Synthesis of New Amidoethanesulfonamides of Betulonic Acid.

New potentially biologically active amidoethanesulfonamides of betulonic acid were synthesized by the acid chloride method via conjugation of betulonic acid with 2-aminoethanesulfonamides as the free bases.

Analysis of Clinical and Biochemical Characteristics of Patients With Genetically Confirmed Familial Hypercholesterolemia in Russian North Western District Residents.

Aim      To compare results of clinical, laboratory, and genetic examination of patients with familial hypercholesterolemia (FHC).Material and methods  112 patients aged 40.2±17.9 years (49 men) were examined. The gene of low-density lipoprotein receptor (LDLR) was analyzed and evaluated using the Dutch Lipid Clinic Network (DLCN) criterion of lipid score ≥6. The LDLR gene mutation was searched for using the conformational polymorphism analysis followed by sequencing of the DNA of isolated LDLR gene exons.Results Mean variables of the blood lipid profile were total cholesterol (C), 10.12±2.32 mmol/l, LDL-C, 7.72±2.3 mmol/l. Corneal arcus was observed in 15 % of patients, tendon xanthomas in 31.8 %, and xanthelasma palpebrarum in 5.3 %. The types of LDLR gene mutations included missense mutations (42.8 %), mutations causing a premature termination of protein synthesis (41.1 %), and frameshift mutations (16.1 %). In the presence of a mutation in exon 4, patients with IHD compared to patients with no IHD had significantly higher levels of total C (10.88±2.08 mmol/l vs. 8.74±1.57 mmol/l, respectively, р=0.001) and LDL-C (8.60±2.14 mmol/l vs. 6.62±1.79 mmol/l, respectively, р=0.005). Patients with IHD compared to patients with no IHD and a mutation in LDLR gene exon 9 had only a higher LDL-C level (8.96±1.53 mmol/l vs. 6.92±1.59 mmol/l, respectively, р=0.022). A differentiated comparison of IHD patients using a logistic regression depending on the identified type of LDLR gene mutation produced formulas for calculating the odds ratio of IHD and myocardial infarction (MI) with adjustments for the patient's age and baseline LDL.Conclusion      The detection rate of the LDLR gene mutations was 42.8 % for missense mutations, 41.1 % for mutations causing a premature termination of protein synthesis, and 16.1 % for frameshift mutations. Blood lipid profiles did not differ between patients from different cities and with different types of LDLR gene mutations. Blood lipid profiles were different in IHD patients depending on the mutation type.

[Correlations between vessel stiffness and biomarkers of senescent cell in elderly patients].

Aim    To study the association between vascular wall stiffness and known markers for accumulation of senescent cells in blood, cells, and tissues of old patients.Material and methods    This study included male and female patients aged 65 years and older who were referred to an elective surgical intervention, that included a surgical incision in the area of the anterior abdominal wall or large joints and met the inclusion and exclusion criteria. For all patients, traditional cardiovascular (CV) risk factors and arterial wall stiffness (pulse wave velocity, PWV) were evaluated. Also, biomaterials (peripheral blood, skin, subcutaneous adipose tissue) were collected during the surgery and were used for isolation of several cell types and subsequent histological analysis to determine various markers of senescent cells.Results    The study included 80 patients aged 65 to 90 years. The correlation analysis identified the most significant indexes that reflected the accumulation of senescent cells at the systemic, tissue, and cellular levels (r>0.3, р<0.05) and showed positive and negative correlations with PWV. The following blood plasma factors were selected as the markers of ageing: insulin-like growth factor 1 (IGF-1), fibroblast growth factor 21 (FGF-21), and vascular endothelium adhesion molecule 1 (VCAM-1). A significant negative correlation between PWV and IGF-1 concentration was found. Among the tissue markers, P16INK, the key marker for tissue accumulation of senescent cells, predictably showed a positive correlation (r=0.394, p<0.05). A medium-strength correlation with parameters of the 96-h increment of mesenchymal stromal cells and fibroblasts and a weak correlation with IL-6 as a SASP (specific senescent-associated secretory phenotype) were noted. Results of the multifactorial linear regression analysis showed that the blood plasma marker, VCAM-1, and the cell marker, 96-h increment of fibroblasts, were associated with PWV regardless of the patient's age.Conclusion    Stiffness of great arteries as measured by PWV significantly correlates with a number of plasma, tissue, and cellular markers for accumulation of senescent cells. This fact suggests PWV as a candidate for inclusion in the panel of parameters for evaluation and monitoring of the biological age during the senolytic therapy.

Synthesis of 2-Aminoethanesulfonamides of Betulinic and Betulonic Acids.

New potentially biologically active sulfonamide derivatives of pentacyclic lupane-type triterpenoids, the sulfonamide group of which was bonded to C-17 of the triterpene skeleton through an amidoethane spacer, were synthesized via conjugation of 2-aminoethanesulfonamides to betulinic and betulonic acids in the presence of Mukaiyama reagent (2-bromo-1-methylpyridinium iodide).

[Sleep-wake cycle regularity and cardiometabolic indicators]. / Regulyarnost' ritma «son ­ bodrstvovanie¼ i kardiometabolicheskie pokazateli.

OBJECTIVE: To evaluate the association between sleep-wake rhythm and cardiometabolic parameters. MATERIAL AND METHODS: 103 participants, aged 25-64 years, underwent actigraphy study with Actigraph Actilife GT3X + device (USA) for 7 days. We assessed actigraphy indicators (physical activity and sleep data), anthropometric indicators, blood pressure and laboratory parameters. Actigraphy data was processed using the nparACT package in the R program with the calculation of nonparametric indicators: intraday variability, interday stability, the average level of lowest activity for five hours (L5) and ten hours with the highest activity (M10), relative amplitude is the ratio of M10/L5. RESULTS: The nonparametric analysis showed an association of the higher night activity with sleep effectiveness, wake after sleep onset, indicators of physical activity. A more stable sleep pattern is associated with more steps, less weight and waist circumference, lower levels of diastolic blood pressure, creatinine and insulin. Increased fragmentation of sleep patterns is associated with increased CRP and increased sedentary time. Participants with higher activity contrast have less waist circumference, hips and body mass index, lower levels of CRP and insulin. CONCLUSIONS: Rhythm and quality of sleep are important parameters associated with cardiometabolic indicators. Stable sleep patterns, higher activity during the day and lower night activity are associated with a more favorable condition of cardiovascular system.

Data on characterization of glass biochips and validation of the label-free biosensor for detection of autoantibodies in human serum.

The data represent in-depth characterization of a novel method for highly sensitive simultaneous measuring in human serum of both critical parameters of autoantibodies: concentration and native kinetics. The latter refers to autoantibody interaction with free, not immobilized, antigen. The method and related biosensors are based on the spectral-correlation and spectral-phase interferometry. The data cover: multi-factor optimization and quantitative characterization of the developed affordable single-used biochips, including X-ray photoelectron spectroscopy (XPS) control of chemical modifications of the surface during fabrication; antibody screening; optimization and verification of protocols for label-free biosensing in human serum; mathematical model for fitting experimental data and calculation of kinetic constants of interaction of autoantibodies with free antigen; comprehensive verification of the method specificity; correlation between the data obtained with the developed biosensor and with enzyme linked immunosorbent assay (ELISA); comparison of analytical characteristics of the developed biosensor with the most advanced label-based methods. The data importance is confirmed by a companion paper (DOI 10.1016/j.bios.2020.112187), which shows that the combination of mentioned autoantibody parameters is promising for more accurate criteria for early diagnostics and efficient therapy of autoimmune disorders. The obtained data can be used in development of a wide range of biosensors, both label-free and based on various labels.

Multiplex label-free biosensor for detection of autoantibodies in human serum: Tool for new kinetics-based diagnostics of autoimmune diseases.

A multiplex label-free biosensor is developed for diagnostics of autoimmune diseases by highly sensitive measuring in human serum both critical characteristics of autoantibody: concentration and native kinetic parameters that reflect autoantibody aggressiveness to the organism's tissues. The biosensor is based on the spectral-correlation interferometry and image processing of a microarray glass biochip, affordable to be single-used in medical applications. Simultaneous 25-min detection and activity characterization of several autoantibodies in the same serum sample have been demonstrated for anti-thyroglobulin (anti-TG) and anti-thyroid peroxidase (anti-TPO) as models. The biosensor offers extremely high sensitivity: limits of detection in serum are 1.7 IU/mL and 6 IU/mL for anti-TPO and anti-TG, respectively. The dynamic range covers the whole range of clinically relevant concentrations of the autoantibodies up to 1000 IU/mL. The developed method of characterization of autoantibody activity by recording the kinetics of their binding with free native antigens is based on autoantibody polyvalency. The measurements in clinical serum samples have shown that the native kinetic parameters are independent of concentration. The proposed biosensor and method of native kinetic registration can be used to develop new criteria for comprehensive diagnostics of autoimmune diseases, based not only on traditional measurements of concentration but also on quantitative evaluation of autoantibody aggressiveness. The developed method can be adapted to other label-free sensors such as those based on the surface plasmon resonance, optical waveguides, etc.

[Office and 44-hour ambulatory blood pressure and central haemodynamic parameters in the patients with end-stage renal diseases undergoing haemodialysis].

AIM: To assess the incidence of blood pressure (BP) control and various phenotypes of BP by comparing the results of office and 44-hour ambulatory brachial and central BP measurement in patients with end-stage renal disease (ESRD) on program hemodialysis (HD). MATERIALS AND METHODS: In 68 patients ESRD receiving renal replacement therapy we evaluated office peridialysis BP and performed 44-hour ambu latory monitoring (ABPM) of brachial and central BP during peridialysis period using a validated oscillometric device BPLabVasotens (OOO "Petr Telegin"). Results were considered statistically significant with p<0.05. RESULTS: The frequency of control of peripheral office BP before the HD session was 25%, after - 23.5%; control of central BP - 48.6% and 49%, respectively. According to office measurement the frequency of systolic-diastolic hypertension was 44.1%, isolated systolic hypertension - 25%, isolated diastolic hypertension - 5.9%. The values of peripheral and central office systolic BP (SBP) before and after HD were not consistent with the corresponding mean and daily SBP levels for 44 hours and for the first and second days of the interdialysis period. The frequency of true uncontrolled arterial hypertension (AH) according to peripheral ABPM was 66.5%, masked uncontrolled AH - 9%. Circadian rhythm abnormalities for 44-h peripheral BP were detected in 77%, for central - in 76%. In 97% of patients agreement between phenotypes of the daily profile of peripheral and central BP was observed. 73% of patients had a significant increase in peripheral and central SBP and pulse pressure (PP) and an increase in the proportion of non-dippers from the 1st to the 2nd day. CONCLUSION: Patients with ESRD on HD were characterized by poor control of BP control and predominance of unfavourable peripheral and central ambulatory BP phenotypes. A single measurement of clinical peripheral and central BP in the peridialysis period was not sufficient to assess the control of hypertension in this population. The 24-h BP profiles in the 1st and 2nd days of interdialysis period had significant differences.

Data on characterization and validation of assays for ultrasensitive quantitative detection of small molecules: Determination of free thyroxine with magnetic and interferometric methods.

The presented data refer to optimization and quantitative characterization of a rapid lateral flow assay based on high-affinity bifunctional ligand and magnetic nanolabels, which was developed for detection of small molecules of thyroid hormones. The results were obtained by several techniques, including the magnetic particle quantification method, spectral-correlation interferometry and spectral-phase interferometry, dynamic light scattering, enzyme linked immunosorbent assay. The long-term stability of "antibody - magnetic nanoparticle" conjugates is shown. The assay specificity is confirmed, and verification of successful combination of magnetic particles and antibodies is demonstrated. The kinetic and equilibrium dissociation constants are determined for interactions between thyroxine and monoclonal antibodies. The obtained data could be used for design of other platforms for detection of small molecules.

Large artery stiffness according to different assessment methods in adult population of St.Petersburg.

OBJECTIVE: The aim of the present study was to assess the prevalence of increased arterial stiffness by different diagnostic methods and its association with cardiovascular risk in Russian population-based cohort. DESIGN AND METHODS: In terms of Russian epidemiological study ESSE-RF a random selection of 452 apparently healthy Saint-Petersburg inhabitants aged 25-65 years was performed. Fasting lipids, glucose and blood pressure measurements were performed. We used 3 diagnostic methods of arterial stiffness assessment: pulse wave velocity by applanation tonometry (SphygmoCor - PWV-S) and pulse wave velocity by volumetric sphygmography (VaSera - PWV-V), and cardio-ankle vascular index (CAVI) by VaSera. RESULTS: 341 (75,4%) had normal parameters of arterial stiffness assessed by all methods. Spearmen's coefficient of correlation and "kappa" coefficient for PWV-S and CAVI were 0,74 and 0,04, for PWV-S and PWV-V - 0,10 and 0,06, for CAVI and PWV-V - 0,28 and 0,03, respectively. There was a significant correlation between cardiovascular risk (defined by SCORE) and PWV-S (r = 0,38, p < 0,001) and a non-significant trend of increasing CAVI along with cardiovascular risk (r = 0,35, p = 0,14). CONCLUSIONS: Different methods of arterial stiffness assessment showed a weak correlation with each other. Carotid-femoral pulse wave velocity detected by applanation tonometry is associated with high cardiovascular risk score and might be considered as better additional risk marker for cardiovascular risk stratification.

Ultrasensitive detection enabled by nonlinear magnetization of nanomagnetic labels.

Geometrically confined magnetic particles due to their unique response to external magnetic fields find a variety of applications, including magnetic guidance, heat and drug delivery, magneto-mechanical actuation, and contrast enhancement. Highly sensitive detection and imaging techniques based on the nonlinear properties of nanomagnets were recently proposed as innovative strong-translational potential methods applicable in complex, often opaque, biological systems. Here we report on the significant enhancement of the detection capability using optical-lithography-defined, ferromagnetic iron-nickel alloy disk-shaped particles. We show that an irreversible transition between strongly non-collinear (vortex) and single domain states, driven by an alternating magnetic field, translates into a nonlinear magnetic response that enables ultrasensitive detection of these particles. The record sensitivity of ∼3.5 × 10-9 emu, which is equivalent to ∼39 pg of magnetic material is demonstrated at room temperature for arrays of patterned disks. We also show that unbound disks suspended in the aqueous buffer can be successfully detected and quantified in real-time when administered into a live animal allowing for tracing of their biodistribution. The use of nanoscale ferromagnetic particles with engineered nonlinear properties opens prospects for further enhancing the sensitivity, scalability, and tunability of noise-free magnetic tag detection in high-background environments for various applications spanning from biosensing and medical imaging to anti-counterfeiting technologies.

Cholesterol Levels in Genetically Determined Familial Hypercholesterolaemia in Russian Karelia.

Familial hypercholesterolaemia (FH) is a rare disease that tends to be diagnosed lately. In Russia, the genetic and phenotypic characteristics of the disease are not well defined. We investigated 102 patients with definite FH. In 52 of these patients (50.9%) genetic analysis was performed, revealing pathogenic mutations of the low density lipoprotein (LDL) receptor gene in 22 patients. We report here five mutations of the LDL receptor gene found in the Karelian FH sample for the first time. The detection rate of mutations in definite FH patients was 42.3%. Two groups of patients with a definite diagnosis of FH according to the Dutch Lipid Clinic Network criteria were compared: the first group had putatively functionally important LDL receptor gene mutations, while in the second group LDL receptor gene mutations were excluded by single-strand conformation polymorphism analysis. Total and LDL cholesterol levels were higher in the group with LDL receptor mutations compared to the mutation-free population. The frequency of mutations in patients with LDL cholesterol > 6.5 mmol/L was more than 3 times higher than that in patients with LDL < 6.5 mmol/L. Total and LDL cholesterol levels and the frequency of coronary heart disease and myocardial infarction were higher in the group with definite FH compared to groups with probable and possible FH. Cholesterol figures in FH patients of different age and sex from the Karelian population were comparable.

[Adipokines Level in Metabolically Healthy Obese Saint-Petersburg Inhabitants (ESSE-RF)].

AIM: to elucidate prevalence of phenotypes of metabolically heathy obesity (MHO) among inhabitants of Saint-Petersburg using various criteria and assess dependence levels of adipokines on obesity phenotype. MATERIAL AND METHODS: Within a framework of epidemiologic study ESSE-RF we formed a random sample of 1600 Saint-Petersburg inhabitants stratified according to gender and age. Examination of participants included anthropometry with measurement of waist circumference CRP and estimation of body mass index (BMI), measurement of arterial pressure, determination of fasting blood glucose, insulin (with calculation of index of insulin resistance -IIR), lipid spectrum, C-reactive protein (CRP), adiponectin, leptin. In subjects with obesity (BMI more or equal 30 kg/m) we used Meigs and Wildman MHO criteria. To Wildman criteria we applied 3 variants of definition of elevated CRP and IIR: 90th percentile among subjects with BMI <25 kg/m (variant 1) or among all participants (variant 2), and (variant 3) definition from publication by E.Oliveros et al. (2014). RESULTS: Obesity (BMI more or equal 30 kg/m) was found in 430 participants. Numbers/rates of MHO according to the Wildman criteria were the following: variant 1 - 49/12% (among them 13/10% men and 36/14% women, =0.15); variant 2 - 85/22% (24/18% men, 61/23% women, =0.13); variant 3 - 59/15% (13/10% men, 46/18% women, =0.02). Portion of MHO according to Meigs criteria was 138/35% (among them 48/36% men, 90/35% women, =0.4). Significant differences in adipokines levels between subjects with MHO and metabolically unhealthy obesity (MUHO) were revealed only among women. There was no difference in leptin level between subjects with MHO and MUHO irrespective of gender. CONCLUSION: Rate of MHO phenotype in a sample of inhabitants of Saint-Petersburg varied from 12 to 35% depending on criteria used. Gender differences in MHO rates were minimal and depended on selected criteria. Elevated adiponectin level among obese women could be presumably related to more favorable metabolic profile.

[Gender Differences of Behavioral Risk Factors in Saint-Petersburg Inhabitants].

BACKGROUND: In developed countries there are significant gender differences in lifetime expectancy that can be explained by behavioral risk factors (RF). OBJECTIVE: The aim of our study was to estimate gender features of behavioral RF in general population of Saint-Petersburg, Russia. METHODS: As a part of all-Russian epidemiology survey ESSE-RF a random sampling of 1600 Saint-Petersburg inhabitants (25-64 y.o.) stratified by age and sex was performed. All participants filled in the questionnaire. Anthropometry (weight, height, body-mass index (BMI), waist circumference (WC) and fasting blood-tests (lipids, glucose by Abbott Architect 8000 (USA)) were performed. RESULTS: There were examined 573 (36%) men and 1027 (64%) women. No gender differences in obesity were found according to BMI criteria--in 178 (31.2%) women and 352 (35.1%) men. Obesity was more often detected in females according to WC criteria: ATPIII--44.1 vs 30.3%; IDF 51.2 vs 66.4% (p < 0.001 for both). Linear regression analysis was performed and age was associated with BMI--1.6 kg/m2/decade, WC in women--5.2 cm/decade and WC in men--2.8 cm/decade, p < 0.001 for all anthropometric parameters. Optimal level of physical activity was equally documented in both genders--540 (61.2%) women and 286 (58.9%) men. Daily intake of sweets was lower in men--228 (39.8%) vs 539 (52.5%) in women (p < 0.001). 810 (50.6%) of trial subjects were non-smokers, 395 (24.7%) were former smokers, and 395 (24.7%) were smokers at the moment of trial. The higher number of female smokers was observed--194 (19.1%). CONCLUSION: A high prevalence of obesity is observed in sample of Saint-Petersburg inhabitants--it is higher among women according to WC criteria regardless of menopause, possibly due to bigger sweets consumption. Males smoke more often and consume less fresh fruits and vegetables which is accompanied by a higher prevalence of hyperglycemia and hypertriglyceridemia.

Development of immunoassays using interferometric real-time registration of their kinetics.

A method for effective development of solid-phase immunoassays on a glass surface and for optimization of related protocols by highly sensitive quantitative monitoring of each assay step has been proposed and experimentally implemented. The method is based on the spectral correlation interferometry (SCI) that allows real-time measuring of the thickness of a biomolecular layer bound to the recognition molecular receptors on the sensor chip surface. The method is realized with compact 3-channel SCI-biosensors that employ as the sensor chips standard cover glass slips without deposition of any additional films. Different schemes for antibody immobilization on a glass surface have been experimentally compared and optimized toward a higher sorption capacity of the sensor chips. Comparative characterization of the kinetics of each immunoassay stage has been implemented with the optimized protocols: i) covalent immobilization of antibody on an epoxylated surface and ii) biotinylated antibody sorption on a biotinylated surface via a high-affinity biotin-streptavidin bond. We have shown that magnetic nanoparticles employed as labels with model detection of cardiac troponin I further amplify the SCI signal, resulting in 100-fold improvement of the detection limit. The developed protocols can also be used with the alternative immunoassay platforms, including the label methods based on registration of only the final assay result, which is the quantity of bound labels.

[Detection of pyrethroids by spectral correlation interferometry].

A label-free method based on spectral correlation interferometry has been developed for highly sensitive detection of pyrethroids by competitive immunoassay on the surface of sensor chips made of widely available microscopy glass cover slips. It is shown that the method allows independent optimization of each step of the sensor surface modification. This fact may be used to increase the efficiency of development of protocols for a wide spectrum of immunoassays that employ glass surface as a solid phase. Detection of 3-phenoxybenzoic acid, which is one of the most stable metabolites of a large number of pyrethroids, on the surface of the optimized sensor chips has been demonstrated on the level of 15 pg/ml. That is 50 times better than the sensitivity of the enzyme-linked immunosorbent assay (ELISA).