IN SILICO PROFILING OF THE NEW ALLOSTERIC MODULATOR OF AMPA RECEPTORS.

The aim of this study is to investigate the computer-aided prediction of pharmacological activity and mechanisms of action of 6-[4-methoxy-3-(1H-pyrazol-1-ylmethyl)benzyl]-1,11-dimethyl-3,6,9-triazatricyclo[7.3.1.1]tetradecane-4,8,12-trione (TCT-9). The compound was designed for modulation of ionotropic glutamate AMPA receptors, and its affinity for the receptor has been earlier proven experimentally. A cognitive stimulating effect of TCT-9 has been shown using a model of freezing behavior in mice. The drug candidate TCT-9 is now under the development process: it is intended for the treatment of cognitive impairments in case of brain injury. Following the existing requirements, the present study was carried out in the framework of secondary pharmacodynamic studies to determine possible off-target effects and interaction of the compound with regulatory signaling and metabolic networks/pathways. In silico study of the TCT-9 binding to pharmacologically significant targets and the new AMPA receptor modulator's effects on signaling pathways was carried out by the analysis of structure-activity relationships. Prediction of biological activity spectra was performed using PASS (Prediction of Activity Spectra for Substances), which estimates the probabilities for more than five thousand biological activities. The PharmaExpert program assessed information on the belonging of the targets predicted by the PASS program to the signaling and metabolic pathways. The prediction results are the basis for the experimental verification of the binding of the TCT-9 to the steroid hormone receptor ERR1 and further studies of the drug activity in animal models of diseases.

[The detection of the 25B-NBOMe derivative of phenylethylamine in the biological material]. / Obnaruzhenie 25B-NBOMe - proizvodnogo fenilétilamina v biologicheskom materiale.

This article is focused on the conditions for the detection and identification of 2-[4-bromo-2.5-dimethoxyl]-N-[(2-methoxyphenyl)methyl] ethamine (25B-NBOMe) and its major metabolites by the combination of the HPLC/MS/MS techniques. The high-resolution mass spectra obtained with the use of a linear ion trap are described. The results of the study give evidence of the possibility for the detection of the analytes within 24 hours after drug consumption and within 3 months after the storage of the biological material of interest in a refrigerator at a temperature of 3-5 °C. The data obtained confirmed high stability of 2-(4-bromo-2.5-dimethoxyl]-N-[(2-methoxyphenyl)methyl] ethamine and its metabolites in the biological tissues.

[The development and validation of the method for the identification of ethyl glucuronide and ethyl sulfate as the markers of the consumption of ethyl alcohol during one's lifetime]. / Razrabotka i validatsiia metodiki opredeleniia étilgliukuronida i étilsul'fata kak markerov prizhiznennogo upotrebleniia étilovogo spirta.

The objective of the present study was the development and validation of the rapid reproducible method for the identification of ethyl glucuronide and ethyl sulfate allowing to store and transport the study specimens without the loss of the substances of interest by placing the samples on the paper. We have developed the validated technique for the detection and quantitative determination of ethyl glucuronide and ethyl sulfate in the cadaveric blood and urine by means of low-resolution tandem mass-spectroscopy with the use of deuterated derivatives of these substances as the internal standards. The low threshold for quantitative determination of both above substances is 50 ng/ml for the blood and 100 ng/ml for the urine. The method is characterized by the accuracy and precision with the coefficient of variation below 15% and the influence of the matrix with the coefficient of variation below 15%. The evaluation of stability of the two analytes in blood when stored in the dry condition on the paper carrier during 2 weeks showed that the coefficient of variation did not exceed 6.4%. The comparative study of ethyl glucuronide and ethyl sulfate in the samples of cadaveric blood and urine containing from 0 to 5.2% of ethyl alcohol was carried out. The methods for the transportation of the biological fluids and for the extraction of ethyl glucuronide and ethyl sulfate placed on the paper carrier (Whatman 903) have been proposed. The possibility has been demonstrated to use ethyl glucuronide and ethyl sulfate as the markers of the consumption of ethyl alcohol during one's lifetime for the purpose of investigation of the putrifactive changes of the blood components.

The origin of elevated levels of circulating DNA in blood plasma of premature neonates.

The relationship between the concentration of cell-free DNA (cfDNA) and the number of proliferating/apoptotic lymphocytes in peripheral blood of premature newborns of different gestation age and full-term newborns was determined. The experiments were performed using fluorescent spectrophotometry (with Hoechst 33342), flow cytometry, and microscopy (Feulgen staining of lymphocytes). It was determined that the lymphocyte population of premature newborns may consist of 4.6% of proliferating and 22.1% apoptotic cells. For full-term newborns, the percentage was 2.5% and 2.9%, respectively. A direct correlation between the concentration of extracellular DNA and the number of proliferating lymphocytes of full-term newborns was ascertained (r= 0.400; P < 0.05). For premature newborns, the concentration of extracellular DNA correlated both with proliferating lymphocytes and apoptotic cells. The results show that premature birth causes the induction in lymphocytes of both apoptosis and proliferation that are accompanied by an increased extracellular DNA concentration in the blood of newborn babies.