Moskaleva, Natalia E

Kozhevnikova M.V., Kakotkina A.V., Korobkova E.O., Kuznetsov I.V., Shestakova K.M., Moskaleva N.E., Appolonova S.A., Belenkov Y.N.

The diagnosis of heart failure with preserved ejection fraction (HFpEF) remains challenging. The use of metabolomics approaches seems promising in speeding up and simplifying the diagnostic process in HFpEF patients, which can lead to earlier treatment initiation and better improvement of patient condition. The aim of this study was to develop a diagnostic panel of metabolites (metabolomic biomarkers) for the detection and diagnosis of HF with preserved ejection fraction. The study included 76 participants with hypertension, 36 of whom were diagnosed with HFpEF. The blood plasma metabolomic profile, including 72 metabolites, was detected using high-performance liquid chromatography combined with mass spectrometry. There were 18 statistically significant differences in concentrations of metabolites and 3 differences in their ratios between HFpEF and hypertension groups. The prognostic model for detecting the possibility of HFpEF included seven metabolites and two ratios: hexadecenoylcarnitine, arginine, trimethylamine-N-oxide, asymmetric dimethylarginine (ADMA), arginine/ADMA ratio, kynurenine, kynurenine/tryptophan, neopterin, and anthranilic acid. The area under the ROC curve was 0.981 ± 0.017. The resulting model was statistically significant (p < 0.001). The metabolomic panel could be considered as an addition to the present HFpEF laboratory diagnostic criteria for blood plasma analysis in clinical practice.

Ageev A.A., Kozhevnikova M.V., Tyurina D.A., Korobkova E.O., Kondratieva T.O., Shestakova K.M., Moskaleva N.E., Markin P.A., Khabarova N.V., Appolonova S.A., Belenkov Y.N.

Aim      To identify metabolomic and structure and function markers of remote left ventricular (LV) remodeling in patients with chronic heart failure (CHF) of ischemic etiology and LV ejection fraction (EF) <50%.Material and methods  This prospective study included 56 patients with 3-4 NYHA functional class CHF of ischemic etiology (mean age, 66±7 years) and 50 patients with ischemic heart disease (IHD) without signs of CHF (69 [64; 73.7] years). Concentration of 19 amino acids, 11 products of kynurenine catabolism of tryptophan, 30 acylcarnitines with different chain lengths were measured in all participants. The metabolites that showed statistical differences between the comparison groups were then used for the analysis. Echocardiography was used to assess LV cavity remodeling at the time of the CHF patient inclusion in the study and after 6 months of follow-up. Predictors of long-term LV cavity remodeling were assessed for this cohort taking into account statistically significant echocardiographic parameters and metabolites.Results Patients with CHF of ischemic etiology, predominantly (81%) had pathological calculated types of LV remodeling (concentric and eccentric hypertrophy, 46 and 35%, respectively). However, this classification had limitations in describing this cohort. In addition, in this group, the concentrations of alanine, proline, asparagine, glycine, arginine, histidine, lysine, valine, indolyl-3-acetic acid, indolyl-3-propionic acid, C16-1-OH, and C16-OH were significantly (p<0.05) lower, and the concentrations of most medium- and long-chain acylcarnitines were higher than in patients with IHD without signs of CHF. The long-term (6 months) reverse remodeling of the LV cavity in CHF of ischemic etiology was influenced by changes in the interventricular septum thickness (hazard ratio, HR, 19.07; 95% confidence interval, CI, 1.76-206.8; p=0.006) and concentrations of anthranilic acid (HR 19.8; 95% CI 1.01-387.8; p=0.019) and asparagine (HR 8.76; 95% CI 1.07-71.4; p=0.031).Conclusion      The presence of an interventricular septum thickness of more than 13.5 mm, anthranilic acid concentrations of higher than 0.235 μM/l, or an asparagine concentration of less than 135.2 μM/l in patients with CHF of ischemic etiology after 6 months of follow-up affects their achievement of LV cavity reverse remodeling. 

Maria F. P., Anastasia V. K., Vita Yu. Z., Ekaterina O. K., Tamerlan N. E., Anton A. A., Ksenia M. S., Natalya E. M., Svetlana A. A., Yury N. B., Maria V. K.

Background — Chronic heart failure with preserved ejection fraction (CHFpEF) develops as a result of many diseases that lead to significant metabolic disorders. Given the heterogeneity of this group of patients, therapeutic options for this syndrome are extremely limited. In this regard, it seems promising to study the metabolomic profile in patients with CHFpEF to identify biomarkers, examine their roles in the pathogenesis of the syndrome, and search for potential targets for targeted therapy. Objective — The study aimed at testing the correlation between the threonine level and the features of the clinical course of CHFpEF. Methods — The study included a total of 154 patients: 82 with CHFpEF, 45 with hypertension and/or coronary artery disease (comparison group), and 27 healthy volunteers (control group). Threonine levels were assessed using high-performance liquid chromatography-mass spectrometry. Results — The threonine concentration was significantly reduced in patients with CHFpEF (1) vs. comparison group (2) and control group (3): p<0.001; p1-3<0.001; p2-3=0.037). A reduction in the threonine level was characteristic for patients with diabetes mellitus vs. patients without it (p=0.029). Conclusion — Given the importance of threonine in energy metabolism and significant changes in its level in various pathophysiological processes, it should be considered as an additional diagnostic and prognostic criterion for CHFpEF. Additional studies are needed to better understand the role of threonine in the pathophysiology of cardiovascular diseases.

Mikhel I.B., Bakhrushina E.O., Petrusevich D.A., Nedorubov A.A., Appolonova S.A., Moskaleva N.E., Demina N.B., Kosenkova S.I., Parshenkov M.A., Krasnyuk I.I., Krasnyuk I.I.

Recently, ribavirin has demonstrated effectiveness in treating glioblastoma through intranasal administration utilizing the nose-to-brain delivery route. Enhancing ribavirin’s bioavailability can be achieved by utilizing intranasal stimuli-responsive systems that create a gel on the nasal mucosa. The research examined thermosensitive, pH-sensitive, and ion-selective polymers in various combinations and concentrations, chosen in line with the current Quality by Design (QbD) approach in pharmaceutical development. Following a thorough assessment of key parameters, the optimal composition of gellan gum at 0.5%, Poloxamer 124 at 2%, and purified water with ribavirin concentration at 100 mg/mL was formulated and subjected to in vivo testing. Through experiments on male rats, the nose-to-brain penetration mechanism of the active pharmaceutical ingredient (API) was elucidated, showcasing drug accumulation in the olfactory bulbs and brain.

Savitskii M.V., Moskaleva N.E., Brito A., Markin P.A., Zigangirova N.A., Soloveva A.V., Sheremet A.B., Bondareva N.E., Lubenec N.L., Tagliaro F., Tarasov V.V., Tatzhikova K.A., Appolonova S.A.

Growing antimicrobial resistance has accelerated the development of anti-virulence drugs to suppress bacterial toxicity without affecting cell viability. Fluorothiazinon (FT), an anti-virulence, type three secretion system and flagella motility inhibitor which has shown promise to suppress drug-resistant pathogens having the potential to enhance the efficacy of commonly prescribed antibiotics when used in combination. In this study we characterized the pharmacokinetics, tissue distribution, bioavailability and excretion of FT in rats and rabbits. FT presented a dose-proportional linear increase in the blood of rats. Tissue distribution profiling confirmed that FT distributes to all organs being substantially higher than in the blood of rats. The bioavailability of FT was higher when administered with starch than with water implying FT should be ideally dosed with food. FT was primarily excreted in the feces in rats and rabbits while negligible amounts are recovered from the urine.

Baskhanova S.N., Savitskii M.V., Moskaleva N.E., Samoylov V.M., Lubenec N.L., Luyksaar S.I., Soloveva A.V., Zolotov C.A., Zigangirova N.A., Appolonova S.A.

An HPLC-MS/MS technique for quantitative determination of the new antibacterial drug fluorothiazinone in human blood plasma using an internal standard was developed and validated. The validation protocol proved the selectivity, accuracy, and precision of the method and the lack of significant carryover, matrix, and sample-dilution effects. The calibration curve was linear in the concentration range 0.1 – 300 ng/mL with a correlation coefficient r2 > 0.9990. The lower limit of quantitation was 0.1 ng/mL. The studied analyte was shown to be stable during storage in a stock solution (for 6 h at room temperature) and in plasma samples (for 6 h at room temperature, after three freeze–thaw cycles, and for 30 d at –70°C) and in an autosampler after sample preparation (for 24 h at +10°C).

Markin S.S., Ponomarenko E.A., Romashova Y.A., Pleshakova T.O., Ivanov S.V., Bedretdinov F.N., Konstantinov S.L., Nizov A.A., Koledinskii A.G., Girivenko A.I., Shestakova K.M., Markin P.A., Moskaleva N.E., Kozhevnikova M.V., Chefranova Z.Y., et. al.

AbstractCardiovascular 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.

Girel S., Markin P.A., Tobolkina E., Boccard J., Moskaleva N.E., Rudaz S., Appolonova S.A.

AbstractThe steroid submetabolome, or steroidome, is of particular interest in prostate cancer (PCa) as the dependence of PCa growth on androgens is well known and has been routinely exploited in treatment for decades. Nevertheless, the community is still far from a comprehensive understanding of steroid involvement in PCa both at the tissue and at systemic level. In this study we used liquid chromatography/high resolution mass spectrometry (LC/HRMS) backed by a dynamic retention time database DynaSTI to obtain a readout on circulating steroids in a cohort reflecting a progression of the PCa. Hence, 60 relevant compounds were annotated in the resulting LC/HRMS data, including 22 unknown steroid isomers therein. Principal component analysis revealed only subtle alterations of the systemic steroidome in the study groups. Next, a supervised approach allowed for a differentiation between the healthy state and any of the stages of the disease. Subsequent clustering of steroid metabolites revealed two groups responsible for this outcome: one consisted primarily of the androgens, whereas another contained corticosterone and its metabolites. The androgen data supported the currently established involvement of a hypothalamic-pituitary–gonadal axis in the development of PCa, whereas biological role of corticosterone remained elusive. On top of that, current results suggested a need for improvement in the dynamic range of the analytical methods to better understand the role of low abundant steroids, as the analysis revealed an involvement of estrogen metabolites. In particular, 2-hydroxyestradiol-3-methylether, one of the compounds present in the disease phenotype, was annotated and reported for the first time in men.

Басханова С.Н., Савицкий М.В., Москалёва Н.Е., Самойлов В.С., Лубенец Н.Л., Луйксаар С.И., Соловьева А.В., Золотов С.А., Зигангирова Н.А., Апполонова С.А.

В результате исследования была разработана и валидирована ВЭЖХ-МС/МС-методика количественного определения нового антибактериального лекарственного вещества фтортиазинон в плазме крови человека с использованием внутреннего стандарта. В ходе валидационной оценки была подтверждена селективность, правильность и прецизионность метода, отсутствие значительного эффекта переноса и эффекта матрицы, а также влияния разбавления образцов. Градуировочная кривая линейна в диапазоне концентраций 0,1 – 300 нг/мл, коэффициент детерминации r2 > 0,9990. Нижний предел количественного определения составил 0,1 нг/мл. Установлено, что исследуемый аналит стабилен при хранении в виде рабочего раствора (в течение 6 ч при комнатной температуре), в образцах плазмы (в течение 6 ч при комнатной температуре, после трех циклов заморозки-разморозки, в течение 30 дней при –70 °C) и после пробоподготовки в автосамплере (при +10 °C в течение 24 ч).

Savitskii M.V., Moskaleva N.E., Brito A., Zigangirova N.A., Soloveva A.V., Sheremet A.B., Bondareva N.E., Lubenec N.L., Kuznetsov R.M., Samoylov V.M., Tagliaro F., Appolonova S.A.

Pseudomonas aeruginosa (PA) infection is commonly associated with hospital-acquired infections in patients with immune deficiency and/or severe lung diseases. Managing this bacterium is complex due to drug resistance and high adaptability. Fluorothiazinon (FT) is an anti-virulence drug developed to suppress the virulence of bacteria as opposed to bacterial death increasing host’s immune response to infection and improving treatment to inhibit drug resistant bacteria. We aimed to evaluate FT pharmacokinetics, quorum sensing signal molecules profiling and tryptophan-related metabolomics in blood, liver, kidneys, and lungs of mice. Study comprised three groups: a group infected with PA that was treated with 400 mg/kg FT (“infected treated group”); a non-infected group, but also treated with the same single drug dose (“non-infected treated group”); and an infected group that received a vehicle (“infected non-treated group”). PA-mediated infection blood pharmacokinetics profiling was indicative of increased drug concentrations as shown by increased Cmax and AUCs. Tissue distribution in liver, kidneys, and lungs, showed that liver presented the most consistently higher concentrations of FT in the infected versus non-infected mice. FT showed that HHQ levels were decreased at 1 h after dosing in lungs while PQS levels were lower across time in lungs of infected treated mice in comparison to infected non-treated mice. Metabolomics profiling performed in lungs and blood of infected treated versus infected non-treated mice revealed drug-associated metabolite alterations, especially in the kynurenic and indole pathways.

Belenkov Y.N., Ageev A.A., Kozhevnikova M.V., Khabarova N.V., Krivova A.V., Korobkova E.O., Popova L.V., Emelyanov A.V., Appolonova S.A., Moskaleva N.E., Shestakova K.M., Privalova E.V.

Background: Progressive myocardial remodeling (MR) in chronic heart failure (CHF) leads to aggravation of systolic dysfunction (SD) and clinical manifestations. Identification of metabolomic markers of these processes may help in the search for new therapeutic approaches aimed at achieving reversibility of MR and improving prognosis in patients with CHF. Methods: To determine the relationship between plasma acylcarnitine (ACs) levels, MR parameters and clinical characteristics, in patients with CHF of ischemic etiology (n = 79) and patients with coronary heart disease CHD (n = 19) targeted analysis of 30 ACs was performed by flow injection analysis mass spectrometry. Results: Significant differences between cohorts were found for the levels of 11 ACs. Significant positive correlations (r > 0.3) between the medium- and long-chain ACs (MCACs and LCACs) and symptoms (CHF NYHA functional class (FC); r = 0.31−0.39; p < 0.05); negative correlation (r = −0.31−0.34; p < 0.05) between C5-OH and FC was revealed. Positive correlations of MCACs and LCACs (r = 0.31−0.48; p < 0.05) with the left atrium size and volume, the right atrium volume, right ventricle, and the inferior vena cava sizes, as well as the pulmonary artery systolic pressure level were shown. A negative correlation between C18:1 and left ventricular ejection fraction (r = −0.31; p < 0.05) was found. However, a decrease in levels compared to referent values of ACs with medium and long chain lengths was 50% of the CHF-CHD cohort. Carnitine deficiency was found in 6% and acylcarnitine deficiency in 3% of all patients with chronic heart disease. Conclusions: ACs may be used in assessing the severity of the clinical manifestations and MR. ACs are an important locus to study in terms of altered metabolic pathways in patients with CHF of ischemic etiology and SD. Further larger prospective trials are warranted and needed to determine the potential benefits to treat patients with CV diseases with aberrate AC levels.

Chuchueva N., Carta F., Nguyen H.N., Luevano J., Lewis I.A., Rios-Castillo I., Fanos V., King E., Swistushkin V., Reshetov I., Rusetsky Y., Shestakova K., Moskaleva N., Mariani C., Castillo-Carniglia A., et. al.

Head and neck cancer (HNC) is the fifth most common cancer globally. Diagnosis at early stages are critical to reduce mortality and improve functional and esthetic outcomes associated with HNC. Metabolomics is a promising approach for discovery of biomarkers and metabolic pathways for risk assessment and early detection of HNC. To summarize and consolidate the available evidence on metabolomics and HNC in plasma/serum, saliva, and urine. A systematic search of experimental research was executed using PubMed and Web of Science. Available data on areas under the curve was extracted. Metabolic pathway enrichment analysis were performed to identify metabolic pathways altered in HNC. Fifty-four studies were eligible for data extraction (33 performed in plasma/serum, 15 in saliva and 6 in urine). Metabolites with high discriminatory performance for detection of HNC included single metabolites and combination panels of several lysoPCs, pyroglutamate, glutamic acid, glucose, tartronic acid, arachidonic acid, norvaline, linoleic acid, propionate, acetone, acetate, choline, glutamate and others. The glucose-alanine cycle and the urea cycle were the most altered pathways in HNC, among other pathways (i.e. gluconeogenesis, glycine and serine metabolism, alanine metabolism, etc.). Specific metabolites that can potentially serve as complementary less- or non-invasive biomarkers, as well as metabolic pathways integrating the data from the available studies, are presented. The present work highlights utility of metabolite-based biomarkers for risk assessment, early detection, and prognostication of HNC, as well as facilitates incorporation of available metabolomics studies into multi-omics data integration and big data analytics for personalized health.

Kozhevnikova M.V., Korobkova E.O., Krivova A.V., Kukharenko A.V., Moskaleva N.E., Shestakova K.M., Mesonzhnik N.V., Ageev A.A., Boldin A.A., Brito A., Appolonova S.А., Privalova E.V., Belenkov Y.N.

Aim. Branched-chain amino acids (BCAAs) have been postulated as potential indicators of cardiovascular risk. The objective of this study was to explore the relationship between plasma BCAAs and different stages of cardiovascular disorders.Material and methods. In our cross-sectional study, plasma BCAAs (valine, leucine and isoleucine) in individuals without cardiovascular diseases (CVDs) (nonCVD group, total n=27, with n=16 healthy, but with metabolic disorders) were compared to patients diagnosed with CVDs [CVD group, total n=109, being n=61 hypertension (n=31 with signs of beginning of myocardial remodeling) and n=48 patients with coronary artery disease (CAD)].Results. The plasma concentration of BCAAs was significantly higher in the group of patients with cardiovascular disease compared with the healthy group (p<0.05 for all amino acids tested): valine concentration was 238.7 [219.6; 267.0] μM in the non-CVD group and 261.2 [233.8; 298.7] μM in the CVD group; leucine concentration was 134.8 [122.4; 153.2] μM and 146.8 [129.0; 166.6] μM, respectively; and isoleucine 72.7 [65.3; 84.4] μM and 81.7 [68.0; 96.2] μM, respectively. Leucine and isoleucine concentration levels were minimal in the healthy participant subgroup and maximal in the IBS patient subgroup. No statistically significant differences in BCAAs concentrations were found in the subgroups without CAD. Significant increases in concentrations were observed in the subgroups of patients with CAD as follows: valine concentration was 256.3 [219.0; 297.9] μM in hypertension group and 261.7 [236.5; 307.5] μM in CAD group; leucine concentration was 141.8 [123.5; 166.6] μM and 154.1 [134.7; 172.7] μM, respectively, and isoleucine 72.8 [65.7; 94.0] μM and 85.7 [74.9; 101.7] μM, respectively. BCAAs profiles in all participants with metabolic disorders had “good” diagnostic accuracy with area under the receiver operating characteristics curve being 0.72, 0.70 and 0.70 for valine, leucine and isoleucine, respectively.Conclusion. BCAAs concentrations are elevated with higher severity of the cardiovascular disorder and exhibit potential as early independent indicators of coronary artery disease.

Shestopalov A.V., Кit O.I., Davydov V.V., Baizyanova Y.M., Zlatnik E.Y., Novikova I.A., Sagakyants A.B., Appolonova S.A., Moskaleva N.E., Rumyantsev S.A.

Introduction. Researchers in the field of oncology have a significant interest in the role of microorganisms in development of malignant neoplasms.Aim. To study the levels of 2-heptyl-3-hydroxy-4-quinolone (PQS) and 2-heptyl-4-quinolone (HHQ) produced by Pseudomonas aeruginosa in the blood of patients with lung cancer and to analyze the relation between their changes and changes in the level of immunoglobulins and vascular endothelial growth factor (VEGF) in the blood of patients with lung cancer.Materials and methods.  PQS and HHQ were quantified in the blood of patients using high performance liquid chromatography. The levels of immunoglobulins G (IgG),  secretory immunoglobulin A (s-IgA),  and VEGF in the blood were determined using ELISA.Results. Analysis have shown that the level of PQS in the blood of patients with lung cancer is 2-fold higher than in the control group. This change is accompanied by a decrease in the level of immunoglobulins IgG, as well as an increase in the content of s-IgA and growth factor VEGF in the blood.Conclusion. PQS level in the blood of patients with lung cancer is elevated creating conditions aggravating the course of the main disease and worsening its prognosis.

Shestakova K.M., Moskaleva N.E., Boldin A.A., Rezvanov P.M., Shestopalov A.V., Rumyantsev S.A., Zlatnik E.Y., Novikova I.A., Sagakyants A.B., Timofeeva S.V., Simonov Y., Baskhanova S.N., Tobolkina E., Rudaz S., Appolonova S.A.

AbstractLung cancer is referred to as the second most common cancer worldwide and is mainly associated with complex diagnostics and the absence of personalized therapy. Metabolomics may provide significant insights into the improvement of lung cancer diagnostics through identification of the specific biomarkers or biomarker panels that characterize the pathological state of the patient. We performed targeted metabolomic profiling of plasma samples from individuals with non-small cell lung cancer (NSLC, n = 100) and individuals without any cancer or chronic pathologies (n = 100) to identify the relationship between plasma endogenous metabolites and NSLC by means of modern comprehensive bioinformatics tools, including univariate analysis, multivariate analysis, partial correlation network analysis and machine learning. Through the comparison of metabolomic profiles of patients with NSCLC and noncancer individuals, we identified significant alterations in the concentration levels of metabolites mainly related to tryptophan metabolism, the TCA cycle, the urea cycle and lipid metabolism. Additionally, partial correlation network analysis revealed new ratios of the metabolites that significantly distinguished the considered groups of participants. Using the identified significantly altered metabolites and their ratios, we developed a machine learning classification model with an ROC AUC value equal to 0.96. The developed machine learning lung cancer model may serve as a prototype of the approach for the in-time diagnostics of lung cancer that in the future may be introduced in routine clinical use. Overall, we have demonstrated that the combination of metabolomics and up-to-date bioinformatics can be used as a potential tool for proper diagnostics of patients with NSCLC.

Staden R.S., Ion C.B., van Staden J.(.

Zhang P., Xia W., Bai Y., Zhang F., Zhang Y., Jiang W., Yuan H.

Bilel S., Corli G., Tiziani E., Chirenti D., Dall'acqua S., Comai S., Ferraro L., Marti M., Beggiato S.

Guo J., Kong L., Tian L., Han Y., Teng C., Ma H., Tao B.

Kullebi B., Alat Ö., Aksakal Ö., Yılmaztürk D., Lafzi A., Şişman T.

ABSTRACTDetailed studies on the embryotoxic and teratogenic effects of synthetic cannabinoids known to be abused are very limited. The present study aimed to evalutate the possible embryotoxic, teratogenic, behavioral, and molecular effects of 4F‐MDMB‐BUTICA, a new generation synthetic cannabinoid, using zebrafish embryos. The zebrafish embryos were exposed to the cannabinoid at 0.15, 0.30, 0.60, 1.20, 2.40, and 4.80 mg/L from 3 to 24 hpf (acute) and 3 to 120 hpf (subacute). No developmental abnormalities and mortality were observed in embryos after acute exposure. Subacute 4F‐MDMB‐BUTICA exposure induced mortality of the embryos with the 120 hpf LC50 and EC50 of 1.932 and 0.960 mg/L, respectively. 4F‐MDMB‐BUTICA also caused embryonic deformities, including spine formation, pericardial edema, impaired blood flow, yolk sac edema, and delayed development. Additionally, subacute cannabinoid exposure induced hypoactivity in response to the stimulus in 120‐hpf larvae. qPCR analyses were performed on a subset of 19 genes associated with specific adverse outcomes. The cannabinoid exposure altered the transcriptional expression levels of apoptosis (casp3a, casp8, ifng1, and tp53) DNA repair (rad51), dopamine (dat, drd1, and drd3), serotonin (5ht1aa, 5ht1a, 5ht1b, and 5ht2c), γ‐aminobutyric acid (gabra1, gat1, abat, and gad1b), and behavior (gnrh3, gnrhr3, and kiss2)‐related genes. In conclusion, the subacute exposure to 4F‐MDMB‐BUTICA induces mortality, developmental toxicity, hypoactivity of larval behavior, and changes in some essential genes in zebrafish. These findings suggest that 4F‐MDMB‐BUTICA may have similar embryotoxic effects in humans.

Carvalho I.A., Silva C.F., Borges K.B.

ABSTRACTA miniaturized self‐assembly pipette tip with restricted access mesoporous polypyrrole solid‐phase extraction, combined with capillary electrophoresis with diode array detection (CE‐DAD), was developed to rapidly extract and determine enalapril from urine samples. The CE‐DAD technique used 50 mmol L−1 phosphate (pH 7) as the background electrolyte, a voltage of 13 kV, a 30 mbar hydrodynamic injection for 4 s, a capillary temperature of 25°C, and a wavelength of 195 nm to achieve a migration time of 6.3 min with satisfactory peak asymmetry and no interfering and/or baseline noise. The factors that influenced the extraction efficiency were evaluated and optimized: 750 µL sample at pH 7.5, 40 mg adsorbent, 250 µL hexane as a washing solvent, and 750 µL acetonitrile as eluent, resulting in recoveries around 74%. Linearity was acceptable in the 100–3000 ng mL−1 range, and the selectivity and accuracy were also suitable. The limits of detection and quantitation were 30 and 50 ng mL−1, respectively. The adsorbent effectively removed 87% of the proteins and may be reused three times. The analytical approach was successfully verified and used to analyze enalapril in urine samples collected from volunteers. Finally, the greenness of the researched technique was assessed using five measures that showed good eco‐friendliness.

Bano A., Vats R., Yadav P., Kamboj M., Bhardwaj R.

Blaurock J., Grunewald S., Engel K.M.

Abstract In contrast to the conventional spermiogram, metabolomics approaches give insights into the molecular composition of semen and may provide more detailed information on the fertility status of the respective donor. Given the intra-individual variability of spermiogram parameters between two donations, this study sought to elucidate the biological variability of the seminal plasma metabolome over an average period of 8 weeks. Two time-shifted semen samples from 15 healthy donors were compared by a targeted metabolomics approach utilizing the Biocrates AbsoluteIDQ p180 kit. Next to intraclass correlation coefficients (ICC), which represent a measure of reliability, coefficients of variation within individuals (CVW) and coefficients of variation between individuals (CVB) were calculated for each metabolite to demonstrate its stability. Furthermore, men were divided into two cohorts, a similar sperm concentration (SSC) and a differing sperm concentration (DSC) cohort, based on the observed variance in sperm concentration between the two semen donations. The ICC was higher in the SSC compared to the DSC cohort. The levels of 18 metabolites, primarily acylcarnitines, varied between the initial and subsequent donations. After subdivision into subgroups, only ornithine and phosphatidylcholine 40:5 exhibited differential levels between the two donations in the SSC group, compared to 14 metabolites in the DSC group. CVB was higher than CVW but both differed between the metabolite subclasses. Biogenic amines were identified as the least reliable analytes over time, exhibiting the highest CVW, compared to sphingomyelins, which demonstrated the highest reliability with the lowest variation. CVB was the highest for ether-bound glycerophosphatidylcholines and the lowest for amino acids.

Kozhevnikova M.V., Kakotkina A.V., Korobkova E.O., Kuznetsov I.V., Shestakova K.M., Moskaleva N.E., Appolonova S.A., Belenkov Y.N.

The diagnosis of heart failure with preserved ejection fraction (HFpEF) remains challenging. The use of metabolomics approaches seems promising in speeding up and simplifying the diagnostic process in HFpEF patients, which can lead to earlier treatment initiation and better improvement of patient condition. The aim of this study was to develop a diagnostic panel of metabolites (metabolomic biomarkers) for the detection and diagnosis of HF with preserved ejection fraction. The study included 76 participants with hypertension, 36 of whom were diagnosed with HFpEF. The blood plasma metabolomic profile, including 72 metabolites, was detected using high-performance liquid chromatography combined with mass spectrometry. There were 18 statistically significant differences in concentrations of metabolites and 3 differences in their ratios between HFpEF and hypertension groups. The prognostic model for detecting the possibility of HFpEF included seven metabolites and two ratios: hexadecenoylcarnitine, arginine, trimethylamine-N-oxide, asymmetric dimethylarginine (ADMA), arginine/ADMA ratio, kynurenine, kynurenine/tryptophan, neopterin, and anthranilic acid. The area under the ROC curve was 0.981 ± 0.017. The resulting model was statistically significant (p < 0.001). The metabolomic panel could be considered as an addition to the present HFpEF laboratory diagnostic criteria for blood plasma analysis in clinical practice.

Ramsay S., Hyvärinen E., González-Arriagada W., Salo T., Ajudarte Lopes M., Mikkonen J.J., Kashyap B., Kullaa A.M.

Abstract Introduction This longitudinal study assessed the salivary metabolic profile in patients with head and neck cancer (HNC) treated with radiotherapy (RT). This study aims to investigate salivary metabolites and biological oral pathways induced by RT. Methods Clinical data and unstimulated whole-mouth saliva (USWMS) were obtained from 45 HNC patients before, during, and one week after the RT. Data was also collected from 30 healthy controls. NMR spectroscopy identified and quantified 24 metabolites. Spearman’s rank correlation analysis and pathway enrichment analysis (MetaboAnalyst 6.0) was performed to check the effect of cancer therapy on the correlation and pathways of different salivary metabolites. Results Of 24 metabolites identified, 17 salivary metabolites showed a consistent decrease in the concentration during and after treatment of HNC patients. The metabolite proline decreased, whereas fucose and 1,2-Propanediol were increased in the saliva causing altered redox balance and abnormal fucosylation in HNC patients compared to controls. Spearman correlation analysis indicated changes between pyruvate and some other metabolites, including alanine, trimethylamine, choline, taurine, and succinate, during RT. Five pathways (Pyruvate metabolism; Glycolysis / Gluconeogenesis; Glycine, serine, and threonine metabolism; Glyoxylate and dicarboxylate metabolism; and Alanine, aspartate and glutamate metabolism) are affected, demonstrating the metabolic dysregulation due to RT. The pyruvate metabolism was overpresented with the high Pathway Impact score. Conclusion Salivary metabolomics analysis revealed significant alterations in the metabolic profile of HNC patients undergoing RT, providing valuable insights into treatment-induced oral pathobiological changes. Alterations in salivary pathways during RT suggest disturbances in redox homeostasis, oxidative stress, and inflammation. The ability to monitor salivary metabolites and pathways non-invasively holds promise to personalized medicine in HNC treatment by enabling early detection of treatment-related toxicities, monitoring treatment response, and tailoring interventions to patient needs.

Martins-Chaves R.R., Bastos V.C., Vitório J.G., Duarte-Andrade F.F., Pereira T.D., Leite-Lima F., Gomes T.E., Lebron Y.A., Moreira V.R., França M.S., Santos L.V., Lange L.C., Macedo A.N., Picossi C.R., Pontes H.A., et. al.

Understanding the early molecular events driving oral carcinogenesis is vital for diagnosing oral squamous cell carcinoma (OSCC) promptly. While metabolic differences between oral leukoplakia (OLK), OSCC, and healthy oral mucosa have been reported, the metabolic changes distinguishing malignant transformed OLKs (MT-OLK) from non-transformed OLKs (NT-OLK) remain unexplored. Here, we examine the metabolomic profiles of 5 cases of MT-OLK and 15 of NT-OLK to identify key predictive molecules using untargeted high-performance liquid chromatography-mass spectrometry. The potentially discriminant compounds were highlighted through a robust statistical analysis workflow, and the dysregulated metabolic pathways were illustrated by enrichment analysis. Seventeen molecular features, primarily lipids—including phospholipids, oxidised lipids, cholesteryl esters, and fatty acids—were identified as discriminants between MT-OLK and NT-OLK across statistical and bioinformatic approaches. Pathway enrichment analysis revealed alterations in lipid metabolism, particularly fatty acid synthesis and degradation, steroid hormone biosynthesis, and glycerophospholipid metabolism. Predictive models showed high accuracy (AUC = 0.88) in distinguishing the two groups. This study suggests that metabolomics has the potential to differentiate between MT-OLK and NT-OLK by identifying candidate biomarkers that may contribute to the understanding of malignant transformation. Validation in larger cohorts is warranted to translate these findings into clinical practice.

Karaduman A.B., Ilgın S., Aykaç Ö., Yeşilkaya M., Levent S., Özdemir A.Ö., Girgin G.

Background/Objectives: Asymptomatic carotid artery stenosis is usually detected by physicians in patients, coincidentally, during an ultrasound examination of the neck. Therefore, measurable biomarkers in blood are needed to define the presence and severity of atherosclerotic plaque in patients to identify and manage it. We hypothesized that biomarkers that indicate pathways related to the pathogenesis of atherosclerosis could be used to identify the presence and severity of atherosclerotic plaque. For this purpose, the levels of participants’ inflammatory and oxidative stress biomarkers were determined. Kynurenine/tryptophan and neopterin levels were measured as relatively new biomarkers of inflammation in this study. Methods: Our study included 57 patients diagnosed with asymptomatic carotid artery stenosis and 28 healthy volunteers. Blood kynurenine and tryptophan levels were measured with LCMS/MS. Blood catalase, total superoxide dismutase (t-SOD), glutathione peroxidase (GPx), malondialdehyde, and neopterin levels were measured using the ELISA assay method. Result: The kynurenine/tryptophan ratio reflecting IDO activity was higher in patients than in healthy volunteers. Decreased tryptophan levels and increased kynurenine and neopterin levels were observed in patients who underwent carotid endarterectomy. In patients, catalase, t-SOD, and malondialdehyde levels were higher, while GPx activity was lower. These differences were found to be more significant in patients who underwent carotid endarterectomy. Conclusions: Increased kynurenine/tryptophan ratio and neopterin levels in patients with asymptomatic carotid artery stenosis were associated with the inflammatory status of the patients. Oxidative stress and inflammatory biomarkers can be considered effective diagnostic and severity indicators for asymptomatic carotid artery stenosis.

Heimann D., Kohnhäuser D., Kohnhäuser A.J., Brönstrup M.

The rise of antimicrobial resistance represents a significant global health threat, driven by the diminishing efficacy of existing antibiotics, a lack of novel antibacterials entering the market, and an over- or misuse of existing antibiotics, which accelerates the evolution of resistant bacterial strains. This review focuses on innovative therapies by highlighting 19 novel antibacterials in clinical development as of June 2024. These selected compounds are characterized by new chemical scaffolds, novel molecular targets, and/or unique mechanisms of action, which render their potential to break antimicrobial resistance particularly high. A detailed analysis of the scientific foundations behind each of these compounds is provided, including their pharmacodynamic profiles, current development state, and potential for overcoming existing limitations in antibiotic therapy. By presenting this subset of chemically novel antibacterials, the review highlights the ability to innovate in antibiotic drug development to counteract bacterial resistance and improve treatment outcomes.

Almeida A.S., Pinho P.G., Remião F., Fernandes C.

New psychoactive substances (NPSs) emerged in the 2000s as legal alternatives to illicit drugs and quickly became a huge public health threat due to their easy accessibility online, limited information, and misleading labels. Synthetic cannabinoids and synthetic cathinones are the most reported groups of NPSs. Despite NPSs being widely studied, due to their structural diversity and the constant emergence of novel compounds with unknown properties, the development of new techniques is required to clarify their mode of action and evaluate their toxicological effects. Metabolomics has been a useful tool to evaluate the metabolic effects of several xenobiotics. Herein, a systematic review was performed, following PRISMA guidelines, regarding metabolomic studies on synthetic cathinones and synthetic cannabinoids to evaluate their effects in cellular metabolism. In the studies, in vivo models were the most employed (86%) and the analysis mostly followed untargeted approaches (75%) using LC-MS techniques (67%). Both groups of NPSs seem to primarily interfere with energy metabolism-related pathways. Even though this type of study is still limited, metabolomics holds great promise as a tool to clarify mechanisms of actions, identify biomarkers of exposure, and explain the toxicological effects of NPSs.

Liu C., Wang C., Jiang J., Bo Y., Nan L., Zhang Y., Zhu K., Wang X., Feng X., Lian X., Qin S.

ObjectiveThere reportedly exists a significant comorbidity between insomnia and neurodegenerative diseases, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD), indicative of a potential link to serum metabolic dysregulation.MethodTo elucidate shared pathophysiological mechanisms between insomnia and AD/PD, we performed comprehensive two-sample Mendelian randomization (MR) analyses, investigating 1,400 serum metabolic characteristics for their causal relationships with the risks of insomnia, AD, widely defined AD (WDAD), and PD. We employed publicly available genetic data; the primary estimate was determined using inverse-variance weighting, supplemented by weighted median, simple mode, weighted mode, and the MR-PRESSO and MR-Egger methods to evaluate heterogeneity and pleiotropy.ResultsThe ratio of N-palmitoyl-sphingosine to N-palmitoyl-sphinganine is linked to higher risks of insomnia (OR = 1.137, 95% CI = 1.015–1.273, p = 0.026) and AD (OR = 1.090, 95% CI = 1.005–1.183, p = 0.037). The acetylcarnitine to propionylcarnitine ratio is a risk factor for insomnia (OR = 1.190, 95% CI = 1.003–1.370, p = 0.016) but has protective effects against AD (OR = 0.868, 95% CI = 0.784–0.961, p = 0.006) and WDAD (OR = 0.892, 95% CI = 0.817–0.973, p = 0.010). Glutamine conjugate of C7H12O2 levels are associated with reduced risk of insomnia (OR = 0.863, 95% CI = 0.749–0.995, p = 0.042) and PD (OR = 0.856, 95% CI = 0.746–0.981, p = 0.026).ConclusionOur findings highlight the crucial role of serum metabolic characteristics in the comorbidity of insomnia with neurodegenerative diseases, providing valuable insights into prospective therapeutic targets and diagnostic markers.

Bozkurt B., Ahmad T., Alexander K., Baker W.L., Bosak K., Breathett K., Carter S., Drazner M.H., Dunlay S.M., Fonarow G.C., Greene S.J., Heidenreich P., Ho J.E., Hsich E., Ibrahim N.E., et. al.

Yang Y., Liu X., Liu X., Xie C., Shi J.

The kynurenine pathway (KP) serves as the primary route for tryptophan metabolism in most mammalian organisms, with its downstream metabolites actively involved in various physiological and pathological processes. Indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO) serve as the initial and pivotal enzymes of the KP, with IDO playing important and intricate roles in cardiovascular diseases. Multiple metabolites of KP have been observed to exhibit elevated concentrations in plasma across various cardiovascular diseases, such as atherosclerosis, hypertension, and acute myocardial infarction. Multiple studies have indicated that kynurenine (KYN) may serve as a potential biomarker for several adverse cardiovascular events. Furthermore, Kynurenine and its downstream metabolites have complex roles in inflammation, exhibiting both inhibitory and stimulatory effects on inflammatory responses under different conditions. In atherosclerosis, upregulation of IDO stimulates KYN production, mediating aromatic hydrocarbon receptor (AhR)-induced exacerbation of vascular inflammation and promotion of foam cell formation. Conversely, in arterial calcification, this mediation alleviates osteogenic differentiation of vascular smooth muscle cells. Additionally, in cardiac remodeling, KYN-mediated AhR activation exacerbates pathological left ventricular hypertrophy and fibrosis. Interventions targeting components of the KP, such as IDO inhibitors, 3-hydroxyanthranilic acid, and anthranilic acid, demonstrate cardiovascular protective effects. This review outlines the mechanistic roles of KP in coronary atherosclerosis, arterial calcification, and myocardial diseases, highlighting the potential diagnostic, prognostic, and therapeutic value of KP in cardiovascular diseases, thus providing novel insights for the development and application of related drugs in future research.

Bakhrushina E.O., Mikhel I.B., Kondratieva V.M., Zubareva I.M., Kosenkova S.I., Belyatskaya A.V., Stepanova O.I., Krasnyuk I.I., Grebennikova T.V., Krasnyuk I.I.

The rapid growth in the prevalence of infectious diseases requires timely action from drug developers. In recent years, the COVID-19 pandemic has demonstrated the unpreparedness of the population for such emergencies. The introduction of modern methods of Design of Experiments (DoE) is required to accelerate the process of drug development and bring a drug to market. The main objective of this study was to develop an ion-triggered in situ system for intranasal delivery of VLP using a Quality by Design approach. Based on a literature review and initial studies, the key QTPP, CQA, CPP, and CMA were identified to develop a novel delivery system for virus-like particles. As a result of the studies on the quality attributes of the developed delivery system, an ion-triggered in situ gel meeting all the specified parameters was obtained using the Quality by Design method.

Belyi S.A., Lukashenko V.I., Kriventsov A.V., Nemkov A.S., Khubulava G.G.

The article presents a clinical case of a patient with severe chronic heart failure of ischemic origin. In 2020, the patient with a long history of ischemic heart disease, as confirmed by clinical data and instrumental examination, was diagnosed with severe cardiomegaly and NYHA class III chronic heart failure. The course of heart failure was aggravated by the presence of arrhythmia in the form of atrial fibrillation. At the first stage, a drug therapy and lifestyle modifications were recommended. In 2021, a beneficial tendency in clinical and instrumental indexes was observed, which made it possible to move on to the surgical stage of treatment. A coronary artery bypass grafting was performed with ablation of the left atrial posterior wall using the “box lesion” technique. A follow-up examination performed a year later showed normalization of the left ventricular dimension and recovery of its contractile function. The symptoms of heart failure regressed to the level of NYHA functional class I; no relapses of atrial fibrillation were detected. The patient continues to receive recommended drug therapy.

Savitskii M.V., Moskaleva N.E., Brito A., Zigangirova N.A., Soloveva A.V., Sheremet A.B., Bondareva N.E., Lubenec N.L., Kuznetsov R.M., Samoylov V.M., Tagliaro F., Appolonova S.A.

Pseudomonas aeruginosa (PA) infection is commonly associated with hospital-acquired infections in patients with immune deficiency and/or severe lung diseases. Managing this bacterium is complex due to drug resistance and high adaptability. Fluorothiazinon (FT) is an anti-virulence drug developed to suppress the virulence of bacteria as opposed to bacterial death increasing host’s immune response to infection and improving treatment to inhibit drug resistant bacteria. We aimed to evaluate FT pharmacokinetics, quorum sensing signal molecules profiling and tryptophan-related metabolomics in blood, liver, kidneys, and lungs of mice. Study comprised three groups: a group infected with PA that was treated with 400 mg/kg FT (“infected treated group”); a non-infected group, but also treated with the same single drug dose (“non-infected treated group”); and an infected group that received a vehicle (“infected non-treated group”). PA-mediated infection blood pharmacokinetics profiling was indicative of increased drug concentrations as shown by increased Cmax and AUCs. Tissue distribution in liver, kidneys, and lungs, showed that liver presented the most consistently higher concentrations of FT in the infected versus non-infected mice. FT showed that HHQ levels were decreased at 1 h after dosing in lungs while PQS levels were lower across time in lungs of infected treated mice in comparison to infected non-treated mice. Metabolomics profiling performed in lungs and blood of infected treated versus infected non-treated mice revealed drug-associated metabolite alterations, especially in the kynurenic and indole pathways.

Belenkov Y.N., Ageev A.A., Kozhevnikova M.V., Khabarova N.V., Krivova A.V., Korobkova E.O., Popova L.V., Emelyanov A.V., Appolonova S.A., Moskaleva N.E., Shestakova K.M., Privalova E.V.

Background: Progressive myocardial remodeling (MR) in chronic heart failure (CHF) leads to aggravation of systolic dysfunction (SD) and clinical manifestations. Identification of metabolomic markers of these processes may help in the search for new therapeutic approaches aimed at achieving reversibility of MR and improving prognosis in patients with CHF. Methods: To determine the relationship between plasma acylcarnitine (ACs) levels, MR parameters and clinical characteristics, in patients with CHF of ischemic etiology (n = 79) and patients with coronary heart disease CHD (n = 19) targeted analysis of 30 ACs was performed by flow injection analysis mass spectrometry. Results: Significant differences between cohorts were found for the levels of 11 ACs. Significant positive correlations (r > 0.3) between the medium- and long-chain ACs (MCACs and LCACs) and symptoms (CHF NYHA functional class (FC); r = 0.31−0.39; p < 0.05); negative correlation (r = −0.31−0.34; p < 0.05) between C5-OH and FC was revealed. Positive correlations of MCACs and LCACs (r = 0.31−0.48; p < 0.05) with the left atrium size and volume, the right atrium volume, right ventricle, and the inferior vena cava sizes, as well as the pulmonary artery systolic pressure level were shown. A negative correlation between C18:1 and left ventricular ejection fraction (r = −0.31; p < 0.05) was found. However, a decrease in levels compared to referent values of ACs with medium and long chain lengths was 50% of the CHF-CHD cohort. Carnitine deficiency was found in 6% and acylcarnitine deficiency in 3% of all patients with chronic heart disease. Conclusions: ACs may be used in assessing the severity of the clinical manifestations and MR. ACs are an important locus to study in terms of altered metabolic pathways in patients with CHF of ischemic etiology and SD. Further larger prospective trials are warranted and needed to determine the potential benefits to treat patients with CV diseases with aberrate AC levels.

Aulia Mardikasari S., Katona G., Budai-Szűcs M., Sipos B., Orosz L., Burián K., Rovó L., Csóka I.

A recommended first-line acute bacterial rhinosinusitis (ABR) treatment regimen includes a high dose of orally administered amoxicillin, despite its frequent systemic adverse reactions coupled with poor oral bioavailability. Therefore, to overcome these issues, nasal administration of amoxicillin might become a potential approach for treating ABR locally. The present study aimed to develop a suitable carrier system for improved local nasal delivery of amoxicillin employing the combination of albumin nanoparticles and gellan gum, an ionic-sensitive polymer, under the Quality by Design methodology framework. The application of albumin nanocarrier for local nasal antibiotic therapy means a novel approach by hindering the nasal absorption of the drug through embedding into an in situ gelling matrix, further prolonging the drug release in the nasal cavity. The developed formulations were characterized, including mucoadhesive properties, in vitro drug release and antibacterial activities. Based on the results, 0.3 % w/v gellan gum concentration was selected as the optimal in situ gelling matrix. Essentially, each formulation adequately inhibited the growth of five common nasal pathogens in ABR. In conclusion, the preparation of albumin-based nanoparticles integrated with in situ ionic-sensitive polymer provides promising ability as nanocarrier systems for delivering amoxicillin intranasally for local antibiotic therapy.

Bakhrushina E.O., Khodenok A.I., Pyzhov V.S., Solomatina P.G., Demina N.B., Korochkina T.V., Krasnyuk I.I.

The development of thermosensitive in situ systems has become widespread and prospective due to the optimal parameters of the phase transition - in the temperature range from room to physiological. Those properties can provide thermosensitive polymers, for example, poloxamers - as the most common. It is worth noting that the addition of active pharmaceutical ingredients (APIs) changes the parameters of in situ systems, but no systematic study of the effect of APIs has been conducted. The aim of this work was to develop a systematic approach to studying the effect of APIs on the in situ rheological properties of poloxamer compositions. The biopharmaceutical classification system (BCS) was chosen as the basis. Accordingly, the following APIs were selected for the experiment: BCS class I - lidocaine hydrochloride and ketorolac tromethamine, class II - ibuprofen and diclofenac, class III - pyridoxine hydrochloride and ribavirin, class IV - furosemide and abiraterone. To create thermoreversible compositions, previously studied for stability combinations of poloxamer 407, poloxamer 188 and PEG 1500 were used. At the stage of preparation of experimental samples formulations with APIs of classes II and IV of BCS were excluded, since the solubilizing ability of poloxamers is not enough to obtain stable combined complexes. In the course of the work, the following results were obtained: BCS class I APIs significantly reduced the phase transition temperature of the matrix of poloxamers 407 and 188, while the addition of PEG 1500 eliminated the effect of APIs on gels; BCS class III APIs practically did not affect the rheological properties of the studied combinations; the phase transition temperature of the gel based on poloxamer 407 did not change with the addition of Class I and Class III APIs. Nevertheless, the obtained results made it possible to reveal the regular behavior of in situ complexes of poloxamer matrices depending on the class of BCS of the API. Further research is required.

Huang L., Wang W., Xian Y., Liu L., Fan J., Liu H., Zheng Z., Wu D.

Intervertebral disc (IVD) degeneration occurred with the increasing age or accidents has puzzled peoples in daily life. To seal IVD defect by injectable hydrogels is a promising method for slowing down IVD degeneration. Herein, we reported a rapidly in situ forming injectable chitosan/PEG hydrogel (CSMA-PEGDA-L) through integrating photo-crosslink of methacrylate chitosan (CSMA) with Schiff base reaction between CSMA and aldehyde polyethylene glycol (PEGDA). The CSMA-PEGDA-L possessed a stronger compressive strength than the photo-crosslinked CSMA-L hydrogel and Schiff-base-crosslinked CSMA-PEGDA hydrogel. This chitosan/PEG hydrogel showed low cytotoxicity from incubation experiments of nucleus pulpous cells. When implanted on the punctured IVD of rat's tail, the CSMA-PEGDA-L hydrogel could well retard the progression of IVD degeneration through physical plugging, powerfully proven by radiological and histological evaluations. This work demonstrated the strategy of in situ injectable glue may be a potential solution for prevention of IVD degeneration.

Narayan S.I., Terre G.V., Amin R., Shanghavi K.V., Chandrashekar G., Ghouse F., Ahmad B.A., S G.N., Satram C., Majid H.A., Bayoro D.K.

Bakhrushina E.O., Dubova A.I., Nikonenko M.S., Grikh V.V., Shumkova M.M., Korochkina T.V., Krasnyuk I.I., Krasnyuk I.I.

The main method of treatment and prevention of endophthalmitis is a combination of intravitreal and topical administration of antibiotics, such as cefuroxime moxifloxacin or vancomycin. However, this method is ineffective due to the rapid elimination of the drug. This problem can be solved with the help of intravitreal in situ injection systems, which are injected with a syringe into the vitreous body and provide prolonged action of the drug at the focus of inflammation. Under the influence of temperature, the liquid drug undergoes a phase transition and turns into a gel after injection. This ensures its prolonged action. The study aimed to develop an intravitreal in situ cefuroxime delivery system for the treatment of endophthalmitis based on a thermosensitive biodegradable composition of poloxamer 407 and hyaluronic acid. A combination of poloxamer Kolliphor® P407, Kolliphor® P188, and PrincipHYAL® hyaluronic acids of different molecular weights was used as a delivery system. The potency of cefuroxime solid dispersion with polyvinylpyrrolidone-10000, polyethylene glycol-400, and polyethylene glycol-1500 in a 1:2 ratio was studied for prolonged action compared to cefuroxime substance. The experimental formulations were studied for the parameters of gelation temperature in a long-term test (4 months), pH, and release of cefuroxime using dialysis bags. To study the distribution parameter in the vitreous body, an in vitro model (1/13) was developed, which was a hollow agar sphere filled with 1% (w/v) polyacrylate gel. For the superior formulations, a HET-CAM test (chorioallantoic membrane test) was performed to determine the absence of irritant effects. According to the study results, a formulation containing a solid dispersion of cefuroxime:PEG-400 (1:2), the matrix of which contained 18% (w/v) Kolliphor® P407 poloxamer, 3% (w/v) Kolliphor® P188 poloxamer, and 0.5% (w/v) hyaluronic acid (1400–1800), was selected. This sample had an average gelation temperature of 34.6 °C, pH 6.7 ± 0.5, and a pronounced prolonged effect. Only 7.6% was released in 3 h of the experiment, whereas about 38% of cefuroxime was released in 72 h. No irritant effect on the chorioallantoic membrane was observed for any formulations studied.

de Figueiredo M., Saugy J., Saugy M., Faiss R., Salamin O., Nicoli R., Kuuranne T., Rudaz S., Botrè F., Boccard J.

Most current state-of-the-art strategies to generate individual adaptive reference ranges are designed to monitor one clinical parameter at a time. An innovative methodology is proposed for the simultaneous longitudinal monitoring of multiple biomarkers. The estimation of individual thresholds is performed by applying a Bayesian modeling strategy to a multivariate score integrating several biomarkers (compound concentration and/or ratio). This multimodal monitoring was applied to data from a clinical study involving 14 female volunteers with normal menstrual cycles receiving testosterone via transdermal route, as to test its ability to detect testosterone administration. The study samples consisted of urine and blood collected during 4 weeks of a control phase and 4 weeks with a daily testosterone gel application. Integrating multiple biomarkers improved the detection of testosterone gel administration with substantially higher sensitivity compared with the distinct follow-up of each biomarker, when applied to selected urine and serum steroid biomarkers, as well as the combination of both. Among the 175 known positive samples, 38% were identified by the multimodal approach using urine biomarkers, 79% using serum biomarkers and 83% by combining biomarkers from both biological matrices, whereas 10%, 67% and 64% were respectively detected using standard unimodal monitoring. The detection of abnormal patterns can be improved using multimodal approaches. The combination of urine and serum biomarkers reduced the overall number of false-negatives, thus evidencing promising complementarity between urine and blood sampling for doping control, as highlighted in the case of the use of transdermal testosterone preparations. The generation in a multimodal setting of adaptive and personalized reference ranges opens up new opportunities in clinical and anti-doping profiling. The integration of multiple parameters in a longitudinal monitoring is expected to provide a more complete evaluation of individual profiles generating actionable intelligence to further guide sample collection, analysis protocols and decision-making in clinics and anti-doping.

Kozhevnikova M.V., Korobkova E.O., Krivova A.V., Kukharenko A.V., Moskaleva N.E., Shestakova K.M., Mesonzhnik N.V., Ageev A.A., Boldin A.A., Brito A., Appolonova S.А., Privalova E.V., Belenkov Y.N.

Aim. Branched-chain amino acids (BCAAs) have been postulated as potential indicators of cardiovascular risk. The objective of this study was to explore the relationship between plasma BCAAs and different stages of cardiovascular disorders.Material and methods. In our cross-sectional study, plasma BCAAs (valine, leucine and isoleucine) in individuals without cardiovascular diseases (CVDs) (nonCVD group, total n=27, with n=16 healthy, but with metabolic disorders) were compared to patients diagnosed with CVDs [CVD group, total n=109, being n=61 hypertension (n=31 with signs of beginning of myocardial remodeling) and n=48 patients with coronary artery disease (CAD)].Results. The plasma concentration of BCAAs was significantly higher in the group of patients with cardiovascular disease compared with the healthy group (p<0.05 for all amino acids tested): valine concentration was 238.7 [219.6; 267.0] μM in the non-CVD group and 261.2 [233.8; 298.7] μM in the CVD group; leucine concentration was 134.8 [122.4; 153.2] μM and 146.8 [129.0; 166.6] μM, respectively; and isoleucine 72.7 [65.3; 84.4] μM and 81.7 [68.0; 96.2] μM, respectively. Leucine and isoleucine concentration levels were minimal in the healthy participant subgroup and maximal in the IBS patient subgroup. No statistically significant differences in BCAAs concentrations were found in the subgroups without CAD. Significant increases in concentrations were observed in the subgroups of patients with CAD as follows: valine concentration was 256.3 [219.0; 297.9] μM in hypertension group and 261.7 [236.5; 307.5] μM in CAD group; leucine concentration was 141.8 [123.5; 166.6] μM and 154.1 [134.7; 172.7] μM, respectively, and isoleucine 72.8 [65.7; 94.0] μM and 85.7 [74.9; 101.7] μM, respectively. BCAAs profiles in all participants with metabolic disorders had “good” diagnostic accuracy with area under the receiver operating characteristics curve being 0.72, 0.70 and 0.70 for valine, leucine and isoleucine, respectively.Conclusion. BCAAs concentrations are elevated with higher severity of the cardiovascular disorder and exhibit potential as early independent indicators of coronary artery disease.

Anker S.D., Usman M.S., Anker M.S., Butler J., Böhm M., Abraham W.T., Adamo M., Chopra V.K., Cicoira M., Cosentino F., Filippatos G., Lund L.H., Moura B., Mullens W., Pieske B., et. al.

Shestakova K.M., Moskaleva N.E., Boldin A.A., Rezvanov P.M., Shestopalov A.V., Rumyantsev S.A., Zlatnik E.Y., Novikova I.A., Sagakyants A.B., Timofeeva S.V., Simonov Y., Baskhanova S.N., Tobolkina E., Rudaz S., Appolonova S.A.

AbstractLung cancer is referred to as the second most common cancer worldwide and is mainly associated with complex diagnostics and the absence of personalized therapy. Metabolomics may provide significant insights into the improvement of lung cancer diagnostics through identification of the specific biomarkers or biomarker panels that characterize the pathological state of the patient. We performed targeted metabolomic profiling of plasma samples from individuals with non-small cell lung cancer (NSLC, n = 100) and individuals without any cancer or chronic pathologies (n = 100) to identify the relationship between plasma endogenous metabolites and NSLC by means of modern comprehensive bioinformatics tools, including univariate analysis, multivariate analysis, partial correlation network analysis and machine learning. Through the comparison of metabolomic profiles of patients with NSCLC and noncancer individuals, we identified significant alterations in the concentration levels of metabolites mainly related to tryptophan metabolism, the TCA cycle, the urea cycle and lipid metabolism. Additionally, partial correlation network analysis revealed new ratios of the metabolites that significantly distinguished the considered groups of participants. Using the identified significantly altered metabolites and their ratios, we developed a machine learning classification model with an ROC AUC value equal to 0.96. The developed machine learning lung cancer model may serve as a prototype of the approach for the in-time diagnostics of lung cancer that in the future may be introduced in routine clinical use. Overall, we have demonstrated that the combination of metabolomics and up-to-date bioinformatics can be used as a potential tool for proper diagnostics of patients with NSCLC.