Non-viral systems for intracellular delivery of genome editing tools.

A hallmark of the last decades is an extensive development of genome editing systems and technologies propelling genetic engineering to the next level. Specific and efficient delivery of genome editing tools to target cells is one of the key elements of such technologies. Conventional vectors are not always suitable for this purpose due to a limited cargo volume, risks related to cancer and immune reactions, toxicity, a need for high-purity viral material and quality control, as well as a possibility of integration of the virus into the host genome leading to overexpression of the vector components and safety problems. Therefore, the search for novel approaches to delivering proteins and nucleic acids into cells is a relevant priority. This work reviews abiotic vectors and systems for delivering genome editing tools into target cells, including liposomes and solid lipid particles, other membrane-based vesicles, cell-penetrating peptides, micelles, dendrimers, carbon nanotubes, inorganic, polymer, metal and other nanoparticles. It considers advantages, drawbacks and preferred applications of such systems as well as suitability thereof for the delivery of genome editing systems. A particular emphasis is placed on metal-organic frameworks (MOFs) and their potential in the targeted intracellular delivery of proteins and polynucleotides. It has been concluded that further development of MOF-based vectors and technologies, as well as combining MOFs with other carriers can result in safe and efficient delivery systems, which would be able to circulate in the body for a long time while recognizing target cells and ensuring cell-specific delivery and release of intact cargoes and, thereby, improving the genome editing outcome.

[Geriatric status of patients over 70 years of age with myocardial infarction and the relationship with the severity of myocardial stress, fibrosis and angiogenesis.]

A prospective study of 92 patients over 70 years old with myocardial infarction was carried out. All patients were diagnosed and determined the severity of geriatric syndromes, including fragility, using specialized scales on the 5th day of hospitalization. The comorbidity was assessed with the calculation of the Charlson Comorbidity Index, levels of NT-proBNP, ST-2, FGF-21, VEGF in blood serum by enzyme immunoassay. In the studied cohort of patients, marked comorbidity, high prevalence of prefragility and fragility in combination with other geriatric syndromes were revealed. Patients over 70 years old with myocardial infarction and fragility showed significantly higher levels of myocardial stress and fibrosis, which is prognostically unfavorable sign for the given group of patients. NT-proBNP and ST-2 levels have significant correlation with major geriatric syndromes and could be the markers of fragility development among elderly and senile patients with myocardial infarction.

[Experience with 13-valent conjugated pneumococcal vaccine in HIV-infected patients].

AIM: Evaluate the effect of PCV13 vaccination on the composition of the microflora of the upper respiratory tract and the immune system in HIV-infected patients. MATERIALS AND METHODS: 100 patients with HIV-infection were included in the study. The patients underwent immunological examination and the collection of biomaterial from the posterior pharyngeal wall for microbiological examination. After obtaining the results of the examinations, PCV13 was intramuscularly administered. 7 days after the introduction of the vaccine, an assessment of adverse events was carried out, after 3 months, microbiological and immunological examinations were repeated. RESULTS: Immediately after the administration of PCV13, 5% of patients felt pain during the administration. Local reactions were reported in 6 patients. One participant showed a rise in temperature to 38.3C over 2 days. Before vaccination, 16 strains of S. pneumoniae were seeded in patients. 3 months after the administration of PCV13, pneumococcus was isolated in 8 patients. 3 months after immunization, the median level of populations and subpopulations of lymphocytes became higher than the pre-vaccination. DISCUSSION: Our results show high risk of pneumococcal infections in HIV-positive patients. A tendency towards a decrease in the level of S. pneumoniae carriage was revealed 3 months after the administration of PCV13. The high level of enterobacteria carriage in HIV-positive patients is noteworthy. There is a pronounced positive effect from the use of PCV13 in HIV-positive patients on cellular factors of the immune system. CONCLUSION: The use of PCV13 is a safe and effective method for the prevention of S. pneumoniae infections.

[The range and potential contribution of irreversibly adsorbed proteins on the surface of artificial implants]. / Spektr i potentsial'naia rol' neobratimo adsorbirovannykh belkov na poverkhnosti iskusstvennykh implantiruemykh materialov.

Protein adsorption is the first stage of the interaction between prosthetic materials with tissues of the body. They undergo conformational changes depending on the chemical composition and the nanotopography surface. Adsorbed proteins induce adhesion and alter the functional state of migrating cells. Plasma samples from patients were incubated with such matrices as titanium, polypropylene or polyester with fluoropolymer coating meshes. Bound peptides were analyzed by electrophoresis. Qualitative analysis of the peptides extracted from the gel was performed by chromatography-mass spectrometry. Quantitative analysis was performed by the MRM method. More than 60 proteins were identified on the analyzed surfaces. Quantitative analysis showed preferential adsorption of vitronectin, albumin, fibrinogen a-chain, C1ѕ component of the complement system. Vitronectin had the maximum relative protein content. Since biocompatibility of the analyzed materials varies considerably this variability may be attributed to conformational changes occurring with vitronectin during its irreversible adsorption.

[Atrial Fibrillation After Coronary Artery Bypass Grafting].

PURPOSE: to identify the role of factors of inflammation and myocardial ischemia in the development of atrial fibrillation (AF) in the early postoperative period after coronary artery bypass grafting (CABG). METHODS: We enrolled in this study 96 patients with ischemic heart disease who had undergone CABG. Patients were divided into 2 groups: (1) without postoperative AF (n=67, 80.6% men, mean age 57.9+/-7.3 years), (2) with early new-onset AF after CABG (n=29, 86% men, mean age 64.0+/-8.4 years). RESULTS: AF occurred on average 4.9+/-3.8 days after surgery. Concentrations of interleukin-10, C-reactive protein, fibrinogen, troponin were not significantly different between two groups both before and after CABG. At multivariate analysis the following parameters were related to development of AF in postoperative period: left atrial dimension >41 mm (odds ratio [OR] 3.6 (95% confidence interval [CI] 1.2-5.8, p=0.0002), interleukin-6 level >68 pg/ml (OR 3.2, 95%CI 1.8-4.7, p=0.009), interleukin-8 level >9.5 pg/ml (OR 2.9, 95% CI 1.4-6.7, p=0.009). CONCLUSION: Our research shows significant increase of interleukin-6 and interleukin-8 in patients with AF after CABG what confirms the role of inflammation factors in development of this complication.

[Assessment of the relationship of inflammation markers, adipokines and parameters of lipid metabolism in patient].

Objective - to assess the changes and the relationship of inflammation markers, adipokine levels and lipid metabolism in overweight and obesity, with signs of the metabolic syndrome. The study involved 62 women aged from 25 to 50 years. Group 1 included women (n=20) with a BMI<27.0, with no signs of metabolic syndrome, group 2 (n=20) - with a BMI>27.0, who were overweight or obese, with no signs of metabolic syndrome, 3 group (n=22) with a BMI>27.0, having overweight or obesity and the presence of any signs of metabolic syndrome. We determined relative and absolute lymphocyte counts and neutrophil granulocytes in the blood, lipid spectrum, blood serum levels of apolipoprotein A-1 (ApoA-1), and apolipoprotein B (apoB), leptin, adiponectin, interleukin-1ß (IL-1ß), interleukin 6 (IL-6), C-reactive protein (CRP). It was noted (p≤0.001) an increase of CRP in the blood of obese women, regardless of the presence of the metabolic syndrome: its concentration in groups 2 and 3 reached 6.69±3.03 and 7.30±3.08 mg/l, respectively, against 2.39±1.66 mg/l in group 1. Increasing BMI was accompanied by (p≤0.01) increase in erythrocyte sedimentation rate (8.1±5.0; 14.3±8.7; 13.9±8.5 mm per hour, respectively, in groups 1, 2 and 3). In women from groups 1 and 3 an increase in triglycerides, accompanied by a decrease in HDL (r=-0.44; p<0.025 and r=-0,57; p<0.001, respectively), and increase of LDL cholesterol (r=1.0; p<0.001) has been shown. The maximum number of positive correlations between indicators of lipid profile and inflammation markers was found in group 1, the minimum in group 3. This analysis showed the presence of chronic inflammation, which was accompanied by various disorders of lipid metabolism, even in the absence of clinical signs of pathology, the severity of which increased with increasing BMI and the development of metabolic syndrome.

[Prevention of inflammatory complications after osteoplastic operations in oral cavity].

It was established that every patient carried out through the oral osteoplastic operation needs immunoprophylaxis to avoid postoperative complications. It doesn't depend on the initial level of the immunity. Asa casual prophylaxis Licopid is recommented. But only one prophylaxis willbenot enough if the anamnesis has any chronical oralfacial disease. The great attention should be taken to the immunocorrection.

[Peripheral blood responses in elderly patients with some prevalent diseases].

The aim of the research was to study blood responses to inflammatory processes in patients of different ages. The subjects of the study were 31 healthy persons and 198 patients, 116 of whom had pneumonia and 82 had chronic pyelonephritis. The patients were divided into three age groups: 16 to 39 years, 40 to 59 years, and older than 60 years. The following parameters were calculated on the basis of peripheral blood leukocyte composition and ESR: leukocyte intoxication index (LII), lymphocyte index (LymI), leukocyte shift index (LSI), leukocyte index (LI), leukocyte to ESR ratio index (LESRRI), lymphocyte-granulocyte index (LGI), total index (TI), neutrophile to lymphocyte ratio index (NLymRI), neutrophile to monocyte ratio index (NMRI), and lymphocyte to monocyte ratio index (LMRI). The majority of the indexes (LII, LymI, LSI, LI, LESRRI, LGI, and LMRI) in acute inflammation processes and some of them (LESRRI, NLymRI, and LMRI) in chronic inflammation altered significantly in all the age groups, which demonstrated that compensatory and adaptive mechanisms remained preserved in the elderly. The were age-dependent differences in blood responses to inflammation: LymI, LI, and LGI were significantly lower in the elderly with chronic pyelonephritis, whereas in Groups 1 and 2 there were no changes; NMRI was decreased in Groups 1 and 2 in chronic pyelonephritis and did not differ from the elderly controls. In acute inflammation, there were no significant differences from the controls in NlymRI in Group 3, while this parameter in Groups 1 and 2 differ from that in the controls. Thus, hematological indices make it possible to assess the work of effectory mechanisms of the immune system, the degree of their compensation in patients belonging to different age groups.