[Dysfunctional High-Density Lipoproteins: Role in Atherogenesis and Potential Targets for Phospholipid Therapy].

In recent years the number of articles on damages of high-density lipoproteins (HDL) properties in patients with atherosclerosis has sharply increased. First, it concerns their ability to accept cholesterol (CH) from macrophages - the basis of antiatherogenic action of HDL. This ability was assessed ex vivo - by activity of cell cholesterol (CH) efflux to HDL or into patient's serum. In many works inverse relationship was shown between CH acceptor capacity of HDL and severity of atherosclerotic disease or frequency of its exacerbations during long-term observation, independent from HDL CH concentration. This led to the emergence of the concept of importance of "not only HDL quantity but also of their quality", i. e. functionality. In this review we consider pathways of cellular CH efflux (mainly mediated by cell proteins), methods used for detection of dysfunctional HDL, and results of relevant studies in various categories of patients. These studies directed to identification of mechanisms of damages of HDL properties by means of analysis of their composition, used various approaches including those of proteomics and lipidomics. However, now there are no proven targets for correction of HDL dysfunctionality. The only factor, that is underlined by many authors, is the significance of HDL phospholipids, which level correlates with activity of cellular CH efflux. This allows to take a fresh look at previously used phospholipid therapy of atherosclerosis. Its mechanism is apparently not lowering of plasma CH, as was previously expected, but the improvement of HDL antiatherogenic properties. For its practical usage it is necessary to elaborate principally novel formulations with high bioavailability of phospholipids - for HDL enrichment by phospholipid and thereby normalization of their ability to remove CH from tissues.

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.

[Splice variants of mRNA of cytochrome P450 genes: analysis by the nanopore sequencing method in human liver tissue and HepG2 cell line]. / Splais-varianty mRNK genov tsitokhromov R450: analiz metodom nanoporovogo sekvenirovaniia v tkani pecheni cheloveka i kletkakh linii HepG2.

The analysis of cytochrome P450 transcripts was carried out by the nanopore sequencing in liver tissue samples of three donors and HepG2 line cells. It has been demonstrated that direct mRNA sequencing with a MinION nanopore sequencer (Oxford Nanopore Technologies) allows one to obtained quantitative profiles for transcripts (and their splice variants) of cytochrome P450 superfamily genes encoding isoforms involved in metabolism of the large (~80%) part of drugs. The splice variant profiles substantially differ for donors. The cytochrome P450 gene expression at the transcript level is significantly weaker in cells of the HepG2 line compared with that in the normal liver tissue. This limits the capability of the direct mRNA nanopore sequencing for studying alternative splicing of cytochrome P450 transcripts in HepG2 cells. Both quantitative and qualitative profiles of the cytochrome P450 gene expression at the transcript level are notably differ in human liver tissue and HepG2 cells.

[Nonimmunogenic staphylokinase in the treatment of acute ischemic stroke (FRIDA trial results)]. / Neimmunogennaya stafilokinaza ­ novyi tromboliticheskii preparat v lechenii ishemicheskogo insul'ta (rezul'taty issledovaniya FRIDA).

AIM OF THE STUDY: To investigate the efficacy and safety of non-immunogenic staphylokinase (NS) compared with alteplase (A) in patients with acute ischemic stroke (AIS) within 4.5 h after symptom onset. MATERIAL AND METHODS: 336 patients with IS within 4.5 h after symptom onset were included in a randomized, open-label, multicenter, parallel-group, non-inferiority comparative trial of NS vs A (168 patients in each group). NS was administered as an intravenous bolus in a dose of 10 mg, regardless of body weight, over 10 s, A was administered as a bolus infusion in a dose of 0.9 mg/kg, maximum 90 mg over 1 hour. The primary efficacy endpoint was a favorable outcome, defined as a modified Rankin scale (mRS) score of 0-1 on day 90. Safety endpoints included all-cause mortality on day 90, symptomatic intracranial haemorrhage, and other serious adverse events (SAEs). RESULTS: At day 90, 84 (50%) patients reached the primary endpoint (mRS 0-1) in the NS group, 68 (41%) patients - in the A group (p=0.10, OR=1.47, 95% CI=0.93-2.32). The difference between groups NS and A was 9.5% (95% CI= -1.7-20.7) and the lower limit of the 95% CI did not cross the margin of non-inferiority (pnon-inferiority<0.0001). There were no significant differences in the frequency of deaths between the groups: on day 90, 17 (10%) patients in the NS group and 24 (14%) in the A group had died (p=0.32). There was a trend towards significant differences in the frequency of symptomatic intracranial haemorrhage: NS group - 5 (3%) patients, A group - 13 (8%) patients (p=0.087, OR=0.37, 95% CI=0.1-1.13). There were significant differences in the number of patients with SAEs: in the NS group - 22 (13%) patients, in the A group - 37 (22%) patients (p=0.044, OR=0.53, 95% CI=0.28-0.98). CONCLUSION: The presented results of the FRIDA trial are the first in the world to use a drug based on NS in patients with IS. It has been shown that a single bolus (within 10 s) administration of NS at a standard dose of 10 mg, regardless of body weight, allows to conduct fast, effective and safe thrombolytic therapy in patients with IS within 4.5 h after symptom onset. In further clinical tials of NS, it is planned to expand the therapeutic window beyond 4.5 h after symptom onset in patients with IS.

[PCR analysis of the expression of chromosome 18 genes in human liver tissue: interindividual variability]. / PTsR-analiz ékspressii genov khromosomy 18 v tkani pecheni cheloveka: mezhindividual'naia variabel'nost'.

Using human chromosome 18 (Ch18) genes as an example, a PCR analysis of the interindividual variability of gene expression in liver tissue was performed. Although the quantitative profiles of the Ch18 transcriptome, expressed in the number of cDNA copies per single cell, showed a high degree of correlation between donors (Pearson correlation coefficients ranged from 0.963 to 0.966), the expression of the significant number of genes (from 13% to 19%, depending on the method of experimental data normalization) varied by more than 4-fold when comparing donors pairwise. At the same time, the proportion of differentially expressed genes increased with a decrease in the level of their expression. It is shown that the higher quantitative variability of low-abundance transcripts is mainly not technical, but biological. Bioinformatic analysis of the interindividual variability of the differential expression of chromosome 18 genes in human liver tissue did not reveal any statistically significant groups of genes related to certain biological processes that indicated a rather transient nature of the interindividual variability of their expression, probably reflecting the response of cells of an individual to specific external stimuli.

[Familial transthyretin amyloidosis].

A case of familial transthyretin amyloidosis with TTR Cys 114 gene polymorphism is described (first in Russia and third in the world). The clinical picture of the proband was dominated by symptoms of autonomous polyneuropathy (orthostatic hypotension, erectile dysfunction, diarrhea, tachycardia, foot dyshydrosis) and of somatic nerve lesions (dumbness, impaired surface and deep sensitivity in the limbs). The patient presented with vitreous body opacity, disturbed eye movements, lateralized sensory symptoms, and difficulty of speech (baryphonia). Electromyographic quantitative autonomous testing and measurement of evoked sympathetic skin potentials confirmed affection of peripheral nerves. Heart ultrasound revealed restrictive amyloid cardiopathy. Histological analysis showed amyloid deposition in the intestines and sural nerve. The proband, his daughter, brother (monozygous twin), and brother's daughter had mutant TTR Cys 114 gene. The brother also had amyloid deposits in the absence of clinical signs of the disease. Analysis of familial medical history demonstrated autosomal dominant inheritance of this mutation in 4 generations. Its possible origin and clinical features of the disease are discussed.

[Thrombolytic therapy of ischemic stroke]. / Tromboliticheskaia terapiia ishemicheskogo insul'ta.

Reperfusion therapy is one of the main treatment strategies of ischemic stroke. The first studies of the efficacy of thrombolytic medications started form the use of streptokinase and fibrinolysin in patients with ischemic stroke in late 50 - early 60 of the XX century in the United States, Soviet Union, and Western Europe. After the development of recombinant tissue plasminogen activator, thrombolysis became one of the main methods of reperfusion in patients with acute ischemic stroke, acute myocardial infarction, or other acute vascular thrombotic events. Later, modified variants of tissue plasminogen activator with prolonged clearance time, high fibrin-selectivity, and bolus delivery were introduced. Another group of thrombolytic agents includes derivatives of flora and fauna - external plasminogen activators, of which streptokinase, staphylokinase, and desmoteplase are most common drugs. These medications are not a structural part of the human organism, and overcoming of immunogenicity while preserving fibrinolytic activity and fibrin specificity is one of the main tasks in applying them in clinical practice.

[Pathomorphology of the thyroid gland (based on emergency biopsy)]. / Patomorfologiia shchitovidnoi zhelezy (po dannym srochnoi biopsii.

The term "struma" denotes any increase of the thyroid volume and weight regardless of its nature. In no other organ there are so frequent polymorphic structures stimulating neoplastic growth. This does not allow for a wide use the diagnosis of early thyroid carcinoma. Differentiation of the thyroid lesions is to a great extent possible due to stereomorphologic and rapid histotopographic study of surgical material with its subsequent embedding into paraffin. Nodular and adenomatoid struma was found in 60%, follicular adenomas in 11% cases among 20,000 operated patients with euthyroidism in a non-general endocrinological department. Thyroid carcinoma represented 5% and autoimmune thyroiditis in 4% while before the operation they were diagnosed in 1% cases only. Microcarcinoma among routine biopsies was revealed every year 10-15 times more rarely than among urgent histotopographic biopsies. Thus it is advisable to use more widely intraoperative biopsies in euthyroid diseases with subsequent comparison to postoperative histological findings.

[Value of rapid biopsy in diagnosis of thyroid cancer]. / Znachenie ékspress-biopsii v diagnostike raka shchitovidnoi zhelezy.

Patients with thyroid carcinoma (ThC) in 67% are erroneously operated on with the diagnosis of nodular struma, this resulting in the recurrences in 30% and mortality and complications reaching 42%. Since 1977 the authors were performing rapid biopsies in all cases of the euthyroid struma and this resulted in the proper diagnosis in 98% of operated patients (48.6% before this). Short-term examination consists of stereo-morphology of the surgical material and the preparation by means of the deep frozen knife, of the histo-topographical section stained by Harris hematoxylin. Microcarcinomas are revealed by this method 30 times more frequently than in routinely treated sections. Short-term biopsy of the thyroid is as necessary as it is in the mammary gland lesions.

[Small thyroid cancer]. / Malyi rak shchitocidnoi zhelezy.

Thyroid carcinoma was detected in 1132 of 32500 patients undergoing surgery because of goiter. Small thyroid carcinoma (STC) was found in 245 patients (21.6%) and is divided into the following subgroups: 1) local (focal) microcarcinoma-a focus of a pronounced proliferation, dysplasia and fibrosis up to 1 mm in diameter; 2) invasive dwarf carcinoma of 5 to 15 mm in diameter which is not detected by clinical and instrumental methods; 3) intermediate STC, 1 to 5 mm in diameter. STC had a papillary-follicular structure in 88%, the rest were of a oncocytic and parafollicular structure.

[Preparation of histotopographic goiter sections and endometrium scrapes for relevant instant diagnosis]. / Prigotovlenie gistotopograficheskikh srezov dliia ékspress- diagnostiki patologii shchitovidnoi zhelezy éndometriia.

Histotopographic examinations of goiters and scrapes taken as a whole are valuable to the emergency pathomorphological diagnosis of thyroid and endometrial diseases essential particularly for the differentiation of adenomas and well differentiated carcinomas. The examination can be carried out only by treating fresh, non-fixed materials with a deeply cooling knife of the routine cold microtome (Model X). Harris' or Ballard's hematoxylins are best used as elective dyes. The whole instant diagnosis fully described in the paper lasts 8 to 12 minutes.

[Tumor necrosis factor in blood plasma and morphofunctional parameters of the hart in patients with chronic heart failure complicated by a course of ischemic heart disease. Dynamics under effect of treatment]. / Faktor nekroza opukholei v plazme krovi i morfofunktsional'nye parametry serdtsa u bol'nykh s khronicheskoi serdechnoi nedostatochnost'iu oslozhnivshei techenie ishemicheskoi bolezni serdtsa. Dinamika pod vliianiem lecheni.

AIM: To examine the course of chronic cardiac failure (CCF), morphofunctional parameters of the heart in correlation with activity of cytokine system; to evaluate effectiveness, tolerance and safety of combined CCF treatment including a cardioselective beta-adrenoblocker bisoprolol in patients with CCF FC I-IV having a high plasm level of tumor necrosis factor alpha (TNF-alpha). MATERIAL AND METHODS: The concentration of TNF-alpha in plasm of 40 patients with IHD complicated by CCF of FC I-IV and ejection fraction under 45% was measured at enzyme immunoassay. The treatment included combined therapy with diuretics, cardiac glycosides, peripheral vasodilators, ACE inhibitors, bisoprolol (group I) or no bisoprolol (group II). RESULTS: CCF patients with TNF-alpha had a significantly reduced IMT, more frequent episodes of CCF decompensation, longer CCF history, lower EF and threshold exercise intensity. In group I patients all these parameters improved. CONCLUSION: CCF patients with elevated plasm levels of TNF-alpha showed a severe course of the disease, low IMT, abnormal myocardial contraction and exercise tolerance. The addition of bisoprolol to combined treatment of the above patients improves their clinical condition, morphological parameters of the heart and exercise tolerance.

[Extraction of cholesterol from erythrocyte ghosts by phospholipids]. / Izvlechenie kholesterina iz tenei éritrotsitov fosfolipidami.

Removal of cholesterol from erythrocyte ghosts by liposomes and micellas of different phospholipids was investigated. It was shown that the positively charged micelles of polyunsaturated phospholipids are the most efficient for cholesterol removal from biological membranes.

LDL-apheresis on affine haemosorbents.

There were synthesized new hemosorbents for selective removal of LDL, based on macroporous silica, immobilised by heparine (sorbent 1) and chytozane-sulphate (sorbent 2). Experimental data showed satisfactory hemo- and biocompatibility of the new sorbents. No significant alterations in biochemical parameters were observed. Sorption cholesterol (Ch) capacity of the sorbents was equal and accounted for 15 mg of Ch per 1 g of the sorbent. Pilot clinical studies in familial hypercholesterolemia (FH) patients have shown the satisfactory hemo- and biocompatibility of the sorbent 1, and its significant Ch withdrawal. We conclude that the hemosorbents can be used in the FH patients treatment.

[Extraction of cholesterol from biological membranes with positively charged micelles of phosphatidylcholine]. / Ekstraktsiia kholesterina iz biologicheskikh membran polozhitel'no zariazhennymi mitsellami fosfatidilkholina.

The content of cholesterol in red cell and platelet membranes was lowered in rabbits with experimental atherosclerosis after intravenous injection of positively charged micelles of soybean phosphatidylcholine. That lowering was accompanied by a reduction in membrane microviscosity, rise of the activity of Na,K- and Ca-ATPases of red cells, and a decrease in the rate of the ADP- and collagen-induced platelet aggregation. Injection of phosphatidylcholine gave rise to an increase in the blood serum content of phospholipids and cholesterol in high density lipoprotein fractions, to a reduction in the content of triglycerides and the atherogenicity index, as well as to the lowering of the microviscosity of high density lipoproteins. The aortal area affected by atherosclerotic lesions was 2 times less in the group of animals given phosphatidylcholine.

[Content of cholesterol extracted from human skin surface--a possible discriminant of atherosclerosis?]. / Kolichestvo kholesterina, ékstragiruemogo s poverkhnosti kozhi cheloveka,- vozmozhnyi diskriminant ateroskleroza?

Basing on the data of Bouissou et al. on an increase of cholesterol content in the skin biopsies in atherosclerosis patients, the cholesterol amount was evaluated after superficial atraumatic rapid treatment of patients palm skin. The optimization of the extraction conditions was performed: the surface skin cholesterol value was constant for individual subjects (variability factor 5%). The range of this value was 1.5-6 Mg/cm2 and it was higher for atherosclerotic patients. No correlations with any conventional plasma lipoprotein parameters were observed. At the same time positive correlation (r = 0.54. p < 0.01) was observed with dyslipoprotein coefficient--(Ch+TG/alpha-Ch). Possible diagnostic significance of this skin cholesterol parameter is discussed.

[Secondary atherosclerotic immunodeficiency]. / Vtorichnyi ateroskleroticheskii immunodefitsit.

A study of peripheral and central mechanisms of secondary immunodeficiency in atherosclerotic patients demonstrated altered structural layout of lymphocyte membranes due to cholesterol accumulation in the cells. This was associated with increased membrane viscosity, suppressed mitotic activity and lower cap formation rate as well as changed intracellular calcium redistribution. Thymic ischemia due to occlusion or stenosis of thymus-supplying vessels was shown to promote age-related transformation of thymic tissue. Experimental simulation of thymic ischemia resulted in an abrupt drop of serum thymic factor in the blood of animals and the development of an immunodeficient condition.