The multifaceted role of proteases and modern analytical methods for investigation of their catalytic activity.

We discuss the diverse functions of proteases in the context of their biotechnological and medical significance, as well as analytical approaches used to determine the functional activity of these enzymes. An insight into modern approaches to studying the kinetics and specificity of proteases, based on spectral (absorption, fluorescence), mass spectrometric, immunological, calorimetric, and electrochemical methods of analysis is given. We also examine in detail electrochemical systems for determining the activity and specificity of proteases.

Bielectrode Strategy for Determination of CYP2E1 Catalytic Activity: Electrodes with Bactosomes and Voltammetric Determination of 6-Hydroxychlorzoxazone.

We describe a bielectrode system for evaluation of the electrocatalytic activity of cytochrome P450 2E1 (CYP2E1) towards chlorzoxazone. One electrode of the system was employed to immobilize Bactosomes with human CYP2E1, cytochrome P450 reductase (CPR), and cytochrome (cyt ). The second electrode was used to quantify CYP2E1-produced 6-hydroxychlorzoxazone by its direct electrochemical oxidation, registered using square-wave voltammetry.

Electrochemical biosensor for trypsin activity assay based on cleavage of immobilized tyrosine-containing peptide.

In this work, we proposed a biosensor for trypsin proteolytic activity assay using immobilization of model peptides on screen-printed electrodes (SPE) modified with gold nanoparticles (AuNPs) prepared by electrosynthetic method. Sensing of proteolytic activity was based on electrochemical oxidation of tyrosine residues of peptides. We designed peptides containing N-terminal cysteine residue for immobilization on an SPE, modified with gold nanoparticles, trypsin-specific cleavage site and tyrosine residue as a redox label.

Biotransformation of phenytoin in the electrochemically-driven CYP2C19 system.

The possibility of the detection of atypical kinetic profiles of drug biotransformation using electrochemical systems based on immobilized cytochromes P450 with phenytoin hydroxylation by cytochrome P450 2C19 (CYP2C19) as an example was evaluated for the first time. For this purpose, we developed an electrochemical system, where one of the electrodes was modified by didodecyldimethylammonium bromide (DDAB) and was used as an electron donor for reduction of heme iron ion of the immobilized CYP2C19 and initiation of the catalytic reaction, while the second electrode was not modified and served for an electrochemical quantitation of 4-hydroxyphenytoin, which is a metabolite of antiepileptic drug phenytoin, by its oxidation peak. It was revealed that the dependence of the rate of 4-hydroxyphenytoin formation on phenytoin concentration is described by the equation for two enzymes or two binding sites indicating the existing of high- and low-affinity forms of the enzyme.

[The interactions of abiraterone and its pharmacologically active metabolite D4A with cytochrome P450 2C9 (CYP2C9)].

Interactions of cytochrome P450 2C9 (CYP2C9) were studied with the antitumor drug abiraterone and its pharmacologically active metabolite D4A, promising as an agent for prostate cancer treatment. It was shown by absorption spectroscopy, that both investigated compounds induced spectral changes of CYP2C9, indicating interactions of the pyridine nitrogen atom with the heme iron ion of the active site of the enzyme, but interactions of the ligands with the enzyme could be mediated by a water molecule bound to the heme iron ion. Based on the spectral changes, the values of dissociation constants (KS) for complexes of abiraterone and D4A with CYP2C9 were calculated as 1.

The Relationship Between Value Orientations and Personal Readiness for Activity in Youth From Russia, Kazakhstan and Latvia.

Background: The development of high-quality human capital is an important objective that involves value orientations, cultural dimensions and psychological characteristics of activity. This article presents a cross-cultural comparison of value orientations and psychological parameters of activity among youth from Russia, Kazakhstan, and Latvia.

Objective: The study addressed three questions: (1) Are there values and attitudes related to the readiness for activity among youth in the three countries? (2) Are there any differences between values and parameters of the psychological system of activity in the Russian, Kazakhstani and Latvian samples? (3) What values and attitudes predict the youth's readiness for activity in each country?

Design: University students from Russia, Kazakhstan and Latvia were invited to participate in the study.

Association of , and Genes Polymorphisms With the Calcium Urolithiasis Development in Russian Population.

Kidney stone disease is an urgent medical and social problem. Genetic factors play an important role in the disease development. This study aims to establish an association between polymorphisms in genes coding for proteins involved in calcium metabolism and the development of calcium urolithiasis in Russian population.

[A complicated case of calcium urolithiasis in a carrier of SLC7A9 gene mutation responsible for cystinuria].

The article describes a clinical case of kidney stone disease (KSD) in a child of 4 y.o. with calcium urolithiasis.

Interactions of galeterone and its 3-keto-Δ4 metabolite (D4G) with one of the key enzymes of corticosteroid biosynthesis - steroid 21-monooxygenase (CYP21A2).

We have investigated interactions of galeterone and its pharmacologically active metabolite - 3-keto-Δ4-galeterone (D4G) - with one of the key enzymes of corticosteroid biosynthesis - steroid 21-monooxygenase (CYP21A2). It was shown by absorption spectroscopy that both compounds induce type I spectral changes of CYP21A2. Spectral dissociation constants (K ) of complexes of CYP21A2 with galeterone or D4G were calculated as 3.

The interactions of a number of steroid-metabolizing cytochromes P450 with abiraterone D4A metabolite: spectral analysis and molecular docking.

The interactions of pharmacologically active 3-keto-Δ4-metabolite of anticancer drug abiraterone (D4A) with steroid-metabolizing cytochromes P450 (CYP51A1, CYP11A1, CYP19A1) was studied by absorption spectroscopy and molecular docking. Both abiraterone and D4A induce type I spectral changes of CYP51A1, one of the enzymes of cholesterol biosynthesis. We have revealed that D4A did not induce spectral changes of CYP11A1, the key enzyme of pregnenolone biosynthesis, unlike abiraterone (type II ligand of CYP11A1).

[Molecular and genetic technologies for the diagnosis of monogenic forms of urinary stone disease: clinical cases].

Kidney stone disease (KSD) is an actual problem of modern health care. By now, more than 80 monogenic forms of urolithiasis have been described. To diagnose such forms of KSD different molecular genetic technologies are used.

[Genetic aspects of primary hyperoxaluria: epidemiology, ethiology, pathogenesis, and clinical signs of the disorder].

Primary hyperoxaluria is a group of rare inherited diseases characterized by impaired oxalate metabolism with the early manifestation of urolithiasis and the development of the chronic kidney disease. The mutations in the AGXT, GRHPR, HOGA1 genes are attributable for different types of primary hyperoxaluria leading to the dysfunction of specific enzymes involved in the oxalate metabolism. The article summary the current data on the epidemiology, genetic and biochemical aspects of pathogenesis of the primary hyperoxaluria types 1-3.

[Genetic aspects of primary hyperoxaluria: diagnostics and treatment].

Primary hyperoxaluria is a group of inherited metabolic diseases characterized by increased formation of calcium-oxalate stones in kidneys with development of nephrolithiasis and chronic kidney disease. The article summarizes the modern information on the diagnostics and treatment of the disorder depending on genotype of the patient (AGXT, GRHPR, HOGA1 genes). The evaluation of the molecular genetic aetiology of the kidney stone disease contributes to the personalized treatment and prevention of the pathology in the patients and their relatives.

Estimation of the inhibiting impact of abiraterone D4A metabolite on human steroid 21-monooxygenase (CYP21A2).

Abiraterone D4A metabolite, the product of 3β-hydroxysteroid dehydrogenase activity toward abiraterone, may serve as a potential antitumor agent for the treatment of prostate cancer. The main adverse effect of abiraterone is the disruption of corticosteroid biosynthesis, and the more pharmacologically active abiraterone D4A metabolite may have the same issues. We therefore estimated the inhibiting impact of the abiraterone D4A metabolite on one of the key corticosteroidogenic enzymes - human steroid 21-monooxygenase (CYP21A2).

[Genetic factors for monogenic forms of calcium urolithiasis].

The article presents pooled results of domestic and international studies investigating genetic aspects of urolithiasis associated with impaired calcium metabolism. The review highlights the importance of early and accurate diagnosis of hereditary diseases associated with kidney stone formation. Of more than 80 currently known monogenic forms of urolithiasis, the authors provide the list of the most significant forms.

[Genetic aspects of urolithiasis].

The article presents summarized results of domestic and international studies of the genetic aspects of urolithiasis. The presented evidence suggests the importance of early and accurate molecular genetic diagnostics of hereditary diseases associated with stone formation for timely treatment and prevention for patients' relatives. Also provided are examples of using molecular genetic diagnostics in urologist's practice for monogenic and multifactorial diseases associated with stone formation.

[The influence of short wavelength ultraviolet radiation on the healing of thermal wounds (an experimental study)].

Unlabelled: The topicality of the problem stated in the title of the this article is attributable to the lack of up-to-date information on the treatment of thermal skin wounds with the use of short wavelength ultraviolet radiation (UVR). The objective of the present study was to evaluate the effectiveness of the courses of short wavelength ultraviolet radiation under experimental conditions using a model of standardized thermal skin wounds in rabbits starting from the proliferative phase of the healing.

Material And Methods: The experiments were carried out on 24 grey rabbits.

[The study of neurotoxicity of toluene in conditions of experimental modeling of prenatal hypoxic damage of the brain].

There was executed the study of the impact of toluene on indices of behavior, cognitive capabilities and bioelectric activity of the brain in white rats with normal course of the period of antenatal development and against background ofprenatal hemic hypoxia Prenatal hypoxia was modeled in pregnant female rats by subcutaneous injection of sodium nitrite in a dose of 50 mg/kg at from the 10 to the 19 day of gestation. At the age of 3 months the males from the obtained offspring were exposed to inhalation exposure of toluene (150 ppm, 4 weeks). After exposure to toluene in animals there was evaluated the pattern of individual behavior, indices of cognitive capabilities and also bioelectric activity of the brain.

[Organization of genetic service in multiprofile hospitals].

The necessity to organize genetic service for patients and their relatives in multiprofile hospitals is discussed. Analysis of long-term experience of the Medical Genetic Department at the First Moscow Medical University yielded main characteristics to be used for the organization of medical genetic service at such hospitals based at independent divisions. Such services ensure more efficacious care than those at the regional level.

[Assessment of schemes for sewage purification from petroleum products, by using various flotation methods].

Petroleum products are the most common pollutants in petroleum refinery wastewater and are freed from the latter by flotation that is one of the most frequently applied physicochemical methods. The existing petroleum refinery OAO "Angara Petroleum Company" scheme for sewage purification from petroleum products, by using pressure flotation and proposed as a competitive purification scheme by applying electrical and impeller flotations underwent a comparative ecologoeconomic analysis. The use of electrical flotation instead of pressure flotation and that of an impeller flotation-electrical flotation system instead of a mechanical purification-pressure flotation one can considerably lower the concentration of petroleum products at the wastewater outlet into the Angara river.

[Use of cycloferon in the treatment of patients with pulmonary tuberculosis with mild clinical manifestations].

Aim: to evaluate the efficacy of cyclopheron used in patients with mild clinical manifestations of focal and infiltrative pulmonary tuberculosis (PT).

Subjects And Methods: Thirty first detected patients with PT, in whom a complex of their etiotropic therapy comprised cyclopheron, and 32 similar patients who received etiotropic agents and placebo were examined. Tuberculosis process activity was confirmed by clinical, X-ray, bacteriological, and immunological studies.

[Induction of synthesis and activation of Penicillium commune alpha-L-rhamnosidase].

For the first time the influence of a number of synthetic porfirin and fluoren derivatives, coordinative germanium substances and surfactants on the biosynthesis and activity of Penicillium commune 266 alpha-L-rhamnosidase has been studied. It is shown that some of porfirin derivatives and coordinative germanium substances may be used as inducers of biosynthesis (143-430%) and also as stimulators of alpha-L-rhamnosidase activity (15-44%). It is more expedient to use biscitratgermanium acid in a complex with asparaginic acid (430%) or threonine (370%) as inducers of biosynthesis of enzyme.

[The first successful bilateral lung transplantation in Russia].

Lung transplantation is one of the most difficult fields of contemporary transplantology. The operation was performed on a 53-year-old female who had been suffering from terminal stage COPD. The period of cold ischemia for the right and left lung was 1 hour 30 minutes and 2 hours 25 minutes respectively.

[Treatment for progressive pulmonary tuberculosis with a complex of stress-limiting drugs].

The study of the performance of major adaptive systems and the parameters of a hemogram in 63 patients with progressive pulmonary tuberculosis has established the stage of anxiety of a distress reaction, which serves as a pathogenetic rationale for the prescription of a complex of stress-limiting drugs, including sodium hydroxybutyrate, glycine, dalargin, and tocopherol acetate. The use of these agents in combination with intensive antituberculous therapy promotes a rapid progression of the stage of anxiety of the adaptation syndrome to the stage of resistance and reduces the time of stabilization of the patients' status and the number of fatal outcomes.