Association of HLA-DRB1 and HLA-DQB1 with red-blood-cell alloimmunization in the Czech population.

BACKGROUND AND OBJECTIVES: Alloimmune antibodies against red-blood-cell (RBC) antigens induced in susceptible individuals (responders) by transfusion, pregnancy or transplantation may have serious clinical consequences. The aim of this study was to investigate association of alloimmunization against selected RBC antigens with HLA-Class II. MATERIALS AND METHODS: A total of 230 responders (106 monoresponders and 124 multiresponders) were enrolled into the study. HLA-DRB1 and HLA-DQB1 variants were determined by PCR-SSO and their frequencies compared between the patients (patient subgroups) and 375 ethnically and regionally matched controls. RESULTS: Development of multiple RBC antibodies was associated with HLA-DRB1*15 and HLA-DQB1*06 allelic groups in the patients, with the relationship being particularly apparent in those with anti-C+D antibodies. Furthermore, DRB1*13 and DQB1*06 were more frequent in multiresponders with anti-E+c antibodies and DRB1*03 and DQB1*02 in those with anti-E+Cw. CONCLUSION: For the first time, we confirmed the association of HLA-DRB1*15 with RBC antibody multiresponder status and found HLA-Class II associations for three frequent RBC antibody combinations. Our data support the concept that HLA restriction plays an important role in the response to RBC alloantigens.

The rs1803274 polymorphism of the BCHE gene is associated with an increased risk of coronary in-stent restenosis.

BACKGROUND: We sought to identify gene polymorphisms that confer susceptibility to in-stent restenosis after coronary artery bare-metal stenting in a Central European population. METHODS: 160 controls without post-percutaneous coronary intervention in-stent restenosis were matched for age, sex, vessel diameter, and diabetes to 160 consecutive cases involving in-stent restenosis of the target lesion within 12 months. Using real time polymerase chain reaction and melting-curve analysis, we detected 13 single-nucleotide polymorphisms in 11 candidate genes - rs1803274 (BCHE gene), rs529038 (ROS1), rs1050450 (GPX1), rs1800849 (UCP3), rs17216473 (ALOX5AP), rs7412, rs429358 (ApoE), rs2228570 (VDR), rs7041, rs4588 (GC), rs1799986 (LRP1) and rs2228671 (LDLR). Multivariable logistic regression was used to test for associations. RESULTS: The rs1803274 polymorphism of BCHE was significantly associated with in-stent restenosis (OR 1.934; 95 % CI: 1.181-3.166; p = 0.009). No association was found with the other studied SNPs. CONCLUSIONS: The A allele of rs1803274 represents a risk factor for in-stent restenosis in Central European patients after percutaneous coronary intervention with bare-metal stent implantation.

[Optimization of antibodies examanination against thrombocytes in pregnant and postpartum women]. / Optimalizace vysetrení protilátek proti trombocytum u tehotných a zen po porodu.

OBJECTIVE: To optimize thrombocyte antibodies examination in pregnant and postpartum women. DESIGN: Retrospective analysis. SETTING: Blood Center, Faculty Hospital and Medical Faculty Ostrava University, Ostrava. METHODS: Analysis and our experience with introducing the new test for thrombocyte antibodies examination. CONCLUSION: The thrombocyte antibodies examination that was introduce in the period 2008-2009 and is the combination of basic and special testing, seems to be optimal for the neonatal thrombocytopenia detection.

Thrombotic thrombocytopenic purpura: incidence of congenital form of disease in north Moravia (region Moravia-Silesia).

Thrombotic thrombocytopenic purpura (TTP) was first described by Eli Moschcowitz in 1924. The pathophysiology of this disease is related to unusual, large multimers of von Willebrand factor in microcirculation, that are insufficiently cleaved by ADAMTS13 protease (a disintegrin-like and metalloprotease with thrombospondin type 1motif,13). Congenital TTP/Upshaw-Schulman syndrome is less frequent than acquired one TTP/HUS (haemolytic-ureamic syndrome). Short characteristic of patients with inherited form of TTP is reported as well as their clinical and laboratory features and management of treatment.

[Therapeutic plasma exchange in the treatment of the hereditary thrombotic thrombocytopenic purpura]. / Moznosti lécby plazmou u pacientu s hereditární formou trombotické trombocytopenické purpury.

BACKGROUND: Therapeutic plasma exchange is the treatment of choice for thrombotic thrombocytopenic purpura (TTP). METHODS AND RESULTS: Patients chronically treated with plasma exchange are frequently exposed to a large number of single plasma donors units, however successful clinical and laboratory improvement is generally achieved. Therapeutic plasma exchange significantly decreased mortality of this disease. Plasma treatment offers two possibilities--plasma infusion or plasma exchange and possibility of different plasma types. Cryoprecipitate-poor plasma was introduced as better form of FFP, however studies presented later did not confirm the therapeutic benefit. Quarantine FFP is widely used for patients with TTP in the Czech Republic; this type is tested frequently for negativity of human immunodeficiency virus, hepatitis B virus, hepatitis C virus and syphilis. Treatment with pathogen inactivated plasma (e.g., solvent detergent plasma, methylen-blue plasma, psoralen treated plasma or riboflavin treated plasma) is not frequently used in the Czech Republic. Authors present different plasma types and their experience with plasma treatment in seven patients with congenital form of TTP. CONCLUSIONS: In five patients therapeutic plasma exchange with quarantine fresh frozen plasma uas used. During each treatment 1.5 of plasma volume was exchanged. Two patients--teenagers were treated at the ex paediatric clinic with plasma infusion in regularly 2 weeks intervals.

High-resolution FISH on super-stretched flow-sorted plant chromosomes.

A novel high-resolution fluorescence in situ hybridisation (FISH) strategy, using super-stretched flow-sorted plant chromosomes as targets, is described. The technique that allows longitudinal extension of chromosomes of more than 100 times their original metaphase size is especially attractive for plant species with large chromosomes, whose pachytene chromosomes are generally too long and heterochromatin patterns too complex for FISH analysis. The protocol involves flow cytometric sorting of metaphase chromosomes, mild proteinase-K digestion of air-dried chromosomes on microscopic slides, followed by stretching with ethanol:acetic acid (3 : 1). Stretching ratios were assessed in a number of FISH experiments with super-stretched chromosomes from barley, wheat, rye and chickpea, hybridised with 45S and 5S ribosomal DNAs and the [GAA]n microsatellite, the [TTTAGGG]n telomeric repeat and a bacterial artificial chromosome (BAC) clone as probes. FISH signals on stretched chromosomes were brighter than those on the untreated control, resulting from better accessibility of the stretched chromatin and maximum observed sensitivity of 1 kbp. Spatial resolution of neighbouring loci was improved down to 70 kbp as compared to 5-10 Mbp after FISH on mitotic chromosomes, revealing details of adjacent DNA sequences hitherto not obtained with any other method. Stretched chromosomes are advantageous over extended DNA fibres from interphase nuclei as targets for FISH studies because they still retain chromosomal integrity. Although the method is confined to species for which chromosome flow sorting has been developed, it provides a unique system for controlling stretching degree of mitotic chromosomes and high-resolution bar-code FISH.