ПРОФИЛАКТИЧЕСКАЯ И КЛИНИЧЕСКАЯ МЕДИЦИНА

 

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Contents 2020-4

PREVENTIVE MEDICINE

  1. . Possibilities of vaccinal prevention ofinfectious diseases in adults
  2. I.G.Bakulin, K.V. Zhdanov, L.Yu. Drozdova, S.M. Kharit, R.S. Kozlov, V.V. Tyrenko, A.A. Zaitsev, G.R. Galstyan, I.A. Karpova, K.D. Ermolenko, L.N. Belousova, G.N. Minkina……………………….4

  3. Hygienic recommendations for the organization and conducting of sanitary and epidemiological investigation of the indoor ammonia
  4. N.O. Barnova, A.V. Meltser, I.Sh. Iakubova,N.T.Gonchar………20

  5. Application of artificial neural networks in preventive and clinical medicine (reliew)
  6. A.V. Golubkov, M.P. Gavrilova………………30

  7. Efficacy of bacteriophages against Pseudomonas aeruginosa biofilms
  8. B.I. Aslanov, L.P. Zueva, A.A. Dolgiy, S.D. Konev, T.A. Grishko ………………………40

  9. Hygienic assessment of pyrolytic incineration technologies for medical waste of the Russian Federation
  10. O.V. Mironenko, L.A. Soprun, O.K. Suvorova, S.N. Noskov, H.K. Magomedov, E.A. Fedorova, A.A. Stepanyan …………………………46

  11. Оn the issue of identification of сryptosporidium by polymerase chain reaction
  12. D.V. Azarov, A.E. Goncharov …………………57

  13. Methodological and practical principles of the formation of a multilevel rehabilitation system for children 0-4 years old as a disabilityprevention
  14. A.V. Kagan, E.V. Plotnikova, Y.V. Gorelik, N.V. Andrushchenko, A.S. Simakhodsky, L.D. Sevostyanova ……………………60

  15. Thesalmonella’s bacteriophages. Regulatory reliew
  16. O.E. Punchenko, E.А. Bereznitskaia, E.I. Ermolenko …………………68

CLINICAL MEDICINE

  1. Multiple primary tumors of the esophagus and lung. Diagnostics and surgical tactics
  2. B.N. Kotiv, K.G. Shostka, I.M. Kuznetsov, A.D. Kucherenko, K.V. Arutyunyan, K.S. Fedorov…….74

  3. The composition of the intestinal microbiota in COVID infection (reliew)
  4. V.P. Novikova, A.V. Polunina ………………………………81

BIOMEDICAL SCIENCES

  1. Reference intervals of p-selectin expression on platelets by flow cytometry
  2. A.I. Ermakov, L.B. Gaikovaya, T.V. Vavilova, O.V. Sirotkina …………………87

  3. Рlatelet vesicules – a potential marker of platelet concentrate quality
  4. V.V. Kishenko, O.V. Sirotkina, S.V. Sidorkevich, T.V. Vavilova …………………93

 

MEDICAL SCIENCE

PREVENTIVE MEDICINE

POSSIBILITIES OF VACCINAL PREVENTION OF INFECTIOUS DISEASES IN ADULTS
I.G.Bakulin1, K.V.Zhdanov2, L.Yu.Drozdova3, S.M.Kharit4,
R.S.Kozlov5, V.V.Tyrenko2, A.A.Zaitsev6, G.R.Galstyan7,
I.A.Karpova8, K.D.Ermolenko4, L.N.Belousova1, G.N.Minkina9
1North-Western State Medical University named after I.I. Mechnikov, Russia, 191015,Saint-Petersburg, Kirochnay street, 41.
2S. M. Kirov Military Medical Academy, Russia,194044, Saint-Petersburg, Academica Lebedeva street, 6.
3 National Medical Research Center for Therapy and Preventive Medicine, Russia, 101990, Moscow, Petroverigsky lane, 10, building 3.
4Research Institute of Childhood Infections of Federal Medical-Biological Agency, Russia, 197022, Saint- Petersburg, Professora Popova street, 9.
5 Smolensk State Medical University, Russia, 214019, Smolensk, Krupskayastreet, 28
6Main Military Clinical Hospital named after N.N. Burdenko, Russia 105229, Moscow, Hospital square, 3.
7National Medical Research Center for Endocrinology, Russia, 117036, Moscow, Dmitry Ulyanov street, 11.
8 Saint-Petersburg Territorial Diabetes Center "City Consultative and Diagnostic Center No. 1", Russia, 194354, Saint-Petersburg,Siqueirosa street, 10A.
9 Moscow State University of Medicine and Dentistry named after A.I. Evdokimov, Russia, 127473, Moscow, Delegatskaya street, 20/1.
Abstract
Article presents a detailed review on a current situation regarding the vaccinal prevention of infectious diseases in adults. On the basis of literature review, the justification of vaccination in adults is provided. The article contains the comparison of schedules of vaccinal prevention in adults performing in various countries worldwide. Particular attention is paid to vaccination for risk patients: patients with chronic diseases, immunodeficiency, military personnel, medical workers, pregnant women. According to analysisresults, it was provided the recommendations on the development of adult vaccination in Russia and expansion of available vaccines.
Key words: vaccination, national immunization schedule, life course immunization.

HYGIENIC RECOMMENDATIONS FOR THE ORGANIZATION AND CONDUCTING OF SANITARY AND EPIDEMIOLOGICAL INVESTIGATION OF THE INDOOR AMMONIA
N.О. Barnova, А.V. Meltser, I.Sh. Iakubova, N.T. Gonchar
North-Western State Medical University named after I.I. Mechnikov, Russia, 191015, Saint-Petersburg, Kirochnaya street, 41

Abstract
Introduction. The problem of indoor air pollution of ammonia of new housing and the appearance of an unpleasant odor required the organization and implementation of control and supervision measures. However, the algorithm for conducting a sanitary and epidemiological investigation and assigning measures to minimize the harmful effects of ammonia released from building and finishing materials has not been developed.
Aim of the study was to develop an algorithm for conducting a sanitary-epidemiological investigation and assigning measures to minimize the harmful effects of ammonia from building and finishing materials.
Materials and methods. Indoor air quality assessment of residential and public newly built buildings was carried out according to the results of field studies. Indoor air research was conducted at the Hygiene and Epidemiology Centre in Saint-Petersburg from 2012 to 2018.
Results. The algorithm of sanitary-epidemiological investigation of air pollution with ammonia in closed rooms of newly built buildings is proposed.
Key words: indoor air, ammonia, ammonia emission, sanitary and epidemiological investigation.

APPLICATION OF ARTIFICIAL NEURAL NETWORKS IN PREVENTIVE AND CLINICAL MEDICINE (RELIEW)
A.V. Golubkov1, M.P. Gavrilova2
1 Main Center for State Sanitary and Epidemiological Surveillance (Special Purpose), Russia, 105005, Moscow, Brigadirsky lane, 13, building 7
2 Saint-Petersburg Scientific-Research Institute for physical culture, Russia, 191040, Saint-Petersburg, Ligovsky prospect, 56, E

Abstract
The relevance of artificial neural networks application in the preventive and clinical medicine is defined by the national goals and strategic development targets set by the Decrees of the President of the Russian Federation, in particular, the achievement of "digital maturity" of key sectors of the economy and social sphere, including healthcare and education. The article presents an analysis of the problems and capabilities of the neural networks for predicting infectious and non-infectious diseases based on a retrospective epidemiological analysis. The development and selection of methods for health risk prediction by using several models based on the technique of artificial neural networks to obtain the most reliable results is proposed. The substantiation of the main directions of further development at the junction of the most relevant national projects - health care and digitalization, in order to increase the efficiency of socially oriented public management systems is presented.
Key words: artificial intelligence, artificial neural networks, prediction of health risk, preventive medicine, public health management.

EFFICACY OF BACTERIOPHAGES AGAINST PSEUDOMONAS AERUGINOSA BIOFILMS
B.I. Aslanov, L.P. Zueva, A.A. Dolgiy, S.D. Konev, T.A. Grishko
North-Western State Medical University named after I. I. Mechnikov, Russia, 191015, Saint-Petersburg, Kirochnaya Street, 41
Abstract
Introduction. It has been proved that many hospital strains, including Pseudomonas aeruginosa, actively use the ability to biofilm formation. The urgency of the problem of antibiotic resistance of hospital acquired P. aeruginosa infection generates the need for alternative antimicrobial agents. Bacteriophages could serve as aneffective antimicrobial. The use of phages to prevent the biofilm formation on invasive devices, in particular on vascular and urinary catheters, is of the great importance.
Aim of the study was to evaluate the efficacy of virulent bacteriophages for the prevention and destruction of biofilms formation by P. aeruginosa hospital strains.
Materials and methods. In were used 14 strains of P. aeruginosa with a high ability of biofilm formation, obtained from different hospitals. The efficacy of five virulent P. aeruginosa bacteriophages from the collection of the laboratory of molecular epidemiology and bacteriophage research was studied. The preventive and destructive activity of bacteriophages on P. aeruginosa biofilms was assessed by evaluation of biofilm optical density.
Results. In most cases of preventive (8 of 14 strains (57.1%)) and destructive (9 of 14 strains (64.3%)) application of bacteriophages on P. aeruginosa biofilms(both single and multiple exposition) the optical density sharply decreased at the beginning and continued to remain at a low level for the entire duration of the experiments for 12 hours.
Conclusion. Both single and multiple exposure of the bacteriophage to P. aeruginosa effectively prevents the biofilmformation. The application of the phages on the already formed biofilms effectively promotes their destruction. The study has shown that bacteriophages may be effective in the prevention and destruction of microbial biofilms.
Keywords: bacteriophage, P. aeruginosa, biofilm, antibiotic resistance, health-careassociated infection.

HYGIENICASSESSMENT OF PYROLYTIC INCINERATION TECHNOLOGIES FOR MEDICAL WASTE IN RUSSIAN FEDERATION
O.V. Mironenko1, 2, L.A. Soprun1, O.K.Suvorova2, S.N. Noskov2,
H.K.Magomedov2, E.A.Fedorova2, A. A.Stepanyan2
1Saint-Petersburg State University, Russia, 199034, Saint-Petersburg, University embankment, 7- 9
2North-Western State Medical University named after I.I. Mechnikov, Russia, 191015,
Saint-Petersburg, Kirochnayastreet, 41.

Abstract
Introduction. The volume of the human accumulated wastes assumes their decontamination only by combustion at specialized plants. Federal Service for Consumer Protection and Human Welfare and environmentalinstitutions should monitor the hygienic assessment to select and evaluate the technology optionsand laboratory control system organization in the construction of the centralized regional enterprises for thermal decontamination of medical waste.
Aim of the study was toprovide thehygienic assessments of the combustion technologies usedin Saint - Petersburg for the decontamination medical waste and to propose a laboratory control program when using these technologies.
Materials and methods.Pollutant emissions from plant combustingmedical wastes, their inventories by instrumental, calculation methods and modeling results.
Results.Based onthe laboratory control data of emissions from waste incinerators and the emission of core components according to a feasibility of priority chemicals for inclusion in the program laboratory control and wildlife management system of regular monitoring in the Russian Federation.
Key words: medical waste, incineration technologies, pyrolysis, polyvinyl chloride, dioxins, fluorine oxides, waste gas treatment, centralized medical waste disposal system.

ON THE ISSUE OF IDENTIFICATION OF CRYPTOSPORIDIUM BY РOLYMERASE CHAIN REACTION
D.V. Azarov, A.E. Goncharov
1North-Western State Medical University named after I.I Mechnikov, Russia, 191015, Saint-Petersburg, Kirochnaya street, 41
2Institute of experimental medicine, Russia, 197376, Saint-Petersburg, Academician Pavlov street, 12
Abstract
Introduction. The present study devoted to assessing the effectiveness of the Cryptosporidium spp. identificationbypolymerase chain reaction (PCR), proposed in the current guidelines for laboratory diagnosis of parasitic diseases and parasitological research.
Materials and methods. A literary search for the primary sources of the technique, bioinformatic analysis of the presented primer sequences and experimental polymerase chain reaction was performed.
Results. It was revealed that the sequences of the forward and reverse primers presented in the guidelines are found in two independent works, in sets of primers intended for the amplification of different regions of the Cryptosporidium genome. It was impossible to successfully reproduce the technique under experimental conditions.
Conclusions. The assessment results do not allow to consider the presented methodology to be effective, which should be taken into account in subsequent guidelines revisions.
Key words: cryptosporidium, cryptosporidiosis, parasitology, guidelines, polymerase chain reaction.

METHODOLOGICAL AND PRACTICAL PRINCIPLES OF THE FORMATION OF A MULTILEVEL REHABILITATION SYSTEM FOR CHILDREN 0-4 YEARS OLD AS A DISABILITYPREVENTION
A.V. Kagan1, E.V. Plotnikova2, Y.V. Gorelik2, N.V. Andrushchenko3,
A.S. Simakhodsky1, L.D. Sevostyanova4
1 Pavlov First Saint Petersburg State Medical University, Russia, 197022, Saint-Petersburg, Leo Tolstoy street, 6-8.
2 Saint-Petersburg Children 's City Multidisciplinary Clinical Center of High Medical Technologies, Russia, 198205, Saint-Petersburg, Avangardnaya street, 14.
3 North-Western State Medical University named after I.I. Mechnikov, Russia, 191015, Saint- Petersburg, Kirochnaya street,41.
4Center for Analysis and Forecast of Maternal and Child Health, Russia, 191023, Saint-Petersburg, Malaya Sadovaya street, 1.
Abstract
Introduction. It was considered the theoretical and practical issues of the used terminology in the projects formation of a multi-level rehabilitation system creation for children aged 0-4 years, as well as methodological and practical principles for goals achievement.
Aim of the study is to create a project for a multi-level rehabilitation system on the basis of  strict methodological principles and practical effectiveness, based on the available materials on the incidence and childhood disability in Saint- Petersburg, treatment results of the childrenaged0-4years in Saint-Petersburg Children 's City Multidisciplinary Clinical Center of High Medical Technologies.
Materials and methods. It was studied the reporting forms of Rosstat No. 19 of children’s clinics and reporting forms of medical and social examination of Rosstat 7-D during 2008-2017, form No. 14 "Information on the activities of medical organization units providing inpatient medical care", registration form No. 030/y "Outpatient observation control card". The present study includes materials from a meeting of the Coordinating Council under the President of the Russian Federation from November 15, 2016, instructions from the Government to the Ministries and Departments from December 21, 2016. Statistics data were obtained from the Petrostat and the Center for Analysis and Forecast of Maternal and Child Health. Statistical processing was carried out using methods of variation statistics (Pearson criterion, angular transformation criterion).
Results. The study was divided into two stages: the first is a traditional assessment of the formation of childhood disability of0-4years old with a description of the diseases structure by which children are recognized as disabled. Herewith, the number of disabled people of the studied age is growing, which does not correlate with the spectrum of pathological lesions in children of the 111health group, as during the treatment there is a dissociation of children in other health groups, and since 2018 only groups of 0-17 years is distinguished by the incidence. In the second stage, it is used a completely different methodological approach, based on the high incidence of children born with extremely low and very low body weight resulted in a high level of disability. An example of the of early specialized multi-stage rehabilitation (habilitation) formation is shown.
Conclusion. The issues of theoretical justification of the methodological approach to the development of a new internal system for the provision of medical and rehabilitation assistance to newborns in a particular children's medical institution are discussed. Practical implementation resulted in the creation of new structures, medical equipment, staff training, and the development of an automated registration and control system. The authors plan to publish the reorganization results in 2022.
Key words: methodology, practical appropriateness, disability structure, stages of medical and rehabilitation assistance.

THE SALMONELLA’S BACTERIOPHAGES.REGULATORY RELIEW
О.Е. Punchenko1,2, Е.А. Bereznitskaia1, E.I. Ermolenko2
1North-Western State Medical University named after I.I. Mechnikov, Russian Federation, 191015, Saint-Petersburg, Kirochnaya Street, 41;
2Institute of experimental medicine, Russian Federation, 197376, Saint-Petersburg, acad. Pavlov str., 12
Abstract
Introduction. Antibiotic resistance of bacteria is one of the major challenges of medicine. Virulent bacteriophage scan be used as an alternative to treatment and prevention. This is especially true for intestinal infections, which are accompanied by the development of dysbiosis.
The aim of the study - analyze the current regulatory documents in the field of health for the use of bacteriophages for infections caused by salmonella.
Results. Salmonella is one of the first places among the causative agents of intestinal infections and foodborne poisoning. In addition to typhoid, paratyphoid, gastroenteritis, they are the cause of Health care associated infections in hospitals. After the disease, the formation of salmonella carriage is possible. Salmonella is extremely resistant to adverse abiotic factors, both physical and chemical; their resistance is increase with the protein. Therefore, Salmonella can survive for a long time in all types of food and pure water. Numerous works describe the resistance of salmonella to chemotherapy and antibiotics, so therapy becomes useless. At the same time, regulatory documents have been approved and are in force in our country that regulate the use of salmonella bacteriophages not only for treatment, but also for emergency prophylaxis of infection.
Conclusion. The use of salmonella bacteriophages is fixed in the law in our country.
Key words: bacteriophage, typhoid fever, salmonella infection.

CLINICAL MEDICINE

MULTIPLE PRIMARYTUMORSOFTHEESOPHAGUSANDLUNG.DIAGNOSTICS AND SURGICAL TACTICS
B.N. Kotiv1, K.G.Shostka2, I.M. Kuznetsov1,A.D. Kucherenko1,
 K.V. Arutyunyan2, K.S.Fedorov1
1 S. M. Kirov Military Medical Academy, Russia, 194044, Saint-Petersburg, AcademicikaLebedev street, 6.
2Leningrad Regional Oncology Dispensary, Russia, 191014, Saint- Petersburg, Liteiny prospect, 37

Abstract
Introduction.The increased number of cancer patients of various localization remains a worldwide tendency. The incidence of polyneoplasia is proportionally increasing.
Aim of the study was to improve the diagnosis and treatment results of patients with multiple primary cancer of the lung and esophagus.
Materials and methods. During 2004-2014, it was observed 17 patients with a preliminary diagnosis of multiple primary cancer of the esophagus and lung. The male to female ratio was 15/2. The average age is 59 ± 6 years. After evaluating the functional operability of the patients, surgical treatment was performed. During simultaneous operations for esophageal and lung cancer, standard esophageal resection (17) with 2F lymph node dissection (abdominal and bilateral mediastinal) and lung resection (lobectomy - 12, pneumonectomy - 3, and wedge resection - 2) were performed.
Results. According to the drugs histological study after surgery, multiple primary cancer of the esophagus and lung was confirmed in 12 patients, in 5 cases esophageal cancer with a single metastasis to the lung was detected. In the postoperative period, complications were noted in 6 cases (27%), in one case a lethal outcome was found. Patients with stage 4 esophageal cancer with lung metastasis (n = 5) lived for less than one year. Patients with multiple primary cancer of the esophagus and lung (n = 12) had a more favorable prognosis: 10 people lived for more than one year, 5 people lived over three years, and 5-year survival was stated in 3 patients.
Conclusions. Differential diagnosis of multiple primary cancer of the esophagus and lung and metastatic lesions is possible only after histological verification (in a significant part of patients (47%) only intraoperatively). In case of synchronous cancer of the esophagus and lung, it is justified to perform simultaneous operations, providing that the staging is correct and the functional operability is assessed correctly. For synchronous cancer of the esophagus and left lung, a two-stage operation is more preferable.
Keywords: multiple primary cancer, synchronous cancer of the esophagus and lung, assessment of functional operability, simultaneous surgery.

THE COMPOSITION OF THE INTESTINAL MICROBIOTA IN COVID INFECTION (RELIEW)
V.P. Novikova, A.V. Polunina
Saint-Petersburg State Pediatric Medical University, Russia, 194100, Saint-Petersburg, Litovskaystreet, 2,

Abstract
The article provides an overview of foreign literature on the intestinal microbiota in coronavirus infection. Due to the fact that in patients with coronavirus infection, from 20% to 50% of cases are affected by the intestines - nausea, vomiting and diarrhea and SARS-CoV-2 RNA can be detected in stool samples, it is interesting to consider the relationship between the virus and the intestinal microbiota, the pathogenesis of this virus. A number of researchers have found significant changes in the fecal mycobiome in patients with coronavirus infection compared to the control group, an increase in its diversity.Others, on the contrary, report a decrease in microbial diversity with coronavirus infection. Clarification is required of the influence of the gut microbiome on the pathogenesis of coronavirus infection and the clinical course of the disease.
Keywords: coronavirus infection, microbiome, intestinal microbiota, SARS-CoV-2.

BIOMEDICAL SCIENCES

REFERENCEINTERVALSOF P-SELECTIN EXPRESSION ON PLATELETS BY FLOW CYTOMETRY
A.I. Ermakov1,2, L.B. Gaikovaya1,2, T.V. Vavilova2, O.V. Sirotkina2,3,4
1North-Western State Medical University named after I.I. Mechnikov, Russia, 191015, Saint-Petersburg, Kirochnayastreet, 41
2Almazov National Medical Research Centre, Russia, 197341, Saint-Petersburg, Akkuratovastreet, 2
3Petersburg Nuclear Physics Institute named by B.P. Konstantinov of National Research Centre «Kurchatov Institute», Russia, 188350, Saint-Petersburg, Gatchina, OrlovaRostcha,1
4Pavlov First Saint-Petersburg State Medical University, 197101, Russia, Saint-Petersburg, Leo Tolstoystreet, 6/8
Abstract
Introduction.Currently,the evaluation of the functional activity of platelets using flow cytometry could be crucial not only in such clinical situations as the diagnostics of hereditary thrombocytopathy, but also in monitoring of the reaction of platelets to antiplatelet drugs.However, the main problem in using this method as a diagnostic test is the standardization of sample preparation, optimization and validation of the test kit and the determination of reference intervals.
The aim of this study was to determine the reference intervals for the expression of P-selectin on the platelet surface by flow cytometry among the healthy adult population of Saint-Petersburg.
Materials and methods.To achieve these goals, a reference group was determined including an equal number of men and women of different age groups from 18 to 65 years. A monoclonal antibody to alinearly associated megakaryocyte-platelet marker (CD61), fluorescently-labeled with fluorescein isothiocyanate (FITC) was selected for platelet identification in functional activity of platelets analysis by flow cytometry. As a platelet activation marker before and after induction, the expression of P-selectin on the platelet surface was measured using a monoclonal antibody CD62P fluorescently labeled with phycoerythrin (PE). ADP solutionwasselectedas the platelet activating inducer in a final concentration of 20 μmol/l.
Results.According to study results, it was found that healthy women have a higher content of the relative number of platelets positive for CD62P (P-selectin) without activation, in comparison with the same indicator for healthy men – 0,70% [0,63; 1,42] and 0,23% [0,10; 0,54], respectively, p = 0,01; in the presence of an inducer of platelet activation, these differences were not revealed. In this regard, the establishedreference intervals for the expression of P-selectin on the surface of platelets outside of activation took into account the gender of the examined persons. Also, for the expression of P-selectin on the surface of stimulated platelets, the coefficient of intraindividual biological variation (CVi) was determined, which was 25% and the coefficient of interindividual biological variation (CVg), equal to 47%.
Conclusion. The standardization of the analysis of functional platelet activity using flow cytometry resulted in the creation of a stable, relatively simple and convenient laboratory test for assessing the expression of P-selectin with acceptable variability within the assay. In addition, this is the first study to determine the reference intervals in a relatively large cohort of the healthy population of Saint-Petersburg depending on gender.
Key words:flow cytometry, P-selectin, platelets, reference intervals.

PLATELET VESICULES – A POTENTIAL MARKER OF PLATELET CONCENTRATE QUALITY
V.V. Kishenko 1, O.V. Sirotkina 1,2,3, S.V. Sidorkevich 1, T.V. Vavilova 1
1Almazov National Medical Research Centre, 197341, Russia, Saint-Petersburg, Akkuratova street, 2
2Petersburg Nuclear Physics Institute named by B.P. Konstantinov of National Research Centre «Kurchatov Institute», Russia, 188350, Saint- Petersburg, Gatchina, Orlova Rostcha,1
3Pavlov First Saint-Petersburg State Medical University, Russia, 197101, Saint-Petersburg, Leo Tolstoy street, 6/8
Abstract
Introduction. Platelet concentrate has a wide clinical use, primarily in hemorrhagic syndrome in patients with thrombocytopenia or thrombocytopathy. Laboratory quality control of platelet concentrate involves assessing the number of platelets, leukocytes and pH value, but these parameters do not characterize the functional state of platelets in platelet concentrate. Processes of platelet activation and vesiculation negatively affect the functional state of platelets and could accelerate the processes of cell apoptosis in platelet concentrate.
The aim of this study was to determine the number of platelet vesicles by flow cytometry as a possible marker for assessing the quality of platelet concentrate during storage.
Materials and methods. The study used platelet concentrate obtained by the method of automatic thrombopheresis. Sampling of 1.6 ml was carried out from the container under sterile conditions on the 2nd and 7th days of storage. The platelet concentrate fractions after differential centrifugation were analyzed on a Cytoflex flow cytometer (Beckman Coulter) using anti-CD41-FITC and anti-CD62P-PE. The processes of platelet activation and vesiculation during storage of platelet concentrate were visualized using low voltage scanning electron microscopy.
Results. On the 7th day of storage, the relative content of platelets in platelet concentrate, carrying P-selectin on their surface, increased 3-10 times indicating cell activation. The number of platelet membrane vesicles and exosomes increased on the 7th day of platelet concentrate storage compared to the 2nd day of storage: 11.78 [9.78-67.93] events/μl versus 3.21 [2.88-5.44] events/μl and 4.35 [3.18-5.87] events/μl versus 0.68 [0.32-1.46] events/μl, respectively (p<0.0078). At the same time, using electron microscopy, the activation of platelets and the separation of vesicles from the terminal part of the pseudopodia were proved on the 7th day of platelet concentrate storage.
Conclusion. The most promising method for assessing the quality of platelet concentrate according to the results of the presented study is the determination of the number of platelet vesicles using flow cytometry.
Key words: platelets, platelet concentrate, vesicles, flow cytometry.

 

 

 

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