A Comparison of the Dynamics of S100B, S100A1, and S100A6 mRNA Expression in Hippocampal CA1 Area of Rats during Long-Term Potentiation and after Low-Frequency Stimulation

The interest in tissue- and cell-specific S100 proteins physiological roles in the brain remains high. However, necessary experimental data for the assessment of their dynamics in one of the most important brain activities, its plasticity, is not sufficient. We studied the expression of S100B, S100A1, and S100A6 mRNA in the subfield CA1 of rat hippocampal slices after tetanic and low-frequency stimulation by real-time PCR. Within 30 min after tetanization, a 2–4 fold increase of the S100B mRNA level was observed as compared to the control (intact slices) or to low-frequency stimulation. Subsequently, the S100B mRNA content gradually returned to baseline. The amount of S100A1 mRNA gradually increased during first hour and maintained at the achieved level in the course of second hour after tetanization. The level of S100A6 mRNA did not change following tetanization or low-frequency stimulation

Subetta Treatment Increases Adiponectin Secretion by Mature Human Adipocytes In Vitro

Purpose. To investigate the mechanism of action in peripheral tissues of novel complex drug containing release-active dilutions of antibodies to the beta subunit of the insulin receptor and antibodies to endothelial nitric oxide synthase (Subetta), which has shown efficacy in animal models of diabetes. Methods. Human mature adipocytes were incubated either with Subetta, with one of negative controls (placebo or vehicle), with one of nonspecific controls (release-active dilutions of antibodies to cannabinoid receptor type I or release-active dilutions of rabbit nonimmune serum), or with dimethyl sulfoxide (DMSO) at 37°C in a humidified incubator at 5% CO2 for three days. Rosiglitazone was used as reference drug. Secretion of adiponectin was measured by quantitative enzyme-linked immunosorbent assay (ELISA). Results. Only Subetta significantly stimulates adiponectin production by mature human adipocytes. Nonspecific controls did not significantly affect adiponectin secretion, resulting in adiponectin levels comparable to background values of the negative controls and DMSO. Conclusion. Increasing adiponectin production in absence of insulin by Subetta probably via modulating effect on the beta subunit of the insulin receptor might serve as one of the mechanisms of the antidiabetic effect of this drug. These in vitro results give first insight on possible mechanism of action of Subetta and serve as a background for further studies

Novel Approach to Activity Evaluation for Release-Active Forms of Anti-Interferon-Gamma Antibodies Based on Enzyme-Linked Immunoassay

<div><p>Selection of a suitable assay to measure the activity of drug agents based on release-active forms of anti-interferon-gamma antibodies (RA forms of Abs) is an important step forward in the investigation of such agents. In this study, the enzyme-linked immunosorbent assay was utilized to examine the effect of RA forms of Abs specific for human interferon gamma on the interaction between monoclonal anti-interferon gamma antibodies and recombinant human interferon gamma. The experimental data and the results obtained by using relevant mathematical analysis showed that such RA forms of Abs are able to modulate the monoclonal antibody interaction with both soluble and immobilized (to the assay plate well) interferon gamma. These data demonstrated the importance of using relatively low concentrations of both soluble and plate-immobilized interferon gamma to detect the effects of RA forms of Abs to interferon gamma on the binding of monoclonal antibodies to interferon gamma. It has been suggested that the observed influence of RA forms of Abs on ‘antibody-antigen’ interaction could be used to detect and analyze the activity of drugs containing RA forms of Abs.</p></div

Dose–Response Effect of Antibodies to S100 Protein and Cannabinoid Receptor Type 1 in Released-Active Form in the Light–Dark Test in Mice

Earlier studies have shown that combination of antibodies to S100 protein and to cannabinoid receptor type 1 in released-active form (Brizantin) may possess anxiolytic properties and decrease nicotine dependence. Released-active form of antibodies is a novel approach that permits to modify natural functions of the target molecule (antigen) under investigation. The aim of the present study was to evaluate the anxiolytic-like effect of Brizantin in the light–dark test in mice, according to its ability to influence the number of entries into the lit compartment and the total time spent there. Three doses of Brizantin (2.5, 5, and 10 mL/kg) were compared with diazepam (1 mg/kg), placebo, and vehicle control. Anxiolytic-like effect of the tested drug was shown to be dose dependent, with an increasing trend from 2.5 to 10 mL/kg. Brizantin in its highest dose significantly increased studied behavioral parameters, although its effect was less pronounced than that of the reference drug diazepam (1 mg/kg)

Linear relationship between mAbs to IFN-gamma dilutions and optical density.

<p>The concentration of IFN-gamma absorbed on the plate is 1 µg/ml. Dilutions of mAbs are on X axis (where 0.00001 means dilution of starting antibodies to 1∶100000 times, 0.00002 means dilution of starting antibodies to 1∶50000 times etc.). Optical densities are average values of three measurements. The error bars are the standard deviations.</p

The influence of absorbed IFN-gamma concentration on detection of RA forms of anti-IFN-gamma antibodies.

<p>The figure shows the influence of the concentration of IFN-gamma adsorbed onto microtiter plates on the mAbs dilution curves observed in the presence of control or RA-antibodies to IFN-gamma (RA Abs to IFNg). Test samples prepared of mAbs, pre-diluted up to 1∶5000–1∶135000 in TBS, and either RA forms of Abs to IFN-gamma (RA Abs to IFNg) or control, mixed at a ratio 1∶4 v/v respectively. 100 µL of each sample was transferred into the wells (3 wells for each sample) and was incubated for 45 minutes at room temperature while shaking (shaker-incubator Dynatech, USA). The remainder of the experiment was conducted in accordance with the standard ELISA protocol. Dilutions of mAbs on X axis are those observed after mixing with RA forms of Abs to IFN-gamma or control (where 0.00001 means dilution of starting antibodies to 1∶100000 times, 0.00002 or 2E-05 means dilution of starting antibodies to 1∶50000 times etc.). The Y-axis displays 490 nm optical densities. The error bars represent the standard deviations of the measurements. The concentrations of IFN-gamma adsorbed on the plate are: A 6 µg/ml B - 2 µg/ml C - 0.7 µg/ml; D - 0.25 µg/ml. RA samples are significantly different from the control (F31/60 = 93.4; p<0.0001) in the case of the C and D curves.</p