PLATELET AGGREGATION ACTIVITY IN PATIENTS WITH ISCHEMIC STROKE, CARRIERS OF A GENETIC POLYMORPHISM OF SOME TOLL - LIKE RECEPTORS

2019 ◽  
pp. 15-23
Author(s):  
Ю.А. Крохалева ◽  
Н.Н. Страмбовская ◽  
Б.И. Кузник
Keyword(s):  
Ischemic Stroke ◽  
Genetic Polymorphism ◽  
Platelet Aggregation ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Toll Like Receptors ◽  
Blood Leukocytes ◽  
Primary Hemostasis ◽  
Subsequent Decrease ◽  
Spontaneous Platelet Aggregation

Цель - изучение генетического полиморфизма толл-подобных рецепторов при ишемическом инсульте и его влияние на агрегационную активность тромбоцитов. Методика. В исследовании приняли участие 115 больных ишемическим инсультом, в возрасте 55-65 лет и 94 сопоставимых по возрасту здоровых респондента. Материалом для молекулярно-генетического анализа служили образцы ДНК, выделенные из лейкоцитов периферической крови. Первичный гемостаз (спонтанная и индуцированная агрегатометрия) изучался на двухканальном лазерном анализаторе агрегации тромбоцитов АЛАТ-2. Результаты показали, что предикторами ишемического инсульта являются аллели: 753Arg TLR2, 249Pro TLR6, 1237C TLR9 и генотипы 753Arg/Arg TLR2, 299Asp/Asp TLR4, 249Pro/Pro TLR6 с отношением шансов от 1,49 до 3,35. Для развития инсульта имеет значение не только качество полиморфных маркеров, но и количество предикторных аллельных вариантов. У больных ишемическим инсультом в острейший период и в период формирования инфаркта мозга отмечается увеличение скорости и степени спонтанной агрегации тромбоцитов с последующим снижением к концу острого периода. При внесении АДФ (5 и 1,25 мкг/мл) и коллагена наблюдается снижение скорости и степени агрегации на 1-е и 10-е сут инсульта с увеличением таковых к 21-м сут. У больных - обладателей двух и более предиктивных аллелей полиморфизма толл-подобных рецепторов в геноме в динамике инсульта отмечаются сравнительно низкие показатели степени и скорости индуцированной агрегации тромбоцитов с увеличением радиуса агрегатов. Aim. To study genetic polymorphism of toll-like receptors in ischemic stroke and the polymorphism effect on the platelet aggregation ability. Methods. The study involved 115 patients with ischemic stroke aged 55-65 and 94 age-matched healthy respondents. DNA samples isolated from peripheral blood leukocytes were used for molecular genetic analysis of TLR2 (Arg753Gln), TLR3 (Phe412Leu), TLR4 (Asp299Gly), TLR6 (Ser249Pro), TLR9 (T - 1237 C). Primary hemostasis (spontaneous and induced aggregatometry) was studied using an ALAT-2 two-channel laser analyzer of platelet aggregation. Results. The study showed that predictors of ischemic stroke included the 753Arg TLR2, 249Pro TLR6, 1237C, and TLR9 alleles and the -753Arg/Arg TLR2, 299Asp/Asp TLR4, and 249Pro/Pro TLR6 genotypes with odds ratios from 1.49 to 3.35. Not only the quality of polymorphic markers but also the number of predictive allelic variants are important for the development of stroke. In patients with ischemic stroke in the acute phase and during the formation of cerebral infarction, increased rate and degree of spontaneous platelet aggregation were observed with a subsequent decrease by the end of acute period. Addition of ADP (5 and 1.25 mg/ml) and collagen decreased the rate and degree of platelet aggregation on days 1 and 10 of stroke with an increase in both by day 21. In patients carrying two or more predictive alleles of the toll-like receptor polymorphism in the genome, relatively low rate and degree values of induced platelet aggregation with increased radius of the aggregates were observed in the dynamics of stroke

LYMPHOCYTE-PLATELET ADHESION IN PATIENTS WITH ISCHEMIC STROKE — CARRIERS OF GENETIC POLYMORPHISM OF SOME TOLL-LIKE RECEPTORS

2018 ◽  
Author(s):  
Ю.А. Крохалева ◽  
Н.Н. Страмбовская ◽  
Б.И. Кузник
Keyword(s):  
Ischemic Stroke ◽  
Genetic Polymorphism ◽  
Platelet Adhesion ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Toll Like Receptors ◽  
Blood Leukocytes ◽  
Arithmetic Average ◽  
Aggregation Activity ◽  
Genetic Index

Цель исследования: изучить феномен лимфоцитарно-тромбоцитарной адгезии и агрегационную активность тромбоцитов (спонтанную и индуцированную) у больных ишемическим инсультом (ИИ) в динамике заболевания с учетом носительства генетического полиморфизма некоторых толл-подобных рецепторов (TLRs) и генетического индекса. Материалы и методы. Обследовано 115 больных ИИ и 94 относительно здоровых резидента. Лимфоцитарно-тромбоцитарный индекс оценивали как среднее арифметическое количество тромбоцитов, присоединившихся к одному лимфоциту. Материалом для молекулярно-генетического анализа TLRs служили образцы ДНК, выделенные из лейкоцитов периферической крови. Результаты. Выявлено, что предикторами ИИ являются аллели -753Arg TLR2, -249Pro TLR6, -1237C TLR9 и генотипы -753Arg/Arg TLR2, -299Asp/Asp TLR4, -249Pro/Pro TLR6 с относительным риском от 1,49 до 3,35. В крови больных ИИ многократно увеличено число лимфоцитарно-тромбоцитарных агрегатов (ЛТА). Заключение. Для развития инсульта имеет значение не только качество полиморфных маркеров, но и количество предикторных аллельных вариантов. Чем выше индивидуальный генетический индекс, тем больше в острейший период острого нарушения мозгового кровообращения в кровотоке образуется ЛТА; их количество снижается до контрольных значений к 21 дню заболевания. Aim: to study the phenomenon of lymphocyte-platelet adhesion (LTA) and platelet aggregation activity (spontaneous and induced) in patients with ischemic stroke (IS) over time the disease taking into account the carriage of genetic polymorphism of some Toll-like receptors (TLRs) and genetic index. Materials and methods. We examined 115 patients with IS and 94 relatively healthy residents. The lymphocyte-platelet index was estimated as the arithmetic average of platelets number that joined one lymphocyte. DNA samples isolated from peripheral blood leukocytes were the material for molecular genetic analysis of TLRs. Results. We revealed that alleles -753Arg TLR2, -249Pro TLR6, -1237C TLR9 and genotypes -753Arg/Arg TLR2, -299Asp/Asp TLR4, -249Pro/Pro TLR6 with a relative risk 1,49–3,35 were predictors of IS. The number of blood lymphocyte-platelet aggregates (LTA) increased manyfold in patients with IS. Conclusion. The qualities of polymorphic markers as well as the number of predictor allelic variants are important for stroke development. In the sharpest period of acute stroke the greater number of LTA in circulation is formed in patients with higher individual genetic index. The number of LTA decreased to control values by 21 day of the disease.

scholarly journals THE MOLECULAR-GENETIC ANALYSIS OF POLYMORPHISM OF IL-4, TNF-α GENES IN PATIENTS WITH LIVER CIRROSIS AND PORTAL HYPERTENSION

2017 ◽  
pp. 110-114
Author(s):  
A. G. Skuratov ◽  
A. N. Lyzikov ◽  
E. V. Voropayev ◽  
O. V. Osipkina
Keyword(s):  
Liver Cirrhosis ◽  
Portal Hypertension ◽  
Genetic Analysis ◽  
Restriction Analysis ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Pathological Process ◽  
Interleukin 4 ◽  
Blood Leukocytes ◽  
Tnf Α

Objective: to assess the association of polymorphism of the markers of interleukin-4 (IL-4) and tumor necrosis factor (TNF-α) in patients with liver cirrhosis with severe pathological process. Material and methods. The research material was DNA isolated from blood leukocytes of the patients. To identify SNP data, the PCR method was chosen with subsequent restriction analysis and electrophoretic detection by horizontal gel electrophoresis and amplification with visualization of the obtained results. Results. The performed molecular and genetic analysis of IL-4 and TNF-α gene polymorphisms made it possible to assume a relation between the frequency of polymorphic genotypes and alleles of the investigated genes and the severity of liver cirrhosis and portal hypertension, which can be used to diagnose and predict the severe course of the disease.

scholarly journals Molecular Genetic Analysis Of C-X-C Motif Chemokine Ligand 9 (CXCL9) As A Novel Biomarker In Atherosclerosis

2021 ◽  
Vol 3 (05) ◽  
pp. 24-30
Author(s):  
Dilbar T. Mirzarakhmetova ◽  
◽  
Yulduz Yusupbaevna Baltaeva ◽  
Keyword(s):  
Adaptive Immunity ◽  
Molecular Genetic ◽  
Gene Signature ◽  
Molecular Genetic Analysis ◽  
Blood Leukocytes ◽  
Innate And Adaptive Immunity ◽  
Chemokine Ligand ◽  
Ifn Γ ◽  
Artery Disease

For almost a century, many have considered lipids as the sine qua non of atherosclerosis. However, in 1856 Rudolf Virchow introduced a theory that inflammation is the driving force of atherogenesis. Recruitment of blood leukocytes to the injured vascular endothelium characterizes the initiation and progression of atherosclerosis and involves many inflammatory mediators, modulated by cells of both innate and adaptive immunity. The pro-inflammatory cytokine, interferon (IFN)-γ derived from T cells, is vital for both innate and adaptive immunity and is also expressed at high levels in atherosclerotic lesions. As such, IFN-γ plays a crucial role in the pathology of atherosclerosis through activation of signal transducer and activator of transcription STAT1.Our study indeed provides evidence that in HMECs STAT1 coordinates a platform for cross-talk between IFNγ and TLR4, and identifies a STAT1-dependent gene signature that reflects a pro-atherogenic state in coronary artery disease (CAD) and carotid atherosclerosis. Taken together, our data indicate that in the presence of appropriate stimuli, HMECs are highly responsive and consistently express Cxcl9. HMECs may therefore provide a better model for in vitro studies of atherosclerosis.

scholarly journals Frequency of Cytokine Gene Polymorphism in Full-Term Newborns with Hypoxic Events

2020 ◽  
Vol 5 (4) ◽  
pp. 21-27
Author(s):  
M. S. Panova ◽  
A. S. Panchenko ◽  
B. S. Pushkarev
Keyword(s):  
Gene Polymorphism ◽  
Brain Damage ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Full Term ◽  
Control Group ◽  
Cytokine Gene ◽  
Cytokine Gene Polymorphism ◽  
Blood Leukocytes ◽  
Intrauterine Hypoxia

Background. An infant brain damage is an extremely urgent problem, this pathology is difficult to prevent, and subsequently it manifests itself with a variety of neurological consequences. Various mechanisms are involved in neurodamage; cytokines, as well as genes that control their activity, are under a great concern today. However, there is little data about their role as predictors of the brain damage among children after hypoxia. Aim of the research. To identify the frequency of cytokine gene polymorphism: interleukin (IL)-1β(C-511T), IL-1β(C3953T), IL-4(C589T), IL-6(C174G), IL-10(C819T), IL-10(G1082A) among newborns with hypoxic events. Materials and methods. The study involved 128 full-term newborn patients with hypoxic events: the first group (n = 48) included newborns who experienced chronic intrauterine hypoxia (CVH), the second group (n = 80) included newborns born in asphyxiation. Control group (52) included babies born without asphyxia and not suffering from CVH. A retrospective analysis of case-records was carried out. The material for molecular genetic analysis was DNA samples isolated from umbilical cord blood leukocytes using DNA Express Blood reagents (Scientific and Production Company LITECH, Moscow). Results. Compared to the control group (p = 0.03) children born in asphyxia had their T allele IL-1β (C-511T) prevailed. The group of newborn who had CVH had their TT genotype (p = 0.04) and the T IL-1β allele (C-511T) (p = 0.01) prevailed compared to the control group. In the same study group while studying the polymorphism of the IL-1β gene, the T allele (p = 0.03) at the point C3953T prevailed, in contrast to the control group. Conclusion. Due to the fact that cytokines are part of a reaction cascade leading to the secondary brain damage, under the action of hypoxia, it was found that among newborns undergoing asphyxia and chronic intrauterine hypoxia the increased frequencies of carriage of IL-1β-511TT and IL-1β-3953TT genotypes, and IL-1β-511T and IL-1β-3953T alleles increase the risk of neurodamage.

scholarly journals EXPERT POSSIBILITIES OF FORENSIC CYTOLOGY INVESTIGATION IN FORENSIC EXAMINATION OF MATERIAL EVIDENCE

2020 ◽  
pp. 738-744
Author(s):  
R. Starovoitova ◽  
T. Lichman
Keyword(s):  
Genetic Analysis ◽  
Human Life ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Blood Leukocytes ◽  
Forensic Examination ◽  
History Of ◽  
Object Of Study ◽  
Material Evidence ◽  
Ultrasonic Water Bath

The article presents the history of the development and implementation of forensic cytological research methods in the practice of conducting forensic examinations of material evidence in the forensic examination bureau of Ukraine. The modern possibilities of forensic cytology methods for solving investigative issues in solving crimes against human life and health are shown in the article. Due to the fact, that the object of study of expert cytologists and expert geneticists is the same substance, namely, nuclear cells of different tissues and organs, blood leukocytes, epithelial cells of saliva, vagina, rectum, sperm, and cells of the outer root vagina hair – forensic cytological studies necessitate the preservation of nuclear cells for further genetic analysis. To this end, the staff of the Forensic Cytology Department of the Main Bureau of Forensic Examination developed a method for the preparation of cytological preparations using an ultrasonic water bath, which allows increasing the number of cellular elements extracted. At the same time, the specialists of the Odesa Regional Bureau of Forensic Examination introduced a technique for conducting molecular genetic analysis of nucleated cells in cytological preparations. Such techniques allow, even with a very small amount of biomaterial, to answer most of the questions posed by the investigator to the expert.

scholarly journals Efficacy of treatment of extensive drug resistant tuberculosis in patients with different genotypes in the biotransformation enzyme genes of CYP2B6 and NAT2

2020 ◽  
Vol 98 (6) ◽  
pp. 40-46
Author(s):  
M. M. Yunusbаevа ◽  
L. Ya. Borodinа ◽  
F. S. Bilаlov ◽  
R. A. Shаripov ◽  
T. R. Nаsibullin ◽  
...  
Keyword(s):  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Patient Treatment ◽  
Control Group ◽  
Drug Resistant ◽  
Blood Leukocytes ◽  
Drug Resistant Tuberculosis ◽  
Polymorphic Loci ◽  
Resistant Tuberculosis ◽  
Efficacy Of Treatment

The objective of the study: to evaluate the efficacy of treatment of extensive drug resistant tuberculosis (XDR-TB) in patients with different genotypes in the biotransformation system NAT2 (rs1041983, rs1799930, rs1799931, rs1801280) and CYP2B6 genes (rs3745274).Subjects and methods. The study involved patients undergoing in-patient treatment at Republican Clinical TB Dispensary in Ufa from 2016 to 2018. XDR TB group included 210 people; the control group included 343 healthy donors. Molecular genetic analysis was performed on DNA samples isolated from peripheral blood leukocytes. Genotyping of polymorphic loci was carried out by kompetitive allele specific PCR (KASP).Results. It was revealed that polymorphic loci rs1799931 of NAT2 gene and rs3745274 of CYP2B6 gene were associated with the risk of developing XDR TB. Regression analysis detected combinations of the predictor genotypes of rs1799931*G/A × rs3745274*G/T and rs1799931*G/G × rs37455274*(G/G+T/T), that significantly reduce efficacy of XDR TB treatment.

scholarly journals Two Novel Compound Heterozygous Pathogenic Variants in P2YR12 gene

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 1154-1154
Author(s):  
Arnaud Dupuis ◽  
Doris Böckelmann ◽  
Patricia Laeuffer ◽  
Katharina Neubauer ◽  
Christian Gachet ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Platelet Aggregation ◽  
Molecular Genetic ◽  
Index Patient ◽  
Molecular Genetic Analysis ◽  
Compound Heterozygous ◽  
Twin Brother ◽  
Pathogenic Variants ◽  
Fibrinogen Binding ◽  
Novel Variants

Abstract Introduction: P2Y12 receptor (P2Y12R) defect is a rare hemorrhagic disorder characterized by mild to moderate bleeding diathesis with easy bruising, mucosal bleedings, and excessive post-operative hemorrhage. It is due to mutations of the P2YR12 gene resulting in impairment of platelet responses to adenosine 5'-diphosphate (ADP) and subsequently to any other agonist as ADP is secreted from the dense granules during platelet activation. The P2Y12R plays a key role in platelet aggregation by amplifying the platelet activation process which is necessary to ensure appropriate primary hemostasis. The activation of the P2Y12R by ADP results in inhibition of adenylyl cyclase (AC) and in activation of phosphoinositide 3-kinase (PIK3), which amplifies and sustains the platelet aggregation response. To date only 12 pathogenic variants in the P2YR12 gene are listed in the Human Gene Mutation Database (HGMD®) (10 missense/nonsense mutations and 2 small deletions). Seven missense mutations are described in the literature in patients suffering from bleeding disorders of various severity. Autosomal recessive (homozygous and compound heterozygous pathogenic variants) as well as autosomal dominant (only one heterozygous pathogenic variant) inheritance has been reported. Patients and Methods: We investigated an 8 year old index patient who came to our outpatient clinic to investigate an increased bleeding tendency. He suffered from easy bruising and reported increased bleeding after tonsillectomy. His twin brother was also affected regarding bleeding disorder. Their father suffered sometimes from epistaxis, their mother and three older siblings were clinically unaffected. Platelet functions were assessed by light transmission aggregometry using citrated PRP. Membrane glycoprotein quantification, fibrinogen binding, von Willebrand factor (VWF) binding and the VASP assay were performed by flow cytometry. Molecular genetic analysis (Sanger sequencing) was performed for all exons including UTR regions and splice sites of the P2RY12-gene (NM_002788.4) for the twins and their parents. Results: The index patient and his brother showed normal values for platelet count, aPTT, INR, FXIII and VWF parameters. Both twins showed impaired platelet aggregation after stimulation with collagen, ADP and epinephrine. For both the index patient and his twin brother ADP (4-20 µM and 4-10 µM respectively) induced aggregation was markedly reduced and rapidly reversible. Flow cytometry revealed severely decreased fibrinogen binding for the index patient and his twin brother after stimulation with ADP (0.25-5.0 μmol/l) compared to control platelets. For CD42a/b (GP Ib/IX-complex), CD41 (GP IIb/IIIa-complex), ristocetin-induced VWF-binding, CD62- and CD63-expression, normal values were measured. The VASP assay revealed a strong defect with a PRI value of patient's platelets as low as 1.2% and 1% respectively (normal range > 69%). Molecular genetic analysis revealed two novel variants in the P2YR12 coding sequence (Tab.1). Conclusion: We have identified two novel variants in the P2YR12 gene with pathogenic predictions in a compound heterozygous state in twin brothers suffering from a chronic bleeding disorder. Flow cytometry analyses (fibrinogen binding) and VASP analyses are valuable tools to diagnose a platelet ADP-receptor defect. Table Table. Disclosures No relevant conflicts of interest to declare.

Effect of Dipyridamole on Spontaneous Platelet Aggregation in Whole Blood Decreases with the Time After Venepuncture: Evidence for the Role of ADP

1987 ◽  
Vol 58 (02) ◽  
pp. 744-748 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
J C Barbenel ◽  
C D Forbes
Keyword(s):  
Platelet Aggregation ◽  
Correlation Coefficient ◽  
Whole Blood ◽  
Blood Platelet ◽  
Inhibitory Effect ◽  
Time Interval ◽  
Adp Receptor ◽  
Spontaneous Platelet Aggregation

SummarySpontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.

Inhibition of Spontaneous Platelet Aggregation by Sulfinpyrazone

1979 ◽  
Vol 42 (02) ◽  
pp. 621-625 ◽  
Author(s):  
G G Nenci ◽  
G Agnelli ◽  
M Berrettini ◽  
P Parise ◽  
E Ballatori
Keyword(s):  
Platelet Aggregation ◽  
Crossover Study ◽  
Double Blind ◽  
Platelet Aggregability ◽  
Initiation Of Therapy ◽  
Spontaneous Platelet Aggregation ◽  
Blind Crossover Study ◽  
Double Blind Crossover Study

SummaryIn a randomized double-blind crossover study in 16 patients with enhanced in vitro spontaneous platelet aggregation, sulfinpyrazone proved to be effective in normalizing platelet aggregability within 4 days after initiation of therapy.

Molecular Characterization and Genetic Diversity Study in F3 Population of Rice

2013 ◽  
Vol 20 (1-2) ◽  
pp. 1-8
Author(s):  
MM Rahman ◽  
L Rahman ◽  
SN Begum ◽  
F Nur
Keyword(s):  
Genetic Distance ◽  
Gene Diversity ◽  
Random Amplified Polymorphic Dna ◽  
Molecular Genetic ◽  
Molecular Genetic Analysis ◽  
Polymorphic Loci ◽  
Rice Genotypes ◽  
High Level ◽  
Genetic Diversity Study ◽  
Diversity Study

Random Amplified Polymorphic DNA (RAPD) assay was initiated for molecular genetic analysis among 13 F3 rice lines and their parents. Four out of 15 decamer random primers were used to amplify genomic DNA and the primers yielded a total of 41 RAPD markers of which 37 were considered as polymorphic with a mean of 9.25 bands per primer. The percentage of polymorphic loci was 90.24. The highest percentage of polymorphic loci (14.63) and gene diversity (0.0714) was observed in 05-6 F3 line and the lowest polymorphic loci (0.00) and gene diversity (0.00) was found in 05-12 and 05-15 F3 lines. So, relatively high level of genetic variation was found in 05-6 F3 line and it was genetically more diverse compared to others. The average co-efficient of gene differentiation (GST) and gene flow (Nm) values across all the loci were 0.8689 and 0.0755, respectively. The UPGMA dendrogram based on the Nei’s genetic distance differentiated the rice genotypes into two main clusters: PNR-519, 05-19, 05-14, 05-12 and 05-17 grouped in cluster 1. On the other hand, Baradhan, 05-9, 05-13, 05-11, 05-5, 05-6, 05-1, 05-4, 05-15 and 05-25 were grouped in cluster 2. The highest genetic distance (0.586) was found between 05-4 and 05-17 F3 lines and they remain in different cluster.DOI: http://dx.doi.org/10.3329/pa.v20i1-2.16839 Progress. Agric. 20(1 & 2): 1 – 8, 2009

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