Tsuchiya T.,Gunma University |
Shimizu H.,Gunma University |
Yamada M.,Gunma University |
Osaki A.,Gunma University |
And 8 more authors.
Clinical Endocrinology | Year: 2010
Background We recently identified a novel anorexigenic protein, nesfatin-1, which is processed from nesfatin/nucleobindin-2 (NUCB2). However, the clinical importance of this protein has not been determined. Objective To investigate its clinical significance in humans, we have established a new specific enzyme-linked immunosorbent assay (ELISA) for human nesfatin-1 in peripheral blood and measured its circulating concentration in healthy subjects. Design The new sandwich-type ELISA method was validated and then used to measure nesfatin-1 levels in plasma samples, under overnight fasting conditions, followed by oral glucose tolerance and meal tests. Patients and measurements A total of 43 nonobese males (age: 24·5 ± 0·6, body mass index (BMI); 21·1 ± 0·3 kg/m2) were recruited to the study for evaluating fasting concentrations of nesfatin-1. In those, fifteen subjects underwent a 75- g oral glucose tolerance test (OGTT) and another 15 underwent a meal test. In addition, fasting concentrations of nesfatin-1 were measured in nine males with high BMI (age: 32·4 ± 3·7, BMI; 37·3 ± 3·8 kg/m2). Results Peripheral concentrations of nesfatin-1 showed a significant negative correlation with BMI, percentage body fat, body fat weight and blood glucose (P < 0·05). Nesfatin-1 concentrations were not significantly changed during OGTT and meal tests. Fasting nesfatin-1 levels were significantly lower in subjects with high BMI compared to nonobese subjects (P < 0·05). Conclusions A new specific and sensitive ELISA for nesfatin-1 was established. Further accumulation of clinical observations is necessary to clarify the role of circulating nesfatin-1 in various metabolic disorders. © 2010 Blackwell Publishing Ltd. Source
Shibayagi Co. | Date: 2014-03-31
The anti-canine N-terminal pro-atrial natriuretic peptide antibodies (anti-canine NT-proANP antibodies) according to the present invention comprise anti-canine NT-proANP recognizing a partial portion or a whole portion of the amino acid sequence 31 to 67 of canine NT-proANP and anti-canine NT-proANP recognizing a partial portion or a whole portion of the amino acid sequence 68 to 98 thereof. The anti-canine NT-proANP antibodies are useful for immunologically measuring canine NT-proANP involved in heart diseases and infections of companion animals such as dogs, such as heart failure, mitral valve insufficiency and filariasis. The immunochromatographic assay using the immunochromatographic means such as immunochromatographic strip is extremely convenient and simple for measuring canine NT-proANP and is used as hand-carried and simple devices.
Kinoshita M.,Teikyo University |
Matsushima T.,Internal Medicine |
Mashimo Y.,Teikyo University |
Kojima M.,Shibayagi Co. |
And 2 more authors.
Experimental Animals | Year: 2010
Abstract: Apolipoprotein B-48 (apoB-48) is produced by the small intestine, is associated with chylomicrons, and appears to be a suitable marker for clinical studies of postprandial dynamics of lipoproteins. It is also associated with cardiovascular risk factors. We have developed an assay system to quantify immuno-reactive apoB-48 in rabbit serum or plasma. A microtiter-plate was coated with monoclonal antibody raised against human apoB-48 C-terminal specific decapeptide that has high homology to the rabbit C-terminal sequence. Appropriate ELISA standard curves were obtained using apoB-48 extracted from rabbit serum by immuno-affinity chromatography. No cross-reactivity was found with apoB-100 in western blot analyses. Intra- and inter-assay CVs were less than 3%. Recovery of rabbit apoB-48 spiked in serum was within 93.4-105%. ApoB-48 levels in healthy controls rabbits fed a normal diet were within the range of 0.903-1.09 μg/ml (mean ± SD: 1.03 ± 0.084 μg/ml). In healthy animals, the blood apoB-48 level was markedly increased by a high fat diet and in the postprandial state in parallel with serum triglyceride. Ezetimibe, cholesterol absorption inhibitor, given orally to rabbits fed on a high fat diet blocked further increase of blood levels of apoB-48 and triglyceride. This method for measuring apoB-48 using the monoclonal antibody is simple, reliable, and suitable for routine analyses. Source
Nishii N.,Tottori University |
Takasu M.,Gifu University |
Kojima M.,Shibayagi Co. |
Hachisu T.,Shibayagi Co. |
And 5 more authors.
Domestic Animal Endocrinology | Year: 2010
A substance interfering with the enzyme-linked immunosorbent assay (ELISA) for feline insulin concentration was investigated in healthy cats. An insulin-binding substance isolated from feline serum showed 2 bands at 25 and 50 kDa in SDS-PAGE, suggesting the presence of immunoglobulin G (IgG). Insulin-binding IgG from healthy cats indeed reduced insulin immunoreactivity in the ELISA for determining insulin concentration. The insulin-binding IgG was polyclonal/polyreactive and showed certain specificity, high affinity, and high binding capacity, which was evaluated by liquid-phase radioimmunoassay with Scatchard plot analysis. Epitope analysis revealed that the insulin-binding IgG showed significant binding at residues A1-5 and B20-30 of the insulin molecule. Removal of the antibodies from serum enabled the determination of serum insulin concentrations by ELISA. Our data indicated that serum from healthy cats contained substantial amounts of natural autoantibodies combined with insulin, and that the antibodies interfered with the heterologous immunoassay for serum insulin concentration. © 2009 Elsevier Inc. All rights reserved. Source