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Biomedical subjects

L Solti

Publications and source records attributed to L Solti.

At least 37 records · Page 2Linked to original sources

Progesterone determination in equine plasma using different immunoassays.

Several assay systems (3H radioimmunoassay (RIA) with and without extraction; microplate enzyme-linked immunoassay (ELISA); qualitative ELISA (tube test)] were used to measure plasma progesterone concentration in mare plasma. The direct RIA showed a close correlation (R = 0.94) with the extraction RIA. The direct RIA and the microplate ELISA were compared in two different studies. In the first study 1155 samples of postpartum mares were used for progesterone determination with both assays. The ELISA resulted in more elevated values both in oestrus and dioestrus (0.19+/-0.3 and 2.44+/-3.62 nmol/l for oestrus, n = 436, and 8.94+/-4.29 and 27.88+/-18.34 nmol/l for dioestrus, N = 719, for the RIA and ELISA, respectively, R = 0.71). The evaluation of individual progesterone profiles has revealed that the microplate ELISA detects the time of ovulation at the same time as it is determined by the RIA and clinical examination. The sensitivity and specificity were calculated for different progesterone threshold values. In the second study including 7 non-pregnant, cycling mares the progesterone concentration of 240 samples was determined by both assays. Basal values (Day 0) obtained with the ELISA were higher (1.57 nmol/l) than those of the RIA (0.2 nmol/l). Both curves reached the same maximum concentration (12.11 and 12.45 nmol/l) 5 days after ovulation. The correlation between the RIA and ELISA values was high (R = 0.90). The tube test was compared to the microplate ELISA as reference using 576 plasma samples of 34 non-pregnant, non-cycling mares included in an ovulation induction study. Of these samples 118 had higher and 458 had lower values than 3.18 nmol/l. In most cases the tube test was in complete agreement with the microplate ELISA. The sensitivity, specificity, + predictive and - predictive values for the tube test were 79.7%, 95.4%, 81.7% and 94.8%, respectively.

Animals↗

Induction of acrosome reaction in dog sperm by calcium ionophore.

The sensitivity of the plasma membrane to calcium ionophore (A23187) challenge was studied in dog sperm using fluorescein lectin staining for the assessment of acrosomal status and viability. Second fraction ejaculates from 5 dogs were washed, resuspended in Ca(2+)-free (EDTA-treated), 50, 100, 500, 1000 and 2000 microM/l Ca(2+)-containing Sp-TALP medium and induced with 50, 250, 500, 1000, 2500 and 5000 nM/l calcium ionophore. Samples were collected from each aliquot after 30 and 60 min of induction to assess the percentage of acrosome reacted sperm cells (AR rate), viability and motility by fluorescein isothiocyanate conjugated peanut agglutinin (FITC-PNA) and ethidium-homodimer combined staining. On each slide, 200 sperm cells were assessed under epifluorescence microscope (x 1250) in a blind manner. The response to ionophore challenge (AR rate, viability, motility) varied with Ca2+ and ionophore concentration in the suspension. A significantly higher AR rate was detected in samples containing 100, 500, 1000 and 2000 microM/L Ca2+ (> 40%) than in that containing 50 microM/L. Acrosome reaction could not be successfully induced in the EDTA-treated sample and in any of the aliquots in which 50, 250 and 500 nM/L ionophore concentrations were used for induction. Motility decreased drastically in all of the treated samples and stopped in that sample where as significant AR rate could be detected. Viability remained high (> 75%) during the incubation and did not differ significantly in the treated and the control groups.

Acrosome↗

A new monoclonal antibody detecting ochratoxin A at the picogram level.

A monoclonal antibody against ochratoxin A was produced after immunization of Balb/c mice with ochratoxin A-BSA. This antibody was of the IgG1 heavy chain subclass with a kappa type light chain. The 50% inhibition value was 0.45 ng ml-1 in a direct competitive ELISA and the detection limit was 42 pg ml-1. This antibody is very specific, cross-reacting only with ochratoxin B (9.3%).

Animals↗

Latent immunization to produce high-affinity monoclonal antibodies to progesterone.

A simple immunization method to obtain high-affinity monoclonal antibodies to progesterone is described in this article. The method is based on the theory of affinity maturation. A long interval between antigen priming and booster ("latent immunization") permits an undisturbed completion of affinity maturation, resulting in the accumulation of memory B lymphocytes with high-affinity Ig receptors, and consequently, in a higher rate of hybridoma clones producing high-affinity antibody after cell fusion. Antibodies obtained after hyperimmunization and latent immunization are compared in a homologous, direct, competitive ELISA. The average numbers of high-affinity antibodies per fusion were 1.3 and 5.7 in the hyperimmunized and latent immunized groups, respectively. There was no significant difference in the specificities between the two immunization groups.

Animals↗

In vitro neutralization of T-2 toxin toxicity by a monoclonal antibody.

A T-2 toxin specific monoclonal antibody, IgG1 K, with a low level of ELISA cross-reactivity to Acetyl T-2, HT-2, and iso T-2 toxins has been produced. The ability of this monoclonal antibody to neutralize the cytotoxicity of T-2 toxin in PHA stimulated cultures of human lymphocytes was determined by the MTT method. The complete neutralization of the toxic effect of 0.02 microM T-2 toxin was obtained with 0.03 microM of MoAb, whereas the 50% neutralizing dose (ND50) was observed at 0.009 microM of MoAb. Partial neutralization was observed with Acetyl T-2 toxin (ND50 = 0.038 microM) and HT-2 (ND50 = 0.94 microM). These results could represent a rational for clinical use of T-2 toxin specific monoclonal antibody in prophylaxis and therapy of T-2 toxemia.

Animals↗

Monoclonal antibody-based enzyme-linked immunosorbent assay of Fusarium T-2 and zearalenone toxins in cereals.

Direct, competitive enzyme-linked immunosorbent assays (ELISAs) with monoclonal antibodies have been developed for quantitative determination of trichothecene T-2 toxin (T-2), and zearalenone (F-2) from different cereals: Among the several extraction solvents tried, 89% acetonitrile with additives was chosen. The extracts were then used without cleanup in the ELISA. With appropriate dilution of the samples (1:25 or 1:50), the matrix effects caused by lipid and/or protein content of the samples can be diminished to the extent that the assay is no longer impaired. The mean recoveries from cereals infected with 100 to 2,000 ng of T-2 and 50 to 500 ng of F-2 per g were 85 and 91%, respectively. The measuring range of the T-2 test is 100 to 2,000 ng/g, and that of the F-2 test is 25 to 400 ng/g. The mean within-assay and interassay coefficients of variation of standard curves are both less than 10%. According to recovery results with artificially infected cereals, our tests proved to be suitable for rapid screening of food and feed samples for the presence of T-2 and F-2 toxins.

Antibodies, Monoclonal↗

Influence of beta-carotene on fertility in rabbits when using embryo transfer programs.

The effect of beta-carotene on reproduction traits in rabbits was studied in 509 (superovulated and normally ovulated) donors and 239 recipients by using embryo/gene transfer performed at 2 different locations. All of the bucks and the half of the females were fed a diet supplemented with 40 mg synthetic beta-carotene (Rovimix((R)))/kg feed. Embryos at the pronucleus stage were collected 19 to 21 hours after induction of ovulation with human chorionic gonadotropin (hCG); they were then microinjected into the male pronucleus and transferred to synchronized recipients. Data were obtained from the time when the donors and recipients were caged, until the pups resulting from the embryo transfers were weaned. Supplemented beta-carotene did not affect most of the 30 traits that were analyzed. However superovulated donors in Project 2 that received the beta-carotene enriched diet had a 14% lighter ovary weight (P<0.05) and less than half of the oocytes were unfertilized (P<0.05). In Project 1 (beta-carotene group) there was a greater number of pups born (36%, P<0.05) and more of these pups were born alive (53%, P<0.05).

Journal Article↗

Treatment of anoestrous mares with a synthetic progestagen, allyloestrenol.

Anoestrous mares were treated with prostaglandin (n = 43) and those that did not respond to prostaglandin (n = 29) with a synthetic progestagen, allyloestrenol, at a dose of 0.05 mg/kg body mass for 12 days. After the cessation of the long-term per os gestagen blockade the animals were checked for heat and, if a preovulatory follicle could be palpated, 2000 IU hCG was administered to induce ovulation. In some animals the plasma 17 beta-oestradiol (E2) and progesterone (P4) levels were also followed up throughout the gestagen treatment and for 10-14 days thereafter. As the favourable oestrus rate (86.2%) and pregnancy rate (65.5%) indicate, the 12-day allyloestrenol treatment seems to be a reliable tool for induction of cycling in mares anoestrous early in the season. These results, supported by the E2 and P4 profiles, suggest that an important reason for anoestrus in mares is the absence of a functional corpus luteum that can be substituted for by exogenous gestagen administration. The timing of ovulation, however, should still be improved.

Allylestrenol↗