scholarly journals One-Step Direct Assay for Mature-type Adrenomedullin with Monoclonal Antibodies

1999 ◽  
Vol 45 (2) ◽  
pp. 244-251 ◽  
Author(s):  
Hideki Ohta ◽  
Tetsuo Tsuji ◽  
Shigeru Asai ◽  
Kazuyuki Sasakura ◽  
Hiroshi Teraoka ◽  
...  

Abstract Adrenomedullin (AM) is a potent hypotensive peptide. Plasma contains mature-type AM (m-AM), which is amidated at the carboxy terminus, and an intermediate, AM-Gly. We developed a one-step two-site IRMA specific for determining human m-AM with monoclonal antibodies. The detection limit was 0.5 pmol/L, and the working range (CV <15%) was 1–300 pmol/L. Dilution of plasma samples showed good linearity. The recovery of added AM was 91–118%. The intra- and interassay imprecision values (CVs) were 4.4–8.2% and 5.5–8.3%, respectively. The assay had no cross-reactivity with AM-Gly or other peptides similar to AM. The mean (± SD) plasma human m-AM concentration of 61 healthy subjects was 1.18 ± 0.65 pmol/L. In conclusion, our IRMA makes it possible to specifically measure m-AM, using a small amount of plasma sample (0.2 mL) by a one-step overnight assay without prior extraction. Our simplified method would be suitable for clinical studies on AM, especially when large numbers of samples must be processed.

1987 ◽  
Vol 115 (2) ◽  
pp. 283-288 ◽  
Author(s):  
K. F. Miller ◽  
R. A. Goldsby ◽  
D. J. Bolt

ABSTRACT Bovine FSH (bFSH) was used to immunize BALB/c mice. Spleen cells were fused to the SP 2/0 cell line to produce hybridomas that secreted monoclonal antibodies to bFSH. One of these antibodies (USDA-bFSH-MC28) was extensively characterized and found to be a gamma 1 with kappa light chains, having extremely low cross-reactivity with other bovine pituitary hormones and with ovine and porcine FSH. The dissociation constant as measured by Scatchard analysis was 4·3 nmol/l, and proved to be in a very useful range for affinity chromatography. In an essentially one-step immunoaffinity chromatography procedure, bFSH was easily isolated in a single chromatographic step from crude anterior pituitary homogenate with better yield and with the same purity as classical chromatographic techniques. J. Endocr. (1987) 115, 283–288


1995 ◽  
Vol 133 (1) ◽  
pp. 33-37 ◽  
Author(s):  
Paloma Gil-del-Alamo ◽  
Katia Saccomanno ◽  
Andrea Lania ◽  
Kim SI Pettersson ◽  
Paolo Beck-Peccoz ◽  
...  

Gil-del-Alamo P, Saccomanno K, Lania A, Pettersson KSI, Beck-Peccoz P, Spada A. Serum levels of β-subunit of chorionic gonadotropin in patients with pituitary tumors. Eur J Endocrinol 1995;133:33–7. ISSN 0804–4643 Many studies have shown that normal and tumoral pituitary is able to synthesize chorionic gonadotropin (CG). The aim of the present work was to investigate the circulating levels of free β-subunit of CG (CG-β) in a large number of patients with pituitary tumors in basal conditions and after thyrotropin-releasing hormone (TRH) injection. The study includes 27 healthy subjects, 23 patients with prolactinoma, 20 with growth hormone-secreting adenoma and 77 with non-functioning pituitary adenoma (NFPA). The CG-β was evaluated using a new one-step immunometric assay employing two monoclonal antibodies directed against epitopes present only on the free CG-β and showing a detection limit of 0.04 U/l and a cross-reactivity with complete CG < 0.01%. In basal conditions, serum CG-β was undetectable in healthy subjects and in the majority of patients, while in seven patients with NFPA and four with prolactinoma the CG-β values ranged between 0.05 and 0.72 U/l. In these 11 patients serum levels of intact CG were found within the normal range (normal range < 5 U/I), while two patients with NFPA and one with prolactinoma had levels of free α-subunit inappropriately high with respect to gonadotropins and thyrotropin. Injection of TRH caused CG-β to increase in two out of 16 patients with NFPA, whereas it was ineffective in 12 healthy subjects and 10 patients with prolactinoma. The present data indicate that detectable level of CG-β not associated with hypersecretion of the intact CG molecule may be observed in about 10% of patients with NFPA or prolactinoma, while abnormal CG-β responses to TRH are observed infrequently in individual patients with NFPA. Paolo Beck-Peccoz, Institute of Endocrine Sciences, Ospedale Maggiore IRCCS, Pad. Granelli, Via F Sforza 35, 20122-Milano, Italy


1984 ◽  
Vol 30 (3) ◽  
pp. 450-451 ◽  
Author(s):  
S Balzan ◽  
A Clerico ◽  
M Grazia del Chicca ◽  
U Montali ◽  
S Ghione

Abstract Reports on the presence of digoxin-like immunoreactive substance(s) (DLIS) in the plasma and urine of several animal species and a few human test subjects prompted us to undertake to confirm the presence of DLIS in plasma and urine of normal persons and to investigate some characteristics of this antibody-DLIS binding. For this purpose we used a modified radioimmunoassay kit involving antidigoxin antibody-coated test tubes and 125I-labeled digoxin to measure DLIS in urine and concentrated plasma of ostensibly healthy subjects. In 12 separate experiments with plasma the mean sensitivity was 5.12 (SD 1.11) pg per tube, expressed as digoxin. There was no significant cross reactivity with human serum albumin in concentrations up to 200 g/L. The mean DLIS value (in digoxin equivalents) for plasma from 24 normal subjects was 33.58 (SD 14.24) ng/L. Its mean concentration in urine from five subjects was 315.00 (SD 91.38) ng/L.


1986 ◽  
Vol 32 (5) ◽  
pp. 797-800 ◽  
Author(s):  
L A Lee ◽  
R A Mooney ◽  
P D Woolf

Abstract Free thyroxin (FT4) measurements by ultrafiltration were compared with results obtained by equilibrium dialysis and commercial RIA methods. In a group of critically ill patients, the mean +/- 2 SD FT4 value by ultrafiltration (26.5 +/- 21.4 ng/L) was higher than in the healthy reference population (19.3 +/- 9.8 ng/L, p less than 0.0001), and correlated well with equilibrium dialysis (23.8 +/- 20 ng/L, r = 0.87). By a two-step RIA method (Clinical Assays), the FT4 value was greater in the critically ill (14.7 +/- 9.6 ng/L) than in the reference population (12.8 +/- 5 ng/L, p less than 0.001) and was also correlated with ultrafiltration results (r = 0.71). FT4 values for the ill patients were lower than for the reference population in two one-step RIA methods (Clinical Assays: 8.6 +/- 7.1 ng/L; Corning Medical: 12.7 +/- 9.4 ng/L), neither of which gave results correlated with ultrafiltration (r = 0.16 and 0.22, respectively). The concentration of free triiodothyronine (FT3), also measured by ultrafiltration, was 4.5 +/- 2.2 ng/L in healthy subjects, 12.5 +/- 14.2 ng/L in hyperthyroidism, 1.7 +/- 2.0 ng/L in hypothyroidism, 2.2 +/- 1.9 ng/L in the critically ill subjects, and 3.8 +/- 0.9 ng/L in pregnancy. Thus, FT4 and FT3 measured by ultrafiltration accurately affects assessment of the thyroid status of the patient except in critical illnesses in which FT3 values are indistinguishable from those in hypothyroidism.


1972 ◽  
Vol 71 (1) ◽  
pp. 24-36 ◽  
Author(s):  
Ariel Gordin ◽  
Pirkko Saarinen

ABSTRACT An account is given of a methodological study of the double-antibody radioimmunoassay of human TSH, using highly purified labelled human TSH as tracer. It was shown that conventional paper electrophoresis was not adequate for studying the purity of labelled human TSH. When polyvinylchloride (Pevikon®) electrophoresis was used, four subfractions could still be separated, even though, on paper electrophoresis, the material seemed to be homogeneous. Only two of the four Pevikon fractions were immunoreactive. Purification of labelled human TSH by Pevikon electrophoresis also improved the sensitivity of the assay. Specific activities of about 100 mCi/mg gave the highest initial binding capacity, produced least damage to the labelled hormone and showed the best stability of the tracer without influencing the sensitivity of the method. In different storage conditions, labelled human TSH was found to be most stable at −20°C and diluted 1/100. Only in pregnancy did the addition of HCG seem necessary. The mean TSH value in healthy subjects was 3.6 ± 1.4 μU/ml (mean±sd) with a range from 1.6 μU/ml to 8.8 μU/ml.


2019 ◽  
Vol 30 (5) ◽  
pp. 585-592 ◽  
Author(s):  
Nicola Montemurro ◽  
Paolo Perrini ◽  
Vittoriano Mangini ◽  
Massimo Galli ◽  
Andrea Papini

OBJECTIVEOdontoid process fractures are very common in both young and geriatric patients. The axial trabecular architecture of the dens appears to be crucial for physiological and biomechanical function of the C1–2 joint. The aim of this study is to demonstrate the presence of a Y-shaped trabecular structure of the dens on axial CT and to describe its anatomical and biomechanical implications.METHODSFifty-four C2 odontoid processes in healthy subjects were prospectively examined for the presence of a Y-shaped trabecular structure at the odontocentral synchondrosis level with a dental cone beam CT scan. Length, width, and axial area of the odontoid process were measured in all subjects. In addition, measurements of the one-third right anterior area of the Y-shaped structure were taken.RESULTSThe Y-shaped trabecular structure was found in 79.6% of cases. Length and width of the odontoid process were 13.5 ± 0.6 mm and 11.2 ± 0.9 mm, respectively. The mean area of the odontoid process at the odontocentral synchondrosis was 93.5 ± 4.3 mm2, whereas the mean one-third right anterior area of the odontoid process at the same level was 29.3 ± 2.5 mm2. The mean area of the odontoid process and its length and width were similar in men and women (p > 0.05). No significant difference was found in the mean area of the odontoid process in people older than 65 years (94 ± 4.2 mm2) compared to people younger than 65 years (93.3 ± 4.4 mm2; p > 0.05).CONCLUSIONSThe authors identified a new anatomical entity, named the Y-shaped trabecular structure of the odontoid process, on axial CT scans. This structure appears to be the result of bone transformation induced by the elevated dynamic loading at the C1–2 level. The presence of the Y-shaped structure provides new insights into biomechanical responses of C2 under physiological loading and traumatic conditions.


2018 ◽  
Vol 15 (1) ◽  
pp. 74-78
Author(s):  
Mohammadali Nazarinia ◽  
Asghar Zare ◽  
Mohammad javad Fallahi ◽  
Mesbah Shams

Background:Systemic sclerosis is a disorder of connective tissue with unknown cause, affecting the skin and internal organs, characterized by fibrotic changes.Objective:To determine the correlation between serum homocysteine level and interstitial lung involvement in systemic sclerosis. </P><P> Materials and Methods: In this case – control study, 59 patients who fulfilled the ACR/EULAR classification criteria for systemic sclerosis and were referred to Hafez Hospital of Shiraz, Iran, were included as the case group. Fifty nine healthy subjects were involved as the control group. Patients were divided into two groups based on interstitial lung involvement and two subtypes, diffuse and limited type. Serum homocysteine, vitamin B12, and folate levels compared between the controls, and cases groups.Results:Of 59 case and control group, 53 (%89.8) were female and the mean age did not differ in both groups (P=0.929). Thirty five (%59.3) patients had interstitial lung involvement and 38(%64.4) had diffuse cutaneous systemic sclerosis. The mean serum homocysteine level was 13.9±6.3 µmol/L in the case and 13.7±9.2 µmol/L in the control group (P=0.86). The mean serum homocysteine level did not differ between the patients with and without interstitial lung involvement (P=0.52). The patients with lung involvement was older than those without lung involvement (P=0.004). Lung disease was more common in diffuse type (P=0.014).Conclusion:In our study, serum homocysteine level did not differ between the patients and healthy subjects. Also, there was no correlation between serum homocysteine level and lung involvement, but lung involvement was more common in older patients and also diffuse subtype.


Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 365
Author(s):  
Fengli Liu ◽  
Yanxin Cao ◽  
Maokai Yan ◽  
Mengxu Sun ◽  
Qingshui Zhang ◽  
...  

Duck viral enteritis is a highly contagious and fatal disease of commercial waterfowl flocks. The disease occurs sporadically or epizootically in mainland China due to insufficient vaccinations. Early and rapid diagnosis is important for preventive intervention and the control of epizootic events in clinical settings. In this study, we generated two monoclonal antibodies (MAbs) that specifically recognized the duck enteritis virus (DEV) envelope glycoprotein B and tegument protein UL47, respectively. Using these MAbs, a colloidal gold-based immunochromatographic assay (ICA) was developed for the efficient detection of DEV antigens within 15 min. Our results showed that the detection limit of the developed ICA strip was 2.52 × 103 TCID50/mL for the virus infected cell culture suspension with no cross-reactivity with other pathogenic viruses commonly encountered in commercially raised waterfowl. Using samples from experimentally infected ducks, we demonstrated that the ICA detected the virus in cloacal swab samples on day three post-infection, demonstrating an 80% concordance with the PCR. For tissue homogenates from ducks succumbing to infection, the detection sensitivity was 100%. The efficient and specific detection by this ICA test provides a valuable, convenient, easy to use and rapid diagnostic tool for DVE under both laboratory and field conditions.


1986 ◽  
Vol 261 (17) ◽  
pp. 7975-7981
Author(s):  
J T Ulrich ◽  
J R Schenck ◽  
H G Rittenhouse ◽  
N L Shaper ◽  
J H Shaper

2021 ◽  
Vol 13 ◽  
pp. 251584142110304
Author(s):  
Emre Aydemir ◽  
Alper Halil Bayat ◽  
Burak Ören ◽  
Halil Ibrahim Atesoglu ◽  
Yasin Şakir Göker ◽  
...  

Purpose: The purpose of this study was to compare the retinal vascular caliber of COVID-19 patients with that of healthy subjects. Methods: This was a prospective case–control study. Forty-six patients who had COVID-19 were successfully treated, and 38 age- and gender-matched healthy subjects were enrolled in this study. Fundus photography was taken using fundus fluorescein angiography (FA; Visucam 500; Carl Zeiss Meditec, Jena, Germany). Retinal vascular caliber was analyzed with IVAN, a semi-automated retinal vascular analyzer (Nicole J. Ferrier, College of Engineering, Fundus Photography Reading Center, University of Wisconsin, Madison, WI, USA). Central retinal artery equivalent (CRAE), central retinal vein equivalent (CRVE), and artery–vein ratio (AVR) were compared between groups. Results: The mean age was 37.8 ± 9.5 years in the COVID-19 group ( n = 46) and 40 ± 8 years in the control group ( n = 38) ( p = 0.45). The mean CRAE was 181.56 ± 6.40 in the COVID-19 group and 171.29 ± 15.06 in the control group ( p = 0.006). The mean CRVE was 226.34 ± 23.83 in the COVID-19 group and 210.94 ± 22.22 in the control group ( p = 0.044). AVR was 0.81 ± 0.09 in the COVID-19 group and 0.82 ± 0.13 in the control group ( p = 0.712). Conclusion: Patients who had COVID-19 have vasodilation in the retinal vascular structure after recovery. As they may be at risk of retinal vascular disease, COVID-19 patients must be followed after recovery.


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