geokas, M.C. (Univ. of California, Davis); Brodrick, J.W.; Johnson, J.H.; Largman, C.

    1977-01-10

    This study demonstrates that a serine endopeptidase of pancreatic origin (elastase 2) circulates in human blood. A specific and highly sensitive radioimmunoassay has been developed for pancreatic elastase 2 in human serum. The inactivation of elastase 2 employed as radioiodinated tracer with an active site-specific reagent (phenylmethanesulfonyl fluoride) was necessary to prevent its binding by serum ..cap alpha../sub 1/-antitrypsin and ..cap alpha../sub 2/-macroglobulin while maintaining its immunoreactivity. The assay is based upon competition of standard human pancreatic elastase 2 with /sup 125/I-labeled phenylmethanesulfonyl elastase 2 for specific antibody binding sites, after which a second antibody precipitation step is used to separate bound from free /sup 125/I-labeled phenylmethanesulfonyl elastase 2. The minimum detectable concentration of elastase 2 was 0.9 ng/ml. The average normal fasting serum level determined was 71 ng/ml, approximately 80-fold greater than the minimum detectable amount.

  11. Radioimmunoassay for nortriptyline and amitriptyline

    International Nuclear Information System (INIS)

    Turner, P.

    1977-01-01

    A description has been given (Aherne, G.W.; Marks, V.; Mould, G.; Stout, G.; Lancet; 1214, June 4 (1977)) of a radioimmunoassay (RIA) technique having advantages for monitoring patient response by estimation of plasma concentrations of nortriptyline and amitriptyline. A wide range of monoamine-reuptake-inhibiting drugs is now used in the treatment of depressive illness. There are other analytical techniques such as gas-liquid chromatography which can be used in the estimation of most of these drugs and their major metabolites, as well as for the identification of some other centrally-acting drugs such as benzodiazepines. The chromatographic technique was able to detect the administration of the wrong drug in one comparative study of two antidepressive drugs. Chromatographic and RIA methods therefore both have their own analytical roles according to the clinical or research problem being investigated. (U.K.)

  12. Radioimmunoassay of bovine growth hormone

    International Nuclear Information System (INIS)

    Dvorak, P.; Becka, S.; Krejci, P.; Chrpova, M.

    1978-01-01

    A radioimmunological method for quantitative determination of bovine growth hormone in blood plasma or serum for routine use was worked out. The antibody is also suitable for radioimmunoassay of bovine growth hormone. The chloramin T and lactoperoxidase methods were used. Standard b-STH isolated by one of the authors was labelled with I-125(Na 125 I, carrier free, Amersham). The I-125-hormone was separated on Sephadex G-50 or G-100 and stored in 0.05M phosphate pH 7.5 with 1% of human serum albumine at 25 deg C. The method was checked on 2500 samples. Rabbit antiserum to b-STH was prepared in sufficient amount to serve also for other laboratories interested in the method. (author)

  13. The radioimmunoassay of steroid glucuronides

    International Nuclear Information System (INIS)

    Chaudhri, B.

    1980-07-01

    A radioimmunoassay method for steroid glucuronides in which the second carboxylated group has been introduced into the steroid structure and used to attach the hapten to the protein. The unequivocal attachment through the second group, rather than through the glucuronide is achieved by leaving the glucuronide protected as its triacetyl methyl-ester (g.a.m.e.) until after the hapten has been coupled to the protein. The hapten, immunogens and antisera to costrone-3-glucuronide have been prepared using a 17-carboxymethyloxime bridge. The antisera raised to these immunogens have been characterised for their ability to discriminate between the free steroid and the steroid glucuronide, and evaluated for use in immunoassay. (author)

  14. Radioimmunoassay for chlorpromazine in plasma

    International Nuclear Information System (INIS)

    Midha, K.K.; Loo, J.C.K.; Hubbard, J.W.; Rowe, M.L.; McGilveray, I.J.

    1979-01-01

    A radioimmunoassay for chlorpromazine in plasma is described. The antiserum was obtained by immunizing rabbits with a conjugate of bovine serum albumin and N-(2-carboxyethyl)desmethylchlorpromazine. It is specific for chlorpromazine and its minor active metabolite, N-desmethylchlorpromazine. Other known active or inactive chlorpromazine metabolites and other psychotropic drugs tested do not cross react with the antiserum. Less than 34 pg of the drug can be detected in 200 μL of plasma. As many as 100 samples can be processed in a day by one technician. Concentrations of chlorpromazine can be measured in 200-μL samples of plasma collected as late as 48 h after a single oral 25-mg dose

  15. Methadone radioimmunoassay: two simple methods

    International Nuclear Information System (INIS)

    Robinson, K.; Smith, R.N.

    1983-01-01

    Two simple and economical radioimmunoassays for methadone in blood or urine are described. Haemolysis, decomposition, common anticoagulants and sodium fluoride do not affect the results. One assay used commercially-available [1- 3 H](-)-methadone hydrobromide as the label, while the other uses a radioiodinated conjugate of 4-dimethylamino-2,2-diphenylpentanoic acid and L-tyrosine methyl ester. A commercially-available antiserum is used in both assays. Normethadone and α-methadol cross-react to a small extent with the antiserum while methadone metabolites, dextropropoxyphene, dipipanone and phenadoxone have negligible cross-reactivities. The 'cut-offs' of the two assays as described are 30 and 33 ng ml -1 for blood, and 24 and 21 ng ml -1 for urine. The assay using the radioiodinated conjugate can be made more sensitive if required by increasing the specific activity of the label. (author)

  16. Radioimmunoassay of human urinary kallikrein

    International Nuclear Information System (INIS)

    Goering, W.

    1980-01-01

    Using a human urinary kallikrein, purified by means of Trasylol sepharose, it has been possible to develop a radioimmunoassay of kallikrein capable of detecting the substance down to a concentration of 0.5 ng/ml. The specific activity of the tracer labelled with 125-iodine using the Chloramine-T method was 30-70 nCi/ng of kallikrein. The antiserum titres for the antikallikrein serum were 1:20.000 up to 1:50.000. Human urine, submandibular and parotid salivae as well as pancreatic secretion in this RIA reacted in the same manner as the kallikrein standard solution. The kallikrein content in urine, as determined by the RIA was between 0 and 300 ng/ml, in the saliva between 400 and 2.000 ng/ml, and in the pancreatic juice between 300 and 12.000 ng/ml. Using human serum, only an incomplete immunological cross-reaction could be achieved. In human liquor as well as in animal preparation, no cross-reacting substances could be detected. (orig.) [de

  17. The radioimmunoassay of steroid glucuronides

    International Nuclear Information System (INIS)

    Chaudhri, R.

    1980-07-01

    A modified radioimmunoassay for steroid glucuronides is described. A second carboxylated group is introduced into the steroid structure and used to attach the hapten to the protein. The unequivocal attachment through the second group, rather than through the glucuronide is achieved by leaving the glucuronide protected as its triacetyl methylester until after the hapten has been coupled to the protein. Subsequently, the glucuronide can be exposed by mild alkaline hydrolysis or alternatively, the protected immunogen can be injected into the rabbits, leaving the animals to deprotect the hapten. The hapten, immunogens and antisera to oestrone-3-glucuronide have been prepared using a 17-carboxymethyloxime bridge. The hapten, immunogen and antisera to pregnanediol-3 α - glucuronide have also been prepared using a 6 β - hemisuccinoxy bridge. A study has been made of the reactive properties of the 6 β-hydroxyl group and its effect on the stereo-specificity of reduction of 4-ene and the 3-oxo group and on the reactivity of the 3 α-hydroxyl group. The antisera raised to these immunogens have been characterised for their ability to discriminate between the free steroid and the steroid glucuronide, and evaluated for use in immunoassay. (author)

  18. Radioimmunoassay of canine growth hormone

    International Nuclear Information System (INIS)

    Eigenmann, J.E.; Eigenmann, R.Y.

    1981-01-01

    A sensitive radioimmunoassay (RIA) for canine growth hormone (GH) was developed. Antibodies were elicited in rhesus monkeys. One antiserum exhibited a working titer at a dilution of 1:500 000. Radioiodination was performed enzymatically employing lactoperoxidase. Logit-log transformation and least squares fitting resulted in straight line fitting of the standard curve between 0.39 and 50 ng/ml. Formation of large-molecular [ 125 I]GH during storage caused diminished assay sensitivity. Therefore [ 125 I]GH was re-purified by gel chromatography. Using this procedure, high and reproducible assay sensitivity was obtained. Tracer preparations were used for as long as 3 months after iodination. Diluted plasma from normal and acromegalic dogs resulted in a dose-response curve parallel to the standard curve. Canine prolactin exhibited a cross-reactivity of 2%. The within-assay coefficient of variation (CV) was 3.8 and the between-assay CV was 7.2%. Mean plasma GH concentration in normal dogs was 1.92 +- 0.14 ng/ml (mean +- SEM.) GH levels in acromegalic dogs were appreciably higher. Insulin-induced hypoglycaemia, arginine and ornithine administration resulted in inconsistent and sluggish GH increment. A better response was obtained by injecting a low dose of clonidine. Clonidine administration to hypopituitary dogs resulted in absent or poor GH increment. (author)

  19. Radioimmunoassay of triiodothyronine in urine

    International Nuclear Information System (INIS)

    Kosowicz, J.; Gembicki, M.; Schneider, E.; Eder, M.

    1977-01-01

    In 21 cases of hypothyroidism, in 39 cases of hyperthyroidism, in 54 healthy subjects, in 23 pregnant women, and in certain internal diseases determinations of triiodothyronine were carried out in urine by radioimmunoassay. Anti-T 3 antibodies were obtained in rabbits and sheep immunized with a complex of bovine albumin with triiodothyronine ester. Labelled triiodothyronine of high specific activity was obtained by iodinating triodothyronine by the chloramine method. Determinations of triiodothyronine were performed in morning urine and the obtained values were calculated for one-hour excretion. In healthy subjects the excretion of T 3 was from 20 to 95 ng/hour, in hyperthyroidism it was significantly raised to from 120 to over 600 ng/hour, while in most cases of hypothyroidism it was decreased. In pregnancy the urinary excretion of T 3 was normal amounting to from 34 to 87 ng/hour, although in most cases the serum T 3 concentration was raised. In cases of anorexia nervosa and in obese starving subjects the excretion of T 3 fell significantly, and similarly low excretion was found in some cases of debilitating diseases and myocardial infarction. (author)

  20. The measurement of platelet activation by radioimmunoassay in asthma

    International Nuclear Information System (INIS)

    Wu Guoxin; Sun Jian; Li Jianyong; Ruan Changgeng

    1992-02-01

    Radioimmunoassay with specific monoclonal antibody was used to evaluate the platelet activation in 14 cases of acute bronchial asthma. The result showed that the number of molecules of alpha-granule membrane protein (GMP-140) which was exposed on the surface of platelet following secretion significantly increased on the surface of platelet and in plasma, while the number of molecules of glycoprotein (GP) I b and GPIII a did not change significantly; the concentration of thromboxane B 2 in plasma was raised, while the concentration of 6-keto-PGF 1a was within the normal limits; the concentrations of β-thromboglobulin (β-TG) and platelet factor 4(PF 4 ) in plasma increased significantly; the number of platelets decreased. These results strongly confirmed that the degree of platelet activation was enhanced during acute asthmatic attack. The significance of platelet activation in the pathogenesis of asthma should be further investigated

  1. Theoretical and practical application of radioimmunoassays

    International Nuclear Information System (INIS)

    Sokolowski, G.; Wood, W.G.

    1981-01-01

    This book describes experiences made not only in developing and producing commercial reagent kits, but also those made in university research and diagnostic laboratories. The fundamental principles (radioactivity, immunoassay) are explained briefly and tersely. Then the components of the radioimmunoassay are described in detail and illustrated by selected practical examples (production of antibodies, separation of bound and free portions, standards). This chapter is followed by systematic instructions for composing a radioimmunoassay. The actual conditions of the complex problem of quality safety is explained, a field in which particularly the authors are to be regarded as pioneers. Finally an outlook indicates that radioactive alternatives, incorporating the principle of immunologic determinations methods, are available or will be developed for at least a few radioimmunoassays. (orig./MG) [de

  2. Serum and cellular biologic tumor markers in testicular cancer

    International Nuclear Information System (INIS)

    Javadpour, N.; Lalehzarian, M.

    1987-01-01

    Several serum and cellular biologic tumor markers in testicular cancer are studied. The role of alpha-fetoprotein and human chorionic gonadotropin in staying testicular tumors is reported and the use of radioimmunoassay is considered. (M.A.C.) [pt

  3. Steroid derivatives and their use in radioimmunoassays

    International Nuclear Information System (INIS)

    Varma, R.K.; Chao, S.T.

    1983-01-01

    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

  4. Radioimmunoassay for C-peptide and proinsulin

    International Nuclear Information System (INIS)

    Heding, L.G.

    1977-01-01

    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.)

  5. A challenging alfa-fetoprotein in a cirrhotic patient.

    Science.gov (United States)

    Gallo, P; Gentilucci, U Vespasiani; Taffon, C; Galati, G; De Vincentis, A; Muda, A Onetti; Picardi, A

    2014-03-01

    A 57-year-old Italian man was admitted to our Hospital for investigation of a progressively raising alfa-fetoprotein (AFP) on the background of chronic hepatitis B infection. At abdominal imaging,liver morphology was suspected for advanced fibrosis but without any focal lesion. Clinical and ultrasonographic examinations were negative for testicular masses. When the patient was screened for gastroesophageal varices, upper intestinal endoscopy did not show signs of portal hypertension, while it revealed a gastric lesion which was histologically characterized as hepatoid adenocarcinoma of the stomach (HAS), with strong immunohistochemical positivity for AFP. The patient underwent subtotal gastrectomy and AFP fell within the normal range. This is a very rare case in which AFP-producing gastric cancer (AFPPGC), in the form of HAS, presented in a patient with chronic liver disease. Physicians should be particularly aware of AFPPGC when following patients with liver disorders due to the common use of AFP in this setting.

  6. Determination of serum IgD radioimmunoassay

    International Nuclear Information System (INIS)

    Fayol, V.; Hartmann, D.J.; Sabbagh, I.; Ville, G.

    1986-01-01

    We describe a sensitive liquid phase radioimmunoassay for serum IgD. Extreme values obtained from 85 control patients sera are 0.2 and 121 mg/l with an arithmetic mean of 25 mg/l. In atopic patients (with high serum IgE levels), arithmetic mean is 47 mg/l [fr

  7. Determination of serum IgD radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Fayol, V.; Hartmann, D.J.; Sabbagh, I.; Ville, G.

    1986-01-01

    We describe a sensitive liquid phase radioimmunoassay for serum IgD. Extreme values obtained from 85 control patients sera are 0.2 and 121 mg/l with an arithmetic mean of 25 mg/l. In atopic patients (with high serum IgE levels), arithmetic mean is 47 mg/l.

  8. Solid phase tube radioimmunoassay for digoxin detection

    International Nuclear Information System (INIS)

    Stellner, K.; Glatz, C.; Linke, R.

    1975-01-01

    A solid phase radioimmunoassay with 125 I is described for cardiac patients. The test for the digoxin determination and the poisoning due to cardiac glycosides can be measured very accurately and carried out easily. In addition, the test determination can be automatically performed in connection with other tests. (GSE/LH) [de

  9. Radioimmunoassay of parathormone in human blood serum

    International Nuclear Information System (INIS)

    Leicht, E.

    1982-01-01

    A radioimmunoassay for immune reactive PTH in human blood serum has been developed and optimized. The method described is capable of largely safely discriminating increased PTH concentrations from normal values. For the group of patients suffering from hypoparathyroidism, the PTH values in the blood serum have been found to be generally lower, but quite an overlapping of lower values and values at the lower normal limit has been stated at the same time. Subsequent to an oral administration of phosphate in an acute experiment, so following a calcium-deficient diet taken over 24 hours, an increase in immune reactive PTH in the serum could be detected. The part of this thesis dealing with the method of treatment thoroughly discussed the various steps of optimization of the assay on the basis of extensive experimental data collected by the author. It presents a detailed assessment of current knowledge on, efficiency of and problems involved in PTH determination by radioimmunoassay. (orig./MG) [de

  10. Radioimmunoassay of deoxynivalenol in wheat and corn

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Y.C.; Zhang, G.S.; Chu, F.S.

    With the availability of antibody against deoxynivalenol triacetate (DON-triacetate), a radioimmunoassay (RIA) for DON in wheat was developed. DON is extracted from the sample with acetonitrile-water defatted with hexane, and then reacted with acetic anhydride to form DON-triacetate. The reaction mixture is loaded onto a C-18 cartridge to remove excess reagents and impurities. Acetylated DON is eluted from the cartridge with 50% methanol in water, and then analyzed by radioimmunoassay utilizing antiserum against DON-triacetate and tritiated DON-triacetate. Overall recovery for DON added to wheat between 50 and 5000 ppb was 86% with a standard deviation of 7% and coefficient of variation of 8%. The limit of detection for DON was about 20 ppb. Analysis of 12 naturally contaminated wheat, corn, and mixed feed samples for DON revealed that RIA results agreed well with thin layer chromatographic analyses performed by other laboratories.

  11. Radioimmunoassay to quantitatively measure cell surface immunoglobulins

    International Nuclear Information System (INIS)

    Krishman, E.C.; Jewell, W.R.

    1975-01-01

    A radioimmunoassay techniques developed to quantitatively measure the presence of immunoglobulins on the surface of cells, is described. The amount of immunoglobulins found on different tumor cells varied from 200 to 1140 ng/10 6 cells. Determination of immunoglobulins on the peripheral lymphocytes obtained from different cancer patients varied between 340 to 1040 ng/10 6 cells. Cultured tumor cells, on the other hand, were found to contain negligible quantities of human IgG [pt

  12. Significance and radioimmunoassay of gastric inhibitory polypeptide

    International Nuclear Information System (INIS)

    Zheng Ping; Zeng Minde; Yuan Jimin

    1995-01-01

    We have established the GIP Radioimmunoassay which has high sensitivity and specificity by labelling with iodogen and purified with HPLC. Using this method, the plasma GIP level was measured in 64 cases of which there are 10 normal individuals, 25 cases of diabetes and 29 cases of liver cirrhosis . The results showed that the plasma GIP level was significantly increased in patients with liver cirrhosis and correlated to degree of glucose tolerance damage

  13. Radioimmunoassay of peptide hormones using polyethyleneglycol, 2

    International Nuclear Information System (INIS)

    Ito, Yoshinori; Kikuchi, Akira; Inabe, Seijiro; Yabuuchi, Shinji; Kihara, Akira

    1975-01-01

    The radioimmunoassay (RIA) of glucagon was performed using 125 I in combination with polyethyleneglycol (PEG). In order to define the concentration of glucagon, the reaction time, the temperature and separation of F/B, the effects of the addition of various amounts of serum were studied. This method was found to be applicable, because of its reliability, simplicity, economy, and the shortness of the second reaction time. (Namekawa, K.)

  14. Radioimmunoassay of measles virus antibodies in SSPE

    International Nuclear Information System (INIS)

    Jankowski, M.A.; Gut, W.; Kantoch, M.

    1982-01-01

    A sensitive radioimmunoassay (RIA) was introduced for detecting measles virus IgG and IgM antibodies. The hyperimmune response to the measles virus could be demonstrated more accurately by RIA than by haemagglutination inhibition (HI). The ratio between RIA and HI antibody titres was decidedly higher in sera and cerebrospinal fluids of patients with subacute sclerosing panencephalitis than in those of other groups tested. (author)

  15. Radioimmunoassay of parathyroid hormone: past and future

    International Nuclear Information System (INIS)

    Yalow, R.S.

    1986-01-01

    In this report on radioimmunoassay of parathyroid hormone (iPTH) it was shown that the rate of disappearance of iPTH from plasma differed markedly in patients with primary hyperthyroidism or those with uremia and secondary hyperparathyroidism and that for each patient the rate of disappearance depended on the antiserum used for assay. The heterogeneity of iPTH in plasma was soon rapidly confirmed in many laboratories. (Auth.)

  16. Simultaneous radioimmunoassay for luteinizing hormone and prolactin

    International Nuclear Information System (INIS)

    Steele, M.K.; Deschepper, C.F.

    1985-01-01

    A combined radioimmunoassay (RIA) for the measurement of the anterior pituitary proteins luteinizing hormone (LH) and prolactin (PRL) is described and compared with individual RIAs for these hormones. The standard curves and the sample values for LH and PRL were identical when determined in a combined or in an individual RIA. This technique may prove useful to a number of laboratories where it is desirable to determine levels of more than one hormone in limited sample volumes

  17. Radioimmunoassays - what is their diagnostic value

    International Nuclear Information System (INIS)

    Panitz, N.

    1983-01-01

    A selection is made of hormones and enzymes which are measurable by radioimmunoassay and their organ-specific function is described. Endocrine function tests together with the stimulating or suppressing substances, reaction in the organism and the examined regulation are listed. Non-radioactive immunological determination methods and the use of monoclonal antibodies in vitro and in vivo with ''tumor imaging'' are indicated. (orig.) [de

  18. A radioimmunoassay for urinary and serum dexamethasone

    International Nuclear Information System (INIS)

    Gilliland, J.M.; Morris, H.A.

    1986-01-01

    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)

  19. 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.

  20. Diagnostic value of α-fetoprotein in hepatocellular carcinoma

    International Nuclear Information System (INIS)

    Hernández Perera, Julio César

    2009-01-01

    In the world the hepatocellular carcinoma (HCC) is the fifth to sixth most common malignancy. An increase in its incidence is looming in Cuba as a result of the population aging and the increase of morbi-mortality due to liver cirrhosis (CH). α-fetoprotein (AFP) by SUMA®, was used in this work as a tumor marker technique in 189 Cirrhotic patients evaluated at the CIMEQ, between January 1999 and September 2005. The main factors associated with an elevation of AFP were HCC, viral cirrhosis, and the age of 50 years or more. Presented CHC 22 (11.64%) patients, who had an average age of 55 years and the predominant sex was male (86,36%). The most important cause associated with this malignant disease was viral CH, mainly the virus of hepatitis C. This tumor marker showed a sensitivity and specificity of 68.18% and 92.17%, respectively. When combined with abdominal ultrasound was increase 86.36% sensitivity and 100% specificity. Elevated AFP levels associated with tumor prognosis factors. It can be concluded that the AFP had value in the diagnosis of HCC. This work is the first of its kind carried out in Cuba, and allows the design of a workable strategy for the research, monitoring and prognosis of HCC, having a wide network of laboratories SUMA®,, with a low cost per each determination. (author)

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

    International Nuclear Information System (INIS)

    Salas Chaves, P.; Villalobos Castro, A.

    1999-01-01

    The present study focuses on the normal mean values of AFP in 124 Costa rican pregnant women. In the women 554 determinations were done between the fifteenth and twentieth week of gestation. For this purpose, and Irma test and normal values were established. The concentration values of AFP were expressed as multiples of the median, considering a value of 2.5 Mm as the normal superior limit and of 0.25 as the inferior limit. The confidence limits of the median were 24-57 Ku/I and the 2.5MM value located between 60-143 Ku/I. (author)

  2. Alpha-fetoprotein, HCV and HBV infections in Nigerian patients with ...

    African Journals Online (AJOL)

    The aim of this study was to determine the relationship of serum AFP with HBV and HCV infections among Nigerian patients with, Primary Hepato-Cellular Carcinoma . Forty-two Nigerian adults comprising of 3 groups – PHCC, Liver cirrhosis (LC) and Controls were recruited into the study. The findings showed that only nine ...

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

    Science.gov (United States)

    ... Down syndrome. Why do I need an AFP test? The American Pregnancy Association says that all pregnant women should be ... Medicine [Internet]. Johns Hopkins Medicine; Health Library: Common Tests During Pregnancy [cited 2017 Jun 5]; [about 3 screens]. Available ...

  4. Quality control of radioiodinated gastrin for radioimmunoassay

    International Nuclear Information System (INIS)

    Ginabreda, M.G.P.; Borghi, V.C.; Bettarello, A.

    1988-07-01

    Radioiodinated human gastrin has been prepared at IPEN laboratory for radioimmunoassay use. This work developed the quality control of this tracer analyzing parameters of the labelling reaction, chromatographic purification and radioimmunoassay. The radioiodination yield obtained in five experiments was reproducible and similar when analyzed on 7% polyaraylamide gel eletrophoresis - PAGE - (mean + - SD of 51.70 + - 10.76%) and by1 25 I incorporation checked through thrichloroacetic acid precipitation - TCA - (57-36 + - 9.69%). Similary, after purification the labelled gastrin revaled high and reproducible purity degree when submitted to PAGE (96.57 + - 1.06%) and CA (94.82 + - 4.20%) analysis. The respective specific activities varied from 62 to 307 uCi/ug, being determined by the self-displacement method, which is based on the immunoactivity of the tracer. In this way, the antibody titers required to bind 50% of the tracer ranged from 1:32.000 to 1:180.000. Consequently, the respective doses producing 50% fall in the maximum response of the radioimmunoassays ranged from 155.0 to 24.0 pmol/1, but remained unchanged for each tracer even after three months of its preparations. The tracers presented very low non-specific binding values (1.78 + - 0.79%), stablespecific binding values (46.49 + - 5.65%) and a good between-assay precision, evaluated by an internal quality control sample (25.71 + - 4.30%) with coefficient of variation of 16.74%). The PAGE analysis of the unlabeled gastrin used in the first and last radioiodination revealed an unique and unaltered component, confirming the quality of the tracers. (author) [pt

  5. Solid phase separation technique for use in radioimmunoassays

    International Nuclear Information System (INIS)

    Tu, J.I.

    1979-01-01

    A radioimmunoassay procedure, and article of manufacture for carrying out that procedure, are disclosed herein. The solid phase separation technique utilized in the radioimmunoassay of this invention utilizes a test tube, the internal surface of which has been coated with two antibody layers

  6. Transfer of estradiol to human milk. [Radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Nilsson, S.; Nygren, K.G.; Johansson, E.D.B.

    1978-11-15

    A radioimmunoassay for the measurement of estradiol in human milk is evaluated. The detection limit was found to be 25 pg of estradiol per milliliter of milk. In milk samples collected from four lactating women during three to four months and from one pregnant and lactating woman, the concentration of estradiol was found to be below the detection limit of the assay. When six lactating women were given vaginal suppositories containing 50 or 100 mg of estradiol, it was possible to estimate the estradiol concentration in milk. A ratio of transfer of estradiol from plasma to milk during physiologic conditions is calculated to be less than 100 : 10.

  7. Solid phase double-antibody radioimmunoassay procedure

    International Nuclear Information System (INIS)

    Niswender, G.D.

    1977-01-01

    The present invention is concerned with the radioimmunoassay (RIA) procedure for assaying body fluid content of an antigenic substance which may either be an antigen itself or a hapten capable of being converted, such as by means of reaction with a protein, to an antigenic material. The present invention is concerned with a novel and improved modification of a double-antibody RIA technique in which there is a first antibody that is specific to the antigenic substance suspected to be present in a body fluid from which the assay is intended. The second antibody, however, is not specific to the antigenic substance or analyte, but is an antibody against the first antibody

  8. Radioimmunoassay in the diagnosis of fascioliasis

    International Nuclear Information System (INIS)

    Tomanek, J.; Willomitzer, J.; Chroustova, E.

    1978-01-01

    The objective of the study was to develop a sensitive and specific radioimmunoassay (RIA) method for the diagnosis of F. hepatica infection that would be convenient to use in examining large numbers of samples. The main problem was the preparation of a suitable antigen or specific fraction, since the crude extract of adult Fasciola hepatica is a heterogeneous mixture of somatic, metabolic and host substances with possible nonspecific antigenic properties. The antigen was a protein fraction separated from crude extract of adult F. hepatica by gel filtration and labelled with 125 iodine. (T.I.)

  9. Radioimmunoassay procedure using a stabilized complex

    International Nuclear Information System (INIS)

    Sultanian, I.V.; Irani, J.H.

    1978-01-01

    An improved radioimmunoassay procedure involves the use of a stabilized complex of labelled antigen and antibody which has an extended shelf life as compared to the same complex absent the stabilizers. Since the time needed to incubate the mixture of labelled antigen and antibody is eliminated, the time for completing the assay is considerably shortened and simplified. The components for carrying out the procedure are packaged in a kit basically including standard antigen for generation of a standard curve, a stabilized labelled antigen-antibody complex and reference serum, if used. A plurality of stabilizers are used in the complex to provide a shelf life of six weeks or more. 10 claims

  10. Determination of carbofuran in water by radioimmunoassay

    International Nuclear Information System (INIS)

    Zhu Guonian; Wu Huiming; Yang Ting; Hu Xiuqing

    2004-01-01

    A competitive radioimmunoassay (RIA) method was established to detect carbofuran in water samples. Compared with traditional analytical methods RIA provided a easy procedure with higher sensitivity. The detective limitation of RIA for carbofuran was proved to be 0.175 ng/ml. A study was performed to test validation of the RIA. In this study carbofuran residues in water samples were detected simultaneously by RIA and by high performance liquid chromatograph (HPLC). The linear correlation coefficient of the test results was measure to be 0.9985. (authors)

  11. Radioimmunoassay of nortestosterone and related steroids

    International Nuclear Information System (INIS)

    Hampl, R.; Starka, L.; Picha, J.; Chundela, B.

    1979-01-01

    A radioimmunoassay of nortestosterone and related steroids, including its principal metabolites, is described and evaluated. Antisera against nortestosterone-17β-hemisuccinate- and nortestosterone-3-carboxymethyloxime-bovine serum albumin were raised in goats. By using a mixture of such antisera with different selectivity, the cross-reactions of several naturally occuring steroids can be reduced. The method can be applied for the detection of nortestosterone in both unprocessed or hydrolyzed urine extracts and also in plasma. It has been used as a screening test for anabolics in doping control. (orig.) [de

  12. Radioimmunoassay of digoxigenin glycosides in Digitalis lanata

    International Nuclear Information System (INIS)

    Lehtola, T.; Hiltunen, R.; Schantz, M. v.; Huhtikangas, A.

    1981-01-01

    Radioimmunoassay (RIA) has been used in the analysis of digoxigenin glycoside contents of crude ethanolic extracts prepared from fresh leaf samples of Digitalis lanata EHRH. The commercial antidigoxin antiserum used in the assay was highly specific against the digoxigenin aglycone. Steroid derivatives with only slight structural differences from digoxigenin remained practically unassayed. The method is highly sensitive, and excellent values for accuracy, precision and correlation were obtained. Digoxigenin glycoside levels of second-year D. lanata leaves were found to be considerably higher than those of first-year leaves. (orig.)

  13. Radioiodinated derivatives for steroid radioimmunoassay. Application to the radioimmunoassay of cortisol

    International Nuclear Information System (INIS)

    Gomez-Sanchez, C.; Milewich, L.; Holland, O.B.

    1977-01-01

    Gamma-emitting steroid tracers for use in the radioimmunoassay of steroids have a number of advantages over the more common tritiated tracers. The steroid derivatives aldosterone-3-(p-hydroxybenzoyl) hydrazone, aldosterone-3-(p-hydroxyphenylpropionyl)hydrazone, deoxycorticosterone-3-(p-hydroxyphenylpropionyl)hydrazone, and cortisol-3-(p-hydroxyphenylpropionyl)hydrazone were synthesized by a one-step procedure and iodinated ([ 125 I]). To illustrate the usefulness of these derivatives, we describe the details of a cortisol radioimmunoassay. The use of the radioiodinated tracer appeared to increase the specificity of the antigen-antibody reaction when compared with [ 3 H]cortisol. The methodology involved in the preparation of the steroid derivatives described above can be extended to other 3-oxo-4-ene-containing steroids, with the advantages of economy, simplicity, and versatility

  14. Autism Spectrum Disorders and Maternal Serum α-Fetoprotein Levels During Pregnancy

    DEFF Research Database (Denmark)

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

    2011-01-01

    Objective: Numerous studies have been trying to disentangle the complex pathophysiology of autism spectrum disorders (ASD). In our study, we explored the potential role of maternal serum (MS) α-fetoprotein (AFP) in the prediction and the pathophysiology of ASD. Methods: A total of 112 patients...

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

    International Nuclear Information System (INIS)

    Froelich, M.; Termorshuizen, W.; Kenter, E.; Molenaar, A.J.

    1977-01-01

    The sensitivity of the radioimmunoassay of steroids is considerably reduced by high blank values which may be derived in part from co-chromatographed standards. Blank levels approach the detection limit of the radioimmunoassay of aldosterone, testosterone and androstenedione when 10,000 dpm (30-35 pg) labelled steroids are used as reference standard. When 20 μg aldosterone, testosterone, or androstenedione is used as standard, blank levels of up to 12,800 pg were measured in the radioimmunoassay. Application of the standards on a separate strip does not improve the results. From the experiments it appeared that contamination took place by transport by the solvent

  16. 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-(1...

  17. Beta-endorphin radioimmunoassay: specificity studies

    Energy Technology Data Exchange (ETDEWEB)

    Colas-Linhart, N.; Perdrisot, R.; Petiet, A.; Bok, B.

    1986-01-01

    This note describes the technical details of a cerebrospinal fluid (CSF) B-Endorphin (B-End) radioimmunoassay (RIA). We used an antiserum raised in rabbits against human B-End which cross-reacs 100% with B-Lipotrophin (B-Lph). Thus, filtration chromatography is used to separate both peptides. The assay is sensitive (limit detection=14 pmoles/l), reproducible (the intra and inter assay coefficients of variation are 5 and 6% respectively). Specificity studies are performed to verify the cross-reactions with other opioid peptides and the non specific reactions with the biological fluid (CSF). In order to evaluate the effects of iodine-containing contrast media on the RIA, additional standard curves were analyzed in the presence of varying concentrations of two contrast materials.

  18. Radioimmunoassay of cholecystokinin in tissue and plasma

    International Nuclear Information System (INIS)

    Jansen, J.B.M.J.

    1984-01-01

    The physiological and pathophysiological role of the pancreas hormone, the polypeptide 'cholecystokinin' (CCK) is not well-established yet. This is due to the lack of specific and reliable radioimmunoassays for CCK. The aim of this thesis is to develop such an assay meeting the requirements of high specificity and sensitivity. Several problems were faced, such as (1) the cross-reactivity of existing antibodies with the stomach hormone gastrin and (2) changes in immunoreactivity caused by the introduction of the labelling isotope 125 I and various labels (prepared according to the Bolton-Hunter method) into the polypeptide. The reliability of the assay for the measurement in human tissue and blood is extensively evaluated, inter alia, in patients with pancreas insufficiency (alcohol, cystic fibrosis) and with coeliac disease. (Auth.)

  19. Radioimmunoassay in basic and clinical pharmacology

    International Nuclear Information System (INIS)

    Patrono, C.; Peskar, B.A.

    1987-01-01

    The subject of the book is the development, validation and application of radioimmunoassay (RIA) techniques for the measurement of a variety of substances in animal and human body fluids. The book discusses methodological and conceptual issues related to the main classes of mediators of drug action and to drugs themselves, as assayed by this particular analytical technique. A number of introductory chapters provide basic information concerning production and characterization of antibodies, labeling techniques, statistical aspects and validation criteria, insight into problems related to the development and validation of RIA for the newly discovered mediator(s). In the following chapters, the emphasis is placed on the technical details relevant to each class of compounds and on specific aspects of their applications to basic and/or clinical pharmacological studies. New developments in this area, such as monoclonal antibodies and non-radioactive labeling techniques, are also covered

  20. Interference of immunoglobulins in two glucagon radioimmunoassays

    International Nuclear Information System (INIS)

    Von Schenck, H.; Grubb, A.O.

    1982-01-01

    Radioimmunoassays of glucagon in plasma may be complicated by interaction with other substances of high molecular mass. Precipitates of such substances with ammonium sulfate showed, after isoelectric focusing, two fractions having glucagon immunoreactivity. One fraction (pI approx.10) evidently is associated with the Fc portion (but not the Fab portion) of purified polyclonal immunoglobulin G (IgG). Equal amounts of purified monoclonal IgG of various subclasses, especially IgG 1, gave different ''glucagon'' readings, suggesting that some IgG may interfere more strongly than others. The other fraction (pI 5-6) appeared less consistently, and on gel chromatography appeared to be slightly larger than IgG. Together these fractions add about 50-100 ng/L to the immunoreactive glucagon values in plasma. Therefore methods in which glucagon is extracted before assay should be used for determining the concentration of glucagon present physiologically

  1. Studies on a radioimmunoassay for human erythopoietin.

    Science.gov (United States)

    Lertora, J J; Dargon, P A; Rege, A B; Fisher, J W

    1975-07-01

    A highly purified erythropoietin (ESF) preparation (12,000 units per milligram of protein) was labeled with Na'125I using the Chloramine-T method. Undamaged immunoreative labeled ESF was separated from the damaged, nonimmunologically receiveESF by Sephadex G-150 fractionation. This undamaged immunreactive ESF was usedin radioimmunoassay for human erythropoietin. Separation of bound from free antigen was acheived using the double-antibody technique. Approximately 55 per cent binding wasobserved at an antiserum dilution of 1:1500. This assay appears to be sensitive enough to detect as little as 0.025 milliunits of the International Reference Preparation erythropoietin. The estimated levels of thid hormone in normal and anemic uremic human subjects suggests that immunoreactive serum erythropoietin levels are elevated above normal in anemia of uremia.

  2. Radioimmunoassay of 17-beta - estradiol in plasma

    International Nuclear Information System (INIS)

    Kyian, T.S.; Wajchenberg, B.L.

    1980-01-01

    A radioimmunoassay technique for the measurement of plasma E 2 was standardized utilizing a highly specific antisera against E 2 [-6(-0-carboximetil)-oxime] BSA without the need of previous chromatographic purification. The anti-E 2 serum was highly specificic, showing high affinity with affinity constants:K 1 =1.62 x 10 12 M -1 and K 2 =2.94x10 11 M -1 , calculated by Scatchard plot. The standard-curve sensitivity was 2 picograms. The method was specific and accurate, showing an intra-assay precision with a mean C.V. of 2.9%, with the inter-assay evaluation showing a mean C.V. of 5.0%. This method was employed to evaluate E 2 secretion during the menstrual cycle in 6 normal female. (Author) [pt

  3. A direct radioimmunoassay for aldosterone in plasma

    International Nuclear Information System (INIS)

    Lun, S.; Espiner, E.A.; Nicholls, M.G.; Yandle, T.G.

    1983-01-01

    This rapid radioimmunoassay for aldosterone is performed directly on 100 microL of unprocessed plasma, with 125 I-labeled aldosterone as the labeled antigen. Researchers use of steroid-free plasma in preparing the standard curve resulted in an overestimate of aldosterone; this problem was overcome by adding to such plasma a mixture of other steroids to provide a constant steroid/aldosterone ratio. Over a wide range of aldosterone concentrations, results agreed well between the present assay and a routine method involving solvent extraction and paper chromatography (r . 0.85), and sensitivity (20 ng/L) and inter- (10.4%) and intra- (3.9%) assay CVs were better with the present assay. This assay is especially useful for multiple samples and (or) when only small-volume samples are available

  4. A sensitive radioimmunoassay of scorpion neurotoxin

    International Nuclear Information System (INIS)

    Tessier, M.; Delori, P.; Bechis, G.; Rochat, H.

    1978-01-01

    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 125 I 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

  5. Radioimmunoassay for progesterone in bovine milk

    International Nuclear Information System (INIS)

    Ruiz, Miriam; Figueredo, Nancy; Castillo, Sonia; Pizarro

    2002-01-01

    A system for the measurement of progesterone in bovine milk by radioimmunoassay has been developed and validated. This assay includes an iodine tracer purified by HPLC, the standard prepared in fat-free milk and an antibody anti-progesterone combined with second antibody. The detection limit of the assay is at 0.2 nmol/L calculated from the maximum binding menus two standard deviations and the precision is satisfactory. In the recovery assay was used 4 milk different samples and the result was 98% of recuperation. The progesterone was determinate in milk samples from post-partum animals taking samples three times per week for 40 days. The assay is simple, rapid and possibility the progesterone measurement without sample dilution, distinguish the cyclic changes of this hormone that reflect the ovarian activity in the animals. (author)

  6. A beta-endorphin radioimmunoassay: specificity studies

    International Nuclear Information System (INIS)

    Colas-Linhart, N.; Perdrisot, R.; Petiet, A.; Bok, B.

    1986-01-01

    This note describes the technical details of a cerebrospinal fluid (CSF) B-Endorphin (B-End) radioimmunoassay (RIA). We used an antiserum raised in rabbits against human B-End which cross-reacs 100% with B-Lipotrophin (B-Lph). Thus, filtration chromatography is used to separate both peptides. The assay is sensitive (limit detection=14 pmoles/l), reproducible (the intra and inter assay coefficients of variation are 5 and 6% respectively). Specificity studies are performed to verify the cross-reactions with other opioid peptides and the non specific reactions with the biological fluid (CSF). In order to evaluate the effects of iodine-containing contrast media on the RIA, additional standard curves were analyzed in the presence of varying concentrations of two contrast materials [fr

  7. Radioimmunoassay of cholecystokinin in human plasma

    International Nuclear Information System (INIS)

    Byrnes, D.J.; Henderson, L.; Borody, T.; Rehfeld, J.F.

    1981-01-01

    A sensitive radioimmunoassay for cholecystokinin (CCK) has been developed. Porcine CCK-33 was labelled by conjugation with 125 I-hydroxyphenyl-propionic acid succinimide ester. Antibodies were raised against porcine CCK-33 covalently coupled to egg albumin. Plasma samples were extracted with 96% ethanol prior to assay. Free and bound hormone were separated by dextran-coated charcoal. The antibodies bound CCK-8 and CCK-33 with equimolar potency. The assay detection limit was 1 pmol/l plasma. Within and between assay coefficients of variation were +-12.7 and 13.0% at mean plasma CCK concentrations of 13.2 and 13.6 pmol/l. The concentration of CCK in 47 normal fasting subjects ranged from undetectable to 22 pmol/l. Ingestion of a mixed meal in 9 normal subjects increased the plasma concentration from 8.3 +- 2.5 S.E. to 24.4 +- 6.5 pmol/l. (Auth.)

  8. Report on the national seminar on radioimmunoassays

    International Nuclear Information System (INIS)

    1979-01-01

    Deliberations of the National Seminar on Radioimmunoassays (RIA) held at Bombay during 16-20 January 1978 are reported. Various aspects of the application of RIA techniques in India were discussed in the seminar. They included the basic requirements of RIA in India, the state-of-the-art of RIA in India, radiation protection in RIA, quality control of RIA, usefulness and limitations of RIA in clinical diagnosis and use of RIA in tropical infectious diseases. Difficulties encountered in the practice of RIA techniques in India were found to be mainly related to availability of 125 I, RIA kits, antisera and hormones. The need for establishing a centralised assay service for RIA was examined and it was concluded that such a step would be premature in the present situation. Recommendations made and guidelines spelt out deal with resources for RIA, research, referral assay service, training and information exchange and dissemination. (M.G.B.)

  9. Radioimmunoassay of luteinizing hormone in hypothyroidism

    International Nuclear Information System (INIS)

    Sobieszczyk, S.

    1975-01-01

    Radioimmunoassay of luteinizing hormone was performed in 18 women with primary hypothyroidism and 15 women with secondary hypothyroidism. The results of determinations were compared with LH values found in healthy women at reproductive age and after menopause. It was observed that in primary hypothyroidism the level of LH is normal, in young women it was from 6 to 25 m IU/ml, while in the postmenopausal period it increased to 70 to 200 m IU/ml. In secondary hypothyroidism due to pituitary hypofunction the LH level is undetectable or lies in the range of lowest values observed in healthy subjects, not exceeding 8 m IU/ml. Determinations of serum LH may be useful for differential diagnosis of primary and secondary hypothyroidism. (author)

  10. Radioimmunoassay for GRF and CRF in humans

    International Nuclear Information System (INIS)

    Stalla, G.K.; Losa, M.; Kaliebe, T.; Stalla, J.; Schopohl, J.; Muller, O.A.; Von Werder, K.

    1987-01-01

    In 1981 the structure of ovine CRF was established. One year later others isolated a 44 amino acid peptide with GH releasing activity from a pancreatic islet cell carcinoma of an acromegalic patient (hp GRF/sup 1-44/). In 1983 the gene of human CRF was cloned and the amino acid sequence of hCRF could be elucidated. It differs in 7 amino acids from oCRF. Many investigators demonstrated the biological activity of these peptides in vitro and in vivo. The aim of the authors study was to establish radioimmunoassays for GRF and hCRF with the synthetic derivates, measure endogenous GFR and CRF and circulating GRF- and CRF-levels after intravenous injection and calculate metabolic clearance rate and half-time of disappearance from serum for both releasing hormones

  11. Determination of plasma oxytocin by radioimmunoassay

    International Nuclear Information System (INIS)

    Ogawa, Satsuki; Fukuchi, Soitsu; Miura, Tadashi

    1978-01-01

    A simple radioimmunoassay was applied to the measurement of oxytocin in human plasma. A high specificity of immunoassay was demonstrated by the fact that large excess of angiotensin I and II, and ACTH did not displace labelled oxytocin from the antibody. Lysine-8-vasopressin and arginine-8-vasopressin showed very little cross-reaction in the assay, possessing only 0.002% of the immunological potency of oxytocin. The specific activity of 125 I-oxytocin was 166 μCi/μg. Adsorption and extraction capacities of Florisil were 96.6 +- 2.1% and 85.7 +- 2.5%, respectively. Intra- and inter-assay variability were 7.2 +- 4.9% and 4.3 +- 2.2%, respectively. The sensitivity of the assay was below 1 pg/tube. Normal levels of plasma oxytocin were 0 - 2.2 pg/ml (n=13) in males and 0 - 10.4 pg/ml (n=10) in females. Plasma oxytocin levels in the 39th and 40th weeks of pregnancy were 27.9 +- 4.14 pg/ml (n=4) and 29.8 +- 17.1 pg/ml (n=13), respectively. The levels increased to 33.1 +- 12.1 pg/ml (n=7) and 37.1 +- 17.5 pg/ml (n=7) in the first and third stages of labor, and decreased to 13.6 +- 5.25 pg/ml (n=6) on the 2nd to 8th day after labor. The radioimmunoassay for oxytocin in plasma is considered to be sufficiently applicable for clinical use. (auth.)

  12. 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)

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

    International Nuclear Information System (INIS)

    Dow, B.C.; Barr, A.; Crawford, R.J.; Mitchell, R.

    1983-01-01

    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.)

  14. Radioimmunoassay of methaqualone in human urine compared with chromatographic methods

    International Nuclear Information System (INIS)

    Mule, S.J.; Kogan, M.; Jukofsky, D.

    1978-01-01

    The 125 I-radioimmunoassay for methaqualone in human urine was evaluated by a comparison with newly modified gas-liquid chromatographic and thin-layer chromatographic methods. The statistically significant sensitivity value for the radioimmunoassay was at 2 μg of methaqualone per liter of urine. The coefficient of variation was 2.88 -+ 0.16% intraassay. There was cross-reactivity only with metabolites of methaqualone, 4'-hydroxymethaqualone being twice as sensitively measured as methaqualone. There was complete agreement between results by radioimmunoassay and by gas-liquid chromatography in 96.7% of the samples analyzed. Only 1.2% of the radioimmunoassay values were false positives, and 2.1% false negatives (phi = 0.8917, P < 0.001). Comparisons between the thin-layer chromatographic data and the gas--liquid chromatographic or radioimmunoassay data showed less agreement because of the 50- to 200-fold higher sensitivity of the latter techniques. Gas--liquid chromatography therefore appears to represent the best reference method for the evaluation of the radioimmunoassay, which appears to be a very sensitive and reliable technique for detecting methaqualone and its metabolites in human urine

  15. Simultaneous use of two prostaglandin radioimmunoassays employing two antisera of differing specificity. II. Relative stability of prostaglandins E1, E2, and F1alpha in cell cultures of BALB/c 3T3 and SV3T3 mouse fibroblasts

    International Nuclear Information System (INIS)

    Ritzi, E.M.; Stylos, W.A.

    1976-01-01

    The relative stability of Prostaglandins (PGs) E1, E2 and F1α in cultures of BALB/c 3T3 and SV3T3 cells has been evaluated using 3 different approaches. First, total recovery of tritium in the ethyl acetate phase following incubation and extraction of PGF1α and PGE1 demonstrated greater stability for PGF1α (88.8 percent) than PGE1 (65.9 percent). Second, analysis of incubated, extracted, tritiated PGs by thin layer chromatography revealed decreases of up to 23 percent in the PGE zone following incubation of 3H-PGE1. With increasing time of incubation, decreases in the PGE zone were accompanied by increase in PGA-like compounds. 3H-PGF1α demonstrated greater stability, having greater than 90 percent recovery of the tritium in the PGF zone. A third approach to the assessment of PG stability in culture was the comparison of the production of individual PGs by radioimmunoassay (RIA). The data obtained by RIA indicated a lag in the increase of PGA and PGB, until an initial rise in PGE was noted, suggesting that PGA and PGB may be secondary products arising from PGE which exhibits only partial stability in culture. By employing two RIAs, one for total PGE and one for PGA and PGB, the composite determination PG [E + (A + B)] can be used to provide a more meaningful determination of PG production because of the instability of the PGs. On the other hand, individual determinations are helpful in assessing the stability of PGEs in cell cultures

  16. 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.

  17. Development of radioimmunoassay for prolactin binding protein

    International Nuclear Information System (INIS)

    Raikar, R.S.; Sheth, A.R.

    1982-01-01

    Using a homogenous prolactin binding protein (PBP) preparations from rat seminal vesicle secretion, a sensitive and specific radioimmunoassay (RIA) for PBP has been developed. The assay was highly specific and showed no cross-reaction with other protein hormones from various species. The antiserum had an affinity constant (Ka) of 2.66 x 10 10 M -1 . The assay sensitivity was in the range of 0.5-1.0 ng of pure PBP per assay tube and the intra- and inter-assay coefficients of variations were 6-8% and 12-14.5% respectively. The overall recovery of PBP to the rat seminal vesicle secretion was 96.8%. Using this RIA, PBP levels in various biological fluids and reproductive tissues were measured. Azoospermic human semen contained significantly higher levels of PBP than normospermic semen. The seminal vesicle of rat exhibited the highest concentration of PBP. Administration of antiserum to PBP to mature male rats resulted in a significant reduction in the weight of ventral prostrate and serum prolactin levels were significantly elevated in these animals suggesting that the antibody raised against the PBP was capable of blocking prolactin receptors. (author)

  18. Chemical Kinetics of Progesterone Radioimmunoassay System

    International Nuclear Information System (INIS)

    Abdel-Fattah, A.A.; Moustsfs, K.A.; El-Kolally, M.T.

    2004-01-01

    Progesterone is one of the steroids secreted by the corpus Iuteum in females during the menstrual cycle, and in a much higher amount by the placenta during pregnancy. It is also secreted in a minor quantities by the adrenal cortex in both males and females. Measurement of serum progesterone represents one of diagnostic values in menstrual disorders and infertility. The progesterone radioimmunoassay is based on the competition between unlabelled progesterone and a fixed quantity of 125 I-labeled progesterone for a limited number of binding sites on progesterone specific antibody. Allowing for a fixed amount of magnetizable immunosorbent to react, the antigen-antibody complex is bound on solid particles which are then separated by magnetic rack, and the radioactivity of the solid phase was counted using gamma counter. In this work, the chemical kinetics of the assay was followed, where the specific rate constant (K) was calculated at 4 degree and 37 degree and the activation energy (E act ) were calculated and the reaction rate was deduced

  19. The radioimmunoassay of human serum inhibin

    International Nuclear Information System (INIS)

    McLachlan, R.; Robertson, D.M.; Healy, D.L.; Findlay, J.K.; De Kretser, D.M.; Burger, H.G.

    1987-01-01

    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

  20. Radioimmunoassay in developing countries: General principles

    International Nuclear Information System (INIS)

    Piyasena, R.D.

    1992-01-01

    Radioimmunoassay (RIA) is probably the most commonly performed nuclear medicine technique. It is an in vitro procedure, where no radioactivity is administered to the patient. But this alone is not the reason for its widespread use. It provides the basis for extremely sensitive and specific diagnostic tests, and its use in present day medicine has brought a virtual information explosion in terms of understanding the pathophysiology of many diseases. The fact that the technology involved is within the technical and economic capabilities of the developing world is evident from the increasing demand for its introduction or expansion of existing services. RIA facilities need not be restricted to urban hospitals, as in the case of in vivo nuclear medicine techniques, but may be extended to smaller district hospitals and other laboratories in peripheral areas. It is also possible to send blood samples to a central laboratory so that a single centre can serve a wide geographical area. There are many laboratories in the industrialized world that receive a major proportion of samples for assay by mail. In recent years, substantial RIA services have been established in many of the developing countries in Asia and Latin America. The International Atomic Energy Agency (IAEA) and World Health Organisation (WHO) have made vital contributions to these activities and have played a catalytic role in assisting member states to achieve realistic goals. In the past five years, more than 250 individual RIA laboratories in developing member states have been beneficiaries of IAEA projects

  1. Radioimmunoassay for human health in developing countries

    International Nuclear Information System (INIS)

    Piyasena, R.D.; Airey, P.L.; Ganatra, R.D.; Nofal, M.

    1989-01-01

    Since first introduced in the early 1960s, radioimmunoassay (RIA) has gained wide acceptance as an analytical method adopted by an increasing number of developing countries as an appropriate technology that can be managed within the capabilities of local infrastructures. An informed estimate would be that there are, at present, more than 500 hospitals, university, or other laboratories in the developing world engaged in RIA on some scale. In the developing world, RIA is used primarily for patient management, but research activity is also increasing as expertise and resources improve. The majority of patient samples processed are in relation to thyroid disorders. However, the technique also is used widely in the investigation of other endocrine conditions and public health problems. Some developing countries have gained the capability to perform radioisotopic microassays in areas of clinical and research importance such as steroid receptor quantification in breast tissue; diagnosis of bacterial and parasitic disorders; investigation of infertility and sterility; narcotic drug abuse; and organ transplantation. 1 fig

  2. Parathyroid hormone: radioimmunoassay and clinical interpretation

    International Nuclear Information System (INIS)

    Hawker, C.D.

    1975-01-01

    A radioimmunoassay for serum immunoreactive parathyroid hormone (iPTH), which has had widespread clinical use for five years, is described in detail. The iPTH results in large groups of patients are reported, and are discussed in relation to the specificity of the assay and in relation to other assays. The assay has excellent precision and is highly proficient in discrimination of groups of patients. Ninety-three percent of 412 patients with surgically proven primary hyperparathyroidism were confidently separated from normal subjects or patients with hypercalcemia owing to other causes, while 86 percent of 160 patients with chronic renal failure and secondary hyperparathyroidism had iPTH values more than 2 S.D. above the normal mean. Results in patients with ectopic hyperparathyroidism were lower than in primary hyperparathyroidism although these groups showed considerable overlap. The antiserum used in this assay for iPTH appears to be specific for the carboxy-terminal region of the secreted or intact form of PTH but recognizes predominantly the secreted form rather than carboxy-terminal fragments believed to be in the circulation. It does not recognize amino terminal fragments. The assay is useful in selective venous catheterization for preoperative localization of hyperfunctioning parathyroid tissue

  3. Radioimmunoassay of penicilloyl groups in biological fluids

    International Nuclear Information System (INIS)

    Wal, J-M.; Bories, G.

    1975-01-01

    Penicilloyl-protein conjugates may be formed in man or animals after therapeutic treatment but cannot be detected by the classical microbiological assay methods used for penicillin. Allergies have been noted in consumers of products coming from animals treated with penicillin (milk particularly). Although these compounds present a risk to public health, they passed unnoticed through hygienic food inspection. Therefore a specific and sensitive method was devised for the assay of these derivatives in biological fluids where they can be present in trace amounts. An iodine-125 labelled conjugate has been prepared. Already usable for the detection of antipenicilloyl antibodies in sera of penicillin allergic patients, it has been used for the development of a radioimmunoassay of penicilloyl groups. Assay is done directly on milk, urine and serum without previous extraction and with a detection limit of a few ppb. It permits a rapid, specific and easy to handle determination in the sera of hospital patients as well as in the inspection of animal products at the slaughter house and at the dairy

  4. Radioimmunoassay for platelet activation specific protein GMP-140 on the platelet surface and in plasma

    International Nuclear Information System (INIS)

    Wu Guoxin; Li Jianyong; Ruan Changgeng

    1991-08-01

    Using monoclonal antibody (McAb) SZ-51 which is specific for an alpha-granule membrane protein (GMP-140) on the surface of human activated platelets, the platelet GMP-140 expression in fixed whole blood was measured by direct radioimmunoassay and GMP-140 microparticles in plasma was measured by sandwich method. The GMP-140 molecules per platelet or milliliter (mL) were calculated for the following subjects; acute myocardial infarction; cerebro thrombosis; diabetic mellitus; asthma attack; epidemic hemorrhagic fever etc.. By comparing with the concentration of thromboxane B 2 (TXB 2 ) and von Willebrand factor (vWF) in plasma, it is confirmed that the measurement of GMP-140 molecules is better than that of TXB 2 and vWF. It is a sensitive and specific method for evaluating the platelet activation degree in vivo. The establishment of this method will be useful to diagnosing the thrombotic disorders and studying the pathogenesis of some other diseases

  5. Enhancement of human adaptive immune responses by administration of a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensisTumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors

    DEFF Research Database (Denmark)

    Lobner, M.; Walsted, A.; Larsen, R.

    2008-01-01

    The effect of consumption of Immulina, a high-molecular-weight polysaccharide extract from the cyanobacterium Arthrospira platensis, on adaptive immune responses was investigated by evaluation of changes in leukocyte responsiveness to two foreign recall antigens, Candida albicans (CA) and tetanus......-alpha, IL-1beta, and IL-6 responses, indicating that it acts by inducing a pro-inflammatory state. Taken together, the data suggest that Immulina causes an age-dependent, temporary enhancement of adaptive immune responses Udgivelsesdato: 2008/6...

  6. The determination of serum prolactin by radioimmunoassay

    International Nuclear Information System (INIS)

    Tantirasdr, K.

    1980-01-01

    The iodination of hPRL with 125 I utilizing chloramine-T technique at temperature 4 degrees C gave 125 I-hPRL having immunoreactivity. 125 I-hPRL could be used as a labelled compound for determination of human prolactin by radioimmunoassay. The assay was performed using 125 I-hPRL 5,000 cpm/tube and anti-hPRL 1 : 400,000 dilution (final) incubated at 25 degrees C for 24 hr. One percent dextran-coated charcoal suspension (0.5 ml) was used for separation of free from bound form. The criteria of reliability of the method was evaluated in terms of accuracy, precision, specificity and sensitivity. Under the condition of the assay, the addition of known amount of hPRL (6-50 ng/ml) to charcoaled serum, the recovery of the added hPRL was found to be 97.3-120.8%. The within and between assay variation were evaluated from replicate measurements of the same samples in the same assay and in three different assays. The percentage coefficients of variation was found to be 7.7-15.8% and 13.1-24.0% for within and between assays respectively. Cross-reactivities of tested several protein hormones indicated that the anti-hPRL was very specific. The sensitivity was found to be 1.90 ng/ml. The method was applied to measure serum prolactin for one cycle of 4 volunteers Thai women having normal menstrual cycle. The mean ± S D of serum prolactin was found to be 15.9 ± 5.6, 21.2 ± 12.7 and 16.4 ± 6.2 ng/ml at follicular phase, mid-cycle and luteal phase respectively

  7. Radioimmunoassay of thyrotropin concentrated from serum

    International Nuclear Information System (INIS)

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

    1978-01-01

    A method for concentrating human TSH (hTSH) from serum for use in RIAs is described. The method takes advantage of the affinity of the plant lectin, concanavalin A, for the carbohydrate portion of the hTSH molecule. The hTSH from 2.5 ml serum was adsorbed to concanavalin A covalently linked to sepharose and then radioimmunoassayed using the hTSH antiserum and hTSH for iodination distributed by the National Pituitary Agency. For the RIA standard curve, the hTSH reference preparation was concentrated from a serum wwith undetectable hTSH in order to correct for recovery and to control for nonspecific effects. The percentage of serum hTSH extracted from 2.5 ml serum with the concentration procedure was 76.6 +- 3.4% (mean +- SD). The coefficient of correlation between serum hTSH, determined with the concentration procedure, and serum hTSH determined without was 0.979 (P < 0.001). Over 95% of normal adult men and women had detectable levels of serum hTSH, ranging from < 0.56 to 4.0 μU/ml. The mean of detectable serum hTSH levels in normal adult women (n = 11) was 1.54 +- 1.03 μU/ml (mean +- SD) and in normal men (n = 9) was 2.02 +- 1.15 μU/ml (mean +- SD). Clinically hyperthyroid patients with diffuse and nodular toxic goiters (n = 8) and patients with hypothyroidism secondary to pituitary disease (n = 6), four of whom were taking replacement doses of thyroid hormone, had undetectable serum hTSH levels. Serum hTSH in patients with primary hypothyroidism uniformly exceeded the normal range. This hTSH concentrating procedure enhances the effective sensitivity and, therefore, the clinical utility of the RIA for hTSH in serum

  8. Determination of hCG-alpha subunit in threatened pregnancy

    International Nuclear Information System (INIS)

    Talas, M.; Pohanka, J.; Fingerova, H.; Janouskova, M.; Krikal, Z.; Prasilova, J.; Zupkova, H.

    1987-01-01

    Radioimmunoassay of the hCG-alpha subunit was made using an antibody anti hCG-alpha serum, highly purified hCG-alpha for 125 I-labelling and the standard hCG-alpha. Sera of healthy pregnant women sampled throughout the whole pregnancies were used to determine x-bar±S.D. of hCG-alpha for 14-day intervals. Included in the study were groups of women with high risk of premature labor, late toxemia of pregnancy, twins and fetal hypotrophy. It was shown that increased hCG-alpha is found in pregnant women in whom signs of late toxemia of pregnancy are combined with high risk of premature labor, or with twin pregnancies, while in those with fetal hypotrophy hCG-alpha is within normal limits. (author). 3 figs., 7 refs

  9. Determination of triiodothyronine in serum by enzyme- and radioimmunoassay

    International Nuclear Information System (INIS)

    Oellerich, M.; Haindl, H.; Medizinische Hochschule Hannover

    1981-01-01

    An evaluation of a heterogeneous enzyme immunoassay for determination of triiodothyronine in serum (Enzymun-Test T 3 , Boehringer Mannheim) is presented. The enzyme immunoassay was compared with the laboratory routine radioimmunoassay. The precision of both assays was satisfactory at triiodothyronine concentrations between 1.0 and 8.0 nmol/l (coefficients of variation from day to day 3 from 96-104% and with the radioimmunoassay from 88-111%. A comparison of the results obtained by Enzymun-Test T 3 and the radioimmunoassay in a series of 103 patients showed a good correlation between both methods. L-thyroxine did not cause a relevant cross-reaction in the enzyme immunoassay. About 20 unknown samples can be analyzed in triplicate by Enzymun-Test T 3 within 260 minutes. (orig.) [de

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

    International Nuclear Information System (INIS)

    Weiner, M.H.

    1980-01-01

    The authors have developed radioimmunoassays to the Candida carbohydrate, mannan, and to an Aspergillus cell wall carbohydrate. They evaluate these radioimmunoassays with sera from rabbit models of disseminated mycoses, and further evaluate the radioimmunoassays for their diagnostic usefulness in a panel of patient sera. (Auth.)

  11. Problems in radioimmunoassay of human lutropin with commercially available regents

    International Nuclear Information System (INIS)

    Hammond, J.E.; Phillips, J.C.; Straight, C.B.; Hammond, M.G.

    1980-01-01

    To evaluate five commercially available reagent sets supplied for the radioimmunoassay of lutropin, we determined whether there was parallelism between the curve given by dilutions of the standards supplied by the manufacturers, by dilutions of a serum pool, and by dilutions of a standard preparation from human pituitaries, LER-907. These studies demonstrated significant analytical problems with three of the five sets. We conclude that each user should carefully evaluate all commercially available radioimmunoassays for lutropin (and, by inference, for other peptide hormones) before use

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

    International Nuclear Information System (INIS)

    Hazra, D.K.; Shukla, A.K.; Arvind, B.; Khandelwal, S.; Singh, R.; Lahiri, V.L.

    1986-01-01

    The logistics of national radioimmunoassay services are analysed in regard to: applications of immunoassays in developing countries; provision of matched reagents; collection, storage and transport of patient samples, particularly from rural areas; development of robust new immunoassay systems, especially those using non-isotopic labels; quality control; and interrelation with international agencies and commercial suppliers. The effect of using different methods and temperatures of sample storage on representative glycoprotein, steroid, thyroid and communicable disease analytes is examined. The need for stratification of assay services (district, state and national/regional levels) for various tests is discussed in relation to their clinical urgency. A trouble shooting guide suited to developing countries has been written. Starting from various symptoms of assay failure, it describes the differential diagnosis of likely causes, lists appropriate tests to confirm the diagnosis and finally suggests remedial action to prevent future similar mishaps or to salvage the assay. (author)

  13. Radioimmunoassay of glicagon secretion in diabetes mellitus

    International Nuclear Information System (INIS)

    Milanova, St.; Koparanova, O.; Milkov, V.; Visheva, N.; Kurtev, I.; Maleeva, A.

    1988-01-01

    Radioimunoassay of glucagon, lh, fsh, aldosterone, cortisol, acth, sth and lth was performed using preparations from the firms SORIN, AMERSHAM and HOECHST. The basal glucagon secretion was estimated in 403 diabetic patients and 84 normal subjects. It was transitorily suppressed by glucose. The alpha-cellular reactivity in diabetic patients was stimulated by insulin-induced hypoglycemia and with arginine and vasopressin. The tested group of diabetic patients had absolute hyperglucagonemia, despite the hyperglycemia, which is an evidence of abnormal alpha-cellular function. The insulin-dependent nature of hyperglucagonemia in diabetes mellitus and the hyperreactivity of glucagon secretion after protein stimulation was demonstrated. The correlation of these results leads to essential diagnostic inferences

  14. Direct radioimmunoassay and immunocytochemical localization of renin in human kidneys

    International Nuclear Information System (INIS)

    Menard, J.; N'Goc, P.W.; Bariety, J.; Guyenne, P.T.; Corvol, P.

    1979-01-01

    Highly specific antibodies to human renin were prepared in rabbits and used for the preparation of a renin-free substrate, the direct radioimmunoassay of renin in plasma and kidneys, and the localization of renin with fluoresceinated antibodies. In a patient with a partially infarcted kidney, plasma renin activity, measured enzymatically by the radioimmunoassay of angiotensin I, was increased, and could be activated by acid. The direct measurement of plasma renin by radioimmunoassay gave identical values before and after acidification. In the ischaemic part of the kidney, renin content was high, both by the enzymatic and the direct method of measurement. It was low in the non-ischaemic part of the kidney. Thus whereas renin has been previously measured as an activity by enzymatic assays, this intra-renal hormone can now be measured as an entity by a specific radioimmunoassay in human plasma and kidney tissue. All afferent and some interlobular arteries of the obsolescent glomeruli were stained with fluoresceinated anti-renin antibodies. In the non-ischaemic area, the juxtaglomerular apparatus did not stain. (author)

  15. Cost/benefit and predictive value of radioimmunoassay

    International Nuclear Information System (INIS)

    Albertini, A.; Ekins, R.P.; Galen, R.S.

    1984-01-01

    The present symposium is organized to discuss the benefits of radioimmunoassay. The discussion includes several aspects: the real diagnostic values of the measurements; the way of organization to maximise the diagnostic reliability and usefulness whilst minimizing the real costs; the prospects existing for the improvement of current methods and, implicitly, of cost/benefit ratios. (Auth.)

  16. A solid-phase-radioimmunoassay for total serum thyroxine

    International Nuclear Information System (INIS)

    Moedder, G.; Sokolowski, G.

    1978-01-01

    A new solid phase radioimmunoassay for total serum thyroxine was evaluated over a longer time under clinical routine conditions and compared with an established test system. The results show up that the T 4 values are precise, reliable and reproducible, the is incomplicate to handle and well suitable for semiautomatic pipetting systems. (orig.) 891 MG [de

  17. A radioimmunoassay for anti-virus antibodies in farm animals

    International Nuclear Information System (INIS)

    Rodak, L.; Smid, B.; Sedlacek, M.

    1978-01-01

    A radioimmunoassay for determination of antibodies to Aujeszky's disease virus in piq serum is described. The results show a number of advantaqes of this method over the routinely employed virus-neutralization test. The possibility of using the RIA method in diagnosing other viral diseases of farm animals is suggested. (authors)

  18. Development of a radioimmunoassay of tetracycline and its derivatives

    International Nuclear Information System (INIS)

    Zitzewitz, A. von.

    1981-01-01

    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 3 H tetracycline hydrochloride were performed for radioimmunoassay reference. The possible application of tetracycline radioimmunoassay in food analysis is discussed. (orig./MG) [de

  19. 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...... role in the pathophysiology of ASD makes AFP a good candidate for further larger scale studies to confirm such an association and to determine whether this pattern is unique to ASD or related to other psychiatric disorders as well....

  20. Elevated maternal serum α-fetoprotein after minor trauma during pregnancy may predict adverse fetal outcomes.

    Science.gov (United States)

    Tanizaki, Shinsuke; Maeda, Shigenobu; Matano, Hideyuki; Sera, Makoto; Nagai, Hideya; Kawamura, Shotaro; Ishida, Hiroshi

    2014-09-01

    We evaluated the relationship between minor trauma during pregnancy and elevated maternal serum α-fetoprotein level. This is a retrospective review of pregnant patients admitted to Fukui Prefectural Hospital with trauma during a 10-year period. Charts were reviewed for maternal age, gestational age, injury characteristics, Injury Severity Score, the presence of abdominal pain, systolic pressure and heart rate on arrival, fetal hemoglobin level, and maternal serum α-fetoprotein (MSAFP) concentration on arrival. Fifty-one pregnant patients with any trauma were treated at Fukui Prefectural Hospital. All patients were hemodynamically stable and had minor trauma. An adverse pregnancy outcome occurred in three patients (5%). One patient's fetus had a left kidney injury. Intrauterine fetal death occurred in two patients. The time from injury to fatal death was 180 minutes in one patient and 18 hours in the other patient. The mean ± SD fetal hemoglobin was 0.57% ± 0.88%. The mean ± SD MSAFP was 511 ng/mL ± 1,263 ng/mL. Three patients with adverse pregnancy outcome had a high MSAFP of greater than 1,000 ng/mL. High level of MSAFP may be a predictor of poor fetal outcome following trauma during pregnancy regardless of the severity of the trauma or the mother's hemodynamic status. Epidemiologic study, level V.

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

    DEFF Research Database (Denmark)

    Fenger, M; Johnsen, A H

    1988-01-01

    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...

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

    International Nuclear Information System (INIS)

    Campo, S.; Nicolau, G.; Pellizari, E.; Rivarola, M.A.

    1977-01-01

    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.) [de

  3. 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).

  4. β2-microglobulin, albumin radioimmunoassay and its clinical appraisal

    International Nuclear Information System (INIS)

    Yu Yuming; Zhu Qiyuan; Cheng Guangliang

    1994-01-01

    The β 2 -MG, Alb RIA kits were made in the Radioisotope Department of China Institute of Atomic Energy. They may be used as new type kits inspecting the function indexes of kidney. In the report, these kits are considered to be of high quality by the data of 15 tests. The serum and urine β 2 -MG and urine Alb values of 82 normal subjects and 210 abnormal subjects of different kidney diseases are measured by radioimmunoassay. The results show that the values of β 2 -MG and Alb values of the patients are significantly higher than that of healthy subjects. It is suggested that the β 2 -MG and Alb values measured by radioimmunoassay are very useful and sensitive function indexes of kidney in the clinical diagnosis of different kidney diseases

  5. Development of antiserum against bovine prolactin for radioimmunoassay

    International Nuclear Information System (INIS)

    Razdan, M.N.; Kaker, M.L.; Galhotra, M.M.

    1980-01-01

    Antiserum against bovine prolactin was raised in male rabbits and tested against crossreacting contaminants by immunodiffusion and immunoelectrophoresis. One sharp precipitin band against prolactin was observed. The antiserum showed a small faint band against bovine growth hormone. No crossreaction was observed against bovine follicle stimulating hormone, bovine luteinizing hormone, bovine thyroid stimulating hormone and normal calf serum. The crossreaction with bovine growth hormone vanished after treating the antiserum with normal calf serum. Antibody titre of the absorbed anti-prolactin serum determined by radioimmunoassay was high, which could bind 31.30% at 1:10,000 final dilution. Maximum binding was not inhibited by the addition of higher amounts of contaminants, suggesting that the absorbed antiserum was immunologically monospecific and could be used for radioimmunoassay work. (author)

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

    International Nuclear Information System (INIS)

    Hampl, R.; Starka, L.; Putz, Z.; Dvorak, P.

    1984-01-01

    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)

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

    International Nuclear Information System (INIS)

    Maguire, K.P.; Burrows, G.D.; Norman, T.R.; Scoggins, B.A.

    1978-01-01

    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

  8. Quantitation of avian RNA tumor virus reverse transcriptase by radioimmunoassay

    International Nuclear Information System (INIS)

    Panet, A.; Baltimore, D.; Hanafusa, T.

    1975-01-01

    A radioimmunoassay was developed that can detect and quantitate 3 ng or more of the avian RNA tumor virus reverse transcriptase. The assay detected no antigenic sites in Rous sarcoma virus α virions or in virions of a murine RNA tumor virus. About 70 molecules of reverse transcriptase were found per virion of avian myeloblastosis virus with this assay or with an assay based on antibody inhibition of enzymatic activity. The assay detected about 270 ng of enzyme per mg of cell protein in virus-producing cells; uninfected cells had much less antigenic material but contained some determinants able to displace radioactive antigen. No additional antigenic determinants on reverse transcriptase could be detected that were not found on the separated α subunit of the enzyme. Although sevenfold less sensitive than enzymatic activity as a measure of reverse transcriptase, the radioimmunoassay can detect antigen using small amounts of protein and in the presence of inhibitors

  9. A specific radio-immunoassay for Gonadotrophin-releasing hormone

    International Nuclear Information System (INIS)

    Hendricks, S.; Millar, R.; Pimstone, B.

    1975-01-01

    A specific antiserum has been made to synthetic gonadotrophin-releasing hormone (GnRH) conjugated to keyhole limpet haemocyanin and appears to be directed against amino acids 6 - 8 of this decapeptide. This has allowed the development of a radio-immunoassay for GnRH sensitive to 5 picograms per tube. Although it is easily measurable in hypothalamic extracts, we have failed to detect GnRH in plasma and urine from normal subjects and menopausal women

  10. A micro-radioimmunoassay for measurement of rat luteinizing ...

    African Journals Online (AJOL)

    A sensitive and reproducible double antibody radio-immunoassay for rat LH measurement, which requires at most 50μl of serum or plasma per tube, is described for use with 125l and rabbit anti-rat LH serum. The assay has a scnsitivity of 16 ng LH/ml, and coefficients of inter-assay and intra-assay variations of +/-6,6% and ...

  11. Semiautomated radioimmunoassay for mass screening of drugs of abuse

    International Nuclear Information System (INIS)

    Sulkowski, T.S.; Lathrop, G.D.; Merritt, J.H.; Landez, J.H.; Noe, E.R.

    1975-01-01

    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. Synthesis of iodobuprenorphine for use in radioimmunoassay. [Analgesic

    Energy Technology Data Exchange (ETDEWEB)

    Debrabandere, L.; Boven, M. van; Daenens, P. (Leuven Univ. (Belgium). Lab. of Toxicology)

    1992-08-01

    The synthesis of [[sup 125]I]iodobuprenorphine is described. This compound has been used as a tracer molecule in the development of a new radioimmunoassay for buprenorphine. The parameters that effect the direct iodination of buprenorphine using sodium [[sup 127]I]iodide, in the presence of either chloramine-T or Iodo-Beads, were studied. The structure of iodobuprenorphine was confirmed by mass spectrometry. (author).

  13. Preparation and characterization of reagents for epitestosterone radioimmunoassay

    International Nuclear Information System (INIS)

    Bilek, R.; Hampl, R.; Starka, L.; Putz, Z.

    1987-01-01

    The preparation of reagents, suitable for radioimmunoassay of 17α-hydroxy-4-androsten-3-one (epitestosterone) in body fluids is described. It includes a simplified synthesis of the immunogen, epitestosterone-3-(0-carboxymethyl)oxime coupled to bovine serumalbumin its characterization and preparation of the radioligand ( 125 I) iodohistaminyl derivative of epitestosterone. The assay protocol for epitestosterone detection in urine avoiding extraction is described and the reliability criteria of the method are given. (author)

  14. Automation of steroid radioimmunoassays for clinical and research purposes

    International Nuclear Information System (INIS)

    Vihko, R.; Hammond, G.L.

    1979-01-01

    In recent years an exponential increase in the research and clinical application of radioimmunoassays has created a demand for manipulative aids, to increase the general efficiency of the technique as well as precision of the determinations. At present a number of systems exist for sample preparation and the automation of assays. However, due to inherent problems of carry-over, priming, cleaning and the necessity for chemically inert components most of these systems tend to be rather inflexible. The instrument of choice ought therefore to be extremely versatile and provide the operator with a wide spectrum of alternatives, in order to optimize the initial capital outlay. Moreover, in the light of technological developments in the field of simultaneous, multisample gamma-counting and data processing, it is anticipated that a new generation of multisample processors will emerge of sufficient flexibility to accommodate the wide variety of assay protocols in present use. In this paper the problems encountered in the development of automated techniques for the radioimmunoassay of steroid hormones are reviewed, and a preliminary description of a versatile modular discrete instrument for the automation of radioimmunoassays is presented, which is based on simultaneous multisample preparation, and subsequent counting and data processing. (author)

  15. A newly developed precise and sensitive radioimmunoassay for clonidine

    International Nuclear Information System (INIS)

    Arndts, D.; Struck, C.J.; Staehle, H.

    1979-01-01

    A new precise and sensitive radioimmunoassay for clonidine has been developed. Synthesis and analysis of the hapten (4-carboxy-clonidine; St 1984) as well as antibody production in rabbits are described in detail. At a final dilution of 1:1000 the resulting immune serum binds 50% of a tritiated clonidine standard containing 1 ng of clonidine. The detection limit of the presented radioimmunoassay for clonidine is 0.1 ng/ml. The coefficient of variation did not exceed 4.3% for any of 7 standard determinations with 5 replicates. There was no relevant crossreactivity of inactive clonidine metabolites apart from 4-OH-clonidine. To avoid any errors from cross-reaction clonidine was selectively and quantitatively extracted into diethylether from unknown plasma samples. Following concentration of the extracts even such low concentrations as 20 pg of clonidine/ml plasma were detectable. With the radioimmunoassay applied in pharmacokinetic studies a maximal clonidine concentration in blood plams of healthy human volunteers was determined to 0.6 ng/ml 1.5 h after oral administration of 150 μg. (orig.) [de

  16. Reference Intervals for Alpha-Fetoprotein (AFP) and Carcinoembryonic Antigen (CEA) in Guangxi Zhuang Ethnic Males from the FAMHES Project.

    Science.gov (United States)

    Lao, Xianjun; Yang, Dongmei; Mo, Zengnan; Gao, Yong; Deng, Yan; Qin, Xue; Li, Shan

    2016-01-01

    Several studies have reported the reference intervals of serum AFP and CEA levels in ethnically diverse populations, but there is a lack of such reference data among Zhuang ethnic males. The aim of this study was to establish the locally validated reference intervals for AFP and CEA in the male population of the Guangxi Zhuang ethnic group. A total of 283 Zhuang ethnic males, aged 22 to 69 years, were included from the Fangchenggang Area Male Health and Examination Survey (FAMHES) project database. The one-sided upper 95th-percentile limit was used to estimate the reference intervals for serum AFP and CEA. The total non-parametric reference intervals for Zhuang ethnic males were reference intervals for serum AFP and CEA values deviated from that reported in previous studies. Age-specific reference intervals should be performed in clinical laboratories to obtain more precise estimations for the clinical conditions of young adults and elderly people.

  17. Detection of α-fetoprotein in human serum using carbon nanotube transistor

    Science.gov (United States)

    So, Hye-Mi; Park, Dong-Won; Lee, Seong-Kyu; Kim, Beom Soo; Chang, Hyunju; Lee, Jeong-O.

    2009-03-01

    We have fabricated antibody-coated carbon nanotube field effect transistor (CNT-FET) sensor for the detection of α-fetoprotein (AFP), single chain glycoprotein of 70 kDa that is normally expressed in the fetal liver, in human serum. The AFP-specific antibodies were immobilized on CNT with linker molecule such as pyrenebutyric acid N-hydroxysuccinimide ester. To prevent nonspecific adsorption of antigen, we performed blocking procedure using bovine serum albumin (BSA). Antibody-antigen binding was determined by measuring electrical conductance change of FET and took an average of thereshold voltage change before and after binding. Also we checked concentration-dependent conductance change in human serum using both p-type SWNT-FETs and n-type SWNT-FETs.

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

    International Nuclear Information System (INIS)

    Nilson, B.; Aakerstroem, B.; Bjoerck, L.

    1986-01-01

    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 alpha 1 -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.)

  19. Specific and sensitive radioimmunoassay for 3-methoxy-4-hydroxyphenylethyleneglycol (MOPEG)

    Energy Technology Data Exchange (ETDEWEB)

    Keeton, T.K. (Univ. of Texas Health Science Center, San Antonio); Krutzsch, H.; Lovenberg, W.

    1981-02-06

    Antibodies that specifically bind the norepinephrine metabolite 3-methoxy-4-hydroxyphenylethyleneglycol (MOPEG) were produced in rabbits after injection of a derivative of MOPEG conjugated with bovine thyroglobulin. A sensitive radioimmunoassay was devised with this antiserum, in which as little as 0.5 nanogram of MOPEG can be accurately measured with a final antibody dilution of 1:180. The antibody appears to be specific for MOPEG, since tritiated MOPEG was not displaced from the antibodies by norepinephrine, epinephrine, dopamine, serotonin, or their major metabolites, including MOPEG-sulfate (333 nanograms each).

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

    International Nuclear Information System (INIS)

    Caso, R.

    1997-01-01

    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

  1. An introduction to radioimmunoassay and related techniques. 3. rev. ed.

    International Nuclear Information System (INIS)

    Chard, T.

    1987-01-01

    This book is designed as an introductory text for the staff of clinical or research laboratories who conduct or intend to conduct radioimmunoassay and related immunoassay techniques, and will also be of great value to the clinicians who make use of such services. The emphasis is on general principles with practical illustrations rather than a detailed and encyclopaedic review of the literature. This 3rd revised edition places more emphasis on alternative labels (non-isotopic immunoassays), the use of labelled antibodies (immunometric assays), and the use of monoclonal antibodies. Both principles and practice are illustrated by diagrams and illustrations. (Auth.)

  2. 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.

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

    International Nuclear Information System (INIS)

    Yin Shihua; Song Shiyun; Li Kelin; Chen Guanglian; Liu Fengmin

    1993-01-01

    Serum β 2 -microglobulin (β 2 -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 (P 2 -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 β 2 -MG is the indicator of too frequent blood donating. The results also showed that the content of β 2 -MG in donor's blood is not a normal reference value

  4. Chromatographic purification of tritiated steroids prior to use in radioimmunoassay

    International Nuclear Information System (INIS)

    Manlimos, F.S.; Abraham, G.E.

    1975-01-01

    The purity of tritiated steroids used as reagents in radioimmunoassay plays an important role in the reliability of the assay. These radioactive reagents should be assessed for purity upon receipt and the purity should be checked periodically afterward. For such purposes, we have used chromatographic purification on Celite microcolumns. By changing the polarity of the stationary and mobile phases, 20 different tritiated steroids with a wide range of polarity could be purified on these microcolumns. This approach is easy, rapid, economical, and reliable. (U.S.)

  5. Radioimmunoassay for determination of thyroglobulin in human serum

    International Nuclear Information System (INIS)

    Rodriguez Cabrera, M.E.; Blanca Fernandez, S.; Baldor Navarro, F.; Rodriguez Gonzalez, J.C.

    1996-01-01

    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 %

  6. Radioimmunoassay of somatostatin: methodological problems and physiological investigations

    International Nuclear Information System (INIS)

    Penman, E.; Wass, J.A.H.

    1981-01-01

    Somatostatin, a tetradecapeptide, has a wide distribution throughout the central nervous system and the gastrointestinal tract and a broad spectrum of biological actions. In order to investigate its various physiological roles in man, a radioimmunoassay was developed for somatostatin in human blood plasma, which is described here. This RIA was used to investigate possible factors influencing somatostatin secretion. Changes in somatin levels produced by changes in insulin, glucagon and growth hormone levels were studied via the response of plasma immunoreactive somatostatin to hormonal stimuli in normal man. The influence of fasting and food consumption was studied; and the site of origin of circulating immunoreactive somatostatin was investigated in patients. (Auth.)

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

    International Nuclear Information System (INIS)

    Quiroga, S.

    1984-10-01

    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

  8. Radioimmunoassay of human muscle carbonic anhydrase III in dystrophic states

    International Nuclear Information System (INIS)

    Heath, R.; Jeffery, S.; Carter, N.

    1982-01-01

    A radioimmunoassay for the human isozyme carbonic anhydrase III (CAIII) has been developed. The assay can detect levels as low as 4μg/l of sample. Plasma CAIII levels in patients suffering from Duchenne muscular dystrophy were found to be up to 39 times greater than in a control group. Urine CAIII levels in patients suffering from Duchenne muscular dystrophy were not significantly different from the levels found in urine from normal adults. Measurement of plasma CAIII levels may be useful in prenatal diagnosis of Duchenne muscular dystrophy, and in investigation of adult skeletal muscle disease. (Auth.)

  9. Preparation and purification of 7-Iodoclonazepam for use in radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Goddard, C.P.; Law, B.; Mason, P.A.; Stead, A.H.

    1986-04-01

    A method is described for the preparation and purification of 7-(/sup 125/I)-Iodoclonazepam (5-(o-Chlorophenyl)-2,3-Dihydro-7-(/sup 125/I)-Iodo-1H-1,4-Benzodiazepin-2-one). The structure was confirmed by mass spectrometry using 7-(/sup 127/I)iodoclonazepam prepared by the same method. 7-(/sup 125/I)-Iodoclonazepam binds well to a benzodiazepine antiserum. Although readily displaced by all the benzodiazepines commercially available in the UK, it is not displaced by structurally related nonbenzodiazepines except at very high concentrations. 7-(/sup 125/I) Iodoclonazepam should therefore be useful for the development of a screening radioimmunoassay (RIA) for benzodiazepines.

  10. Studies on digoxin derivatives in different biological media by radioimmunoassay

    International Nuclear Information System (INIS)

    Barmasch, Martha; Perez, L.N.; Altschuler, Noe.

    1978-05-01

    The fundamental aim of this study was to develop a methodology to be applied to pharmacological studies, directed to demonstrate the passage of digitalic compounds through the blood-brain barrier. This study was a comparative one between β methyl digoxin (βMD) and digitoxin (Dt). A methodology of radioimmunoassay was developed for different biological media: serum (S), plasma (P) and cerebro spinal fluid (CSF). In addition, the immunochemical behaviour (affinity, displacement, etc.) of βMD was studied when reacted with digoxin (D) and digoxin-antisera, supplied by the commercial kits utilized in these studies. (author) [es

  11. Radioimmunoassay test system for detection of anti-insulin antibodies

    International Nuclear Information System (INIS)

    Dudko, N.V.; Piven', N.V.; Ibragimova, G.V.; Kasatkin, Yu.N.

    1995-01-01

    A radiodiagnostic test system has been developed and commercial kit for radioimmunoassay of anti-insulin antibodies in human blood serum created. Clinical trials of the kit in patients (150 diabetics with types 1 and 2 condition) and normal subjects (n=100) demonstrated the possibility of using this kit for the detection of preclinical forms of diabetes and for distinguishing groups at risk of diabetes among children and adults, for the detection of insulin resistance, for the differential diagnosis of diabetes, and for monitoring the efficacy of insulin therapy. 9 refs.; 1 tab

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

    International Nuclear Information System (INIS)

    Stracke, H.

    1978-01-01

    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.) [de

  13. Radioimmunoassay and radioenzymatic assay of a new aminoglycoside antibiotic, netilmicin

    International Nuclear Information System (INIS)

    Broughton, A.; Strong, J.E.; Pickering, L.K.; Knight, J.; Bodey, G.P.

    1978-01-01

    A radioimmunoassay and a radioenzymatic assay for netilmicin, a new aminoglycoside, were developed in our laboratories to assist in the study of the pharmacology of the drug and establish values for use in its monitoring. The assays are sensitive, precise, and rapid, giving results that correlate (r = 0.90) with each other and with those of a microbiological assay in which Klebsiella pneumoniae is used as the test organism. Preliminary pharmacological studies show the drug to have a biological half-life of 135 min, which is comparable to that for other aminoglycosides

  14. Radioimmunoassay of total testosterone in urine of men

    International Nuclear Information System (INIS)

    Bilek, R.; Chundela, B.; Hampl, R.; Starka, L.

    1985-01-01

    The results are described and evaluated of the radioimmunoas radioimmunoassay of total testosterone in the urine in a sample of five men of the ages between 18 and 21 years. The levels of testosterone were related to creatinine in the urine in order to eliminate the effect of urine dilution. The analysis of the results showed that there exists a statistical difference between the levels of total testosterone related to creatinine in dependence on the time of sampling. The average value with standard deviation of total testosterone was 109 +- 41 ng testosterone/1 ml urine, 24.07 +- 8.33 pmol testosterone/1 μmol creatinine. (author)

  15. Comparison of radioimmunoassay and gas chromatographic mass spectrometric assay for d-amphetamine

    International Nuclear Information System (INIS)

    Powers, K.H.; Ebert, M.H.

    1979-01-01

    Quantification of low levels of psychotropic drugs (10 -7 to 10 -9 g ml -1 ) in small volumes of plasma requires sensitive and accurate methods. Validation of these methods is best achieved by comparing results obtained using several techniques. In this study, amphetamine levels in plasma were measured using gas chromatography mass spectrometry and radioimmunoassay. Correlation of the results obtained by the two methods was found to be positive and high (R = 0.9822). The average coefficient of variation between assays for gas chromatography mass spectrometry was 5.8% and for radioimmunoassay was 12.3%, while the average coefficient of variation within assays for gas chromatography mass spectrometry was 4.9% and for radioimmunoassay 6.9%. Although gas chromatography mass spectrometry was 1.9 times more sensitive than radioimmunoassay, for most purposes, the convenience of the radioimmunoassay method outweighs the technical superiority of gas chromatography mass spectrometry. (author)

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

    International Nuclear Information System (INIS)

    Matsuhashi, Kazuo

    1974-01-01

    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 3 H-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.)

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

    International Nuclear Information System (INIS)

    Fleury, B.

    1981-10-01

    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 [fr

  18. A specific and sensitive radioimmunoassay for human choriogonadotropin

    International Nuclear Information System (INIS)

    Muralidhar, K.; Chaudhuri, G.; Lippes, J.; Bahl, O.P.

    1983-01-01

    A specific and sensitive radioimmunoassay for human choriogonadotropin (hCG) has been developed using rabbit antiserum to chemical analogs of beta subunit of human chorionic gonadotropin prepared by controlled reduction and S-alkylation of its disulfied linkages. The assay was highly specific for hCG as the binding of [ 125 1]-hCG to the antibody was not affected by standard human lutropin, by human male serum, postpartum serum from women, serum from post-menopausal women and human menopausal gonadotropin (Pergonal). The assay was highly sensitive, the minimal detection limit in terms of highly purified hCG (L-129, 12.500 IU/mg) being 1 ng/ml or 0.2 ng/tube (or 12 mlU/ml in terms of WHO 2nd international reference preparation of hCG). Using this assay we were unable to detect any immunoreactive hCG in human tissues like lung, liver and colon. The high specificity, sensitivity, accuracy and reproducibility of the assay make this a highly desirable radioimmunoassay for human choriogonadotropin. (orig.)

  19. Secretin-radioimmunoassay, physiology and pathophysiology in man

    International Nuclear Information System (INIS)

    Haecki, W.H.

    1978-01-01

    Production of antibodies to secretin for radioimmunoassay is straightforward. Secretin is iodinated by weak oxydation with lactoperoxydase and subsequent purification by ionexchange chromatography (Sephadex C25). The specific activity of fresh label is between 650 and 900 mCi x mol -6 . The label is highly purified and may be used in radioimmunoassay for several months. In order to eliminate plasma interference sepharose-beads with covalently coupled secretin antibodies are used to produce secretin-free standard plasma samples. Delay in the separation of plasma from fresh blood samples can lead to erronous results, even to falsely elevated secretin levels. - Duo-denal acidification only leads to physiological increases of secretin plasma levels. This may happen by intraduodenal instillation of acid, or by an acidic oral drink, or to a lesser extent after a meal. Secretin is distributed throughout the plasma volume and has a short halflife of around 3 minutes. Impaired release of secretin is found in children with coeliac disease. The role of secretin in peptic ulcer however is not clear. Chronic pancreatitis and renal insufficiency are without effect on plasma secretin levels. (orig.) [de

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

    International Nuclear Information System (INIS)

    Eder, W.

    1982-01-01

    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.) [de

  1. Radioimmunoassay of prostaglandin F in plasma of pregnant women

    International Nuclear Information System (INIS)

    Albert, P.

    1980-01-01

    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 3 H-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) [de

  2. Radioimmunoassay of chorionic gonadotropin in the diagnosis of ectopic pregnancy

    International Nuclear Information System (INIS)

    Skalba, P.; Grochal, A.; Kokocinska, D.; Parylak, H.

    1977-01-01

    The applicability of radioimmunoassay of the chorionic gonadotropins (HCG) was studied considering the diagnosis of ectopic pregnancy. The radioimmunoassay technique was presented basing on personal anti HCG serum and HCG preparation (Serono). The technique of separation, based on solid phase was described. The technique is not fully specific, permits, nevertheless to determine the serum hormone concentrations ranging over 0.4 μg per ml. The investigations were carried in 26 cases to tubal pregnancy and in 22 unpregnant women - where basing on the initial diagnosis and clinical observations, ectopic pregnancy was suspected. Moreover the serum HCG was established in 64 women in recent intrauterine pregnancy - lasting 3 to 8 weeks. The mean serum HCG concentration in cases of ectopic pregnancy was 5.4 μg per ml and was significantly lower than in cases of 4 weeks lasting intrauterine gestation. The presence of HCG was demonstrated in all cases of real ectopic gestations, it was absent in the compared group of the suspected, but unpregnant, subjects. The paper suggests that the presented method can be applied for the differential diagnosis of ectopic pregnancy and permits to plan the further clinical procedures. (author)

  3. Radioimmunoassay for mammalian type C viral reverse transcriptase

    International Nuclear Information System (INIS)

    Krakower, J.M.; Barbacid, M.; Aaronson, S.A.

    1977-01-01

    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

  4. 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.

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

    Science.gov (United States)

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

    2009-01-01

    AIM: To investigate the expression of α-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 reverse-transcriptase 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 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. PMID:19360917

  6. Protein microarray with horseradish peroxidase chemiluminescence for quantification of serum α-fetoprotein.

    Science.gov (United States)

    Zhao, Yuanshun; Zhang, Yonghong; Lin, Dongdong; Li, Kang; Yin, Chengzeng; Liu, Xiuhong; Jin, Boxun; Sun, Libo; Liu, Jinhua; Zhang, Aiying; Li, Ning

    2015-10-01

    To develop and evaluate a protein microarray assay with horseradish peroxidase (HRP) chemiluminescence for quantification of α-fetoprotein (AFP) in serum from patients with hepatocellular carcinoma (HCC). A protein microarray assay for AFP was developed. Serum was collected from patients with HCC and healthy control subjects. AFP was quantified using protein microarray and enzyme-linked immunosorbent assay (ELISA). Serum AFP concentrations determined via protein microarray were positively correlated (r = 0.973) with those determined via ELISA in patients with HCC (n = 60) and healthy control subjects (n = 30). Protein microarray showed 80% sensitivity and 100% specificity for HCC diagnosis. ELISA had 83.3% sensitivity and 100% specificity. Protein microarray effectively distinguished between patients with HCC and healthy control subjects (area under ROC curve 0.974; 95% CI 0.000, 1.000). Protein microarray is a rapid, simple and low-cost alternative to ELISA for detecting AFP in human serum. © The Author(s) 2015.

  7. 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...... (ACTH)-(1-39), ACTH-(1-14) and alpha MSH immunoreactivity]. alpha MSH and ACTH-(1-14) were only present in non- or mono-acetylated forms. Only large forms of gamma 1MSH and gamma 2MSH were present in partly glycosylated states. The hinge peptides were amidated to an extent two to three orders...

  8. Buffett's Alpha

    DEFF Research Database (Denmark)

    Frazzini, Andrea; Kabiller, David; Heje Pedersen, Lasse

    Berkshire Hathaway has realized a Sharpe ratio of 0.76, higher than any other stock or mutual fund with a history of more than 30 years, and Berkshire has a significant alpha to traditional risk factors. However, we find that the alpha becomes insignificant when controlling for exposures to Betting-Against-Beta...... in publicly traded stocks versus wholly-owned private companies, we find that the former performs the best, suggesting that Buffett's returns are more due to stock selection than to his effect on management. These results have broad implications for market efficiency and the implementability of academic...

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

    International Nuclear Information System (INIS)

    Lahlou-Kassi, A.; Lakhdissi, H.

    1984-01-01

    The report summarizes the principal applications of progesterone radioimmunoassay (P 4 -RIA) in Morocco for evaluating and improving fertility in cattle, sheep and camels. In cattle, P 4 -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. P 4 -RIA is an important tool in fertility improvement programmes involving induction and synchronization of oestrus and testing for early pregnancy. In sheep, P 4 -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 P 4 -RIA could be used for early pregnancy testing. (author)

  10. Radioimmunoassay of hair for determining opiate-abuse histories

    International Nuclear Information System (INIS)

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

    1979-01-01

    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

  11. Radioimmunoassay for prostatic acid phosphatase in human serum. Methodologic aspects

    International Nuclear Information System (INIS)

    Pradalier, N.; Canal, P.; Pujol, A.; Fregevu, Y.; Soula, G.

    1982-01-01

    We propose a double antibody radioimmunoassay for human prostatic acid phosphatase (PAP) in serum for diagnosis and management of prostatic adenocarcinoma under treatment. The antigen is purified from human prostatic fluid by a gel-filtration on Sephadex G 100 followed by affinity chromatography on Con A Sepharose. A specific antibody is raised in rabbits and purified by immunoadsorption with a female serum. The described technique offers both radioisotopic sensibility and immunologic specificity. Physiological values determined in the serum of 125 healthy males are below 2 ng/ml. No significative differences are observed with age. The proposed technique also shows significant differences between values evaluated for benign prostatic hyperplasia and prostatic adenocarcinoma [fr

  12. Production of biological reagents for radioimmunoassay second antibody

    International Nuclear Information System (INIS)

    Borghi, V.C.; Silva, S.R. da; Bellini, M.H.; Lin, L.H.

    1992-02-01

    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)

  13. Development of a heterologus radioimmunoassay for human prolactin (hprl)

    International Nuclear Information System (INIS)

    Kothari, K.K.; Meera, V.; Soman, P.D.; Lal, R.; Sivaprasad, N.

    1991-01-01

    The development of a heterologous radioimmunoassay for the measurement of human prolactin in serum is reported. The assay uses the less expensive ovine prolactin for the preparation of 125 I labelled prolactin and anti prolactin serum while the human prolactin in used as standard. The assay is based on a double antibody- polyethylene glycol separation system for the separation of the antibody bound antigen from the free antigen. The optimized assay has a preincubation of the antibody with the cold antigen for 18 hours followed by a post incubation of 3 hours with the tracer. The sensitivity of this assay is 2ng/mL. The inter and intra assay variations are less than 9%. The values of the serum samples analysed by this method compares well with immunoradiometric assay. (author). 12 refs. , 6 figs., 2 tabs

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

    International Nuclear Information System (INIS)

    Roser, J.F.; Chang, Y.S.; Papkoff, H.; Li, C.H.

    1984-01-01

    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

  15. The milk samples collected with filter paper for progesterone radioimmunoassay

    International Nuclear Information System (INIS)

    Zhang Jiahua; Geng Guoxia; Zhang Huaiyu

    1985-01-01

    The cow milk was collected with filter paper treated with ethanol during eastrus-day (0 day) and 22th and 24th day after mating. Then it was dried and stored in room temprature until analysis for progesterone by means of radioimmunoassay. The sensitivity is 13.62 pg/bule (n = 4), the coefficients of variation within a group and between groups are 8.8% (n = 10) and 16.65% (n = 8) respectively, and the recovery is 91.23% (n = 4). The average progesterone level for 22th and 24th day in the pregnant cows (6.28 +- 1.28 ng/ml) was much higher than that in the non-pregnant cow (2.00 +- 1.18 ng/ml), the difference being significant (P < 0.001). The judgement based on progesterone level (5 pregnant and 5 non-pregnant cows) faily agreed with the clinical diagnosis

  16. Farm application of radioimmunoassay technology in dairy cattle management

    International Nuclear Information System (INIS)

    Alejandrino, A.L.; Asaad, C.O.; De Vera, A.C.; Deocaris, C.C.; Ignacio, L.M.; Herrera, M.S.; Mateo, A.B.

    1996-01-01

    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)

  17. Radioimmunoassay data processing program for IBM PC computers

    International Nuclear Information System (INIS)

    1989-06-01

    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

  18. Provision of radioimmunoassays for reproductive hormones in developing countries

    International Nuclear Information System (INIS)

    Sufi, S.B.; Donaldson, A.; Jeffcoate, S.L.; Goncharov, N.P.

    1986-01-01

    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)

  19. Radioimmunoassay for chlorpromazine in plasma. [/sup 3/H tracer technique

    Energy Technology Data Exchange (ETDEWEB)

    Midha, K.K.; Loo, J.C.K.; Hubbard, J.W.; Rowe, M.L.; McGilveray, I.J.

    1979-01-01

    A radioimmunoassay for chlorpromazine in plasma is described. The antiserum was obtained by immunizing rabbits with a conjugate of bovine serum albumin and N-(2-carboxyethyl)desmethylchlorpromazine. It is specific for chlorpromazine and its minor active metabolite, N-desmethylchlorpromazine. Other known active or inactive chlorpromazine metabolites and other psychotropic drugs tested do not cross react with the antiserum. Less than 34 pg of the drug can be detected in 200 ..mu..L of plasma. As many as 100 samples can be processed in a day by one technician. Concentrations of chlorpromazine can be measured in 200-..mu..L samples of plasma collected as late as 48 h after a single oral 25-mg dose.

  20. Radioimmunoassay for human plasma 8-arginine-vasopressin

    International Nuclear Information System (INIS)

    Conte-Devolx, B.; Rougon-Rapuzzi, G.; Millet, Y.

    1977-01-01

    A radioimmunoassay for human plasma vasopressin (AVP) which permits the estimation of antidiuretic hormone (ADH) level as low as 0.8pg/ml, was developed. The average plasma level of AVP after overnight water restriction was found to be 14.3pg/ml (sd=4.4pg/ml) in normal subjects. They provoked a hypersecretion of ADH by the intravenous injection of 1-2mg of nicotine. In 11 volunteer normal subjects this stimulation by nicotine provoked ADH hypersecretion which reached a maximum between 2nd and 15th minutes after injection. In 3 cases of diabetes insipidus, nicotine injection did not induce ADH hypersecretion: in 1 case of potomania this response was weak; in 2 cases of syndrome of inappropriate ADH secretion, AVP plasma levels were elevated and the response after nicotine stimulation was exaggerated [fr

  1. Radioimmunoassay of paralytic shellfish toxins in clams and mussels

    International Nuclear Information System (INIS)

    Yang, G.C.; Imagire, S.J.; Yasaei, P.; Ragelis, E.P.; Park, D.L.; Page, S.W.; Carlson, R.E.; Guire, P.E.

    1987-01-01

    Shellfish contaminated with paralytic shellfish poisons (PSP) compromise human health. The threat of this contamination results in enormous economic losses in the recreational and commercial exploitation of shellfish resources in the affected areas. Most states deal with the PSP problem either by prohibiting the collection of shellfish during certain time periods or by instituting monitoring programs. The only recognized method of analysis for PSP that is currently and routinely used in monitoring programs is the time-of-death mouse bioassay. Several attempts to develop simple and highly specific biochemical assays for the detection and quantitation of the PSP toxins have been reported. More recently, much improved immunoassays have been developed. To evaluate the validity and usefulness of the immunoassay for the determination of PSP toxins, the authors have used extracts of shellfish gathered from Maine and Connecticut to compare the results of the mouse bioassay and HPLC methods with the radioimmunoassay developed previously

  2. Measurement of plasma canine C peptide by radioimmunoassay

    International Nuclear Information System (INIS)

    Besch, W.; Woltanski, K.P.; Fischer, U.; Kohnert, K.D.; Ziegler, M.

    1985-01-01

    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)

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

    International Nuclear Information System (INIS)

    Elbanna, I.M.; Ragab, M.T.

    2000-01-01

    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

  4. An improved radioimmunoassay for urinary Tamm-Horsfall glycoprotein

    International Nuclear Information System (INIS)

    Dawnay, A.B. St. J.; Thornley, C.; Cattell, W.R.

    1982-01-01

    A rapid specific radioimmunoassay has been used to measure Tamm-Horsfall glycoprotein (TH glycoprotein) in urine, and the method described. The apparent concentration increased with increasing dilution of urine in water, reaching a plateau at 1 in 20. This increase was greater the higher the osmolality and TH glycoprotein concentration and the lower the pH of the original sample. The apparent concentration of TH glycoprotein in neat or diluted urine was not affected by freezing or by storage at 4 0 C or room temperature for at least 2 days. A physiological range for the urinary excretion rate was established as 22-56 mg/24h, (considerably higher than the amount present in serum) based on samples from 29 individuals with normal renal function, as defined by their creatinine clearance. There was no significant correlation between serum concentrations of TH glycoprotein and its urinary excretion rate, nor between urinary excretion rate and creatinine clearance. (author)

  5. Recombinant-derived chicken growth hormone used for radioimmunoassay

    International Nuclear Information System (INIS)

    Proudman, J.A.

    1984-01-01

    The use of recombinant-derived chicken growth hormone (rcGH) in an avian growth hormone (GH) radioimmunoassay (RIA) procedure is described. Antiserum to turkey GH bound 125 I-labeled rcGH, and unlabeled rcGH or turkey GH displaced binding in a dose-related manner. The dose-response curves of sera and pituitary extract from chickens and turkeys were parallel to the rcGH standard curve. Sera from hypophysectomized (hypox) chickens and turkeys produced no dose-response and did not inhibit binding of labeled rcGH. Recovery of rcGH added to hypox sera was quantitative. Modification of the homologous turkey GH RIA protocol of Proudman and Wentworth (1) to use rcGH made possible either an increase in assay sensitivity or a 3-day reduction in incubation time

  6. High molecular somatostatin, an interfering factor in radioimmunoassay

    International Nuclear Information System (INIS)

    Diel, F.; Schneider, E.; Baumann, H.

    1977-01-01

    Cyclic Tyr 1 -somatostatin (Tyr 1 -SRIF) is radioiodinated by the lactoperoxidase method. Purification is achieved by Sephadex G-25 adsorption chromatography. Specific anti-SRIF serum (FA1) has been raised in rabbits. A dose response curve is obtained in the range of 5 - 5,000 pg per tube using an antiserum dilution of 1:2,000. There is little cross-reaction with linear somatostatin and none with ocytocin, (lys-, arg-) vasopressin, valinomycin, polymyxin, insulin, glucagon, human growth hormone (hGH), and thyrotropin-releasing hormone (TRH). For recovery tests, extraction procedures are necessary. Thin-layer chromatography (TLC) and polyacrylamide-disc-electrophoresis (Disc-PAGE) are performed to identify the presumed high molecular 125 I-Tyr 1 -SRIF associate. This high molecular associate may represent an interfering factor in the radioimmunoassay for cyclic SRIF. (orig./AJ) [de

  7. Engineering of radioimmunoassay (RIA) IP10.1

    International Nuclear Information System (INIS)

    Hari Nurcahyadi

    2010-01-01

    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)

  8. Radioimmunoassay for phencyclidine: application to kinetic analysis in the rat

    International Nuclear Information System (INIS)

    Ward, D.P.; Trevor, A.J.

    1980-01-01

    We report the development of a radioimmunoassay for phencyclidine (PCP) that is simple, rapid and sensitive to 0.5 ng/ml. Antibodies were raised in rabbits against the hapten, N-succinyl-3-aminophencyclidine. These antibodies proved to be very specific for PCP and exhibited less than 4% cross reactivity with the drug's two major metabolites. The assay was used for kinetic analysis of PCP in the rat following subcutaneous injection of 5 mg/kg of the drug. Serum and brain tissues were analyzed for PCP and the respective half lives were calculated to be 36 and 29 min for the α phase and 130 and 121 min for the β phase. The accuracy of the method was verified by concomitant assay of a number of kinetic samples by gas chromatography employing a nitrogen-phosphorus detector

  9. Performance of various mathematical methods for calculation of radioimmunoassay results

    International Nuclear Information System (INIS)

    Sandel, P.; Vogt, W.

    1977-01-01

    Interpolation and regression methods are available for computer aided determination of radioimmunological end results. We compared the performance of eight algorithms (weighted and unweighted linear logit-log regression, quadratic logit-log regression, Rodbards logistic model in the weighted and unweighted form, smoothing spline interpolation with a large and small smoothing factor and polygonal interpolation) on the basis of three radioimmunoassays with different reference curve characteristics (digoxin, estriol, human chorionic somatomammotropin = HCS). Great store was set by the accuracy of the approximation at the intermediate points on the curve, ie. those points that lie midway between two standard concentrations. These concentrations were obtained by weighing and inserted as unknown samples. In the case of digoxin and estriol the polygonal interpolation provided the best results while the weighted logit-log regression proved superior in the case of HCS. (orig.) [de

  10. Effect of radioimmunoassay procedures on therapeutic drug monitoring

    International Nuclear Information System (INIS)

    Kampa, I.S.

    1985-01-01

    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

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

    International Nuclear Information System (INIS)

    Gylstorff, B.

    1982-01-01

    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) [de

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

    International Nuclear Information System (INIS)

    Knyazev, Yu.A.; Bespalova, V.A.; Vakhrusheva, L.L.; Kirbasova, N.P.; Severtseva, V.V.

    1984-01-01

    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

  13. Radioimmunoassay of aldosterone and its level in plasma and urine

    International Nuclear Information System (INIS)

    Putz, Z.; Hampl, R.; Veleminsky, J.; Starka, L.

    1981-01-01

    A method of plasma and urine aldosterone radioimmunoassay is described and evaluated. Highly specific antisera were obtained through rabbit immunization using partly aldosterone-18,21-dihemisuccinate-BSA, partly aldosterone-3-carboxymethyl-oxime-BSA, free of the 18-derivative. The use of a more specific antiserum against the 3-derivative permitted the determination to be performed immediately from the biological specimen extract. Hydrolysis with sulphuric acid in the presence of dichloromethane proved the most appropriate of the different techniques of releasing aldosterone-glucuronoside for the determination of the total urine hormone. The normal values (average +- 2 S. D.) of plasma aldosterone were 0.19 +- 0.067 nmol/l, those of the free urine hormone 0.918 +- 0.458 nmol/day, and the total urine aldosterone 15.3 +- 5.8 nmol/day. (author)

  14. The use of diatomite microcolumns for the chromatographic separation of steroids prior to radioimmunoassay

    International Nuclear Information System (INIS)

    Abraham, G.E.

    1975-01-01

    Since most anti-steroid antibodies are not usually specific enough to permit direct radioimmunoassay of these compounds, chromatographic purification is required to achieve reliability Celite microcolumns were used for the separation of a wide range of steroids prior to radioimmunoassay. Depending on the polarity of the steroids to be separated, various mixtures of solvents are used as stationary and mobile phases. This system is rapid, easy, economical and reliable

  15. Miniaturized technique for the rapid extraction of plasma glucagon for radioimmunoassay

    International Nuclear Information System (INIS)

    Flatt, P.R.

    1981-01-01

    Radioimmunoassay of glucagon is hampered by interference from non-specific factors introduced into the incubation mixture by the plasma sample. The proteolytic destruction of glucagon must be minimized by using proteolytic enzyme inhibitors or plasma extraction techniques. The present report, which describes a miniaturized technique for the quantitative extraction of glucagon from plasma, is aimed at increasing the popularity of this approach to the plasma glucagon radioimmunoassay. (Auth.)

  16. Comparative studies on the determination of alphafetoprotein by enzyme immunoassay and by radioimmunoassay

    International Nuclear Information System (INIS)

    Haller, G.; Linneke, P.; Voss, P.; Jeske, W.

    1987-01-01

    Alphafetoprotein (AFP) was determined in serum of pregnant women in the tenth till sixteenth week of pregnancy by means of two enzyme immunoassays (Enzymun-Test AFP, Boehringer Mannheim, FRG and AFP EIA 'Dessau' 1000, Research Institute for Vaccine Dessau, GDR) and a radioimmunoassay (Radioimmunoassay Kit, AFP-PR, CIS, France). Parallel determinations in sera of 438 patients, who had come to surveillance for the first consultation were estimated. A comparison between the methods showed a good correlation. (author)

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

    Science.gov (United States)

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

    2011-06-28

    To investigate the role of α-fetoprotein (AFP), a cancer-associated fetal glycoprotein, in glucocorticoid-induced precocious maturation in rat colon. 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. 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-d-old 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. GCs promote the development of rat colon. AFP appears to be involved, in

  18. Evaluation of Patients With Hepatocellular Carcinomas That Do Not Produce α-Fetoprotein.

    Science.gov (United States)

    Agopian, Vatche G; Harlander-Locke, Michael P; Markovic, Daniela; Zarrinpar, Ali; Kaldas, Fady M; Cheng, Elaine Y; Yersiz, Hasan; Farmer, Douglas G; Hiatt, Jonathan R; Busuttil, Ronald W

    2017-01-01

    Serum α-fetoprotein (AFP) is a biomarker for hepatocellular carcinomas (HCCs) associated with a more aggressive tumor phenotype and inferior outcomes after a liver transplant (LT). Data on the outcomes for patients with HCCs that do not produce AFP are limited. To compare characteristics and outcomes among LT recipients with radiographically apparent HCC lesions with AFP-producing tumors or with tumors that do not produce AFP (hereafter referred to as non-AFP-producing tumors), and to identify factors influencing recurrence in LT recipients with non-AFP-producing tumors. Retrospective analysis at a university transplant center of 665 adults with HCC who underwent an LT during the period from 1989 to 2013. Of the 665 LT recipients, 457 (68.7%) had AFP-producing tumors, and 208 (31.3%) had non-AFP-producing tumors (the maximum AFP level before an LT was ≤10 ng/mL). Dates of study analysis were from August 2015 to June 2016. Liver transplant. Recurrence-free survival and recurrence rates. Patients with non-AFP-producing tumors had radiographic tumor characteristics similar to those of patients with AFP-producing tumors, but, pathologically, they had fewer lesions (25% vs 35% with >2 lesions; P = .03), smaller cumulative tumor diameters (4.2 vs 5.0 cm; P = .02), fewer microvascular (17% vs 22%) and macrovascular (2% vs 9%) invasions (P 2 tumors [HR, 4.98; 95% CI, 1.72-14.4; P = .003]; cumulative diameter [1.70 per log SD; 1.12-2.59; P 2 tumors [4.39; 1.32-14.6; P = .02]; cumulative diameter [2.32 per log SD; 1.43-3.77; P = .001]; microvascular [3.07; 1.02-9.24; P = .05] and macrovascular invasion [8.75; 2.15-35.6; P = .002]). Nearly one-third of patients with radiographically apparent HCC have non-AFP-producing tumors that have more favorable pathologic characteristics, lower posttransplant recurrence, and superior survival compared with patients with AFP-producing tumors. Posttransplant HCC recurrence for patients with non

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

    Directory of Open Access Journals (Sweden)

    Huixiao Hong

    2016-03-01

    Full Text Available 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.

  20. Development and clinical application of human gastrin radioimmunoassay

    International Nuclear Information System (INIS)

    Ginabreda, M.G.P.; Borghi, V.C.; Bettarello, A.

    1988-08-01

    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 125 I-gastrin, standard gastrin (zero to 500 pmol/l) or unknown samples with the antiserum for 4 days at 4 0 C. 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) [pt

  1. A radioimmunoassay for lignin in plant cell walls

    Energy Technology Data Exchange (ETDEWEB)

    Dawley, R.M.

    1989-01-01

    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 {beta}-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 {eta}g/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. {sup 125}I 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 NaClO{sub 2} delignification. The RIA was found to be less sensitive than expected, so several avenues for improving the method are discussed.

  2. The studies on aldosterone secretion rate by radioimmunoassay

    International Nuclear Information System (INIS)

    Takenouchi, Takahiko

    1974-01-01

    The aldosterone secretion rate was measured by radioimmunoassay in 12 normal subjects and 47 hypertensive patients. The values ranged from 25.0 to 60.2 ng/day with a mean of 39.6+-10.7 (S.D.) in 8 normal males and from 30.2 to 85.4 ng/day with a mean of 62.6+-26.8 (S.D.) in 4 normal females. The mean value in 21 cases with benign essential hypertension was found to be within normal range. No significant difference was found in the aldosterone secretion rate between the group of benign essential hypertension with suppressed renin activity and with nonsuppressed renin activity. Metabolic clearance rate in benign essential hypertension was observed to be within normal range. The aldosterone secretion rate in a few cases of benign essential hypertension failed to increase normally in response to sodium restriction (<50 mEq/day). The values in 11 cases of primary aldosteronism were found to be clearly higher, when compared with the values of normal subjects and subjects with benign essential hypertension. High values were found in patients suffering from malignant hypertension and in 3 with unilateral renal artery stenosis. The values in cases of bilateral renal artery stenosis, of pheochromocytoma, and of acromegaly with hypertension were within normal range. Low values in the aldosterone secretion rate were found in 3 cases each of 17α-hydroxylase deficiency and Cushing's syndrome. (JPN)

  3. Radioimmunoassay for progesterone in human saliva during the menstrual cycle

    Energy Technology Data Exchange (ETDEWEB)

    Luisi, M.; Franchi, F.; Kicovic, P.M.; Silvestri, D.; Cossu, G.; Catarsi, A.L.; Barletta, D.; Gasperi, M. (Pisa Univ. (Italy))

    1981-10-01

    A sensitive, specific and accurate radioimmunoassay of progesterone in human saliva is described, using /sup 3/H. The assay had a sensitivity of 8 pg/tube and blanks were negligible. The intra- and inter-assay coefficients of variation were 5.2 and 9.4%, respectively. The mean recovery from 60 samples was 93.2 +- 6.3%. Results obtained from nine healthy, normally menstruating women showed that salivary progesterone rose from the 4th day before ovulation to a mean peak (+- SD) of 1.14 +- 0.17 ng/ml on the 8th day after ovulation, followed by a gradual decline. Correlation of salivary and simultaneously obtained plasma progesterone levels was good (r = 0.47; P < 0.001), although the maximum percent increase in salivary progesterone was more than 10 times greater than that of plasma progesterone. Salivary progesterone is thought to reflect the unbound fraction of plasma progesterone and this non-invasive technique can be used for serial investigations in which frequent samplings are required.

  4. Radioimmunoassay in the diagnosis of atypical form of thyrotoxicosis

    Energy Technology Data Exchange (ETDEWEB)

    Livshits, G.Ya.

    1984-11-01

    Fifty-six patients with ''unmotivated'' disorder of the cardiac rhythm were examined. A combined radionuclide study including a study of iodoabsorptive function with a standard technique, thyroid visualization and determination of the thyroxin, triiodothyronine level in the blood serum with the radioimmunoassay using standard diagnostic kits, was conducted, Latent thyroid hyperfunction was revealed in 24 patients (42.8%). Study of iodoabsorptive function revealed pathological changes in 8 patients only, whereas raaioimmunoassay revealed a significant elevation of the peripheral thyroid hormone level as compared to that of the control group in 24 patients. The conclusion is that patients with ''unmotivated'' disorder of the cardiac rhythm often suffer from latent thyrotoxicosis which is the main etiological factor and trigger mechanism of arrhythmias. In such a situation they are the only clinical symptom of thyrotoxicosis that makes it possible to regard this form of disease as monosymptomatic. The early detection of the cause of cardiac rhythm disorder and the prescription of pathogenetic thyrostatic therapy resulted in the return of the cardiac cycle rate to normal in all the patients with sinus tachycardia and prevented relapses of the paroxysmal forms of rhythm disorder.

  5. Labelling and standardizing some pituitary hormones for radioimmunoassay

    International Nuclear Information System (INIS)

    Kim, Y.S.

    1976-11-01

    Optimum conditions for efficient 125 I labelling of human follicle stimulating hormone (FSH) and human chorionic gonadotropin (HCG) using chloramine-T have been established for radioimmunoassay (RIA). The amount of the hormone, chloramine-T, 125 I, and the reaction time were, respetively, controlled evaluating the yield and the bindability of the labelled hormone to its antibody. To measure the bindability, the labelled hormone was incubated together with its antibody for a definite temperature. In the separation of the free hormone (F) from the antibody bound (B), a double antibody technique was applied comparing with the chromatoelectrophoresis. For the efficient separation of the labelled hormone, two methods of separation such as gel filtration and gel electrophoresis were compared in the sensitivity and in the immunological activity points of view. Experiments for the production of HCG antibody were also conducted. The produced antisera were tested in two ways; i.e., the incubation test with the labelled hormone, and the Ouchterlony test. Using the produced anti-HCG serum and the purchased anti-FSH serum, standard dose-response curves were plotted correlating with the international standard preparation of the hormones

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

    International Nuclear Information System (INIS)

    Vaudry, H.; Vague, P.; Dupont, W.; Leboulenger, F.; Vaillant, R.

    1975-01-01

    A radioimmunoassay technique has been developed for measuring frog plasma corticotropin (ACTH) without prior extraction. Using synthetic porcine ACTH as a reference standard, 131 I-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 131 I-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

  7. Radioimmunoassay of angiotensin-II: methodology and standardization.

    Science.gov (United States)

    Lall, S B; Peshin, S S; Seth, S D

    1995-12-01

    The study was planned to set up and standardize the radioimmunoassay of Ang-II and to validate the procedure in terms of precision, sensitivity, specificity and recovery. The application of the developed assay was studied in normal healthy volunteers and in patients of renovascular hypertension (RVHT) and renal hypertension (RH). Synthetic human Ang-II was coupled to BSA by carbodimide condensation to get the hapten carrier conjugate which was injected in rabbits to raise the antibodies. The titre of 1:250 showed significant binding (56.79%) and was used for the assay. The sensitivity of the assay was 2 pg/ml and cross-reactivity with analogues of Ang-II was minimum. Mean Ang-II levels in normal subjects was 16 +/- 3.6 pg/ml. In patients of RVHT and RH, the peripheral blood Ang-II levels were found to be 876 +/- 8.6 and 108 +/- 2.3 pg/ml respectively. In patients of unilateral RVHT, renal vein Ang-II levels of the affected side were significantly higher than the unaffected side (P < 0.001). The results indicate that unextracted samples can be successfully utilized to estimate Ang-II levels.

  8. An Interlaboratory study of lipid effects on steroid radioimmunoassay

    International Nuclear Information System (INIS)

    Bolelli, G.F.; Franceschetti, F.; Mimmi, P.; Malvano, R.; Pilo, A.; Zucchelli; Rota, G.

    1986-01-01

    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)

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

    International Nuclear Information System (INIS)

    Vihko, R.; Hammond, G.L.; Pakarinen, A.; Viinikka, L.

    1977-01-01

    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.) [de

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

    International Nuclear Information System (INIS)

    Wassom, D.L.; Loegering, D.A.; Gleich, G.J.

    1979-01-01

    Guinea pig eosinophil major basic protein (MBP) was measured by radioimmunoassay (RIA) using 131 I-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)

  11. Radioimmunoassay of erythropoietin: analytical performance and clinical use in hematology.

    Science.gov (United States)

    Schlageter, M H; Toubert, M E; Podgorniak, M P; Najean, Y

    1990-10-01

    We report here the performance of a recently commercialized radioimmunoassay kit for determining erythropoietin (EPO) in serum or plasma. The lower detection limit of the method was 3 U/L. Precision, analyzed by the variation coefficients between different assay runs and in the same experiment, was always less than 10%; accuracy was assessed by recovery and dilution tests. In anemic patients (hematocrit 18-39%), the concentration of EPO was logarithmically related to hematocrit. A relatively large dispersion of the results was noted, as reported by others with various RIAs. Patients with severe renal failure demonstrated a very low EPO value, whatever the degree of their anemia. In some chronic anemias resulting from malignancy, EPO concentrations were also relatively low. In the polycythemia vera group, the EPO mean was below normal for greater than 95% of the patients, whatever their clinical stage (first evaluation, relapse, or remission). In contrast, 91% of the patients with pure erythrocytosis had a normal or increased EPO value, even when the etiology was unknown. Measurement of EPO concentration may be useful for the clinical differentiation of myeloproliferative disorders and, subsequently, for their prognosis and choice of treatment.

  12. Radioimmunoassay of antidiuretic hormone. Application to rats and to man

    International Nuclear Information System (INIS)

    Fressinaud-Masdefeix, Philippe.

    1976-06-01

    The general principles of antidiuretic hormone secretion are known from kidney function explorations and the use of biological methods. Radioimmunoassay of the hormone should contribute towards a better understanding of this secretion in the fields of both physiology and pathology. After a review on antidiuretic hormone and the methods used so far for its investigation, part two of this work concentrates on the development of our own radioimmunological system applied to this hormone. The system is specific, though interference in the measurement from metabolite(s) of the hormone cannot be absolutely ruled out; reproducible, as witnessed by the 'inter-test' variation coefficient of 8%; and sensitive, since the hormone is easily measured in the urine, when secretion is slowed down. In spite of its sensitivity however the hormone determination in the plasma is not possible for a small sample volume because of the difficulty of obtaining high-affinity specific antibodies. To our knowledge only one plasma ADH determination is in common use to date. Part three is devoted to the application of this technique in rats and man, and the results obtained [fr

  13. Radioimmunoassay in the diagnosis of atypical form of thyrotoxicosis

    International Nuclear Information System (INIS)

    Livshits, G.Ya.

    1984-01-01

    Fifty-six patients with ''unmotivated'' disorder of the cardiac rhythm were examined. A combined radionuclide study including a study of iodoabsorptive function with a standard technique, thyroid visualization and determination of the thyroxin, triiodothyronine level in the blood serum with the radioimmunoassay using standard diagnostic kits, was conducted, Latent thyroid hyperfunction was revealed in 24 patients (42.8%). Study of iodoabsorptive function revealed pathological changes in 8 patients only, whereas raaioimmunoassay revealed a significant elevation of the peripheral thyroid hormone level as compared to that of the control group in 24 patients. The conclusion is that patients with ''unmotivated'' disorder of the cardiac rhythm often suffer from latent thyrotoxicosis which is the main etiological factor and trigger mechanism of arrhythmias. In such a situation they are the only clinical symptom of thyrotoxicosis that makes it possible to regard this form of disease as monosymptomatic. The early detection of the cause of cardiac rhythm disorder and the prescription of pathogenetic thyrostatic therapy resulted in the return of the cardiac cycle rate to normal in all the patients with sinus tachycardia and prevented relapses of the paroxysmal forms of rhythm disorder

  14. Precision of radioimmunoassays by various types of data processing systems

    International Nuclear Information System (INIS)

    Ujiie, Morimichi

    1979-01-01

    The following are the result of the investigation of the precision of radioimmunoassays by various types of data processing systems presently performed in Japan. Chose 50 facilities, where 10 facilities of clinical laboratories were included. Tested 334, 485 cases per month, which corresponds to 40% of the total number of the cases treated all over the country. It was known that the data of 74% of all facilities and 94% of all tests were being processed by computing machines. In 66 times of data processing of insulin and T3 measurements, 32 were performed by the hyperbola method, 11 by the log-logit curve method of secondary degree, 9 by the logistic curve method and 8 by the log-logit curve method of first degree. Practically in more than 90% of all cases were processed by these 4 types of curve methods. In insulin measurements of many standard points (8 points), favourable results could be obtained by the logistic curve method. On the contrary, in T3 measurements of fewer standard points (5 points), the results from the method of log-logit curve of secondary degree were good. Favourable results of both insulin and T3 measurements could be obtained by the hyperbola method. While, in the insulin measurement by the method of log-logit curve of first degree, the values were a bit lower in the higher and lower concentration ranges, and 3 -- 4% higher than the actual values in the middle concentration range. (author)

  15. The radioimmunoassay of clomipramine (Anafranil-Geigy): a tricyclic antidepressant

    International Nuclear Information System (INIS)

    Read, G.F.; Riad-Fahmy, D.

    1977-01-01

    A radioimmunoassay has been developed for the tricyclic antidepressant, clomipramine (Anafranil-Geigy) which allows accurate determination of plasma levels without a pr-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 didesmethyclomipramine 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 G.C. or G.C./M.S. 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/week per technician) make the assay ideal for supervision of patient compliance and routine assay of samples generated in large clinical trials. (orig.) [de

  16. Specific radioimmunoassay of human. beta. -endorphin in unextracted plasma

    Energy Technology Data Exchange (ETDEWEB)

    Wiedemann, E. (Univ. of California, Berkeley); Saito, T.; Linfoot, J.A.; Li, C.H.

    1979-09-01

    With an antiserum against human ..beta..-endorphin (..beta..-EP) crossreacting <2% with human ..beta..-lipotropin (..beta..-LPH) by weight we have developed a radioimmunoassay that can detect 1 pg ..beta..-EP in diluted raw plasma. In a.m. fasting plasma of 14 normal subjects ..beta..-EP ranged from <5 to 45 pg/ml. ..beta..-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, ..beta..-EP was undectectable after dexamethasone and increased after metyrapone administration and insulin-induced hypoglycemia. ..beta..-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, ..beta..-EP behaves like a hormone responding to the same stimuli as ACTH and ..beta..-LPH and blood appears to contain enzymes both generating and destroying immunoreactive ..beta..-EP.

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

    International Nuclear Information System (INIS)

    Schmid, M.

    1981-01-01

    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) [de

  18. Solid phase radioimmunoassays for human C-reactive protein

    International Nuclear Information System (INIS)

    Shine, B.; Beer, F.C. de; Pepys, M.B.

    1981-01-01

    Two new, rapid and sensitive radioimmunoassays for human C-reactive protein (CRP) have been established using antiserum coupled to magnetizable cellulose particles, which facilitate phase separation. A single antibody method, using solid phase anti-CRP, provides a sensitivity of 50 μg/l with a 1-h incubation time and intra- and inter-assay coefficients of variation of 10%. A double antibody method, using fluid phase rabbit anti-CRP serum and solid phase sheep anti-rabbit IgG serum, provides a sensitivity of 3 μg/l with an overnight incubation and intra- and inter-assay coefficients of variation of 10%. Among 468 sera from normal adult volunteer blood donors the median CRP concentration was 800 μg/l, interquartile range 340-1700 μg/l and range 70-29,000 μg/l. Ninety percent of samples contained less than 3 mg/l and 99% less than 10 mg/l. Low levels (14-650 μg/l) of CRP were detected both in amniotic fluids and in cerebrospinal fluids. (Auth.)

  19. Detection of myocardial infarct extension by CK-B radioimmunoassay

    International Nuclear Information System (INIS)

    Rothkopf, M.; Boerner, J.; Stone, M.J.; Smitherman, T.C.; Buja, L.M.; Parkey, R.W.; Willerson, J.T.

    1979-01-01

    Myocardial infarct extension after the acute event was defined as a second rise in the myocardial isoenzyme of serum creatine kinase (CK-B) after the initial return of CK-B to normal values. In 43 patients with acute myocardial infarct, CK-B was measured by radioimmunoassay every 12 hrs for 14 days. Nineteen patients had anterior transmural myocardial infarcts (AMI), 14 had inferior transmural myocardial infarcts (IMI), and 10 had subendocardial myocardial infarcts (SEMI). Infarct extension as detected by a second rise in serum CK-B occurred in six patients (32%) with AMI, two (14%) with IMI, and two (20%) with SEMI. Four patients with AMI also had clinically evident infarct extension. In the other six, the infarct extension was undetected clinically. The measurement of serum CK-B values with a quantitative and sensitive assay suggests that myocardial infarct extension occurs more commonly than clinically recognized, but the frequency of extension may be less than that reported in patients in whom precordial mapping and total serum CK values were measured to identify this phenomenon

  20. Laboratory training manual on radioimmunoassay in animal reproduction

    International Nuclear Information System (INIS)

    1984-01-01

    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

  1. An Evaluation by TSH Radioimmunoassay on Familial Thyroid Disorders

    International Nuclear Information System (INIS)

    Kim, Ji Yeul

    1989-01-01

    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 131 I or 99m Tc 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.

  2. Development and validation of a new radioimmunoassay for parathyrin (PTH)

    International Nuclear Information System (INIS)

    Kao, P.C.; Jiang, N.S.; Klee, G.G.; Purnell, D.C.

    1982-01-01

    A radioimmunoassay for human parathyrin has been developed and characterized with whole-molecule (residues 1-84) human parathyrin and with the 1-34, 44-68,and 53-84 amino acid residue fragments of it. The antiserum used reacted with the whole molecule and with the 44-68 and 53-84 fragments, but not with the 1-34 fragment. Parathyrin concentrations in the serum of 118 normal subjects and of 112 patients with surgically proved primary hyperparathyroidism were determined with this assay. The mean results were 39 (SD 13) μL-Eq/mL for the normals and 111 (SD 77) μL-Eq/mL for the patients with primary hyperparathyroidism (p < 0.0005). The upper 95% confidence limit of the normal range was 60 μL-Eq/mL. For 54 patients with primary hyperparathyroidism, the preoperative values for serum parathyrin, calcium, and phosphate-but not creatinine-were statistically different from the postoperative values (paired t-test, p <0.0005). Normal subjects showed significant (p <0.0005) differences in serum calcium concentrations but not in parathyrin concentrations, compared with concentrations found in cancer patients and patients who had thiazide induced hypercalcemia. Phosphate concentration in serum, although not a specific indicator of disease, is a valuable clue to the diagnosis of primary hyperparathyroidism

  3. Radioimmunoassay of antidiuretic hormone in human urine. Applications

    International Nuclear Information System (INIS)

    Zebidi, Abdelkrim.

    1977-10-01

    This work is devoted mainly to the development of a radioimmunological system of antidiuretic hormone (ADH) determination in the urine and its physiological and pathological applications. The radioimmunological method thus replaces the biological measurement of antidiuretic hormone in the urine. This new technique was not possible until specific arginine vasopressin antibodies were obtained and a labelled hormone was prepared according to the criteria set for a radioimmunoassay. The labelled hormone is lysine vasopressin (greater stability). Although 125 I-LVP has lost most of its biological activity the molecule keeps all its immunological properties, behaving in the same way as non-iodinated synthetic LVP towards anti-LVP antibodies. Once specific antivasopressin antibodies and immunologically competent labelled hormone were available, conditions were defined for the radioimmunological ADH test in the urine. This technique, relatively easy to use, allows twenty samples to be measured simultaneously. With this sensitive, specific and reproducible method, it is thus possible to estimate the urinary ADH excretion rates from a 20 ml volume of urine after previous extraction on amberlite CG 50. This extraction method is aimed at both concentrating the hormone and eliminating non-specific interferences. The hormone extraction yield is about 92%+-8 [fr

  4. An improved method for estradiol-17B radioimmunoassay

    International Nuclear Information System (INIS)

    El-Banna, I.M.; El-Asrag, H.A.; Gamal, M.H.

    1991-01-01

    This work describes an improved radioimmunoassay (RIA) of serum estradiol-17 B (E) using locally generated immuno-chemicals. Estradiol hemisuccinate (E -3-H S) was prepared and conjugated to bovine serum albumin (BSA). The obtained conjugate; E 3-H S: BSA, hadλ max at 280 mu and the steroid BSA molar ratio was 25:1. The immunogen was injected subcutaneously in New Zealand rabbits and large amount of antiserum was harvested with 1 : 10500 antibody titre. The antibody cross reactions with estrone (E ), estriol (E ) and progesterone (P) were determined. Blood samples were collected from cycling Osemi ewes during follicular phase, pregnant ewes near term and daily from a cycling ewe over two consecutive estrous cycles. Serum samples were analysed for E both directly and after diethyl ether extraction (DE). The higher E values were found in the direct assay for pregnant ewes. The direct serum minnature RIA system, described herein, was found to be specific, sensitive, precise and economic.5 fig. 2 tab

  5. 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 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.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.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.An efficient HCC tissue-specific RNAi system (AFP-Cre/LoxP-shRNA was successfully established. The system provides a usable tool for HCC-specific RNAi

  6. Identification and subcellular localization of a 21-kilodalton molecule using affinity-purified antibodies against. cap alpha. -transforming growth factor

    Energy Technology Data Exchange (ETDEWEB)

    Hazarika, P.; Pardue, R.L.; Earls, R.; Dedman, J.R.

    1987-04-07

    Monospecific antibodies were generated against each of six different peptide sequences derived from rat and human ..cap alpha..-transforming growth factor (..cap alpha..-TGF). The affinity-purified antibody to the 17 amino acid carboxyl-terminal portion of the molecule proved most useful in detecting ..cap alpha..-TGF. When used in a peptide-based radioimmunoassay, it was possible to measure nanogram quantities of native ..cap alpha..-TGF in conditioned cell culture media. When used to analyze cell lysate, these antibodies specifically recognized a 21-kilodalton protein species. Indirect immunofluorescence localization procedures revealed a high concentration of ..cap alpha..-TCF in a perinuclear ring with a diffuse cytoplasmic distribution. These results suggest that a precursor form of ..cap alpha..-TGF has a cellular role beyond that of an autocrine growth factor.

  7. 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.

  8. Monitoring the reproductive performance in awassi ewes using progesterone radioimmunoassay

    International Nuclear Information System (INIS)

    Zarkawi, M.

    1999-01-01

    Jugular serum progesterone (P-4) concentrations were assessed using radioimmunoassay (RIA) kits for about 1 year in 16 local Awassi ewes aged about 1 year and weighing on average 37.4 ±4.9 (range 29.5-44.0 kg). Average pre-mating live weight of animals was different (P 0.05) between animals in average basal pre-mating P-4 concentration, maximum P-4 concentration during the luteal phase in non-pregnant ewes and in the length of the gestation period average basal pre-mating P-4 concentration was 0.62±0.44 (range 0-1.7 nmol l -1 ). Average maximum P-4 concentration during the luteal phase in non-pregnant ewes was 11.0±3.3 (range 5.8-18 nmol l -1 ). Length of gestation period varied from 149 to 155 days with an average of 152±2.3 days. Maximum P-4 concentration during gestation was different (P -1 ). The accuracy of pregnancy diagnosis at around 21 days pot-mating using RIA was 100%. It was concluded that Awassi sheep in Syria have a long breeding season that could start as early as April and last through September. Therefore, ewes might be included in oestrous synchronisation programmes to give three lambs in 2 years. It was also concluded that P-4 concentration under 3.18 nmol l -1 is indicative of either anoestrous or follicular and early luteal phase of the oestrous cycle in Awassi ewes kept in Syria and that RIA could be used for P-4 determination and successful early diagnosis of pregnancy in Awassi sheep. (author)

  9. Quality control of radioimmunoassay reagents for T4

    International Nuclear Information System (INIS)

    Wayan Radiatning, S.

    1987-01-01

    Quality control of radioimmunoassay reagents for T4. A program of quality control testing has been carried out for 125 I-T4, T4 standards and T4 antisera. 125 I-labelled T4 has been tested for its specific activity, radiochemical purity using a Sephadex G-25 column, immunological activity, based on the immunological reaction between labelled antigen and excess T4 antibody, and non-specific binding. The useful shelf-life of the labelled compound was determined by monitoring the decrease in radiochemical purity and immunological activity, and the increase in non-specific binding. T4 standards were calibrated by means of T4 RIA kit manufactured by DPC (Diagnostic Products Corporation). A test on parallelism was also performed using hyperthyroid sera. T4 antisera were evaluated with respect to titre, avidity and specifity. The test results on 125 I-T4 show a specific activity varying between 1830-2020 uCi/ug, a radiochemical purity above 90%, an immunological more than 80% and a non-specific binding of less than 5%. The standard curve for T4 was found to coincide well with the standard curve of the DPC kit and parallel with the curve for hyperthyroid sera. The titre of T4 antisera obtained was 1:300, the avidity was about 4.8 x 10 7 and the cross-reaction for T3 was 1.6%. It can be concluded from the experimental results, that the 125 I-T4, T4 standards and T4 antisera prepared meet the requirements for the manufacture of T4 kits. (author). 5 refs.; 14 figs

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

    International Nuclear Information System (INIS)

    Zarkawi, M.; Al-Masri, M. R.

    1998-12-01

    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)

  11. 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.

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

    International Nuclear Information System (INIS)

    Delgado S, B.; Lavalley E, C.; Ruiz J, A.; Garcia F, C.; Zamorano A, F.

    1991-12-01

    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)

  13. A method of quality control in continuous for the thyroid hormones radioimmunoassay

    International Nuclear Information System (INIS)

    Savoie, J.C.; Kawadry, G.; Leger, F.A.; Hantour, Z.; Baulieu, J.L.

    1980-01-01

    The exploration of the thyroid function requires nowadays the T4, T3 and TSH radioimmunoassays. These assays, performed several times each week, raise the problem as to how control the quality of every new series and of every result. The authors describe the principle features of a program devised to automatically control 'in continuous' the quality of the radioimmunoassays. This program was developed on a SIMIS 3-INFORMATEK system. It is derived from Rodbard's work, uses Logit-Log transformation and ensures the consistent interseries control of many characteristic parameters. No series nor individual results are exploited i.e. edited without prior control of quality a decision from which is made to validate or reject. This program is but one solution -among many others possible- corresponding to a necessity: the need presently felt and perhaps to-morrow requested for some permanent quality control of the results of radioimmunoassays [fr

  14. Quantitative determination of albumin in urine (microalbuminuria) by electroimmunoassay and radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Zanazzi, M.; Sverzellati, E.; Scarpioni, L.; Pantano, E.; Cerri, L.; Cecchettin, M.

    1986-10-01

    Electroimmunoassay and radioimmunoassay are two methods specifically designed for rapid and accurate analysis of subclinical amounts of proteins in urine. These assays are very sensitive to detect low concentrations of albumin in urines (17-75 mg/24 hrs) and to discover patients with microalbuminuria. Electroimmunoassay and radioimmunoassay are similar for precision and accuracy. Electroimmunoassay results can be obtained within few hours (3 hrs) and a remarkable advantage is the low cost of the equipment and the reagents. Radioimmunoassay results can be obtained within 2 hours. This assay is useful to perform multiple assays in big laboratories, but electroimmunoassay is much more convenient for little assays, when there are few urine samples to investigate.

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

    International Nuclear Information System (INIS)

    Meriadec, B.; Roubertie, P.

    1979-01-01

    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.)

  16. Expression and possible role of 20alpha-hydroxysteroid dehydrogenase in the placenta of the goat.

    Science.gov (United States)

    Jayasekara, Walimuni Samantha Nilanthi; Yonezawa, Tomohiro; Ishida, Maho; Yamanouchi, Keitaro; Nishihara, Masugi

    2005-04-01

    20Alpha-hydroxysteroid dehydrogenase (20alpha-HSD) catalyzes the conversion of progesterone to its inactive form 20alpha-dihydroprogesterone (20alpha-OHP). 20Alpha-HSD is expressed in the murine placenta, suggesting a role, yet unidentified, played by this enzyme during the course of pregnancy. To elucidate the possible roles of 20alpha-HSD during pregnancy, 20alpha-HSD gene expression in the placenta was examined by Northern blot analysis, and progestin (progesterone and 20alpha-OHP) concentrations in the maternal and fetal sera and the amniotic fluid were measured by radioimmunoassay in pregnant Shiba goats. The expression of 20alpha-HSD mRNA was observed in the placenta and the intercaruncular part of the uterus during mid to late pregnancy. Analysis by in situ hybridization revealed that 20alpha-HSD mRNA was mainly localized in the endometrial epithelium of the caruncle side of the placenta. Considerable enzyme activity of 20alpha-HSD was also detected in the cytosolic fraction of the placenta and intercaruncular part of the uterus. Although concentrations of progesterone and 20alpha-OHP in the maternal serum showed similar profiles, progesterone levels in the fetal serum stayed extremely low throughout the pregnancy. The 20alpha-OHP concentration in the fetal serum was always higher than that in the maternal serum. In the amniotic fluid, the concentrations of both progesterone and 20alpha-OHP remained at very low levels throughout the pregnancy. These results support the notion that 20alpha-HSD protects the fetus from the cytotoxic effects of progesterone, and thereby maintains the normal development of the fetus.

  17. Coulomb correction to elastic. alpha. -. alpha. scattering

    Energy Technology Data Exchange (ETDEWEB)

    Bera, P.K.; Jana, A.K.; Haque, N.; Talukdar, B. (Department of Physics, Visva-Bharati University, Santiniketan-731235, West Bengal, India (IN))

    1991-02-01

    The elastic {alpha}-{alpha} scattering is treated within the framework of a generalized phase-function method (GPFM). This generalization consists in absorbing the effect of Coulomb interaction in the comparison functions for developing the phase equation. Based on values of scattering phase shifts computed by the present method, it is concluded that the GPFM provides an uncomplicated approach to rigorous Coulomb correction in the {alpha}-{alpha} scattering.

  18. Development of a heterologous radioimmunoassay for the measurement of human prolactin in serum

    International Nuclear Information System (INIS)

    Vieira, J.G.H.; Russo, E.M.K.; Lengyel, A.M.J.; Maciel, R.M.B.

    1981-01-01

    The development of a heterologous radioimmunoassay method is described for the measurement of human prolactin in serum. Antisera were produced in rabbits using ovine prolactin as the immunogen; the best antiserum produced enebled the development of a simple and rapid method, with sufficient sensitivity (2ng/ml). The cross reactivity with human growth hormone and with placental lactogen was inferior to 0.03%. The labelling of human prolactin was done using the Lactoperoxidase method, followed by a column chromatography purification in a Sephadex G-100. Using 153 sera, the described method was compared with a homologous radioimmunoassay for human prolactin, and no significant difference was shown. (Author) [pt

  19. Development and characterization of a homologous radioimmunoassay for deer mouse (Peromyscus maniculatus bairdii) prolactin

    International Nuclear Information System (INIS)

    Marr, G.A.; Colosi, P.; Desjardins, C.; Talamantes, F.

    1983-01-01

    A highly specific and sensitive homologous radioimmunoassay has been developed for the secreted form of prolactin from the deer mouse Peromyscus maniculatus bairdii. Peromyscus serum and pituitary homogenates displayed paralled dilution response curves, and no cross reaction was seen with either mouse prolactin, mouse growth hormone or rat prolactin. The assay was sensitive to 25 picograms per tube and the intra- and inter-assay coefficients of variation were 5 and 3.6%, respectively. In addition, the authors have demonstrated that Peromyscus prolactin does not show parallel displacement in a homologous radioimmunoassay utilized for measuring prolactin in the common laboratory mouse

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

    Energy Technology Data Exchange (ETDEWEB)

    Vladutiu, A.O.; Sulewski, J.M.; Pudlak, K.A.; Stull, C.G.

    1982-11-19

    A young woman with amenorrhea had a consistently positive pregnancy test result (serum radioimmunoassay measurement of ..beta..-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.

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

    International Nuclear Information System (INIS)

    Vladutiu, A.O.; Sulewski, J.M.; Pudlak, K.A.; Stull, C.G.

    1982-01-01

    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

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

    International Nuclear Information System (INIS)

    Huet, S.

    1984-01-01

    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 [fr

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

    International Nuclear Information System (INIS)

    Agthe, O.

    1980-01-01

    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.) [de

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

    International Nuclear Information System (INIS)

    Pinto, H.; Werner, R.S.; Lerario, A.C.; Toledo e Souza, I.T. de; Wajchenberg, B.L.; Pieroni, R.R.

    1976-01-01

    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 [pt

  5. Polyacrylamide gel electrophoretic preparation of labelled and non-labelled peptides for radioimmunoassay

    International Nuclear Information System (INIS)

    Besch, W.; Woltanski, K.P.; Keilacker, H.; Kohnert, K.D.

    1986-01-01

    Radioiodinated polypeptide hormones, such as insulin, glucagon, human growth hormone, and human C-peptide are employed for radioimmunoassays for investigation of hormonal alterations in states of disturbed carbohydrate metabolism. Iodination was performed using chloramine T. Iodination products of these polypeptide hormones and, for preparation of standard material, native human C-peptide from cadaver pancreases were fractionated by polyacrylamide gel electrophoresis at pH 8.9. Disc electrophoresis in 24 cm long gel rods resulted in stable tracers with high specific activity as well as homogeneous standard material being highly suitable for radioimmunoassays. (author)

  6. CA 125 radioimmunoassay for the diagnosis and assessment of the effectiveness of therapy of ovarian cancer

    International Nuclear Information System (INIS)

    Sagan, D.L.; Chebotareva, Eh.D.; Evtushenko, G.V.

    1989-01-01

    The paper is concerned with the results of CA-125 radioimmunoassay in 163 patients (584 tests) with benign and malignant ovarian diseases and other small pelvic diseases which were diagnosed before operation as ovarian tumors. CA-125 radioimmunoassay can be used in combination with other methods for investigation of patients with suspected ovarian cancer and for differential diagnosis of malignant ovarian tumors to distinguish them from other malignant tumors of the small pelvis. The investigation of CA-125 indices in the course of disease was indicative of the effectiveness of antitumor therapy

  7. Intrahepatic expression of interferon alpha & interferon alpha ...

    African Journals Online (AJOL)

    kemrilib

    Alpha m-RNA while 30% only expressed Interferon Alpha Receptor m-RNA. Responders and non-responders to Interferon therapy ... expression of IFN Alpha Receptor mRNA. Regardless of the response to interferon, histological .... generation reverse hybridisation, line probe assay. (Inno-LiPA HCV II; Innogenetics, Ghent,.

  8. Production and use of thyroxine antisera in radioimmunoassay technique

    International Nuclear Information System (INIS)

    Abbas, Sumaia Hussein

    2000-06-01

    This study describes the production of antisera from sheep and its use in the determination of thyroxine hormone (T 4 ) level in serum using radioimmunoassay (RIA) technique. In this study two local sheep (Ovis aris) were subjected to immunization against human T 4 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-T 4 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 T 4 assay conditions including incubation temperature and reaction time were done. Tests for T 4 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 T 4 concentrations measured using the

  9. Measurement of plasma 11-deoxycorticosterone levels by radioimmunoassay in man

    International Nuclear Information System (INIS)

    Fukuchi, Soitsu; Nakajima, Katsuo; Takenouchi, Takahiko; Nishisato, Koji

    1974-01-01

    A radioimmunoassay procedure has been developed to measure 11-deoxycorticosterone (DOC) in human peripheral plasma. DOC-oxime was coupled with porcine gamma globulin and antibodies produced in rabbits. One to 3 ml of plasma, with 1, 2 3 H-DOC added for recovery, was extracted with dichloromethane and purification achieved by a silica gel column and by one paper chromatograph. After overnight incubation of the antibody-steroid mixture at 4 0 C, bound and free fractions were separated using ammonium sulfate. The mean recovery of 3 H-DOC, after extraction and chromatography, was 84.6 +- 7.4%. The method showed adequate specificity, precision and accuracy. Normal plasma DOC levels were found to be 4.4 +- 2.5 ng/100 ml (n=8). Plasma DOC levels were almost normal (0.3 - 26.8 ng/100 ml) in fifteen patients with benign essential hypertension. The mean level of 8.1 +- 8.2 ng/100 ml obtained in hypertensive patients with suppressed plasma renin activity, was not significantly different from normal. Plasma DOC showed a high level, 3.0 - 30.5 (11.4 +- 7.5) ng/100 ml, in 9 patients with primary aldosteronism. Four out of 8 patients with Cushing's syndrome were found to have elevated plasma DOC levels. Higher levels of 21.2 +- 15.8 ng/100 ml were found in 5 patients with adrenal hyperplasia than those of 12.3 +- 8.0 ng/100 ml in 3 with adrenal adenoma. Plasma DOC levels were high, 113 - 176 ng/100 ml, in 2 patients with 17α-hydroxylase deficiency. ACTH administered to 5 subjects produced a mean increase in plasma DOC from 4.8 to 25.8 ng/100 ml. Angiotensin II infused at a rate of 10 ng/kg/min for 30 min into 4 subjects did not increase mean plasma DOC. Similarly, dietary sodium restriction or postural change did not increase plasma DOC. (auth.)

  10. The radioimmunoassay of plasma oxytocin in pregnancy and at parturition

    International Nuclear Information System (INIS)

    Kogure, Satohisa

    1976-01-01

    The titer of the antiserum obtained by inoculating oxytocin-bovine serum albumin antigen into mature male rabbits was 1:64,000 in the final dilution, and the rate of conjugation with labeled-oxytocin was 34%. The cross reaction rate of the antiserum was 0.01% or below, both for lysine vasopressin and arginine vasopressin. The sensitivity of the radioimmunoassay using this antiserum was 5μIU. When a known quantity of oxytocin was added to plasma for measurement, the mean recovery rate was about 93%. The blood oxytocin concentration was 6.1+-2.5μIU/ml (ml omitted hereafter) in the early stage of pregnancy, 12.5+-6.0μIU in the middle stage, and 27.0+-7.5μIU in the terminal stage. The oxytocin concentration in the maternal blood was 34.1+-4.9μIU in the second stage, the concentration in the umbilical arterial blood immediately after delivery 3.5+-5.3μIU, and that in the umbilical venous blood 30.0+-4.2μIU. In cases of caesarean section not in labor, the oxytocin concentration in the maternal blood was 27.1+-6.6μIU, that in the umbilical arterial blood 25.1+-5.4μIU, and that in the umbilical venous blood 25.4+-5.4μIU. In cases of caesarean section in labor, the oxytocin concentration in the maternal blood was 37.1+-7.1μIU, that in the umbilical arterial blood 31.4+-6.7μIU, and that in the umbilical venous blood 27.0+-7.8μIU. The half-life of the oxytocin in the peripheral blood in cases injected with oxytocin was 5-10 minutes. When prostaglandin F2α was administered intravenously, the oxytocin concentration in the peripheral blood was increased in cases in labor. (Chiba, N.)

  11. Radioimmunoassay of inhibin: serum responses to unilateral and bilateral orchidectomy

    International Nuclear Information System (INIS)

    Schanbacher, B.

    1988-01-01

    An overnight double antibody RIA using a rabbit antiserum to porcine inhibin alpha-chain [Tyr30] (1-30) NH2 [pI alpha(1-30)], radioiodinated pI alpha(1-30), and a preprecipitated second antibody complex has been developed to measure inhibin concentrations in sera and other biological fluids. The assay is accurate, precise (intraassay coefficient of variation, 4.8%), sensitive (25 pM; 2.5 fmol/tube), and specific for inhibin. The synthetic reference standard pI alpha(1-30) produced a displacement curve that paralleled intact male ovine and bovine sera, crude bovine follicular fluid, and a partially purified porcine follicular fluid reference preparation (WHO/NIH 86/690). Bilateral castration of prepubertal and postpubertal ram lambs resulted in a rapid decrease in serum inhibin concentrations and a subsequent increase in serum FSH. Inhibin levels were high in prepubertal lambs (approximately 375 pM), but these levels were not sustained near the time of puberty (approximately 180 pM). Intensive sampling by jugular venipuncture after castration indicated a 50% drop in circulating inhibin levels within 2 h of testes removal with chronic castrate levels (approximately 75 pM) achieved by 6 h postcastration. A rapid fall in circulating levels of inhibin was also observed after unilateral castration, but these values stabilized within hours to levels intermediate (i.e. approximately 200 pM) to those of intact and bilateral castrate rams. Hemicastrates exhibited a more subtle rise in serum FSH after testis removal, with FSH and inhibin levels of prepubertal hemicastrates returning to mature intact ram values by 15 weeks of age. Serum inhibin levels remained low and FSH levels high at 14 days in unilateral castrate postpubertal rams. Inhibin immunoreactivity increased abruptly in castrate ewes and rams injected iv with 5 ml bovine follicular fluid

  12. Radioimmunoassay of inhibin: serum responses to unilateral and bilateral orchidectomy

    Energy Technology Data Exchange (ETDEWEB)

    Schanbacher, B.

    1988-11-01

    An overnight double antibody RIA using a rabbit antiserum to porcine inhibin alpha-chain (Tyr30) (1-30) NH2 (pI alpha(1-30)), radioiodinated pI alpha(1-30), and a preprecipitated second antibody complex has been developed to measure inhibin concentrations in sera and other biological fluids. The assay is accurate, precise (intraassay coefficient of variation, 4.8%), sensitive (25 pM; 2.5 fmol/tube), and specific for inhibin. The synthetic reference standard pI alpha(1-30) produced a displacement curve that paralleled intact male ovine and bovine sera, crude bovine follicular fluid, and a partially purified porcine follicular fluid reference preparation (WHO/NIH 86/690). Bilateral castration of prepubertal and postpubertal ram lambs resulted in a rapid decrease in serum inhibin concentrations and a subsequent increase in serum FSH. Inhibin levels were high in prepubertal lambs (approximately 375 pM), but these levels were not sustained near the time of puberty (approximately 180 pM). Intensive sampling by jugular venipuncture after castration indicated a 50% drop in circulating inhibin levels within 2 h of testes removal with chronic castrate levels (approximately 75 pM) achieved by 6 h postcastration. A rapid fall in circulating levels of inhibin was also observed after unilateral castration, but these values stabilized within hours to levels intermediate (i.e. approximately 200 pM) to those of intact and bilateral castrate rams. Hemicastrates exhibited a more subtle rise in serum FSH after testis removal, with FSH and inhibin levels of prepubertal hemicastrates returning to mature intact ram values by 15 weeks of age. Serum inhibin levels remained low and FSH levels high at 14 days in unilateral castrate postpubertal rams. Inhibin immunoreactivity increased abruptly in castrate ewes and rams injected iv with 5 ml bovine follicular fluid.

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

    International Nuclear Information System (INIS)

    Vaudry, Hubert; Leboulenger, Francois; Dupont, Willy; Vaillant, Rene

    1975-01-01

    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 [fr

  14. Method for the optimum formation of angiotensin I from angiotensinogen and radioimmunoassay for angiotensin I

    International Nuclear Information System (INIS)

    Pagnucco, R.G.; Murty, D.R.; Muse, R.J.

    1978-01-01

    The invention deals with a method to optimize the formation of angiotensin I from angiotensinogens by optimizing the temperature, pH value and buffer system. Furthermore, the invention concerns a radioimmunoassay method to determine angiotensin I. Iodine 125 labelled angiotensin I is preferably used as tracer. (VJ) [de

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

    International Nuclear Information System (INIS)

    Castro, A.; Ibanez, J.; DiCesare, J.L.; Adams, R.F.; Malkus, H.

    1978-01-01

    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

  16. Therapeutic drug monitoring by radioimmunoassay: Determination of aminoglycoside antibiotics and vancomycin in plasma

    International Nuclear Information System (INIS)

    Glaubitt, D.; Drechsler, H.J.; Knoch, K.; Siafarikas, K.

    1984-01-01

    Radioimmunoassay of aminoglycoside antibiotics (gentamicin, tobramycin, netilmicin) or vancomycin in plasma may considerably aid to assess their appropriate dosage and, if necessary, to rapidly adjust it to the assumed requirement. Thus the dosage of the antibiotic is kept large enough as to lead to the desired therapeutic result but not as high as to cause side effects. (orig.)

  17. A novel radioimmunoassay of 7-oxo-DHEA and its physiological levels

    Czech Academy of Sciences Publication Activity Database

    Kazihnitová, H.; Zamrazilová, L.; Hill, M.; Lapčík, O.; Pouzar, Vladimír; Hampl, R.

    2007-01-01

    Roč. 72, č. 4 (2007), s. 342-350 ISSN 0039-128X Grant - others:GA MZd(CZ) NR7815 Institutional research plan: CEZ:AV0Z40550506 Keywords : 7-oxo-dehydroepiandrosterone * radioimmunoassay * GC-MS * serum levels Subject RIV: CC - Organic Chemistry Impact factor: 2.143, year: 2007

  18. Preparation, purification and characterization of 125I-glucagon used for radioimmunoassay

    International Nuclear Information System (INIS)

    Ruehlmann, C.; Poege, A.; Stollmaier, P.; Akademie der Wissenschaften der DDR, Leipzig. Zentralinstitut fuer Isotopen- und Strahlenforschung)

    1988-01-01

    A method for the radioactive labelling of glucagon with Na 125 I with the help of chloramine T for the use in radioimmunoassay is described. The purification of 125 I-glucagon with polyacrylamide gel electrophoresis, the characterization of the tracer, its stability and the immunological properties are described. (author)

  19. Comparison of luteinizing hormone and human chorionic gonadotrophin determinations by biological, radioreceptor and radioimmunoassay methods

    International Nuclear Information System (INIS)

    Lange, E.; Paszko, Z.; Padzik, H.

    1980-01-01

    A comparison of the radioreceptor method of assaying LH and HCG with the biological and radioimmunoassay methods is carried out on several preparations of gonadotrophins: human pituitary luteinizing hormone, human menopausal gonadotrophin and human chorionic gonadotrophin. The radioreceptor method better reflects the biological properties of hormones than the radioimmunological method. (author)

  20. Simultaneous detection of Hepatitis B surface antigen and its antibody by radioimmunoassay

    International Nuclear Information System (INIS)

    Crouzat-Reynes, Gerard; Perigois, Francois; Lecureuil, Michel; Lejeune, Bernard

    1981-01-01

    The authors describe an original radioimmunoassay which allows the simultaneous detection of hepatitis B surface antigen and its antibody in a biological sample. Antigen and antibody are indiscriminately detected in a first step and then distinguished in a second step using the same reagents [fr

  1. Filter radioimmunoassay, a method for large-scale serotyping of Neisseria meningitidis.

    NARCIS (Netherlands)

    S. de Marie (Siem); J.H.J. Hoeijmakers (Jan); J.T. Poolman; H.C. Zanen

    1984-01-01

    textabstractA simple and rapid filter radioimmunoassay method can be used to serotype meningococcal strains on a large scale. The technique consists of simultaneous inoculation of 96 strains on nitrocellulose filters. The resulting colonies can be processed in situ, by extraction and fixation,

  2. Elevated serum free thyroxine by thyroxine analog radioimmunoassays in euthyroid patients with familial dysalbuminemic hyperthyroxinemia

    International Nuclear Information System (INIS)

    Rajatanavin, R.; Fournier, L.; DeCosimo, D.; Abreau, C.; Braverman, L.E.

    1982-01-01

    A study was done to ascertain whether the serum free T4 measured by free T4 radioimmunoassay kits would, like equilibrium dialysis, be normal in patients with familial dysalbuminemic hyperthyroxinemia. Five free T4 radioimmunoassay kits were used to measure free T4 in serum samples from 19 patients with familial dysalbuminemic hyperthyroxinemia and 20 healthy volunteers. Values (mean +/- SE) for T4, free T4 index, and free T4 (equilibrium dialysis) in these normal subjects and patients with familial dysalbuminemic hyperthyroxinemia, respectively, were as follows: T4, 8.1 +/- 0.2 and 18.3 +/- 0.7 μg/dL; free T4 index, 3.1 +/- 0.1 and 7.3 +/- 0.3 μg/dL; free T4, 1.4 +/- 0.1 and 1.2 +/- 0.1 ng/dL. The following free T4 radioimmunoassay methods were used: antibody coated microfine silica, microencapsulated antibody, two-step antibody-coated tube, and one-step 125 I-T4 analog (2 kits). The present findings in patients with familial dysalbuminemic hyperthyroxinemia and previous observations in ill euthyroid patients suggest that serum free T4 measured by some radioimmunoassay methods must be interpreted with caution in these two clinical situations

  3. 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.

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

    Science.gov (United States)

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

    1983-03-01

    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.

  5. Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2)

    DEFF Research Database (Denmark)

    Orskov, C; Holst, J J

    1987-01-01

    Gene-sequencing studies have shown that the glucagon precursor contains two additional glucagon-like sequences, the so-called glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). We developed radio-immunoassays against synthetic peptides corresponding to these sequences. Antisera were raised in rabb...

  6. Conditions and limits of serum LH radioimmunoassay in normal, hypophysectomised or castred rats

    International Nuclear Information System (INIS)

    Andre, M.; Boucher, D.; Thieblot, L.

    1976-01-01

    Serum LH was measured by radioimmunoassay (NIAMD Kits) free and linked hormones were separated by double antibodies method. Influence of concentration on antibody-hormone complex is studied. Hypophysectomised rats serum does not modify results. The standard (rat LH-RPl) has the same action as serum LH. Rat serum LH contents are measured in normal or castred rats [fr

  7. 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.

  8. Simultaneous breast cancer and hepatocellular carcinoma in a male patient with alcoholic liver cirrhosis and a normal serum alpha-fetoprotein level

    Directory of Open Access Journals (Sweden)

    Li-Chun Kao

    2017-06-01

    Laparoscopic cholecystectomy and an excision of the left breast mass were performed. The breast mass was pathologically confirmed to be infiltrating ductal carcinoma not otherwise specified and with predominantly cystic degeneration. The patient received chemotherapy with tamoxifen for the breast cancer, and transarterial chemoembolization for the HCC. Concomitant breast cancer and HCC in a male patient were rarely reported in the literature. Herein, we have discussed the possible relationship of these two disease entities.

  9. Differential vascular endothelial growth factor A protein expression between small hepatocellular carcinoma and cirrhosis correlates with serum vascular endothelial growth factor A and alpha-fetoprotein.

    Science.gov (United States)

    Corradini, Stefano Ginanni; Morini, Sergio; Liguori, Francesca; Carotti, Simone; Muda, Andrea Onetti; Burza, Maria Antonella; Siciliano, Maria; Molinaro, Antonio; Cantafora, Alfredo; Blotta, Ida; Merli, Manuela; Berloco, Pasquale; Rossi, Massimo; Attili, Adolfo Francesco; Gaudio, Eugenio

    2009-01-01

    Drugs with antivascular endothelial growth factor A (anti-VEGF-A) action are under clinical evaluation with encouraging results in advanced hepatocellular carcinoma (HCC). The relative VEGF-A protein expression in non-advanced HCC and in the cirrhotic non-tumoral tissue in the same patient, a variable that could be important for treatment efficacy, has been investigated with conflicting results, only using the cirrhotic tissue surrounding the neoplasm (CS). We measured, for the first time, VEGF-A expression in non-advanced HCC and in the respective CS and cirrhotic tissue at a distance from the tumour (CD), in 24 patients who underwent liver transplantation. VEGF-A protein was more expressed (Pprotein expression in HCC was higher than in the corresponding CD in 83% of cases and AFP and serum VEGF-A corrected for the platelet count positively correlated with the differential VEGF-A protein expression between HCC and CD. Our data provide a rationale for clinical trials involving anti-VEGF-A treatments in patients with non-advanced HCC, and suggest that serum AFP and VEGF-A are variables to be taken into account in these studies.

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

    International Nuclear Information System (INIS)

    Kaminogawa, S.; Shimoda, M.; Kurisaki, J.; Yamauchi, K.

    1989-01-01

    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

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

    Science.gov (United States)

    Kaminogawa, S; Shimoda, M; Kurisaki, J; Yamauchi, K

    1989-05-01

    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.

  12. Radioimmunoassay measurement of urinary ligandin excretion in nephrotoxin-treated rats

    International Nuclear Information System (INIS)

    Bass, N.M.; Kirsch, R.E.; Tuff, S.A.; Campbell, J.A.; Saunders, J.S.

    1979-01-01

    A radioimmunoassay (RIA) was used to establish normal urine excretion of ligandin, a renal tubular protein, in rats, and to study the pattern of ligandin excretion after nephrotoxin administration. Validation of RIA measurement of ligandin in urine is described. A normal range of 2-46 ng/h was obtained for ligandin excretion in rats (n = 24). The time course of ligandin excretion in HgCl 2 and K 2 Cr 2 0 7 treated rats correlated with the histological sequence of damage to the pars recta and pars convoluta of the proximal tubule respectively. Results confirm that renal ligandin is confined mainly to the pars recta of the proximal tubule and small quantities of the protein are present in the pars convoluta. Radioimmunoassay provides a sensitive and specific means for measuring ligandin in urine and is a valuable tool for the detection and study of renal tubular damage. (author)

  13. The application of α1-MG radioimmunoassay in the diagnosis of kidney diseases

    International Nuclear Information System (INIS)

    Liu Guifen; Wang Jianchun; Zhang Qingfeng; Liu Shuying; Han Wensheng; Li Zhihong

    1996-01-01

    A new method for the diagnoses of kidney diseases with α 1 -microglobulin (α 1 -MG) radioimmunoassay is developed. 276 healthy persons' serum and urine are determined by α 1 -MG radioimmunoassay. The result shows that the serum normal number is 54.22 +-22.02 mg/l and the urine 5.57 +- 2.63 mg/l. In contrast, the counterparts of 358 patients with different diseases are also examined. The result shows that there is a remarkable difference between urine α 1 -MG content of patients suffering from such diseases as gestation hypertension, renal insufficiency, nephritis and that of the healthy group (P<0.01). The method provides a stable, sensitive index for the diagnoses of kidney diseases and the examination of kidney function. It can be widely used in the clinical diagnosis

  14. Free light chains of immunoglobulins in normal serum and urine determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Soelling, K.

    1975-01-01

    A radioimmunoassay for determination of free light polypeptide chains from human immunoglobulins in serum and urine is described. The free light chains were isolated from the regular immunoglobulins by gel filtration on Sephadex G-100 columns. In the radioimmunoassay the free and antibody-bound 125 I-labelled light chains were separated by gel filtration through Sephadex G-200 columns. The mean concentration of free light chains in serum from 20 normal subjects was 10.6 mg/l for lambda chains and 13.2 mg/l for kappa chains. The 24-hr urinary excretion was 1.1mg for lambda chains and 3.2 mg for kappa chains. (auth)

  15. Development and application of radioimmunoassay and enzyme immunoassays in microbiological and immunological diagnosis. 4

    International Nuclear Information System (INIS)

    Struy, H.; Horeschi, G.; Wolffgang, H.; Morenz, J.

    1982-01-01

    Indirect solid phase radioimmunoassays and enzyme immunoassays were developed for the quantitative determination of low concentrations of IgG, IgA, IgM, and IgE. PVC blister and microtiterplates of polystyrene with adsorptively bound purified immunoglobulins were used as carrier. The determination of the immunoglobulins was accomplished with a uniform system of indicators, which consists of class-specific antihuman immunoglobulin sera from rabbits and of labelled ( 125 I and horseradish peroxydase, resp.) antirabbitglobulin from sheep. The developped competitive radioimmuno- and enzyme immunoassays allows the determination of concentrations of classes of immunoglobulins down to nearly 1 ng/ml, and the indirect sandwich solid phase radioimmunoassay determinations of IgE down to about 1 IU/ml. The values of IgG determined in liquors and those of IgE found in sera correspond very well to values which were measured with commercial kits. (author)

  16. Radioimmunoassay methods for the determination of L-triiodo-thyronine and thyroxine

    International Nuclear Information System (INIS)

    1976-01-01

    An improved, simplified radioimmunoassay method for the in vitro determination of L-triiodo-thyronine in unextracted blood serum is described which involves the use of a combination reagent constituted by a buffered solution containing radioactive L-triiodothyronine and an inhibitor for inhibiting the binding of L-triiodothyronine to thyroxine-binding globulin. Optionally the reagent may also include an antiserum containing an antibody capable of immunoreactivity with L-triiodothyronine. Packaged test kits for use in conveniently carrying out the radioimmunoassay are also provided. Certain salts of 8-anilino-1-naphtalene sulfonic acid, which may be regarded as purified forms of the acid, are preferably employed as inhibitors for inhibiting binding of L-triiodothyronine to thyroxine-binding globulin

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

    International Nuclear Information System (INIS)

    Verdeja I, C.E.

    1994-01-01

    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 (T 4 ), Thyrotropin (TSH) and Triyodotironine (T 3 ) 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)

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

    International Nuclear Information System (INIS)

    Chebotareva, Eh.D.; Shishkina, V.V.; Korolev, V.I.; Sklyar, S.Yu.; Kukot', V.A.

    1989-01-01

    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

  19. Radioimmunoassay screening and GC/MS confirmation of whole blood samples for drugs of abuse

    Energy Technology Data Exchange (ETDEWEB)

    Spiehler, V.R.; Sedgwick, P.

    From 1981 to 1984, an average of 300 radioimmunoassay screens on whole blood were performed each week in the authors laboratory. Most samples were screened for opiates phencyclidine and its analogs, barbiturates, and cocaine or its metabolite benzoylecgonine. A commercially available radioimmunoassay was used with modifications to facilitate screening of whole blood. Increasing sample size increased the sensitivity of the assay. Changing reagent concentration (1:1 dilution), incubation time, sample matrix (water, urine, or blood), or fraction counted (precipitate or supernatant) did not affect the utility of the standard curve or the sensitivity of the assay. All positive results for phencyclidine, opiates, cocaine, and related compounds were confirmed by GC/MA. Barbiturate positives were confirmed by UV spectrophotometry.

  20. Increased specificity of plasma glucagon radioimmunoassay by use of acetone extraction

    International Nuclear Information System (INIS)

    Schenck, H. von

    1983-01-01

    The specificity of glucagon determinations by radioimmunoassay with two pancreatic-type glucagon antisera has been determined by means of gel permeation chromatography before and after treatment of human plasma with acetone and ethanol. Acetone precipitates the high molecular mass substances which interfere with the antisera directed against the carboxyl-terminal region of the glucagon molecule and applied in the radioimmunoassay, and which are apparent in the void volume and cross-reacting precursor fragments, while pancreatic glucagon is extracted to slightly more than 50%. Hence acetone treatment is suggested as a suitable means of obtaining specific glucagon results in single samples. Ethanol also precipitates the high molecular mass substances, but partly extracts the precursor fragments in addition to the glucagon. Ethanol treatment is therefore suggested for chemical diagnosis of the glucagonoma syndrome in which the tumor often produces precursor fragments. (Auth.)

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

    International Nuclear Information System (INIS)

    Scheidt, C.

    1982-01-01

    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) [de

  2. The role of carbohydrates in the radioimmunoassay of human low-molecular-mass kininogen

    International Nuclear Information System (INIS)

    Turpeinen, U.; Kaerkkaeinen, T.

    1985-01-01

    The immunoreactivity of human low-molecular-mass kininogen from Cohn plasma fraction IV was investigated after deglycosylations and carbohydrate modifications by radioimmunoassay using the conformation-specific antiserum. Removal of all sialic acids, 44% of amino sugars and 63% of neutral sugars did not alter the immunoreactivity of the protein but the periodate-treated concanavalin A fractions showed strikingly diminished immunoreactivity. A conformational change could account for the observed effect of periodate on the decreased reactivity of the protein in radioimmunoassay. Externally added carbohydrates had no effect on immunoreactivity. The results suggest that the carbohydrate part of kininogen is not involved in the immunoreactivity although it accounts for the observed lectin-binding heterogeneity. (Auth.)

  3. Radioimmunoassay determination of antigenic concordance among hemagglutinins of vaccine and epidemic influenza virus strains

    Energy Technology Data Exchange (ETDEWEB)

    Blokha, V.V.; Yamnikova, S.S.; Karpovich, L.G.; Yakhno, M.A.; Zakstel' skaya, L.Ya.

    Radioimmunoassay studies were conducted on the antigenic concordance of hemagglutinin of influenza A H3N2 viruses, to determine the suitability of vaccine strains in engendering immunity against viruses circulating in nature. Specifically, the inhibition studies involved the hemagglutinins of the A/Victoria/35/72 vaccine strain, the proposed vaccine strain A/Khabarovsk/15/76, and the RK-5 recombinant strains, containing antigenic determinants of viruses isolated in the 1972-1976 epidemic period (A/Victoria/3/75, A/Leningrad/173/75, A/Victoria/112/76). The results showed that A/Victoria/35/72 is becoming less important as a vaccine, but that RK-5 and A/Khabarovsk/15/76 can provide significant immunity with respect to influenza viruses circulating in 1975-1976. These observations point to the usefulness of radioimmunoassay in assessing the suitability of influenza A viruses for vaccine production. 12 references, 2 figures.

  4. Radioimmunoassay of renin-angiotensin-aldosterone in patients with adrenal tumors

    International Nuclear Information System (INIS)

    Slavnov, V.N.; Yakovlev, A.A.; Yugrinov, O.G.; Gandzha, T.I.

    1983-01-01

    The results are presented of a study of the renin-angiotensin-aldosterone system in 89 patients with aldosteronoma, corticosteroma, pheochromocytoma and hypertension. Radioimmunoassay was used to measure aldosterone concentration and renin activity in the peripheral blood and blood from vena cava inferior, the renal and adrenal veins, the circadian cycle of their content and the responsiveness of the glomerular zone of the adrenal cortex and the juxtaglomerular renal system under the influence of lasix intake and the change over from a horizontal into vertical position. Patients with adrenal tumors have shown disorders of renin-angiotensin-aldosterone function. Radioimmunoassay of the renin-angiotensin-aldosterone system promotes early detection of adrenal tumors in the general population of patients with hypertension and can be used for control over therapeutic efficacy

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

    International Nuclear Information System (INIS)

    Rauch, P.; Fukal, L.; Brezina, P.; Kas, J.

    1988-01-01

    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 B 1 /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

  6. Radioimmunoassay for Zearalenone and Zearalanol in Human Serum: Production, Properties, and Use of Porcine Antibodies

    OpenAIRE

    Thouvenot, Daniel; Morfin, Robert F.

    1983-01-01

    To produce antigens susceptible to raise antibodies for resorcylic acid lactones, the 6′-carboxymethyloxime derivatives of zearalenone and zearalanone were bound to bovine serum albumin. Pigs could be immunized by using these antigens, the best titer in antibodies being obtained with the zearalenone antigen. The porcine antibodies were specific for the resorcylic acid lactones of structural resemblance with zearalenone. This specificity made the antibodies usable for a radioimmunoassay of zea...

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

    Energy Technology Data Exchange (ETDEWEB)

    Rauch, P.; Fukal, L.; Brezina, P.; Kas, J.

    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 ..mu..g aflatoxin B/sub 1//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.

  8. Radioimmunoassay of ferritin, CEA and prolactin for assessment of prognosis and efficacy of breast cancer therapy

    International Nuclear Information System (INIS)

    Sklyar, S.Yu.; Chebotareva, Eh.D.; Ganul, V.L.; Korolev, V.I.

    1987-01-01

    The levels of tumor markers - CEA, deritin and prolactin (the hormone of the anterior pituitary gland) were studied in the blood serum of 178 breast cancer patients in the menopause during combined therapy using a radioimmunoassay. This combination was shown to be informative for assessment of prognosis of disease and efficacy of antitumor therapy which was confirmed by clinical and x-ray findings and observation of a tumor process in breast cancer patients over a 5-year period

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

    International Nuclear Information System (INIS)

    Viana, F.A.B.; Pessoa, J.M.; Biondini, J.

    1991-01-01

    Serum levels of triiodothyronine (T 3 ) and thyroxine (T 4 ) 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 T 3 (P 4 (P<0.05). (author). 10 refs, 1 tab

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

    International Nuclear Information System (INIS)

    Mesquita, C.H. de; Silva, C.P.G. da; Hamada, M.M.

    1985-11-01

    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 - 125 I 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) [pt

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

    International Nuclear Information System (INIS)

    Toledo e Souza, I.T. de; Giannella Neto, D.; Wajchenberg, B.L.

    1988-05-01

    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) [pt

  12. Use of a rapid hCG-beta-subunit radioimmunoassay in acute gynaecological emergencies

    International Nuclear Information System (INIS)

    Seppala, M.; Tontti, K.; Ranta, T.; Stenman, U.-H.; Chard, T.

    1980-01-01

    Chorionic gonadotrophin (hCG) was measured by a rapid and sensitive radioimmunoassay in 100 female patients with lower abdominal pain attending an emergency department. Of the 24 subjects with a positive result, 22 had evidence of pregnancy (ectopic or uterine). Of the 76 patients with negative results, 3 had some evidence of pregnancy. This test may be of considerable practical value in the differential diagnosis of acute abdominal emergencies in women of reproductive age. (author)

  13. Clarification of the technic of intrinsic factor radioimmunoassay for the cat and the rat

    International Nuclear Information System (INIS)

    Perrin, M.O.; Nicolas, J.P.; Dubrasquet, M.

    1983-01-01

    Cats and rats secrete very small quantities of gastric juice (GJ). In order to measure the Intrinsic Factor (IF) in GJ of animals certain modifications are necessary. It is very important to select with great care pernicious anemia sera which will be used to cross react with the animals' IF to determine their IF level by radioimmunoassay. The technic described here enables the dosage of low quantities of IF with a 7 per cent coefficient of variation [fr

  14. A simplified radioimmunoassay for digoxin determination using a 125-I-labelled solid-phase kit

    International Nuclear Information System (INIS)

    Doering, W.; Bluemel, E.

    1978-01-01

    Our experience with a commercially available kit (Radioimmunoassay DIGOXIN, Boehringer, Mannheim) using ( 125 J)-labelled digoxin and antibody-coated tubes is reported. This simplified method requires only two pipetting steps per sample and results can be obtained in 70 min. The intra- and interassay coefficient of variation ranged between 7% and 8%. The specific digoxin antibody antibody gave no clinical relevant cross-reactions with spironolactone or prednisone ( [de

  15. Three magnetic particles solid phase radioimmunoassay for T4: Comparison of their results with established methods

    International Nuclear Information System (INIS)

    Bashir, T.

    1996-01-01

    The introduction of solid phase separation techniques is an important improvement in radioimmunoassays and immunoradiometric assays. Magnetic particle solid phase method has additional advantages over others, as the separation is rapid and centrifugation is not required. Three types of magnetic particles have been studied in T 4 RIA and the results have been compared with commercial kits and other established methods. (author). 4 refs, 9 figs, 2 tabs

  16. Standardization of the method for measurement of plasma estrone by radioimmunoassay

    International Nuclear Information System (INIS)

    Vilanova, M.S.V.; Moreira, A.C.; Sala, M.M. de; Sa, M.F.S. de

    1994-01-01

    The present paper has as objective standardize a radioimmunoassay method for measurement of plasma estrone. Ethyl ether was used for plasma extraction. The sensitivity (Minimal detectable dose) was 3,7 pg/tube; the reproducibility (inter assay error) was 8,6%; the precision (intra assay error) was 4,1%. As a biological control the plasma estrone was ml) and in 24 patients with polycystic ovarian syndrome (median = 77,9 pg/ml). (author). 6 refs, 2 figs, 2 tabs

  17. Anti-DNA antibody in serum measured by radioimmunoassay (Farr technique)

    International Nuclear Information System (INIS)

    Manthorpe, R.; Palit, J.; Bendixen, G.

    1978-01-01

    Anti-DNA antibody determination in serums is increasingly used because it supports the diagnosis of systemic lupres erythematosus (SLE) with high selectivity. The present work evaluates several of the technical variables of the Farr radioimmunoassay for anti-DNA antibody determination and describes a recommendable procedure, emphasizes the most important sources of error and gives the range of normal blood donors and a group of hospital patients without SLE. (author)

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

    International Nuclear Information System (INIS)

    Baltabaev, T.S.

    1989-01-01

    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 T 3 , T 4 , 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

  19. Comparison of the methods for tissue triiodothyronine T(3) extraction and subsequent radioimmunoassay

    International Nuclear Information System (INIS)

    Takaishi, M.; Miyachi, Y.; Aoki, M.; Shishiba, Y.; Asahi Life Foundation, Tokyo

    1978-01-01

    Although there have been various reports on tissue T 3 concentration, the examination of the quality of radioimmunoassay has not been available. In the present study, we tried to determine whether the available methods for T 3 extraction are adequate for the various methods of T 3 radioimmunoassays used. T 3 was extracted from liver by ethanol extraction or by acid butanol extraction (Flock's method) and the extract was applied to radioimmunoassay either by Seralute T 3 column, ANS-double antibody or the ANS-charcoal method. The values of T 3 were compared with those obtained by isotope-equilibration method. The dilution curve of ethanol extract was not parallel with that of the standard in ANS-charcoal or ANS-double antibody technique. When the extract was tested by Seralate method, the dilution curve was parallel to the standard, whereas the T 3 value obtained with this method was two-fold higher than that with the isotope equilibration technique. The analysis of the ethanol extract suggested that the lipid extracted by ethanol interfered with the assay. The acid butanol extract when tested either by the ANS-double antibody or Seralate method, showed parallelism to the standard curve and gave T 3 values almost identical with those by the isotope-equilibration method. When tested by ANS-charcoal method, the dilution curve of the acid butanol extract was not parallel to the standard. Thus, to obtain reliable results, tissue extraction by Flock's method and subsequent T 3 radioimmunoassay by either ANS-double antibody or Seralate T 3 method are recommended. (author)

  20. An improved radioimmunoassay method for the detection of IgG antibodies against cytomegalovirus

    International Nuclear Information System (INIS)

    Berry, N.J.; Grundy, J.E.; Griffiths, P.D.

    1986-01-01

    The non-specific binding seen with human sera in a radioimmunoassay for the detection of IgG antibodies specific for CMV can be reduced greatly by using a murine monoclonal antibody as a radiolabelled detecting antibody. Such non-specific binding formerly obtained with a polyclonal detecting antibody was due to the binding of the polyclonal reagent to factors on the solid phase other than IgG molecules. (Auth.)

  1. Ovulation following gonadotrophin treatment and determination of serum estradiol and progesterone by radioimmunoassay

    International Nuclear Information System (INIS)

    Vesper, B.; Lisse, K.; Ittrich, G.

    1979-01-01

    The use of gonadotrophins to release ovulation is indicated in women with urgent desire for children and with negative response to clomiphene therapy. 69 patients treated with 100 therapeutic gonadotrophin series were examined. Estrogens and progesterone were determined by radioimmunoassay. The treatment resulted in ovulation in 54 women, 12 patients developed an ovarian reaction with an increase of estrogen, and in 34 cases no ovarian response was detectable. 6 women became pregnant during gonadotrophin therapy and further 2 after treatment

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

    International Nuclear Information System (INIS)

    Poland, R.E.; Rubin, R.T.

    1981-01-01

    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

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

    International Nuclear Information System (INIS)

    Lando, V.S.

    1992-01-01

    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)

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

    International Nuclear Information System (INIS)

    Bernhard, U.

    1979-01-01

    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) [de

  5. Development of an enzyme-radioimmunoassay for the measurement of dopamine in human plasma and urine

    International Nuclear Information System (INIS)

    Faraj, B.A.; Walker, W.R.; Camp, V.M.; Ali, F.M.; Cobbs, W.B. Jr.

    1978-01-01

    An enzyme-radioimmunoassay for the measurement of dopamine is described. It is based on the incubation of plasma or urine in the presence of catechol-0-methyltransferase and S-adenosylmethionine. The 0-methylated dopamine metabolite formed (3-0-methyldopamine) was characterized by radioimmunoassay. As little as 0.5 ng of dopamine can be detected. The assay was found to be specific, since no cross-reactivity was noted for several compounds related to dopamine. The enzyme-radioimmunoassay of dopamine was used to determine the concentrations of dopamine in urine and plasma of normal volunteers. In this group, urinary dopamine averaged 182.1 +- 2.2 μg/24 hr, and the plasma concentration 0.211 +- 0.052 ng/ml. However, in children wPth neuroblastoma, there was a several-fold increase over controls in the average urinary and plasma levels of dopamine (8,500 μ/24 hr and 2.3 ng/ml). The assay was also used to monitor blood levels of dopamine following the administration of L-dopa and dopamine to patients with cardiomyopathy

  6. Creatine kinase radioimmunoassay and isoenzyme electrophoresis compared in the diagnosis of acute myocardial infarction

    International Nuclear Information System (INIS)

    Homburger, H.A.; Jacob, G.L.

    1980-01-01

    We compared, in 116 patients, the relative usefulness of results of tests for creatine kinase B-isoenzymes, as measured by radioimmunoassay, and the MB isoenzyme, as measured by electrophoresis, in diagnosis of acute myocardial infarction. The radioimmunoassay was specific for isoenzymes of creatine kinase containing the B subunit. All patients with acute transmural infarcts had positive test results by both techniques, but concentrations of B-isoenzymes were more frequently above normal than were MB bands in the case of patients with acute subendocardial infarcts and in the case of all patients with acute myocardial infarcts from whom sera were collected more than 24 h after onset of chest pain. Concentrations of B-isoenzymes also were increased, even when MB bands were not electrophoretically detectable in specimens from several patients without documented acute myocardial infarcts. These abnormal results presumably were caused by increased concentrations of the BB isoenzyme in serum. Accordingly, an increased concentration of B-isoenzymes had less diagnostic specificity and predictive value for acute myocardial infarction than did a detectable MB band. Results of isoenzyme electrophoresis were more reliable for establishing this diagnosis, but the results of radioimmunoassay were more reliable for excluding it in patients with chest pain as the primary symptom

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

    Energy Technology Data Exchange (ETDEWEB)

    Bagshaw, A.F.; Whicher, J.T. (Bristol Royal Infirmary (UK)); Bhoola, K.D.; Lemon, M.J.C. (Bristol Univ. (UK). Medical School)

    1984-05-01

    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 Na/sup 125/I 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 ..mu..g 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.

  8. A photoelectrochemical immunosensor for detection of α-fetoprotein based on Au-ZnO flower-rod heterostructures

    Energy Technology Data Exchange (ETDEWEB)

    Han, Zhizhong, E-mail: zzhan@fjmu.edu.cn; Luo, Min; Chen, Li; Chen, Jinghua; Li, Chunyan, E-mail: cyli65@126.com

    2017-04-30

    Highlights: • Au NPs were deposited onto ZnO FRs by cyclic voltammetry methods. • Au NPs were used to enhance light absorption and photo-induced charge separation. • The PEC immunosensor based on Au-ZnO FRs presented a higher photocurrent. • The detection limit of this novel PEC immunosensor was 0.56 pg mL{sup −1} for AFP. - Abstract: In this work, a novel label free photoelectrochemical (PEC) immunosensor has been developed for the detection of α-fetoprotein (AFP). The immunosensor was based on Au-ZnO flower-rods (FRs) heterostructure, where Au nanoparticles (NPs) were firstly electrodeposited by cyclic voltammetry methods. Scanning electron microscopy (SEM), X-ray diffraction (XRD), Mott-Schottky plot (MS), UV–vis diffuse reflectance spectrum and fluorescence emission spectrum were used for the characterizations of Au-ZnO FRs. The results demonstrated that Au NPs not only obviously enhanced the visible light absorption of ZnO FRs due to surface plasmon resonance (SPR) but also improved the separation of photo-generated electron-hole pairs. Therefore, the photocurrent of Au-ZnO FRs was increased under simulated sunlight. The photocurrent was reduced after the specific antibody-antigen immune reaction. And the photocurrent decrement was linear with the logarithm of AFP antigen concentration in the range from 0.005 ng mL{sup −1} to 50 ng mL{sup −1} with a low detection limit of 0.56 pg mL{sup −1} (S/N = 3). The PEC immunosensor also exhibited high anti-interference property and acceptable stability. This work would provide a promising photoelectrochemical strategy for the detection of other proteins in clinical diagnosis.

  9. The determination of $\\alpha_s$ by the ALPHA collaboration

    CERN Document Server

    Bruno, Mattia

    2016-01-01

    We review the ALPHA collaboration strategy for obtaining the QCD coupling at high scale. In the three-flavor effective theory it avoids the use of perturbation theory at $\\alpha > 0.2$ and at the same time has the physical scales small compared to the cutoff $1/a$ in all stages of the computation. The result $\\Lambda_\\overline{MS}^{(3)}=332(14)$~MeV is translated to $\\alpha_\\overline{MS}(m_Z)=0.1179(10)(2)$ by use of (high order) perturbative relations between the effective theory couplings at the charm and beauty quark "thresholds". The error of this perturbative step is discussed and estimated as $0.0002$.

  10. Identification of human basic fetoprotein as glucose-6-phosphate isomerase by using N- and C-terminal sequence tags and terminal tag database.

    Science.gov (United States)

    Kuyama, Hiroki; Yoshizawa, Akiyasu C; Nakajima, Chihiro; Hosako, Mutsumi; Tanaka, Koichi

    2015-08-10

    Human basic fetoprotein (BFP), found in fetal serum and tissue extracts as well as in extracts of various cancer tissues, has long been known as a marker protein for cancers; however, the primary sequence has not yet been reported. This paper describes the identification of BFP using the N- and C-terminal amino acid sequence tags (Ac-AALTRDPQFQ and QQREARVQ, respectively) clarified by mass spectrometry-based methods, and a terminal tag database (ProteinCarta). In this study, BFP was identified as glucose-6-phosphate isomerase (G6PI_HUMAN). Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Lyman Alpha Control

    CERN Document Server

    Nielsen, Daniel Stefaniak

    2015-01-01

    This document gives an overview of how to operate the Lyman Alpha Control application written in LabVIEW along with things to watch out for. Overview of the LabVIEW code itself as well as the physical wiring of and connections from/to the NI PCI-6229 DAQ box is also included. The Lyman Alpha Control application is the interface between the ALPHA sequencer and the HighFinesse Wavelength Meter as well as the Lyman Alpha laser setup. The application measures the wavelength of the output light from the Lyman Alpha cavity through the Wavelength Meter. The application can use the Wavelength Meter’s PID capabilities to stabilize the Lyman Alpha laser output as well as switch between up to three frequencies.

  12. New ALPHA-2 magnet

    CERN Multimedia

    Anaïs Schaeffer

    2012-01-01

    On 21 June, members of the ALPHA collaboration celebrated the handover of the first solenoid designed for the ALPHA-2 experiment. The magnet has since been successfully installed and is working well.   Khalid Mansoor, Sumera Yamin and Jeffrey Hangst in front of the new ALPHA-2 solenoid. “This was the first of three identical solenoids that will be installed between now and September, as the rest of the ALPHA-2 device is installed and commissioned,” explains ALPHA spokesperson Jeffrey Hangst. “These magnets are designed to allow us to transfer particles - antiprotons, electrons and positrons - between various parts of the new ALPHA-2 device by controlling the transverse size of the particle bunch that is being transferred.” Sumera Yamin and Khalid Mansoor, two Pakistani scientists from the National Centre for Physics in Islamabad, came to CERN in February specifically to design and manufacture these magnets. “We had the chance to work on act...

  13. Alpha Shapes and Proteins

    DEFF Research Database (Denmark)

    Winter, Pawel; Sterner, Henrik; Sterner, Peter

    2009-01-01

    We provide a unified description of (weighted) alpha shapes, beta shapes and the corresponding simplicialcomplexes. We discuss their applicability to various protein-related problems. We also discuss filtrations of alpha shapes and touch upon related persistence issues.We claim that the full...... potential of alpha-shapes and related geometrical constructs in protein-related problems yet remains to be realized and verified. We suggest parallel algorithms for (weighted) alpha shapes, and we argue that future use of filtrations and kinetic variants for larger proteins will need such implementation....

  14. Targeted Alpha Therapy: From Alpha to Omega

    International Nuclear Information System (INIS)

    Allen, Barry J; Clarke, Raymond; Huang Chenyu

    2013-01-01

    This review covers the broad spectrum of Targeted Alpha Therapy (TAT) research in Australia; from in vitro and in vivo studies to clinical trials. The principle of tumour anti-vascular alpha therapy (TAVAT) is discussed in terms of its validation by Monte Carlo calculations of vascular models and the potential role of biological dosimetry is examined. Summmary of this review is as follows: 1. The essence of TAT 2. Therapeutic objectives 3. TAVAT and Monte Carlo microdosimetry 4. Biological dosimetry 5. Preclinical studies 6. Clinical trials 7. What next? 8. Obstacles. (author)

  15. Effect of twenty minutes of aerobic exercise on in vivo platelet release in moderately trained females: radioimmunoassay of platelet factor 4 and beta-thromboglobulin

    International Nuclear Information System (INIS)

    Rudmann, S.V.

    1986-01-01

    Circulating blood platelets serve an important role in the physiological process of hemostasis. Physical exercise has been documented to result in alterations in many hemostatic parameters including platelet size, number and function. Most published research data support the hypotheses that both hemostasis and fibrinolysis become activated as a consequence of various levels of physical exercise. The purpose of this study was to determine the effect of twenty minutes aerobic exercise on platelet activation in vivo. Platelet activation in vivo is associated with the release of platelet granular contents. Platelet alpha granules contain two platelet specific proteins: platelet factor 4 (PF4) and beta-thromboglobulin (BTG). Elevated plasma levels of these proteins are a specific marker of in vivo platelet activation. Subjects were moderately trained female volunteers between the ages of 22 and 40 years. Subjects were exercised or twenty minutes on a bicycle ergometer at workloads that represented 65 to 75% of their functional capacity. Blood specimens were drawn within five minutes of exercise. Plasma samples from exercise and control subjects were assayed for PF4 and BTG using a sensitive competitive-binding radioimmunoassay procedure. The mean plasma levels of both proteins were significantly greater in the exercising subjects when compared with the non-exercising controls. Data from this study support the following research hypotheses: BTG plasma levels will be significantly higher in exercising subjects than in non-exercising controls, and PF4 plasma levels will be significantly higher in exercising subjects than in non-exercising controls

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

    International Nuclear Information System (INIS)

    Peig Ginabredra, M.G.

    1989-01-01

    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 125 I 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 4 0 C. 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) [pt

  17. Radioimmunoassay for human myoglobin: methods and results in patients with skeletal muscle or myocardial disorders

    International Nuclear Information System (INIS)

    Miyoshi, K.; Saito, S.; Kawai, H.; Kondo, A.; Iwasa, M.; Hayashi, T.; Yagita, M.

    1978-01-01

    A sensitive and specific radioimmunoassay has been developed for the measurement of serum Mb. Immunization of rabbit with human Mb yielded anti-Mb antibody which was purified by affinity chromatography. Human hemoglobin, CK, and the component of serum per se did not appear to cross-react with the antibody. Mb was radiolabeled by the chloramine T method. The radioimmunoassay method could detect as little as 0.3 ng of Mb and was not affected by hemolysis. Information is also given on precision, recovery, and specimen preservation. Mb levels could be detected in all of 120 normal adults, and the values ranged between 1 and 28 ng/ml (mean, 13.1 +- 6.1). No sex difference was observed. Levels were markedly elevated in all the patients with progressive muscular dystrophy, especially in the Duchenne type at the level of 40 to 1700 ng/ml. It was also noticed that about 70% of female gene carriers of Duchenne type had a slightly increased Mb level. An elevated serum Mb was also noted in polymyositis. In every case of acute myocardial infarction, serum Mb levels were increased, peak values ranging from 175 to 4400 ng/ml and averaging 1162 +- 287.9 Mb levels were elevated faster and peaked earlier (within 6 to 12 hr after the attack) than serum CK activity and returned to nearly normal range within 3 to 4 days. The increase in serum Mb was also noticed in shock and surgery. These data indicate that radioimmunoassay of Mb is a useful test for judging the myolytic state of myogenic myopathies and for early detection of myocardial infarction

  18. beta. -endorphin-like and. alpha. -MSH-like immunoreactivities in human milk

    Energy Technology Data Exchange (ETDEWEB)

    Ferrando, T.; Rainero, I.; De Gennaro, T.; Oggero, R.; Mostert, M.; Dattola, P.; Pinessi, L. (Univ. of Turin (Italy))

    1990-01-01

    We measured with radioimmunoassay the {beta}-endorphin-like and {alpha}-MSH-like immunoreactivities in milk and plasma of 8 lactating women. Mean {beta}-endorphin concentrations ({plus minus} SD) were 16.6 {plus minus} 6.7 fmol/ml in milk and 9.9 {plus minus} 4.1 fmol/ml in plasma. {alpha}-MSH concentrations were 39.4 {plus minus} 15.5 pg/ml in milk and 18.2 {plus minus} 8.4 pg/ml in plasma. The concentrations of both peptides in milk were significantly higher than in plasma. No significant correlation between milk and plasma concentrations of these peptides was found.

  19. Radioimmunoassay and chromatographic similarity of circulating endogenous gonadotropin releasing hormone and hypothalamic extracts in man. [/sup 125/I tracer technique

    Energy Technology Data Exchange (ETDEWEB)

    Mortimer, C.H. (St. Bartholomew' s Hospital, London); McNeilly, A.S.; Rees, L.H.; Lowry, P.J.; Gilmore, D.; Dobbie, H.G.

    1976-10-01

    A highly sensitive radioimmunoassay for the gonadotropin releasing hormone has been developed in order to study its physiological importance in man. In view of the expected low concentrations in peripheral blood, large volumes of human plasma were extracted by two different methods and the characteristics of the radioimmunoassayable material compared with those of synthetic decapeptide and extracts of human hypothalami. The results indicate that radioimmunoassayable gonadotropin releasing hormone is present in some human plasmas but the plasma concentrations are less than 2.5 pg/ml. Peripheral levels were more consistently measurable in women at midcycle and after the menopause. The hormone was undetectable in the plasma of normal men, human cerebrospinal fluid, and fetal cerebral tissue, but was present in fetal hypothalami.

  20. A radioimmunoassay method for the rapid detection of Candida antibodies is experimental systematic candidiasis

    International Nuclear Information System (INIS)

    Huang, Y.; Berry, W.; Cooper, H.; Zachariah, Y.; Newman, T.

    1979-01-01

    Rabbits were employed as experimental models to evaluate a solid-phase radioimmunoassay (RIA) method for the diagnosis of systematic candidiasis. Ten rabbits were incubated subcutaneously to mimic superficial candidiasis and were found to produce no antibodies to Candida as determined by both immunodiffusion and RIA procedures. However, 94 per cent of 18 rabbits systematically infected by intravenous injection of Candida cells were observed to produce antibody as assessed by the RIA technique. These data encourage further tests with human sera and the continued development of this RIA procedure as a useful tool in the early serodiagnosis of systematic candidiasis. (Auth.)

  1. Direct radioimmunoassay for the detection of barbiturates in blood and urine

    International Nuclear Information System (INIS)

    Mason, P.A.; Law, B.; Pocock, K.; Moffat, A.C.

    1982-01-01

    A radioimmunoassay has been developed for the detection of barbiturates in blood and urine without any pre-treatment of the sample. It is based on a radioiodinated derivative of 4-hydroxyphenobarbitone which allows use of relatively simple gamma-counting procedures. The assay can detect therapeutic levels of barbiturates in very small amounts (50 μl) of blood and urine samples. It is cheap, rapid, simple to perform and is broadly specific for the barbiturate class of drugs to the exclusion of related drugs. The assay is, therefore, very well suited to the task of screening large numbers of samples for the presence of barbiturates. (author)

  2. Numerical proceessing of radioimmunoassay results using logit-log transformation method

    International Nuclear Information System (INIS)

    Textoris, R.

    1983-01-01

    The mathematical model and algorithm are described of the numerical processing of the results of a radioimmunoassay by the logit-log transformation method and by linear regression with weight factors. The limiting value of the curve for zero concentration is optimized with regard to the residual sum by the iterative method by multiple repeats of the linear regression. Typical examples are presented of the approximation of calibration curves. The method proved suitable for all hitherto used RIA sets and is well suited for small computers with internal memory of min. 8 Kbyte. (author)

  3. Presence of melatonin in various cat brainstem nuclei determined by radioimmunoassay

    International Nuclear Information System (INIS)

    Sallanon, M.; Touret, M.; Claustrat, B.

    1982-01-01

    Microdissected samples of juvenile cat brain tissue were assayed for melatonin content using a double antibody radioimmunoassay. Immunoreactive melatonin was consistently detected, albeit in variable amounts, in pineal, habenula, the region of the nucleus gracilis, gigantocellular reticular formation of the pons and medulla oblongata. Among the negative areas were raphe nuclei, substantia nigra dn locus caeruleus. These findings suggest that melatonin may play a role in some structures of the central nervous system outside the pineal-hypothalamo-pituitary axis. This immunoreactive melatonin could reflect a local synthesis, or a tissular uptake of melatonin from blood or cerebrospinal fluid. (author)

  4. A rapid solid-phase radioimmunoassay for human plasma follicle-stimulating hormone

    International Nuclear Information System (INIS)

    Lovesey, A.C.

    1980-01-01

    The measurement of plasma levels of human follicle-stimulating hormone (FSH) has proved to be of value for the study of the hypothalamic-hypophyseal-gonadal axis, greatly facilitating the diagnosis and management of problems relating to the menopause and infertility. A solid-phase radioimmunoassay for human FSH has been developed. This system is characterised by high precision, is economical, and is considerably faster to operate than conventional double antibody systems used in the hospital assay service. Reference values for plasma FSH in various endocrine states are recorded and discussed. (author)

  5. A radioimmunoassay for antibodies against surface membrane antigens using adhering cells

    International Nuclear Information System (INIS)

    Tax, A.; Manson, L.A.

    1976-01-01

    A radioimmunoassay using cells adhering to plastic is described. In this assay, A-10 mammary carcinoma attached to the surface of plastic in microtiter plates were permitted to bind antibody and the bound antibody was detected with purified rabbit 125 I-antimouse-Fab. The bound radioactive material was eluted with glycine-HCl buffer (pH 2.5), and the acid eluates were counted in a gamma counter. This assay can be used to detect cytolic or noncytolic antibody to cell surface antigens in studies with any tumor or normal cell that will adhere to a solid surface

  6. Results of the HPL determination using a new enzymatic immunoassay - comparison with a commercial radioimmunoassay

    International Nuclear Information System (INIS)

    Leis, D.; Schneider, B.; Keller, A.; Braun, S.

    1982-01-01

    An enzymatic immunoassay recently developed by the firm Organon, Oss, Netherlands, to determine HPL in the serum of pregnant women was compared to a widely used radioimmunoassay marketed by the firm Amersham-Buchler, Braunschweig. The values determined showed a good correlation. The accuracy of measurement was within the generally accepted range for immunological assays of this kind. The assay procedures are comparable with regard to the expenditure of time and work. A great advantage of the EIA is due to the absence of radioactive substances, which permits this test to be performed in any laboratory equipped with a centrifuge and photometer. (orig.) [de

  7. Evaluation of a radioimmunoassay for the determination of anti-native DNA antibodies

    International Nuclear Information System (INIS)

    Johanet, C.; Soulie, E.; Absalon, Y.B.; Ocwieja, T.; Abuaf, N.

    1991-01-01

    The anti-native DNA antibodies were measured by a radioimmunoassay (RIA) type Farr assay in the sera from 648 patients: 108 with active or inactive systemic lupus erythematosus (SLE), 181 with clinical symptoms of another connective tissue disease, 171 with liver diseases, 29 with different pathology and 159 normal sera were obtained from a blood bank. The anti-DNA kit has been calibrated against the first international units/ml. This assay has proved to be sensitive and specific, and appears to be reliable for the diagnosis and follow-up of SLE patients. The authors propose a new reference cut-off level higher than producer's one [fr

  8. Monoclonal antibodies to human chorionic gonadotropin and their application to two-site sandwich radioimmunoassay

    International Nuclear Information System (INIS)

    Mizuchi, A.; Iio, M.; Miyachi, Y.

    1984-01-01

    Monoclonal antibodies were prepared against human chorionic gonadotropin (HCG). One monoclonal antibody recognized a conformational determinant expressed only on native HCG molecule and another monoclonal antibody had the specificity for the epitopes located on the β-subunit of HCG. Monoclonal antibodies reacting with different antigenic determinants on the HCG molecule were used to develop a simplified 2-site sandwich radioimmunoassay in which one monoclonal antibody was immobilized and another labeled with 125 iodine. This assay was highly specific for HCG and there was no cross-reactivity with α,β-subunit of HCG, luteinizing hormone and follicle stimulating hormone. (Auth.)

  9. Radioimmunoassay of class-specific antibodies to Streptococcus mutans in monkey serum and saliva

    International Nuclear Information System (INIS)

    Walker, J.; Colman, G.; Huges, M.

    1979-01-01

    A radioimmunoassay (RIA) has been developed to measure class-specific antibodies to Streptococcus mutans in the serum and saliva of monkeys (Macaca fascicularis). Antihuman immunoglobulin antibodies purified by affinity chromatography on immobilised monkey immoglobulins and labelled with 125 I were employed. Formalised cells of S. mutans and an extract of culture supernatant adsorbed to polystyrene wells were used as solid-phase antigens. The coefficients of variation of IgG, IgA, and IgM assays were less than or equal to 10% for both antigen systems. It is shown that this RIA is a sensitive, reproducible and quantitative method. (Auth.)

  10. Prenatal sex determination by radioimmunoassay of testosterone with and without chromatography of the amniotic fluid

    International Nuclear Information System (INIS)

    Distler, W.; Boniver-Ollmann, U.; Tigges, J.; Terinde, R.; Claussen, U.

    1979-01-01

    Amniotic fluid testosterone measured by radioimmunoassay (RIA) without chromatography (immunoreactive testosterone) seems not to be a definitive test for prenatal sex determination in all cases. In this study testosterone (T) levels measured by RIA with chromatography of the amniotic fluid samples were compared with immunoreactive testosterone (iT) values, to determine the predictive accuracy of the two methods. In 111 amniotic fluid samples between 15 and 19 weeks of gestation iT and T were measured parallelly. There are significant differences between iT- and T-means of both sexes (p [de

  11. Diagnosis application of ACTH radioimmunoassay in diseases of hypothalamus, hypophysis and adrenal axis

    International Nuclear Information System (INIS)

    Moreira, A.C.; Foss, M.C.; Iazigi, N.

    1988-01-01

    The diagnostic value of 900-1,100 am plasma ACTH radioimmunoassay were studied in 10 patients with Cushing's disease before and after treatment, three patients with Cushing's syndrome with adrenal tumours, one Nelson's syndrome patient; 13 patients with Addison's disease and 12 patients with hypo-pituitarism. Twenty-seven normal subjects were controls. The measurement of basal plasma ACTH gave good differentiation between: a. pituitary Cushing's disease from adrenal tumors; b. Addison's disease from hypo-pituitarism. However this assay has a limited value for the differentiation between Cushing's disease from normal subjects and it is often unhelpful in the differential diagnosis of hypo-pituitarism from normal subjects. (author)

  12. Radioimmunoassay for the middle region of human parathyroid hormone: comparison of two radioiodinated synthetic peptides

    Energy Technology Data Exchange (ETDEWEB)

    Sharp, M.E.; Marx, S.J. (National Institute of Arthritis, Diabetes, and Digestive and Kidney Diseases, Bethesda, MD (USA))

    1985-01-01

    Two synthetic peptides were evaluated to develop radioligands for midregion-specific radioimmunoassay (RIA) of human parathyroid hormone (hPTH). Both radioligands were tested using three anti-PTH sera of proven clinical utility. While each of these midregion-directed antisera showed unique specificity, they all reacted with high affinity with both radioligands and none of them discriminated significantly between the two synthetic midregion peptides. Analysis of data on the relation of serum calcium and hPTH midregion immunoreactivity showed a useful separation of groups (all nonazotemic) with primary hyperparathyroidism, secondary hyperparathyroidism, primary hypoparathyroidism and secondary hypoparathyroidism.

  13. Radioimmunoassay for vasopressin: plasma levels after stimulation by I-V nicotine injection

    International Nuclear Information System (INIS)

    Conte-Devolx, B.; Rougon-Rapuzzi, G.; Millet, Y.

    1976-01-01

    A radioimmunoassay for vasopressin which permits the estimation of plasma levels as low as 0.25μIU/ml was developed. The average plasma level of ADH after overnight water restriction was found to be 5.20μIU/ml. A hypersecretion of ADH was provoked by the intravenous injection of 1-2mg of nicotine. In 10 volunteer subjects this stimulation by nicotine provoked ADH secretion which reached a maximum between 2 and 15 minutes after injection; the hormone plasma level reached an average value of 21.3μIU/ml [fr

  14. A sensitive radioimmunoassay of parathyroid hormone in human serum using a specific extraction procedure

    International Nuclear Information System (INIS)

    Christensen, M.S.

    1976-01-01

    A radioimmunoassay of human parathyroid hormone (PTH) is described. PTH was extracted from serum by adsorption to and elution from microfine silica, causing a 3.2-fold greater hormone concentration in extract than in serum. The extraction procedure eliminated from the assay unspecific interference caused by serum factors. The detection limit was 10 pg bovine PTH. The concentration of PTH was measurable in serum from 95 % of normal subjects (mean, 68 pg/ml; S.D., 18), was undetectable in sera from hypoparathyroid patients, and elevated in 96 % of sera from patients with primary hyperthyroidism. The data presented suggest that the assay mainly measures PTH(1-84). (Auth.)

  15. Prospective study of radioimmunoassay for antibodies against neutrophil cytoplasm in diagnosis of systemic vasculitis

    Energy Technology Data Exchange (ETDEWEB)

    Savage, C.O.S.; Winearls, C.G.; Jones, S.; Marshall, P.D.; Lockwood, C.M.

    1987-06-20

    The diagnosis and management of Wegener's granulomatosis and microscopic polyarteritis are complicated by the lack of specific diagnostic tests. The diagnostic performance of a solid-phase radioimmunoassay, which detects the autoantibodies against neutrophil cytoplasm present in these disorders, was assessed in a prospective study of patients with suspected vasculitis and/or rapidly progressive nephritis. The assay had a sensitivity and specificity of 96% when carried out in combination with a specific inhibition stage and indirect immunofluorescence staining of alcohol-fixed normal neutrophils.

  16. Radioimmunoassay of CA 19-9 tumor marker in the diagnosis of thyroid cancer

    International Nuclear Information System (INIS)

    Markov, V.V.; Slavnov, V.N.; Komissarenko, I.V.; Kovpak, N.A.; Kovalenko, A.E.; Guda, B.B.

    1999-01-01

    Applicability of determining carbohydrate antigen CA 19-9 content in blood serum, tissue extracts, and thyroid tumor aspiration biopsy samples to the differential diagnosis of benign and malignant tumors of thyroid is studied. Radioimmunoassay was used for measurements. It is shown that determination of marker CA 19-9 in blood serum is not informationally capable for the differential diagnosis of thyroid tumors. Considerable increase in CA 19-9 concentration was found in tumor aspiration biopsy samples from patients with malignant tumors this fact can be used for preoperative diagnosis of thyroid cancer [ru

  17. Use of radioimmunoassay procedures for the determination of sex hormones in animal tissues

    International Nuclear Information System (INIS)

    Hoffmann, B.

    1983-01-01

    Radioimmunoassay methods for the determination of sex steroids and other compounds with sex hormone-like activities in various edible animal tissues and endocrine glands have been developed. Reliability of these methods, allowing quantification in a range of 10 -11 M, has been adequately demonstrated. When applied to monitoring residues of anabolic sex hormones in edible tissues of veal calves, physiological baseline levels of some endogenous ''anabolic'' steroids (like testosterone, oestrogens) were established; in the case of xenobiotics residues at the scheduled time of slaughter could be quantified (trenbolone) and a regulatory method to implement the ban of diethylstilbestrol was introduced. (author)

  18. Use of radioimmunoassay procedures for the determination of sex hormones in animal tissues

    Energy Technology Data Exchange (ETDEWEB)

    Hoffmann, B. (Institut fuer Veterinaermedizin des Bundesgesundheitsamtes (Robert von Ostertag-Institut), Berlin (Germany, F.R.))

    1983-07-01

    Radioimmunoassay methods for the determination of sex steroids and other compounds with sex hormone-like activities in various edible animal tissues and endocrine glands have been developed. Reliability of these methods, allowing quantification in a range of 10/sup -11/ M, has been adequately demonstrated. When applied to monitoring residues of anabolic sex hormones in edible tissues of veal calves, physiological baseline levels of some endogenous ''anabolic'' steroids (like testosterone, oestrogens) were established; in the case of xenobiotics residues at the scheduled time of slaughter could be quantified (trenbolone) and a regulatory method to implement the ban of diethylstilbestrol was introduced.

  19. Radioimmunoassay of gastrin level in duodenal ulcer, atrophic gostritis and Addison-Biermer's disease

    International Nuclear Information System (INIS)

    Hasik, J.; Kozal, H.; Kosowicz, J.; Hansz, J.

    1975-01-01

    Radioimmunoassay of gastrin level in the blood was performed in 20 controls, 12 patients with duodenal ulcer, 13 patients with atrophic gastritis and 14 patients with Addison-Biermer's disease. Gastrin level in the serum of the patients with duodenal ulcer did not differ significantly from that of controls. In atrophic gastritis and particularly in Addison-Biermer's disease gastrin level was found to be several times higher. This is probably a result of chronic gastrin secretion stimulation which is normally inhibited by gastric juice. (author)

  20. Prospective study of radioimmunoassay for antibodies against neutrophil cytoplasm in diagnosis of systemic vasculitis

    International Nuclear Information System (INIS)

    Savage, C.O.S.; Winearls, C.G.; Jones, S.; Marshall, P.D.; Lockwood, C.M.

    1987-01-01

    The diagnosis and management of Wegener's granulomatosis and microscopic polyarteritis are complicated by the lack of specific diagnostic tests. The diagnostic performance of a solid-phase radioimmunoassay, which detects the autoantibodies against neutrophil cytoplasm present in these disorders, was assessed in a prospective study of patients with suspected vasculitis and/or rapidly progressive nephritis. The assay had a sensitivity and specificity of 96% when carried out in combination with a specific inhibition stage and indirect immunofluorescence staining of alcohol-fixed normal neutrophils. (author)

  1. Specific radioimmunoassay of HCG and its α and β subunits: methods and results

    International Nuclear Information System (INIS)

    Reuter, A.M.; Schoonbrood, J.; Franchimont, P.

    1976-01-01

    To create antisera that are specific for the radioimmunoassay of HCG and its subunits, the antisera are neutralized by incubation with LH or HCG. For each RIA system the inhibition curves of HCG and its subunits LH, FSH, TSH and STH are obtained. The 125 I labelled hormones HCG, α and β subunits and LH were chromatographed over a Sephadex G 100 column. Serum of menopausal and pregnant women were chromatographed in the same way and the fractions subjected to RIA. HCG and its subunits were determined by RIA in the sera of patients with different kinds of cancer

  2. A radioimmunoassay for the detection of diethylstilboestrol and related stilbenes in biological fluids

    International Nuclear Information System (INIS)

    Hallahan, Cornelius; McGarry, Yvonne; Collins, J.D.

    1985-01-01

    A radioimmunoassay for the measurement of the synthetic anabolic agent diethylstilboestrol (DES) is described. It is based on a commercially available antiserum and a tritiated derivative of DES. The method can detect low concentrations of residues (less than 0.5 ng/ml) in small samples (0.05 to 0.2 ml) of biological fluids. DES was measured in plasma, bile and urine obtained from a calf slaughtered 22 days after subcutaneous implantation of 24 mg DES. The assay described is suitable as a rapid screening procedure for identifying animals treated with stilbene substances. (author)

  3. Isolation of a somatomedin binding protein from human preterm amniotic fluid: development of a radioimmunoassay

    International Nuclear Information System (INIS)

    Drop, S.L.S.

    1983-01-01

    This thesis investigates the nature and biological behaviour of a somatomedin binding protein, identified in preterm amniotic fluid (AF). For that purpose a double antibody radioimmunoassay was developed. Purified AF binding protein (AFBP) was iodinated by the chloramine-T method, and dilutions of partially purified AFBP were designated as the standard, with the results expressed in μg equivalent protein/ml. The sensitivity of the assay was improved by adoption of the nonequilibrium procedure. AFBP values were twice as high in preterm AF as in term AF. (Auth.)

  4. Application of pregnancy specific β1 glycoprotein-radioimmunoassay (SP1-ria) in obstetrics and gynecology

    International Nuclear Information System (INIS)

    Zhang Weiyuan

    1988-01-01

    Serum SP 1 values of 395 patients were determined by radioimmunoassay. It was found that SP 1 was apparently absent from the blood of normal men and nonpregnant women, increased with gestational stage in pregnant women, and was highest in late pregnancy. In high-risk pregnancy, SP 1 was lower than normal pregnamey group (PC0.01) could also be detected in ectopic pregnancy. In patients with benign and malignant hydatidiform mole, and choriocarcinoma, AP 1 level decreased with malignant degree. The authors suggest that measurement of SP 1 levels is a valuable index for monitoring high-risk pregnancy, diagnosing ectopic pregnancy and following-up trophoblastic cell diseases

  5. Radioimmunoassay of pharmacologically active compounds: applications to nicotine and its metabolites

    International Nuclear Information System (INIS)

    Langone, J.J.; Vunakis, H. van

    1979-01-01

    Since nicotine is the major tobacco alkaloid, its role in cardiovascular disease, cancer and other physiological effects of cigarette smoking has been the subject of intensive investigation for many years. As part of this investigation into the biochemical and physiological effects of nicotine, the authors developed sensitive and specific radioimmunoassays for the parent alkaloid, a major metabolite cotinine, and for γ-(3-pyridyl)-γ-oxo-N-methylbutyramide (oxoamide). The oxoamide is a minor metabolite derived from cotinine. The authors describe the development of these RIA's and demonstrate their use in determining levels of these compounds in physiological fluids of smokers and in studying metabolic processes in vitro. (Auth.)

  6. Preparation of quality control samples in radioimmunoassay for thyroid stimulating hormone (TSH)

    International Nuclear Information System (INIS)

    Ali, O.M.

    2006-03-01

    To days, the radioimmunoassay is becomes the best technique to analysis different concentrations of substance, especially in medical and research laboratories. Although the specificity of RIA techniques, the quality controls must takes place to give good results as possible. In this dissertation i prepared quality control samples of thyroid stimulating hormone (TSH), to use it in RIA techniques and to control the reliability results of those laboratories which used these methods. We used China production kits of RIA method to determine the level of hormone (low-normal-high) concentration. Statistical parameters were used to drown the control chart of the mean to these data.(Author)

  7. Preparation of quality control samples for thyroid hormones T3 and T4 in radioimmunoassay techniques

    International Nuclear Information System (INIS)

    Ahmed, F.O.A.

    2006-03-01

    Today, the radioimmunoassay becomes one of the best techniques for quantitative analysis of very low concentration of different substances. RIA is being widely used in medical and research laboratories. To maintain high specificity and accuracy in RIA and other related techniques the quality controls must be introduced. In this dissertation quality control samples for thyroid hormones (Triiodothyronine T3 and Thyroxin T4), using RIA techniques. Ready made chinese T4, T3 RIA kits were used. IAEA statistical package were selected.(Author)

  8. Radioimmunoassay for the middle region of human parathyroid hormone: comparison of two radioiodinated synthetic peptides

    International Nuclear Information System (INIS)

    Sharp, M.E.; Marx, S.J.

    1985-01-01

    Two synthetic peptides were evaluated to develop radioligands for midregion-specific radioimmunoassay (RIA) of human parathyroid hormone (hPTH). Both radioligands were tested using three anti-PTH sera of proven clinical utility. While each of these midregion-directed antisera showed unique specificity, they all reacted with high affinity with both radioligands and none of them discriminated significantly between the two synthetic midregion peptides. Analysis of data on the relation of serum calcium and hPTH midregion immunoreactivity showed a useful separation of groups (all nonazotemic) with primary hyperparathyroidism, secondary hyperparathyroidism, primary hypoparathyroidism and secondary hypoparathyroidism. (Auth.)

  9. Computer assessment of the informative value of radioimmunoassays for the diagnosis of stomach cancer

    International Nuclear Information System (INIS)

    Ozherel'ev, A.S.; Tkacheva, G.A.; Glazkova, T.G.

    1987-01-01

    Solution rules in the form of differential-diagnositc tables with coefficients of weight and threshold values of 16 different signs were obtained with the help of a computer EC-assisted algorithm of a generalized picture. These signs included tumor antigens, cyclonucleotides and hormones determined in the blood serum of healthy individuals, stomach cancer patients and patients with diseases of nonmalignant origin by means of radioimmunoassay. These signs being of low informative value by themselves, in a combined use provided an opportunity to differentiate between ''stomach cancer-not cancer'' with a sufficiently high degree of significance

  10. Sample preparation for measuring cow's progesterone in radioimmunoassay technique application

    International Nuclear Information System (INIS)

    Nuning Duria; Budi Santoso; Nuniek Lelananingtiyas; Wiranto Budi Santoso

    2010-01-01

    Sample preparation for measuring cow's progesterone in radioimmunoassay technique application has been done. Preparation includes the preparation of samples of cow's milk samples that has been carried out artificial insemination on day 0, 11 and 21. Then the standard making of progesterone in the form of fresh milk or skim milk (non fat) that has been removed its progesterone and through a series of chemical processes. The last is the preparation work on samples using the RIA kit I 125 . With this milk samples preparation, samples can be counted using RIA counter so that concentration of progesterone hormone can be determined accurately. (author)

  11. Heterotypic reactions in a radioimmunoassay for coxsackie B virus specific IgM.

    Science.gov (United States)

    Pugh, S F

    1984-01-01

    IgM antibody capture radioimmunoassays were developed to detect coxsackie virus B1-B5 specific IgM. Specific IgM was detected in sera from all patients with coxsackie B virus infections proved by isolation; however, sera from 13/32 patients with rising neutralising antibody titres were negative in the assay. Frequent heterotypic responses were seen among the positive sera. Thirty seven patients with other enterovirus infections were also studied, and sera from 15 of these patients reacted in the assay, showing that heterotypic coxsackie B IgM responses occur not only in coxsackie B virus infections but also in other enterovirus infections. PMID:6323548

  12. Alpha1-antitrypsin deficiency

    DEFF Research Database (Denmark)

    Stolk, Jan; Seersholm, Niels; Kalsheker, Noor

    2006-01-01

    The Alpha One International Registry (AIR), a multinational research program focused on alpha1-antitrypsin (AAT) deficiency, was formed in response to a World Health Organization recommendation. Each of the nearly 20 participating countries maintains a national registry of patients with AAT defic...... epidemiology, inflammatory and signalling processes, therapeutic advances, and lung imaging techniques....

  13. Alpha clustering in nuclei

    International Nuclear Information System (INIS)

    Hodgson, P.E.

    1990-01-01

    The effects of nucleon clustering in nuclei are described, with reference to both nuclear structure and nuclear reactions, and the advantages of using the cluster formalism to describe a range of phenomena are discussed. It is shown that bound and scattering alpha-particle states can be described in a unified way using an energy-dependent alpha-nucleus potential. (author)

  14. Plasma concentrations of alpha-melanocyte-stimulating hormone are elevated in patients on chronic haemodialysis.

    Science.gov (United States)

    Airaghi, L; Garofalo, L; Cutuli, M G; Delgado, R; Carlin, A; Demitri, M T; Badalamenti, S; Graziani, G; Lipton, J M; Catania, A

    2000-08-01

    Clinical and/or laboratory signs of systemic inflammation occur frequently in patients undergoing long-term haemodialysis. It is likely, therefore, that a compensatory release of endogenous anti-inflammatory molecules occurs to limit host reactions. The aim of the present research was to determine if the potent anti-inflammatory peptide alpha-melanocyte-stimulating hormone (alpha-MSH), a pro-opiomelanocortin derivative, is increased in plasma of haemodialysis patients. Because endotoxin and cytokines induce alpha-MSH in vivo and in vitro, we also measured plasma concentrations of endotoxin, interleukin-6 (IL-6), and tumour necrosis factor alpha (TNF-alpha), and the two circulating products of activated monocytes, nitric oxide (NO) and neopterin. Thirty-five chronic haemodialysis patients, 20 patients with chronic renal failure not yet on dialysis, and 35 normal controls were included in the study. In the haemodialysis group, blood samples were obtained before and at the end of a dialysis session. Plasma alpha-MSH was measured using a double antibody radioimmunoassay, and IL-6, TNF-alpha, and neopterin using specific enzyme-linked immunosorbent assays. Plasma nitrites were determined by a colorimetric method, and endotoxin with the quantitative chromogenic LAL (limulus amoebocyte lysate) method. Mean plasma alpha-MSH was higher in haemodialysis patients than in control subjects, with the peptide concentrations being particularly elevated in dialysed patients with detectable endotoxin. High alpha-MSH concentrations were observed in the pre-dialysis samples, with no substantial change at the end of the dialysis session. Plasma concentrations of IL-6, TNF-alpha, neopterin, and NO were generally elevated in chronic haemodialysis patients and there was a negative correlation between circulating alpha-MSH and IL-6. In patients with renal failure not yet on dialysis, mean plasma alpha-MSH was similar to that of normal subjects. alpha-MSH is increased in the circulation of

  15. Applications of radioimmunoassay techniques in endocrine studies. Part of a coordinated programme on in vitro assay techniques

    International Nuclear Information System (INIS)

    Marusic, E.T.

    1977-04-01

    The final report consists of a description of the research done, a reference to where the abstract of the results obtained was published, and a list of the corresponding publications and of those in preparation. Work has been done on radioimmunoassay techniques for measuring plasma corticoid values, plasma renin activity (with own reagents), and clinical and research studies measuring aldosterone, corticosterone, and cortisol in control and hypophysectomized patients. Diagnosis of primary aldosteronism has been initiated by radioimmunoassay techniques. Another study concerned the mechanism of K ions on aldosterone production

  16. Insulin and C-peptide in human brain neurons (insulin/C-peptide/brain peptides/immunohistochemistry/radioimmunoassay)

    International Nuclear Information System (INIS)

    Dorn, A.; Bernstein, H.G.; Rinne, A.; Hahn, H.J.; Ziegler, M.

    1983-01-01

    The regional distribution and cellular localization of insulin and C-peptide immunoreactivities were studied in human cadaver brains using the indirect immunofluorescence method, the peroxidase-antiperoxidase technique, and radioimmunoassay. Products of the immune reactions to both polypeptides were observed in most nerve cells in all areas of the brain examined. Immunostaining was mainly restricted to the cell soma and proximal dendrites. Radioimmunoassay revealed that human brain contains insulin and C-peptide in concentrations much higher than the blood, the highest being in the hypothalamus. These findings support the hypothesis that the 'brain insulin' is - at least in part - produced in the CNS. (author)

  17. Genetics Home Reference: alpha thalassemia

    Science.gov (United States)

    ... Facebook Twitter Home Health Conditions Alpha thalassemia Alpha thalassemia Printable PDF Open All Close All Enable Javascript to view the expand/collapse boxes. Description Alpha thalassemia is a blood disorder that reduces the production ...

  18. Dynamic observation of transforming growth factor-alpha content in plasma of pediatric patients with peptic ulcer disease

    International Nuclear Information System (INIS)

    Zhou Mingxiong; Zhang Xinlu

    2001-01-01

    Objective: To elicit the relationship between transforming growth factor alpha (TGF-α) and the pathogenesis as well as healing process of peptic ulcer disease (PUD) in pediatric patients. Methods: The levels of TGF-α in plasma were measured by radioimmunoassay in 57 Children with PUD. Results: TGF-α levels of plasma at active stage of peptic ulcer were significantly lower than those at healing stage as well as in controls (P 0.05). Conclusion: There is an abnormal secretion of TGF-α in PUD patients. Changes of TGF-α release might play a role in the pathogenesis of PUD

  19. Radioimmunoassay measurements of serum thyrotropin in patients with hypothalamic-pituitary and thyroid diseases

    International Nuclear Information System (INIS)

    Miyai, Kiyoshi; Azukizawa, Mizuo; Azukizawa, Hisako; Hosokawa, Mitsuko; Nishi, Keiko

    1975-01-01

    Serum TSH was measured by means of double antibody radioimmunoassay using a commercial Kit Daiichi, in 21 normal subjects, 200 patients with thyroid disease and 130 patients with hypothalamic-pituitary diseases. Serum TSH concentrations in normal subjects were <2 to 8 μU/ml which rose to 8-40 μU/ml after administration of 500 μg TRH intravenously. Serum TSH was undetectable and did not respond to TRH in all untreated patients and in some euthyroid patients with Graves' disease after treatment. Undetectable TSH and no response to TRH were also observed in most patients with a hyperfunctioning thyroid nodule and those with subacute thyroiditis in the acute phase. In some patients with Hashimoto's thyroiditis and all patients with adult myxedema and cretinism, serum TSH levels were increased and showed hyperresponse to TRH. The ratio of bioassay and radioimmunoassay potency estimates for TSH in sera obtained before TRH was not statistically different from that obtained after TRH administration to patients with primary hypothyroidism. Elevated serum TSH was promptly decreased by the administration of thyroid hormone to the patients. More than 50% of patients with pituitary adenoma, acromegaly and craniopharyngioma showed normal basal TSH and no or low response of TSH to TRH. Administration of TRH failed to stimulate a rise in serum TSH in 2 sisters with isolated TSH deficiency with cretinism. Basal TSH was undetectable and showed delayed response to TRH in patients with anorexia nervosa. (auth.)

  20. Radioimmunoassay of total and specific E immunoglobulin. Variations with different parameters in allergic subjects

    International Nuclear Information System (INIS)

    Le Sellin, James.

    1977-05-01

    Owing to the work of 'Ishizaka' and of Johansson and Bennich the discovery of E immunoglobulin (IgE) marks an important stage in the diagnosis of atopic hypersensitivity. However certain limits in the analysis methods must be known to allow better interpretation of the results. The radioimmunoassay of total and specific IgE is still one of the most precise. Our work is divided into two parts: - the first is devoted to a general review on IgE; - the second reports our own study carried out on 371 allergic subjects. Each of these patients was analysed for total IgE by the PRIST radioimmunological method (paper radioimmuno sorbent test) and for IgE specific to one or more antigens by the RAST method (radioallergo sorbent test). A radioimmunoassay of total and specific IgE must be interpreted as a function of clinical data. While the assay can never become systematic it may be used to confirm or back up an atopic hypersensitivity diagnosis in cases where the clinical signs are inadequate [fr

  1. Measurement of IgG-blocking antibodies: development and application of a radioimmunoassay

    International Nuclear Information System (INIS)

    Sobotka, A.K.; Valentine, M.D.; Ishizaka, K.; Lichtenstein, L.M.

    1976-01-01

    A radioimmunoassay for measuring blocking antibodies has been developed. We used the ragweed antigen E system to show that the same blocking antibodies (IgG) measured by inhibition of antigen-induced leukocyte histamine release were precipitated in the binding assay (r/sub s/ = 0.96 p less than 0.001), thus validating a widely applicable technique for measuring blocking antibodies. Binding of phospholipase-A (Phos-A), the major allergen in honey bee venom, was also shown to correlate significantly with inhibition of histamine release. Hymenoptera (insect) hypersensitivity was used as a model to demonstrate application of the binding assay. Sera obtained from patients undergoing whole body extract therapy contained negligible amounts of specific blocking antibodies. Significantly higher blocking antibody titers to both whole honey bee venom and Phos-A were measured in sera drawn from patients immunized with whole venom. The use of the binding radioimmunoassay should facilitate management of allergic disease processes in which blocking antibodies are thought to be protective

  2. Development of a sensitive solid-phase radioimmunoassay for melanin-concentrating hormone

    Energy Technology Data Exchange (ETDEWEB)

    Eberle, A.N.; Baumann, J.B.; Girard, J. (Univ. Children' s Hospital, Basel (Switzerland)); Baker, B.I.; Kishida, M. (Univ. of Bath (England))

    1989-01-01

    A two-step solid-phase radioimmunoassay for melanin-concentrating hormone (MCH) was developed for direct determination of the hormone in plasma samples. To this end, synthetic MCH was coupled to bovine thyreoglobulin and the complex was injected into rabbits. Specific antisera of high titer were obtained which did not crossreact with other hormones. The IgGs were chemically linked to immunobeads, an acrylamide/acrylic acid polymer matrix. In the first step, plasma MCH was immunoextracted by incubation of diluted plasma samples with anti-MCH immunobeads. In the second step, the washed polymer was incubated with radioiodinated MCH tracer for titration of non-occupied sites. This procedure made it possible to determine as little as 4 pg MCH per ml of plasma. Application of the radioimmunoassay to plasma levels of black or white background-adapted trout showed a marked difference in circulating MCH: while trout on a black background contained a mean value of 29 {plus minus} 5.6 pg/ml, animals on a white background had 106 {plus minus} 19 pg/ml.

  3. Development of a radioimmunoassay for the determination of buprenorphine in biological samples

    Energy Technology Data Exchange (ETDEWEB)

    Debrabandere, L.; Boven, M. Van; Daenens, P. (Louvain Univ. (Belgium))

    1993-02-01

    The development of a specific and sensitive radioimmunoassay for the detection of buprenorphine in urine samples is described. With minor adjustments, the assay was also applied to the analysis for buprenorphine in plasma samples. The 2-diazobenzoic acid derivative of buprenorphine has been prepared as a hapten. The immunization of rabbits with the hapten-bovine serum albumin conjugate resulted in the production of antibodies, which cross-reacted with N-dealkylbuprenophine up to about the 90% level. The antibodies showed very low cross-reactivities with the 3-O-glucuronides and with the structural analogue etorphine. The assay was mainly used to prescreen for buprenorphine in urine samples of persons suspected of Temgesic misuse and to determine buprenorphine in plasma samples. A linear calibration graph for buprenorphine was obtained after logit-log regression. The spiking recovery study showed a linear regression. Intra-and inter-assay relative standard deviations were < 4.35 and < 6.36%, respectively. A comparison study of the high-performance liquid chromatographic determination (X) to the radioimmunoassay (Y) resulted in the following regression equation for the urine samples: Y = 1.44 + 1.64 X (n = 32; r 0.910) and Y = 0.007 + 1.58 X (n = 10; r = 0.930) for plasma specimens. The minimum detectable dose of the immunoassay was calculated to be 10 pg ml[sup -1] (Student's t-distribution, p 0.01, degrees of freedom = 8). (Author).

  4. A new unextracted-sample radioimmunoassay method for hepatic endogenous nuclear L-tri-iodothyronine content

    International Nuclear Information System (INIS)

    Yagura, T.; Walfish, P.G.

    1982-01-01

    Endogenous L-tri-iodothyronine content in an hepatic nuclear extract was measured by a new unextracted-sample radioimmunoassay method using 8-anilinonaphthalene-1-sulphonic acid to inhibit the L-[ 125 I]tri-iodothyronine binding to the nuclear L-tri-iodothyronine receptor within the extract. The amount of endogenous L-tri-iodothyronine was 10-40 pg/0.2 ml of hepatic nuclear extract from euthyroid rats, compared with less than 3.125 pg/0.2ml from thyroidectomized rats. The results obtained were compared with a Sephadex G-25 column extracted-sample radioimmunoassay method and showed a good agreement. The values for the endogenous L-tri-iodothyronine content were utilized to correct for the L-tri-iodothyronine concentration within the binding assay mixture in order to accurately determine by Scatchard analysis the binding characteristics of the nuclear L-tri-iodothyronine receptor. The validity of the correction for endogenous L-tri-iodothyronine was demonstrated by using a nuclear extract from a thyroidectomized rat which was preincubated with a small known amount of L-tri-iodothyronine before determining the nuclear L-tri-iodothyronine receptor binding characteristics. It is concluded that the necessity and validity of using endogenous L-tri-iodothyronine corrections in the Scatchard analytical computations of the nuclear L-tri-iodothyronine receptor binding characteristics has been demonstrated, being particularly more important for affinity constant than maximum binding capacity. (author)

  5. Radioimmunoassay of insulin by the double antibody method. Evaluation of the technique in practice

    International Nuclear Information System (INIS)

    Wojckikowski, C.; Zakoslki, W.

    1974-01-01

    The aim of this study was to develop a simplified procedure for the radioimmunoassay of insulin by the double antibody method. Anti-insulin serum was obtained by immunizing guinea pigs with porcine insulin suspended in Freund's incomplete adjuvant supplemented with Hemophilus pertussis vaccine. After two injections, sera from four guinea pigs diluted 1:10,000 bound more than 70% of the 125 I-insulin added. Under the conditions of the method, the insulin binding reagent bound 43%+-5% (n=94) of added 125 I-insulin. Accuracy of the method in simultaneous series for insulin concentrations of 27 μU/ml and 49 μU/ml was +-5% and +-7% respectively; and in a nonsimultaneous series of determinations for insulin concentrations of 21 μU/ml and 77 μU/ml, +-11% and +-8%. Recovery of insulin added to human plasma was 104%. Concentrations of insulin in the plasma of healthy fasting subjects ranged from 5-22 μU/ml (m=13+-5 μU/ml; n=21). The results were in complete agreement with results obtained by the use of the Amersham radioimmunoassay insulin kit. (author)

  6. Non-chromatographic radioimmunoassay for serum dehydroepiandrosterone using a mixture of antisera

    International Nuclear Information System (INIS)

    Sekihara, H.; Ohsawa, N.

    1974-01-01

    A simplified method for evaluating serum dehydroepiandrosterone (DHEA) without chromatography was developed, using mixtures of two different anti-DHEA antisera, anti-3β-hydroxy-Δ 5 antiserum and anti-11-deoxy-17-ketosteroid antiserum, in which cross-reactivity of each antiserum is reduced to a negligible amount. Serum (20 μl) was extracted with 1 ml of n-hexane. One milliliter of 80 percent methanol was added to the n-hexane extract, which was stirred and centrifuged. The n-hexane layer was discarded, and the methanol layer was evaporated to dryness. The residue was incubated with an antiserum mixture containing DHEA-7α- 3 H, pepsin-treated human immune serum globulin and bovine serum albumin. Ammonium sulfate was used to separate free from bound DHEA-7α- 3 H. The accuracy, precision, sensitivity, and specificity were satisfactory. Good agreement was found between the serum DHEA levels obtained by the present radioimmunoassay and those obtained by radioimmunoassay with paper chromatography, making this method suitable for routine use. (U.S.)

  7. Radioimmunoassay of lysergic acid diethylamide (LSD) in serum and urine by using antisera of different specificities

    International Nuclear Information System (INIS)

    Ratcliffe, W.A.; Fletcher, S.M.; Moffat, A.C.; Ratcliffe, J.G.; Harland, W.A.; Levitt, T.E.

    1977-01-01

    We raised high-titre antisera to two LSD-bovine serum albumin conjugates, one linked via the indole nitrogen, the other via the amide side-chain. The antisera were specific for different parts of the LSD molecule, as demonstrated by cross-reactivity studies with LSD, its metabolites, ergot alkaloids, and closely related compounds. The antisera were used to develop a double-antibody radioimmunoassay with a detection limit of about 0.4 μg of LSD per liter of unextracted urine or serum. We saw no nonspecific interference by urine, serum, or from a series of commonly used drugs. There was good correlation between immunoassay values obtained with the two antisera (r = 0.91). However, the antiserum linked via the indole nitrogen gave consistently higher results for samples from persons who had taken LSD, owing to greater cross-reactivity with LSD metabolites. Radioimmunoassay by use of two such antisera is a more specific screening procedure for LSD abuse than has been available previously. In addition, antisera cross-reacting with LSD metabolites allow measurement of these compounds, for which there is no satisfactory method at the concentrations found in biological fluids in man

  8. Radioimmunoassay of human thyrotropin (TSH): an improvement on microtiter plate method

    International Nuclear Information System (INIS)

    Romaldini, J.H.; Rodrigues, H.F.; Tanaka, L.M.; Teixeira, V.L.; Reis, L.C.F.

    1980-01-01

    A methodological improvement is presented of solid phase (microtiter 'U' plates) radioimmunoassay of human TSH. The high concentration (1:1 x 10 4 ) of anti-TSH serum was adsorbed by microtiter cups. The bound and free 125 I-TSH was separated by simple washng with distilled water. The assay was completed within 48 hours. Minimal labeled TSH damage was achieved by a) repurification of 125 I-TSH by a high Sephadex G-100 (1.5 x 70 cm) column on the day ofssay; b) satisfactory low specific activity (67+- 27.5 uCi/ug); c) stoage with 0.2% BSA-PBS (1:5 dilution) at-80 0 C up to 4 weeks. Maximal binding was 59 +- 13%. The minimum detectable amount of TSH in this assay was 0.13 uU/cup. The microtiter plate radioimmunoassay for TSH was reproducible and precise (interassay variability 11.6 to 18.8%, interassay variability 12 to 22.6%). The recovery of standard TSH was 101+- 16% (r=0,939). Results obtained from the clinical application of the method are related. (Author) [pt

  9. Epitope mapping of alpha-transforming growth factor: evidence of an immunodominant region

    International Nuclear Information System (INIS)

    Hazarika, P.; Dedman, J.R.

    1988-01-01

    Antisera were produced in rabbits and sheep against both full-length synthetic rat alpha-transforming growth factor and peptides corresponding to the carboxy-terminal 17 amino acids. These antisera were used to develop a peptide based radioimmunoassay of alpha-TGF. All antisera reacted only with a restricted region of the alpha-TGF corresponding to the 8 residues (43-50) at the carboxy-terminus: Cyslt. slash43, Glult. slash44, Hislt. slash45, Alalt. slash46, Asplt. slash47, Leult. slash48, Leult. slash49, Alalt. slash50. A series of synthetic peptides presenting deletions or substitutions of amino acids in this carboxy-terminal region were tested for competition with 125 I-alpha-TGF. All changes in the above peptide sequence resulted in a marked reduction in competition. All of the polyclonal antisera demonstrated similar specificity whether they were produced against the 50 amino acid, full-length alpha-TGF, against shorter 17 amino acid and 8 amino acid carboxy-terminal sequences

  10. Epitope mapping of alpha-transforming growth factor: evidence of an immunodominant region

    Energy Technology Data Exchange (ETDEWEB)

    Hazarika, P.; Dedman, J.R.

    1988-01-01

    Antisera were produced in rabbits and sheep against both full-length synthetic rat alpha-transforming growth factor and peptides corresponding to the carboxy-terminal 17 amino acids. These antisera were used to develop a peptide based radioimmunoassay of alpha-TGF. All antisera reacted only with a restricted region of the alpha-TGF corresponding to the 8 residues (43-50) at the carboxy-terminus: Cyslt. slash43, Glult. slash44, Hislt. slash45, Alalt. slash46, Asplt. slash47, Leult. slash48, Leult. slash49, Alalt. slash50. A series of synthetic peptides presenting deletions or substitutions of amino acids in this carboxy-terminal region were tested for competition with /sup 125/I-alpha-TGF. All changes in the above peptide sequence resulted in a marked reduction in competition. All of the polyclonal antisera demonstrated similar specificity whether they were produced against the 50 amino acid, full-length alpha-TGF, against shorter 17 amino acid and 8 amino acid carboxy-terminal sequences.

  11. Alpha Thalassemia (For Parents)

    Science.gov (United States)

    ... the body has a problem producing alpha globin Beta thalassemia : when the body has a problem producing beta ... Transfusion Blood Test: Hemoglobin Electrophoresis Sickle Cell Disease Beta Thalassemia Blood All About Genetics Prenatal Genetic Counseling Genetic ...

  12. Preparation of 2-[125I] iodohistamine-labelled Δ8-tetrahydrocannabinol-11-oic acid for use in cannabinoid radioimmunoassay

    International Nuclear Information System (INIS)

    Law, B.; Mason, P.A.; Moffat, A.C.; King, L.J.

    1982-01-01

    A simple method is described for the preparation of 2-[ 125 I]iodohistamine-labelled Δ 8 -tetrahydrocannabinol-11-oic acid with high specific activity for use in radioimmunoassay. This compound is produced in high yield and shows excellent radiochemical stability when stored at 4 0 C. (author)

  13. Competitive binding radioassays for 1α-25(OH)2 vitamin D; comparative evaluation of two receptor assays and a radioimmunoassay

    International Nuclear Information System (INIS)

    Jallet, P.; Bidet, M.; Audran, M.

    1985-01-01

    The performances of a 1α,25-dihydroxy vitamin D assay using the cytosol receptor of bovine thymus gland were evaluated and compared to the results obtained with an assay based on cytosol receptor of chicK intestine and with a radioimmunoassay [fr

  14. ALPHA-2: the sequel

    CERN Multimedia

    Katarina Anthony

    2012-01-01

    While many experiments are methodically planning for intense works over the long shutdown, there is one experiment that is already working at full steam: ALPHA-2. Its final components arrived last month and will completely replace the previous ALPHA set-up. Unlike its predecessor, this next generation experiment has been specifically designed to measure the properties of antimatter.   The ALPHA team lower the new superconducting solenoid magnet into place. The ALPHA collaboration is working at full speed to complete the ALPHA-2 set-up for mid-November – this will give them a few weeks of running before the AD shutdown on 17 December. “We really want to get some experience with this device this year so that, if we need to make any changes, we will have time during the long shutdown in which to make them,” says Jeffrey Hangst, ALPHA spokesperson. “Rather than starting the 2014 run in the commissioning stage, we will be up and running from the get go.&...

  15. Progesterone radioimmunoassay

    International Nuclear Information System (INIS)

    Stroufova, A.; Kozlova, J.

    1976-01-01

    RIA methods of determining progestorone using the SORIN kit made in Italy and the NEN kit made in Great Britain were compared. Plasma extraction, the initial sample size for examination and recovery, the range of calibration curves of the two kits, the variation coefficient, the values of the blank sample and the values of progesterone determined in the normal menstrual cycle are discussed in detail. Variation coefficient: Sorin (calibration curve 9 to 11%, biological material 9.2%), NEN (calibration curve 12%, biological material 27.8%); determinations of progesterone levels during a normal menstrual cycle were 10 to 15% higher with the NEN kit; tritiated samples were measured with a 30 to 37% efficiency using the SORIN kit and 15 to 18% efficiency using the NEN kit. The turbidity and later sediment which formed during the determination of steroid hormones in biological materials after the addition of the scintillation solution did not reduce the efficiency of measurement. Priority is given to the SORIN kit. (L.O.)

  16. Expression of triplicated and quadruplicated alpha globin genes in sheep.

    Science.gov (United States)

    Vestri, R; Pieragostini, E; Yang, F; di Gregorio, P; Rando, A; Masina, P

    1991-01-01

    In the sheep alpha alpha alpha globin gene haplotype, the three genes display from the 5' to the 3' end the percentage efficiencies of about 30:14:6, as indicated by the amounts of the three types of alpha chain produced in the alpha alpha alpha/alpha alpha alpha homozygotes. The 3' gene in the alpha alpha alpha alpha haplotype appears to have an efficiency around 1%, as suggested by analysis of one quadruple alpha homozygote. Moreover, the total outputs of the alpha alpha alpha as well as of the alpha alpha alpha alpha haplotypes do not substantially differ from that of the common alpha alpha haplotype.

  17. Post-treatment levels of α-fetoprotein predict long-term hepatocellular carcinoma development after sustained virological response in patients with hepatitis C.

    Science.gov (United States)

    Tada, Toshifumi; Kumada, Takashi; Toyoda, Hidenori; Kiriyama, Seiki; Tanikawa, Makoto; Hisanaga, Yasuhiro; Kanamori, Akira; Kitabatake, Shusuke; Yama, Tsuyoki; Tanaka, Junko

    2017-09-01

    The rate of hepatocellular carcinoma (HCC) development is reportedly lower in patients with chronic hepatitis C virus (HCV) who have achieved a sustained virological response (SVR) than in patients who were unresponsive to therapy. However, the development of HCC is sometimes observed in patients with SVR. Therefore, we clarified the predictive power of clinical factors for HCC incidence in patients with SVR using receiver operating characteristic (ROC) curve analysis that takes time dependence into account. A total of 571 patients with HCV who achieved SVR with interferon-based therapy were enrolled. Univariate and multivariate Cox proportional hazards models and time-dependent ROC curves were used to analyze clinical factors associated with the development of HCC. Twenty-four patients developed HCC during the follow-up period (median duration, 9.0 years). The 5-, 10-, 15-, and 20-year cumulative incidence rates for HCC were 1.7%, 4.8%, 5.8%, and 6.6%, respectively. Multivariate Cox proportional hazards models showed that older age (hazard ratio [HR], 3.648), male sex (HR, 7.560), lower platelet count at 24 weeks after the end of treatment (SVR24) (HR, 3.939), and higher α-fetoprotein (AFP) at SVR24 (HR, 3.630) were independently associated with HCC development. In addition, time-dependent ROC analysis showed that, compared to platelet count at SVR24, AFP at SVR24 had higher predictive power for HCC incidence approximately 7 years after SVR. Elevated AFP at SVR24 is a risk factor for HCC in patients with HCV, even those who achieve SVR. α-Fetoprotein is a good predictor of HCC development. © 2016 The Japan Society of Hepatology.

  18. Use of progesterone radioimmunoassay techniques for improving reproduction performances in dary cattles

    International Nuclear Information System (INIS)

    Benyoucef, M.T.; Benabdelaziz, A.; Khelili, R.

    1990-07-01

    This report deals with the first measurements of progesterone, by radioimmunoassay (P4-PIA) in blood samples, to assess fertility in livestock (cattle) in Algeria, by improvement of the level of reproduction management. For this purpose, the results obtained throughout first analysis of P4 on a sample of 15 bovines females, composed by 12 heifers and 3 cows confirm the efficiency of the techniques in improving dary cattle reproduction performances. Ovarian activity can be determined for cows at the moment of post partum and for heifers before breeding as well as, pregnancy confirmation on breeding heifers. It is interesting to extend the experiments using these techniques to a higher number of livestocks (breeds and species) and to develop new research subjects in this field

  19. Development and validation of an improved radioimmunoassay for serotonin in platelet-rich plasma

    International Nuclear Information System (INIS)

    Gow, I.F.; Williams, B.C.; Edwards, C.R.W.

    1987-01-01

    A radioimmunoassay (RIA) using a 125 I-tracer is described for measurement of serotonin (5-hydroxytryptamine, 5-HT) in human platelet-rich plasma (PRP). Antisera were raised against 5-HT-succinamate conjugated to bovine albumin and, to improve assay sensitivity, the analyte was made chemically similar to the immunogen by conversion to N-acetylserotonin prior to assay, using the specific amino reagent N-acetoxysuccinimide. The assay shows good correlation with a high-pressure liquid chromatography (HPLC) reference method, indicating that no significant cross-reactions were detected. With this improved assay, it is possible to analyse up to 100 samples/day, compared with 10-20 samples/day by HPLC. (Auth.)

  20. Iodine-125 radioimmunoassay for the direct detection of benzodiazepines in blood and urine

    Energy Technology Data Exchange (ETDEWEB)

    Goddard, C.P.; Stead, A.H.; Mason, P.A.; Law, B.; Moffat, A.C.; McBrien, M.; Cosby, S.

    1986-05-01

    A radioimmunoassay (RIA) for the direct detection of benzodiazepines in blood and urine is described. It is based on a commercially available antiserum and an easily synthesised radio-iodinated derivative of clonazepam that allows the use of relatively simple gamma-counting procedures. The assay can detect low therapeutic levels of all of the benzodiazepines currently available in the UK in 50-..mu..l samples of blood and urine (1-50 ng ml/sup -1/, depending on the drug); no prior sample preparation is required. It is inexpensive, rapid, simple to perform and is broadly specific for the benzodiazepine class of drugs. The assay offers a most suitable means of screening large numbers of samples of forensic interest for the presence of the benzodiazepines.