A monoclonal antibody specific for zearalenone (ZEN) was produced by fusion of mouse myeloma cells (NS-1) and splenocytes isolated from BALB/c mice that had been immunized with a novel type of immunogen. The immunogen was prepared by coupling 5-aminozearalenone, which was synthesized from ZEN in two steps, with bovine serum albumin through the amino group a t the C-5 position of the compound. The anti-ZEN monoclonal antibody (Ab 7-1-144) thus obtained belonged to the IgGl subclass with X light chain. The association constant of the antibody for ZEN was 1.1 X los L/mol. Crossreaction studies showed that the antibody was highly specific for ZEN. The cross-reactivities of this antibody for zearalanone, ZEN 4-methyl ether, a-zearalenol, and ZEN 2,4-dimethyl ether were 4.0,2.5, 0.9, and 0.3%, respectively, of that found for ZEN. Practically no cross-reactivity was observed with P-zearalenol, a-and 0-zearalanols, and trichothecene mycotoxins including T-2 toxin, nivalenol, anddeoxynivalenol. An indirect enzyme-linked immunosorbent assay (ELISA) based on the competitive binding principle was developed for the detection of ZEN using Ab 7-1-144. The response range for ZEN in the present study was between 0.3 and 100 ng/mL (15 and 5000 picograms per assay). The binding inhibition by ZEN was nearly linear in this range.
Antibodies against nivalenol (NIV) tetraacetate (Tetra-Ac-NIV) were prepared by immunizing rabbits with a hemisuccinate derivative of 8-hydroxy-3,4,7,15-tetraacetyl-12,13-epoxytrichothece-9-en conjugated to bovine serum albumin. A radioimmunoassay system with one of these sera was developed to measure NIV contamination in barley. The detection limit for Tetra-Ac-NIV was about 0.5 ng/ml. The relative crossreactivities of the antiserum with Tetra-Ac-NIV, acetyl T-2 toxin, and scirpenol triacetate, which were determined by the competitive radioimmunoassay, were 1, 0.78, and 0.56, respectively. Other derivatives showed no cross-reactivity. For the determination of NIV in a barley sample, NIV was extracted from the sample with acetonitrile-water (7:3), defatted with hexane, and then acetylated with acetic anhydride to form Tetra-Ac-NIV. The reaction mixture was loaded onto a C18 cartridge to remove excess reagents and impurities. Tetra-Ac-NIV was eluted from the cartridge with 50% methanol in water, and the eluate was subjected to radioimmunoassay. Analysis of six naturally contaminated barley samples for NIV revealed that radioimmunoassay results agreed well with gas chromatographic analyses.
Three monoclonal antibodies were obtained by the fusion of mouse myeloma cells with splenocytes isolated from BALB/c mice that had been immunized with 8-hydroxy-3,4,7,15-tetraacetyl-nivalenol hemiglutarate covalently bound to bovine serum albumin. These anti-nivalenol tetraacetate monoclonal antibodies were of the IgG type and highly specific to nivalenol tetraacetate, with an apparent association constant of about 10(8)M-1. The relative cross-reactivities of one monoclonal antibody with nivalenol tetraacetate, acetyl T-2 toxin, and scirpenol triacetate were found to be 1.0, 0.02 and 0.03, respectively. Other derivatives showed no cross-reactivity at all. An indirect enzyme-linked immunosorbent assay (ELISA) based on the competitive binding principle was developed using the antibody from clone D18.102.59. The sensitivity of the system was about 0.1 ng of nivalenol tetraacetate per assay. Comparison of nivalenol levels detected in naturally contaminated barley samples by competitive indirect ELISA and gas chromatography (GC) showed good agreement, indicating that the antibody is useful for the measurement of nivalenol in naturally contaminated cereals and grains.
Introduction: Chiangmai is one of the provinces in Northern Thailand. The Aging population has increased rapidly since family planning commenced in 1972. For the surveillance of aging, we started a health investigation of the elderly over 60 years of age in the suburban area of Chiangmai since 1997 with the Nippon Medical School Team. Materials and Method: Medical examination was carried out for the elderly (male = 34, female = 73) at Department of Family Medicine, Chiangmai University, Chiangmai Thailand in August 2002. Chula Mental Test (CMT) was used for cognitive function and Geriatric Depression Scale was used for evaluation of depressive condition. Results: The major health problems of this group of elderly in Chiangmai Province were visual acuity abnormality (98.1%), anemia (66.4%), hypertension (37.4%), dental caries (29%), bone and joint disease (23.4%), cardiovascular disease (11.2%), gingivitis (11.2%), diabetes mellitus (9.3%), digestive system disease (8.4%), and presbycrusis (8.4%). The prevalence of cognitive impairment and depression were 0.9 and 2.8%, respectively. The prevalence of high cholesterol, triglyceride, and LDL cholesterol were 43.9, 29.0, and 15.9%, respectively. The prevalence of high BUN, creatinine, and uric acid were 17.8, 11.2, and 29.9%, respectively. The prevalence of high GOT, GPT, gamma GT, ZTT, and TTT were 4.7, 1.9, 53.3, 35.5, and 21.5%, respectively. The prevalence of low hemoglobin was 66.4%. Conclusion: There was a high prevalence of anemia and hyperlipidemia in this group of elderly in Chiangmai Province. The analysis of background factors and further investigation should be carried out continuously.
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