A novel preliminary metabolomic panel for IHD diagnostics and pathogenesis.

Cardiovascular disease (CVD) represents one of the main causes of mortality worldwide and nearly a half of it is related to ischemic heart disease (IHD). The article represents a comprehensive study on the diagnostics of IHD through the targeted metabolomic profiling and machine learning techniques. A total of 112 subjects were enrolled in the study, consisting of 76 IHD patients and 36 non-CVD subjects. Metabolomic profiling was conducted, involving the quantitative analysis of 87 endogenous metabolites in plasma. A novel regression method of age-adjustment correction of metabolomics data was developed. We identified 36 significantly changed metabolites which included increased cystathionine and dimethylglycine and the decreased ADMA and arginine. Tryptophan catabolism pathways showed significant alterations with increased levels of serotonin, intermediates of the kynurenine pathway and decreased intermediates of indole pathway. Amino acid profiles indicated elevated branched-chain amino acids and increased amino acid ratios. Short-chain acylcarnitines were reduced, while long-chain acylcarnitines were elevated. Based on these metabolites data, machine learning algorithms: logistic regression, support vector machine, decision trees, random forest, and gradient boosting, were used for IHD diagnostic models. Random forest demonstrated the highest accuracy with an AUC of 0.98. The metabolites Norepinephrine; Xanthurenic acid; Anthranilic acid; Serotonin; C6-DC; C14-OH; C16; C16-OH; GSG; Phenylalanine and Methionine were found to be significant and may serve as a novel preliminary panel for IHD diagnostics. Further studies are needed to confirm these findings.

[Assessment of systemic inflammation activity, myocardial structure and functional features, their relationship in patients with multiple myeloma, receiving bortezomib therapy].

AIM: To study the dynamics of calculated indices [neutrophil-lymphocyte ratio (NLR); systemic inflammation index (SIV)] and biomarkers of systemic inflammation [interleukin-1ß (IL-1ß); high-sensitivity C-reactive protein (hsCRP)], parameters of the structure-and-function state of the myocardium and intracardiac hemodynamics, and their relationship in patients newly diagnosed with multiple myeloma (MM) at the onset of the disease and after 6 courses of chemotherapy (CT) containing the proteasome inhibitor bortezomib. MATERIAL AND METHODS: This prospective study included 30 patients aged 63.8±10.0 years diagnosed with MM; 17 (56.7 %) of them were men. The following tests were performed for all patients: measurement of IL-1ß and hsCRP, calculation of the inflammation indexes NLR and SIV, transthoracic echocardiography before and after 6 courses of bortezomib-containing CT. At the time of study completion, 9 patients dropped out due to reasons not related to cardiovascular complications of CT. RESULTS: The antitumor therapy was associated with increases of immune-inflammation indexes: NLR increased from 1.54 [1.02; 1.83] to 2.9 [1.9; 4.35] (p=0.009) and SIV increased from 402.95 [230.5; 534.0] to 1102.2 [453.1; 1307.9] (р=0.014). IL-1ß increased from 5.15 [4.05; 5.77] to 6.22 [5.66; 6.52] pg/ml remaining within the reference range (p=0.142) whereas hsCRP decreased from 1.02 [0.02; 2.71] to 0.02 [0.02; 0.82] IU/l (p=0.138). Statistically significant changes in parameters of heart remodeling and clinical picture of cardiovascular complications were not observed. A correlation analysis showed significant inverse correlations of hsCRP with left ventricular ejection fraction (LV EF) (r= -0.557; p=0.003), the number of plasma cells (PC) with LV EF (r= -0.443; p=0.023), and a direct correlation of the number of PC with hsCRP (r=0.433; p=0.022). CONCLUSION: During the study, the accepted criteria for cardiotoxicity of bortezomib-containing chemotherapy in patients with MM, were not met. The identified correlations between the level of markers for acute inflammation, indexes of intracardiac hemodynamics, and the immediate MM substrate may indicate the role of chronic low-intensity inflammation in the pathogenesis of myocardial remodeling in patients with MM. This necessitates further studies on larger samples of patients to assess the prognostic significance.

Comparative analysis of tryptophan and downstream metabolites of the kynurenine and serotonin pathways in patients with arterial hypertension and coronary artery disease.

Aim    To compare serum concentrations of tryptophane (Trp) and its metabolites in subjects with no cardiovascular disease (CVD) and patients with СVD, including arterial hypertension (AH) and ischemic heart disease (IHD).Material and methods    This study included 131 participants; 58 participants (11 of them with documented peripheral atherosclerosis) were included into the AH group, 46 participants were included into the IHD group, and 27 participants with no signs of CVD were included into the control group. Plasma concentrations of Trp and its metabolites were measured by high-performance liquid chromatography in combination with a triple quadrupole analyzer.Results    Comparison of the three study groups revealed significant differences in concentrations of Trp (р=0.029), kynurenine (p<0.001), kynurenine/Trp ratio (p<0.001), quinolinic acid (р=0.007), kynurenic acid (р=0.003), serotonin (p<0.001), and 5­hydroxyindoleacetic acid (5­HIAA) (р=0.011). When the AH group was subdivided into subgroups without and with documented peripheral atherosclerosis, the intergroup differences remained for concentrations of kynurenine, kynurenine/Trp ratio, quinolinic acid, kynurenic acid, serotonin, and 5­HIAA. Also, correlations were found between concentrations of Trp metabolites and laboratory and instrumental data, primarily inflammatory markers. Conclusion    Analysis of serum concentrations of Trp and its metabolites in CVD patients showed increases in kynurenine, kynurenine/Trp ratio, quinolinic acid, kynurenic acid, and 5­HIAA along with decreases in concentrations of Trp and serotonin in the groups of AH, AH with documented peripheral atherosclerosis, and IHD.

[Determination of zaleplon and clobazam by high performance liquid chromatography - high resolution tandem mass spectrometry using Orbitrap technology]. / Opredelenie zaleplona i klobazama metodom vysokoeffektivnoi zhidkostnoi khromatografii ­ tandemnoi mass-spektrometrii s vysokim razresheniem s ispol'zovaniem tekhnologii Orbitrap.

THE AIM OF THE STUDY: Was to develop a method for the qualitative and quantitative determination of benzodiazepine receptor agonists (zaleplon and clobazam), taking into account different chemical structures, by high-performance liquid chromatography-high-resolution tandem mass spectrometry using Orbitrap technology for the purposes and tasks of forensic medical examination. A method has been developed for the identification and quantitative determination of zaleplon and clobazam by high-performance liquid chromatography-high-resolution tandem mass spectrometry using Orbitrap technology. The proposed identification method can be used in the future to form a database of domestic mass spectra for potent and narcotic substances.

[The study of tryptophan metabolite concentrations in blood serum and fecal extracts from obese children]. / Izuchenie soderzhaniia metabolitov obmena triptofana v syvorotke krovi i kalovykh ékstraktakh u detei s ozhireniem.

We found that changes in the concentrations of tryptophan metabolites in the blood serum and in the intestinal contents are one of the mechanisms for the formation of metabolic coupling in the system "macroorganism-intestinal microbiota", which undergoes significant changes in the development of obesity. Although blood kynurenine remained basically unchanged in obese children we found an increase in some of its serum metabolites: anthranilic, kynurenic and xanthurenic acids. It is noteworthy that in the analysis of fecal matter in obese children, revealed a 2-fold increase in the level of kynurenine while the concentration of kynurenine pathway metabolites corresponded to the level of the group of healthy children. This may indicate the metabolic activation of the microbiota associated with the intestinal mucosa. This is also supported by the absence of statistically significant differences in the concentration of indole in healthy children and in obese children in fecal analyses, and a significant increase in the concentration of indole-3-lactate and indole-3-acetate in the blood serum of obese children.

[The organization of chemical toxicological service and medical examination of persons for condition of alcohol drunkenness, fact of consumption of drug and other psychoactive substances in the Khanty-Mansi Autonomous Okrug-Yugra].

The article presents the analysis of organization of chemical-toxicological service and in the Khanty-Mansi Autonomous Okrug-Yugra. The introduction of three-level system for provision of addiction medical care of population and implementation of the principle of zoning made it possible to organize three-level system of chemical-toxicological service that permitted to achieve equal access to chemical-toxicological and addiction care services in all territories of the region. The investments in chemical-toxicological laboratories, supply of modern equipment, advanced training of laboratory specialists, cooperation with leading research centers and professional communities outside of the region allowed timely and accurately identify psychoactive substances, prompt response in cases of mass poisoning with novel psychoactive substances, and quick introduction of advanced methods of chemical-toxicological examination to the medical organizations of the large industrial northern region of Russia.

[Metabolomic profiling in patients with metabolic syndrome].

OBJECTIVE: To identify biomarkers, which are most specific for patients with metabolic syndrome (MS) using metabolomic profiling. MATERIALS AND METHODS: Metabolomic profiling of patients with MS and comparison of their profile with the profile of volunteers was performed using high-performance liquid chromatography-mass-spectrometry. RESULTS: The metabolomic profile of MS patients differed in several amino acids, including choline, cysteine, and serine and in the acylcarnitine group (р<0.05 for all comparisons). CONCLUSION: The metabolites most specific for MS patients were identified. Increased concentrations of a combination of amino acids and carnitines can be considered as possible additional risk factors for cardiovascular diseases.

[Fast HPLC-MS/MS screening method for identification of illegal drugs including new psychoactive substances and their metabolites in human urine]. / Bystraya VEZhKh-MS/MS skriningovaya metodika opredeleniya v moche cheloveka zapreshchennykh narkoticheskikh sredstv, vklyuchaya novye psikhoaktivnye veshchestva, a takzhe ikh metabolity.

Aim of this study was to develop a fast screening method for identification of illegal drugs including new psychoactive substances and their metabolites in human urine by means of liquid chromatography mass-spectrometry HPLC-MS/MS screening method for identification of 138 psychoactive substances in urine was established, validated and tested. High-resolution mass-spectrometry was used for confirmation of the developed approach. The developed method was tested on 50 positive urine samples containing the target compounds, which proved reliability and accuracy of the established screening method.

[Authentication study of the preparation 'Avastin' by means of chemical-toxicological analysis]. / Issledovanie podlinnosti preparata 'Avastin' metodami khimiko-toksikologicheskogo analiza.

Falsification and use of low-quality drugs of biological origin creates a threat to public health. To a greater extent, costly drugs, including bevacizumab, are exposed to similar abuses. Timely determination of cases of forgery or the improper clinical use of monoclonal antibody preparations is one of the necessary measures that can be taken to limit the risks and preserve the health of patients. This paper presents the results of the investigation of the bevacizumab preparation 'Avastin', which was withdrawn from ophthalmic clinical practice in the course of the investigation. We compared the qualitative and quantitative composition of the drug samples, which were determined using commonly available methods of chemical and toxicological analysis.

[Comparison of sample preparation techniques in chemical-toxicological analysis of urine for the presence of narcotic drugs, psychotropic substances and their metabolites]. / Sravnenie protsedur probopodgotovki pri khimiko-toksikologicheskom issledovanii mochi na nalichie narkoticheskikh sredstv, psikhotropnykh veshchestv i ikh metabolitov.

The sample preparation procedures within undirected chemical-toxicological urine analysis for GCMS and HPLC-MS/MS methods are presented. These methods are intended for analytical diagnosis of the presence of narcotic drugs, psychotropic substances and their metabolites in humans.

[Metabolomic Profiling of Patients With Cardiovascular Diseases].

Cardiovascular diseases (CVD) are the main cause of death worldwide. A broad study of the pathogenetic mechanisms of the CVD onset and progression has led to understanding of the importance of endothelial dysfunction (ED) in these processes. During recent years intensive work has been conducted in the direction of searching for markers of ED. Metabolomics is an intensively advancing approach to early diagnostics of diseases. Metabolomic analysis based on mass spectrometry allows to study complete metabolic profiles and their deviations resulting from changes in expression of genes and RNA, protein activity, or environmental factors. Metabolomic analysis has already demonstrated significant results in the solving of different scientific and clinical problems. It appears to be a promising method for detecting early biomarkers of CVD. Various aspects of application of metabolomic profiling in the field of cardiovascular diseases are discussed in this article.

[Effect of mexidol on 6beta/free hydroxycortisol ratio. Possibility of CYP3A4 activation].

The influence of mexidol (2-ethyl-6-methyl-3-oxypyridine) after a single peroral administration on the levels of 6beta-hydroxycortisol (6beta-OHC) and free cortisol (FC) in human urine has been evaluated. The 6beta-OHC/FC ratio is increased (approximately 2.96 +/- 0.76 times) against the basal 6beta-OHC/FC ratio during the first 24 hour after drug administration. Data analysis on the second and third day after mexidol administration did not show evident changes in 6beta-OHC/FC ratios. It is suggested that CYP3A4 activation after mexidol administration occurred only during active drug biotransformation and excretion and ceased after full excretion from the human body.

[Experimental pharmacokinetics of the new neurotensine-derived antipsychotic drug dilept].

The metabolism and pharmacokinetics of a new neurotensine-derived dipeptide drug dilept (N-caproyl-L-prolyltyrosine methyl ester) and its tentative metabolites after intravenous and peroral administration of the parent drug and its tabletized form in rats have been studied by HPLC-ESI(-)-MS/MS method. It is established that unchanged dilept (detected in the blood plasma for no less than 30 min) as well as N-caproyl-L-proline and N-caproyl-L-prolyl-L-tyrosine (both detected in the blood over more than 4 h) are the major metabolites in the bloodstream upon peroral administration of the drug. The proposed structures of metabolites were confirmed by countersynthesis. Dilept and N-caproyl-L-prolyl-L-tyrosine penetrate through the blood - brain barrier. The drug is rapidly absorbed, distributed, and metabolized in the rat organism. Peroral administration of dilept in rats in the form of tablets (at a dose of 200 mg/kg) resulted in a significant increase in intestinal absorption, as evidenced by a 22% improvement in the bioavailability, whereas dilept alone showed an absolute bioavailability of less than 1%.

[HPLC-MS determination of 2-ethyl-6-methyl-3-oxypyridine].

An HPLC-ESI-MS method has been developed for determining 2-ethyl-6-methyl-3-oxypyridine (EMO) in human urine upon peroral administration of this substance in form ofmexidol. Various sample preparation (extraction) procedures were tested and compared for evaluating the recovery and matrix effect. Solid-phase extraction procedure followed by derivation with dansyl chloride is proposed as a method of choice. The recovery of analyte was 48.1 +/- 3.4%, and the matrix effect was 99.4 +/- 4.1%. The MS and MS/MS spectra of EMO and its dansyl derivatives are presented and interpreted. The analyses were performed using a mass spectrometer of the ion trap type with electrospray ionization at atmospheric pressure, operating in the regime of positive ion detection.

[Doping in sports].

Drugs used by athletes for the improvement of results are described and classified with respect to chemical structure and pharmacological action. The main groups of drugs treated as doping are considered and the WADA requirements to prohibited preparations are formulated. The main effects produced by drugs on the athletes and animals (race horses, fight dogs, etc ) are described and the measures of therapy against side effects are outlined.