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Background The efficacy and safety of wilate (human von Willebrand factor/coagulation factor VIII) in patients with von Willebrand disease (VWD) has been demonstrated in clinical trials. Here, we present real-world data on the use of wilate for the routine care of patients with VWD. Objectives The objectives of this observational, prospective, phase 4 study were to evaluate the safety, tolerability, and effectiveness of wilate in on-demand treatment of bleeding episodes (BEs), long-term prophylaxis, and surgical prophylaxis among patients with any type of VWD. Methods Patients were enrolled at 31 study centers in 11 countries and followed for up to 2 years. Safety endpoints included adverse drug reactions (ADRs) and drug tolerability. Effectiveness was assessed using annualized bleeding rates (ABRs) during prophylaxis and predefined criteria for the treatment of BEs and surgical prophylaxis. Results A total of 111 patients (76 [68%] female) including 41 (37%) children were treated with wilate. Twenty-five patients received prophylaxis, 29 on-demand treatment, and 62 surgical prophylaxis. Tolerability was rated by patients as “excellent” for 96.2% of 6,497 infusions. No unexpected ADRs or thrombotic events were reported. Median ABR during prophylaxis was 1.9. Effectiveness was assessed as “excellent” or “good” by patients and investigators for 100% of BEs treated on-demand, 98% (patient rating) and 99% (investigator rating) of breakthrough BEs, and 99% of surgical procedures (investigator rating). Conclusion wilate was safe, well tolerated, and effective for the prevention and treatment of bleeding in pediatric and adult VWD patients in a real-world setting.

Background and Aims: Modern stereotactic body radiotherapy (SBRT) techniques and systems that use online image guidance offer frameless radiotherapy of spinal tumors and the ability to control intrafraction motion during treatment. These systems allow precise alignment of the patient during the entire treatment session and react immediately to random changes in this alignment. Online tracking data provide information about intrafractional changes, and this information can be useful for designing treatment strategies even if online tracking is not being used. The present study evaluated spine motion during SBRT treatment to assess the risk of verifying patient alignment only prior to starting treatment. Methods: This study included 123 patients treated with spine SBRT. We analyzed different locations within the spine using system log files generated during treatment, which contain information about differences in the pretreatment reference spine positions by CT versus positions during SBRT treatment. The mean spine motion and intra/interfraction motion was evaluated. We defined and assessed the spine stability and spine significant shifts (SSHs) during treatment. Results: We analyzed 462 fractions. For the cervical (C) spine, the greatest shifts were in the anterior-posterior (AP) direction (2.48 mm) and in pitch rotation (1.75 deg). The thoracic (Th) spine showed the biggest shift in the AP direction (3.68 mm) and in roll rotation (1.66 deg). For the lumbar-sacral (LS) spine, the biggest shift was found for left-right (LR) translation (3.81 mm) and roll rotation (3.67 deg). No C spine case exceeded 1 mm/1 deg for interfraction variability, but 7 of 54 Th spine cases exceeded 1 mm interfraction variability for translations (maximum value, 2.5 mm in the AP direction). The interfraction variability for translations exceeded 1 mm in 2 of 24 LS spine cases (maximum value, 1.7 mm in the LR direction). Only 13% of cases had no SSHs. The mean times to SSH were 6.5±3.9 min, 8.1±5.9 min, and 8.8±7.1 min for the C, Th, and LS spine, respectively, and the mean recorded SSH values were 1.6±0.66, 1.43±0.33, and 1.46±0.47 mm/deg, respectively. Conclusion: Positional tracking during spine SBRT treatments revealed low mean translational and rotational shifts. Patient immobilization did not improve spine shifts compared with our results for the Th and LS spine without immobilization. For the most precise spine SBRT, we recommend checking the patient's position during treatment.

BACKGROUND Chronic wounds and their problematic healing is a widely discussed topic in all branches of medicine. In recent years, vacuum therapy appears to be a very successful non-invasive method supporting the healing of these wounds. The aim of this paper is to demonstrate the possibility of utilizing a vacuum system in the orofacial area where other conservative and surgical procedures have failed. CASES The case reports demonstrate the use of vacuum therapy in non-healing postoperative wounds in cancer patients. CONCLUSION Vacuum therapy has limited use in the orofacial area, but based on our experience, we can conclude that it has a very positive effect on the healing of chronic wounds. Thanks to this treatment, it was possible to reduce the frequency of dressings and significantly shorten the length of hospital stay. Despite these advantages, however, it is necessary to adhere to the conditions for the application of vacuum treatment.

BACKGROUND There is no universally accepted opinion on the use of granulocyte transfusions collected using apheresis (GTA) in neutropenic patients and severe infection. PATIENTS AND METHODS The efficacy and safety of GTAs transfused at a single center over 10 years were analyzed retrospectively. GTAs were harvested from voluntary unrelated donors after priming with methylprednisolone using continuous apheresis and hydroxyethylstarch as sedimentation agent. RESULTS 41 patients with neutropenia and hematologic malignancy (15 females and 26 males aged 22-69 (median 45.5)) were given a median 3.5 GTAs per patient (range: 1-17) containing a median 1.39×1010 granulocyte/GTA (range: 0.65-2.81). The indications for GTA use were soft tissue inflammation, sepsis, and pneumonia in 30, 22, and 14 cases, respectively. After GTA complete (30 patients: 73.2%) or partial (6 patients: 14.6%) healing of the infection was achieved. The success rate was 91.7% in soft tissue infections, 66.7% in invasive fungal infections, and 68% in sepsis. Septic shock (documented in 12 cases) was associated with a poor response (P

Hereditary thrombotic thrombocytopenic purpura (hTTP) is a rare thrombotic microangiopathy characterized by severe congenital ADAMTS13 deficiency and recurring acute episodes causing morbidity and premature death. Information on the annual incidence and severity of acute episodes in patients with hTTP is largely lacking. This study reports prospective data on 87 patients from the Hereditary TTP Registry (clinicaltrials.gov #NCT01257269) for survival, frequency, and severity of acute episodes from enrollment until December 2019. The 87 patients, followed up for a median of 4.2 years (range, 0.01-15 years), had a median age at overt disease onset and at clinical diagnosis of 4.6 years and 18 years (range, 0.0-70 years for both), respectively. Forty-three patients received regular plasma prophylaxis, whereas 22 did not, and treatment changed over time or was unknown in the remaining 22. Forty-three patients experienced 131 acute episodes, of which 91 (69%) occurred in patients receiving regular prophylaxis. This resulted in an annual incidence of acute episodes of 0.36 (95% confidence interval [CI], 0.29-0.44) with regular plasma treatment and of 0.41 (95% CI, 0.30-0.56) without regular plasma treatment. More than one-third of acute episodes (n = 51) were documented in children 40 years (1.18 [95% CI, 0.88-1.55] vs 0.14 [95% CI, 0.08-0.23]). The prophylactic plasma infusion regimens used were insufficient to prevent acute episodes in many patients. Such regimens are burdensome, and caregivers, patients, and their guardians are reluctant to start regular plasma infusions, from which children particularly would benefit.

Purpose We evaluated the efficiency and toxicity of stereotactic hypofractionated boost in combination with conventionally fractionated radiotherapy in the treatment of advanced floor of the mouth cancer. Methods Thirty-seven patients with advanced stage of the floor of the mouth cancer, histologically confirmed squamous cell carcinoma (p16 negative) ineligible for surgical treatment, were indicated for radiochemotherapy or hyperfractionated accelerated radiotherapy (HART). The radiotherapy protocol combined external beam radiotherapy (EBRT) and a stereotactic hypofractionated boost to the primary tumor. The dose delivered from EBRT was 70–72.5 Gy in 35/50 fractions. The hypofractionated boost followed with 10 Gy in two fractions. For the variables—tumor volume, stage and grade a multivariate analysis was performed to find the relationship between overall survival, local progression and metastasis. Toxicity was evaluated according to CTCAE scale version 4. Results After a median follow-up of 16 months, 23 patients (62%) achieved complete remission. The median time to local progression and metastasis was 7 months. Local control (LC) at 2 and 5-years was 70% and 62%, respectively. Progression-free survival (PFS) and overall survival (OS) were 57% and 49% at 2 years and 41% and 27% at 5 years, respectively. Statistical analysis revealed that larger tumors had worse overall survival and a greater chance of metastasis. Log-Rank GTV > 44 ccm (HR = 1.96; [95% CI (0.87; 4.38)]; p = 0.11). No boost-related severe acute toxicity was observed. Late osteonecrosis was observed in 3 patients (8%). Conclusion The combination of EBRT and stereotactic hypofractionated boost is safe and seems to be an effective option for dose escalation in patients with advanced floor of the mouth tumors who are ineligible for surgical treatment and require a non-invasive approach.

Background It remains unclear why only some patients form alloantibodies against foreign RBC antigens. Transfusion of red blood cell (RBC) products and pregnancy are the most relevant causes of immunization against RBC alloantigens. Here we investigated the relationship between RBC alloantibodies, Rh phenotype, and HLA phenotype among patients with multiple RBC alloantibodies Methods In a group of 124 multi-responders—including both pregnant women and transplant recipients—we analyzed the distribution of HLA-Class II variants in subgroups of multi-responders to RBC alloantigens according to their Rh status. Results As expected, the RhD-negative phenotype was overrepresented in our alloimmunized group (49.2%) compared to in the general population. Importantly, HLA-DRB1*15 carriers were significantly overrepresented among D-negative multi-responders compared to D-positive multi-responders (Pc = 0.045). Furthermore, the linked HLA-DRB1*13, HLA-DQB1*06, and HLA-DQA1*01 variants were more frequent in individuals with the DCCee phenotype than in other RhD-positive phenotypes. Conclusion Our present findings showed that RBC multispecific alloimmunization was associated with particular HLA-Class II variants based on Rh status.

Monoclonal antibodies represent a standard part in the treatment of oncologic patients, but their efficacy in multiple myeloma used to be unsatisfactory. Daratumumab monotherapy was approved by the American FDA in 2015, after unprecedented results were obtained in a heavily pre-treated group of patients. In 2016 daratumumab was approved in combination with lenalidomide and dexamethasone, or bortezomib and dexamethasone, for the treatment of myeloma patients who have received at least one prior therapy.The toxicity of the drug is low, and is dominated by infusion-related reactions in more or less half of patients. The development as well as the management of these sometimes urgent reactions is described in depth in this review. As multiple myeloma is characterized by the presence of paraprotein (monoclonal antibody) and CD38 is a ubiquitous antigen, several unexpected complications have been reported during the administration of the drug. In this review, we aim to describe and offer some solutions for the complications that may be encountered during daratumumab treatment, such as interference with serum protein electrophoresis and immunofixation assays that may confuse the assessment of the hematological response, interference with blood compatibility testing that may cause a delay in the delivery of compatible transfusions, and difficulties that may occur in flow cytometric analysis of minimal residual disease. Because of the high activity of daratumumab and its expected widespread use, clinicians should be aware of its side effects and their management. It is also very important to inform colleagues in clinical laboratories about the initiation of daratumumab treatment in particular patient.Key words: multiple myeloma - daratumumab - infusion related reaction - flow cytometry - transfusionThis work was supported by the Czech Ministry of Education, Youth and Sports (project no. IRP- 201550) and by the Czech Ministry of Health (15-29667A).The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Accepted: 22. 8. 2016Submitted: 12. 5. 2016.

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.

Congenital thrombotic thrombocytopenic purpura is an autosomal recessive inherited disease with a clinically heterogeneous course and an incompletely understood genotype-phenotype correlation. In 2006, the Hereditary TTP Registry started recruitment for a study which aimed to improve the understanding of this ultra-rare disease. The objective of this study is to present characteristics of the cohort until the end of 2017 and to explore the relationship between overt disease onset and ADAMTS13 activity with emphasis on the recurring ADAMTS13 c.4143_4144dupA mutation. Diagnosis of congenital thrombotic thrombocytopenic purpura was confirmed by severely deficient ADAMTS13 activity (≤10% of normal) in the absence of a functional inhibitor and the presence of ADAMTS13 mutations on both alleles. By the end of 2017, 123 confirmed patients had been enrolled from Europe (n=55), Asia (n=52, 90% from Japan), the Americas (n=14), and Africa (n=2). First recognized disease manifestation occurred from around birth up to the age of 70 years. Of the 98 different ADAMTS13 mutations detected, c.4143_4144dupA (exon 29; p.Glu1382Argfs*6) was the most frequent mutation, present on 60 of 246 alleles. We found a larger proportion of compound heterozygous than homozygous carriers of ADAMTS13 c.4143_4144dupA with overt disease onset at < 3 months of age (50% vs. 37%), despite the fact that ADAMTS13 activity was

BACKGROUND Hereditary thrombotic thrombocytopenic purpura, also called Upshaw-Schulman syndrome (USS), is a rare disease caused by genetic mutations in the ADAMTS13 gene, which severely decrease the activity of ADAMTS13, a metalloprotease that cleaves von Willebrand factor multimers (VWF). Genotypically identical patients can show great phenotypic diversity. OBJECTIVES Comparison of selected laboratory parameters and ADAMTS13 pharmacokinetics among patients with USS was performed. PATIENTS/METHODS Six patients with USS on prophylactic plasma therapy have been reviewed, retrospectively. Blood counts, lactate dehydrogenase (LDH), and ADAMTS13 activity at various time-points before and after different treatment cycles were evaluated. RESULTS ADAMTS13 recovery and pharmacokinetics were affected by treatment modality, and also reflected the patients' comorbidities and their current physiological and clinical condition. CONCLUSIONS Our present findings support a multifactorial contribution to treatment efficacy, and confirm the importance of adaptability and individualization of USS therapy. Therapeutic plasma exchange even in hereditary TTP is an option that can in some patients prolong intervals between plasma administration.

Background. Granulocyte apheresis is a safe and effective method for granulocyte collection. We present a five year experience (2006-2010) of the Blood Center, Faculty Hospital Ostrava, Czech Republic. Donor granulocyte transfusion is one treatment option for haemato-oncology patients with severe neutropenia complicated by bacterial/fungal infections unresponsive to standard antibiotic/antifungal treatment. In this study, we describe the experiences of the Blood Centre at the Faculty Hospital in Ostrava of granulocyte apheresis. Methods and Results. A total of 149 granulocyte units were collected for 33 patients from the Department of Haematooncology, University Hospital Olomouc, over a 5-year period (2006-2010). Information on donor selection, laboratory screening, mobilization regimen and granulocyte yield was obtained and analyzed. All mandatory screening tests for infection markers, AB0 and Rh and abnormal erythrocyte antibodies were performed. The cytomegalovirus status of the donors was not investigated. Steroids were the only mobilization regimen used, and were generally well accepted. The mean granulocyte yield was 12.6×10 9 /per transfusion unit (range 5.4-30.3). All granulocyte concentrates were irradiated and transfused according to AB0 Rh compatibility within 24 h after collection. Conclusion. Based on our experience, granulocytapheresis is a safe and effective method for obtaining granulocytes but the yield can be significantly influenced by other variables. From the recipients’ perspective, the use of donor granulocytes supports an effective therapeutic modality.

EuBIS, the European blood inspection system, is a collaborative effort of 27 representatives from 19 Member States (MS), including governmental institutions, blood establishments and competent authorities. EuBIS has been initiated by co-funding through the Public Health Programme (Directorate General Sanco) of the European Commission bringing together regulators (competent authorities) and manufacturers (blood establishments) to jointly develop criteria and standards aimed to assist: 1 blood establishments in need to optimize their quality system and self-inspection process and to prepare for regulatory inspections by competent authorities and 2 competent authorities, who wish to use the manual and training guide as a reference for the implementation process of legislative requirements related to regulatory inspections. These standards and criteria have been published in a EuBIS Manual and a EuBIS Training Guide. The structure of the EuBIS training guide follows critical aspects of quality management to be addressed to achieve good/best practice. For each of the quality related critical points, a criterion description, examples evidence and references to relevant international standards (GMP, PIC/S and CoE/EDQM CD-P-TS) and legislative requirements based on the European blood directives are given. Using ‘fishbone’ diagrams, quality indicators for risk assessment can be easily developed in a systematic fashion covering all relevant activities of a blood establishment. Chapter 3·5 refers to Blood collection, testing and processing with detailed subchapters on blood collection, processing and validation, labelling and release of blood and blood components. Additional quality requirements are described in Chapter 3·3 for Premises (Collection, Processing, Storage and Distribution), Chapter 3·4 Equipment and Chapter 3·8 non-conformance. Based on these standards and criteria, quality management systems have been nationally adapted to include reporting systems for risk analysis, prevention and acceptance based on deviations of processes, non conforming products and/or adverse events as well as non-compliances from audits and regulatory inspections.

Transfusion-related acute lung injury (TRALI) is a severe life-threatening complication of blood transfusion, characterized by acute lung injury developing within 2-6 h of transfusion. However, TRALI is difficult to diagnose, and the initial report or suspicion of TRALI depends on close collaboration between clinical departments and transfusion centres. A total of 17 adverse post-transfusion reactions were reported to the Blood Centre of the University Hospital Ostrava as suspected TRALI between 2005 and 2010. We report two cases of serious TRALI with different pathogenetic mechanisms.

Background and Objectives The Public Health Programme (Directorate General Sanco) gave impetus to the need for equivalent recognition of inspections of blood establishments among European Member States (MS) through the development and implementation of commonly accepted criteria and standards by funding the European Blood Inspection Project (EuBIS) (HYPERLINK “http://www.eubis-europe.eu”http://www.eubis-europe.eu). EuBIS is a collaborative effort of 27 representatives from 19 MS, including governmental institutions, blood establishments and competent authorities. It has cooperative working partnerships with five organisations, three projects, and is supported by the European Blood Alliance (EBA). Materials and Methods EuBIS is the first project that has brought together regulators (competent authorities) and manufacturers (blood establishments) to jointly develop criteria and standards aimed to assist: • blood establishments in need to optimise their quality system and self-inspection process • blood establishments to prepare for regulatory inspections by competent authorities • competent authorities, which wish to use the manuals and training guide as a reference for the implementation process of legislative requirements related to regulatory inspections. Results These standards and criteria have been finalised and published in a EuBIS Manual and Training Guide. The manual and training guide are available through the EuBIS homepage. In addition, EuBIS has developed educational material and to be used for seminars and courses. Based on regulatory standards applicable within the European Union (EU), the manual and training guide give references to commonly used internationals standards, such as GMP standards, the PIC/S guideline and the CoE (EDQM) guide. Thus, the EuBIS material should also be useful for those working in the field of quality management in other countries/areas world wide. In order to give practical assistance, the structure of the EuBIS guide follows critical aspects of quality management systems to be addressed in order to achieve good practice. For each of these critical points, a criterion description, examples evidence and references to relevant standards and legislative requirements are given. Conclusion The EuBIS manuals have been used by its partners organising national and international seminars, experimental and regulatory audits assisting the national implementation and further harmonisation of quality standards. In some member states elements of EuBIS have been adapted to national or local quality management systems. The implementation of good practice will improve the quality of blood and ensure that patients receiving blood transfusions are given safe blood.

Introduction: Congenital thrombotic thrombocytopenic purpura (cTTP) is an autosomal recessive disease characterised by acute episodes of thrombotic microangiopathy. cTTP has a clinically heterogeneous course. Currently there are more than 150 disease-causing ADAMTS13 gene mutations reported, though a genotype-phenotype correlation of this ultra-rare disease remains incompletely understood. Characterization of mutations of the ADAMTS13 gene will be useful for carrier detection and genetic counselling. In 2006, the Hereditary TTP Registry (ClinicalTrials.gov NCT01257269; www.ttpregistry.net) was initiated in order to achieve this objective by identifying possible triggers of acute episodes of TTP and to document individual clinical courses and treatment requirements. Methods: The Hereditary TTP Registry is an international cohort study. Individual data were analysed from 121 cTTP patients, who were enrolled between 2006 and the end of 2017. Diagnosis of cTTP was confirmed by a severely deficient ADAMTS13 activity Results: The cTTP cohort consists of 54 and 52 patients from Europe and Asia (90% from Japan), respectively, 13 patients from the Americas and 2 patients from Africa. For 69% of the patients regular treatment was reported, which consisted predominantly of fresh frozen plasma infusion (84%). More compound heterozygous (n=76) than homozygous mutation carriers (n=45) were included. The female to male ratio was 1:1, and the median age at first diagnosis was 25.8 years (range from 0 to 81.3 years). Prior arterial thromboembolic events (mainly myocardial infarction, stroke, TIA) were present in 27% of patients at enrolment and at the age of 50 years more than half of the patients had experienced at least one such event. In the 121 patients, we identified 98 different ADAMTS13 mutations (missense 57; nonsense 12; frameshift due to deletions or insertions 21; splice site 8). The most frequent observed mutations were c.4143_4144dupA (exon 29; p.Glu1382Argfs*6; n=58 alleles), c.3178C>T (exon 24; p.Arg1060Trp; n=13), and c.577C>T (exon 6; p.Arg193Trp; n=11). Nineteen mutations have not been previously reported (8 missense, 3 splice site, 2 nonsense, and 6 frameshift mutations). Mutations are located across the whole ADAMTS13 gene and affect all domains (there was no "hotspot"). We did not find a correlation of the mutations with clinical or laboratory features, except for ADAMTS13 c.4143_4144dupA. For this mutation, the age at initial diagnosis was younger (median age 4.1 years, range 1.8 - 52.1) in compound heterozygous compared to homozygous patients (median 23.3 years, range 0.7 - 47.5,) (p=0.040). Patient numbers were equally distributed (p = 0.51); 16 patients were compound heterozygotes and 21 were homozygotes. All patients were of Scandinavian or Central European ancestry. For the other assessed clinical parameters, no significant difference between compound heterozygotes and homozygotes was observed. Conclusion: At present, the Hereditary TTP Registry is the largest cohort on cTTP, to our knowledge. The large number of different mutations as well as confounding factors, including multi-ethnic and geographical factors in our internationally compiled patient cohort make unravelling the genotype-phenotype correlation in cTTP challenging. Yet, for c.4143_4144dupA, we can conclude that homozygous patients with ADAMTS13 c.4143_4144dupA are clinically less severely affected than compound heterozygotes due to the later onset of the disease in homozygotes, whereas the clinical characteristics are similar between compound heterozygotes and homozygotes. Why homozygous carriers of c.4143_4144dupA, despite having no ADAMTS13 activity in plasma, may have a rather late onset of acute TTP remains to be further elucidated. Disclosures Friedman: Shire: Membership on an entity's Board of Directors or advisory committees; CSL Behring: Membership on an entity's Board of Directors or advisory committees. Knöbl:Ablynx: Consultancy, Other: Member of Advisory Board. Schneppenheim:CSL Behring: Consultancy; SHIRE: Consultancy. Kremer Hovinga Strebel:Ablynx: Other: Advisory Board; SHIRE: Other: Advisory Board, Research Funding.

The European blood legislation has defined several key quality elements to achieve Good Manufacturing Practice (GMP) in the field of blood transfusion. During the recent years, the blood legislation is in the process of implementation throughout its member states. Following the Directive 2002/98/EC, Directive 2005/62/EC has given further requirements for quality-management systems to be fulfilled by blood establishments. In addition, GMP/Good Laboratory Practice (GLP) and ISO standards are used inter alia by blood establishments. In order to support the implementation of the blood legislation, the European Public Health Work Plan (2005/2007) has cofunded two projects, led by the German Red Cross and supported by the European Blood Alliance, delivering a common European Standard Operating Procedure (SOP) methodology (EU-Q-Blood-SOP) and criteria and standards for the inspection of blood establishments (EUBIS). The EU-SOP manual will assist blood establishments in preparing for the inspection of their services related to the implementation of quality relevant elements required by the EU Directive 2002/98/EC and its technical annexes. The standards and criteria for inspection of blood establishments will cross-reference existing quality standards to the directive requirements and define requirements for the structure of quality-management systems based on the directive 2002/98/EC and its technical annexes. Based on these requirements, inspection standards and criteria are developed to assist in the independent assessment of quality systems established by individual blood establishments. These assessments are done in relation to the requirements defined by the European Union legislation on blood, in order to safeguard the quality of blood and to achieve continuous improvement of its quality throughout Europe.

Inherited factor VII (FVII) deficiency is the most frequently observed rare bleeding disorder. The clinical symptoms are extremely variable, and patients may be asymptomatic or may present with life-threatening bleeding disorders. FVII activity (FVII: C) values of 10-15% are considered safe for maintenance of hemostasis, and replacement therapy is recommended for some types of surgical procedures. Twenty-three adult patients with FVII:C deficiency are followed-up in the Ostrava Haemophilia Treatment Centre. Eleven patients underwent a total of fourteen invasive procedures between 2008 and 2013. In terms of replacement therapy, nine patients received plasma-derived FVII and five patients received activated recombinant FVII. None of the patients had excessive blood loss during surgery, and there were no bleeding or other complications during post-operative treatment. In addition, there were no thromboembolic events related to the use of replacement therapy.

Background: Patients with chronic kidney disease (CKD) have significant disorders of bone and mineral metabolism. In addition, they can also develop other bone disorders including osteoporosis. This study evaluated the bone mineral density (BMD) of patients at the start of hemodialysis treatment as well as the relationship between BMD and possible risk factors or biochemical markers. Methods: The study was performed in 82 patients (28 females, 54 males). BMD was measured by dual-energy X-ray absorptiometry (DXA) at the lumbar spine and the proximal femur. Results: We found a high prevalence of 25-hydroxyvitamin D deficiency (96%; mean levels 30.0 ± 17.7 nmol/L) and a reduction of BMD in comparison with gender- and age-matched normal population values at the total hip (Z-score = -0.31 ± 1.11) and the femoral neck (Z-score = -0.48 ± 1.16), but not at the lumbar spine (Z-score = 0.68 ± 1.81). The prevalence of T-scores ≤ -2.5 SD in the group of patients over 50 years was 52.0% in females and 33.3% in males. BMD positively correlated: with male gender and calcium levels at all measured sites, with age at the lumbar spine and with weight or BMI at the proximal femur. Conclusion: CKD patients at the start of hemodialysis treatment had a high prevalence of low T-score values, corresponding to values for osteoporosis in the general population. BMD at the proximal femur was below the expected average for age and gender, but at the lumbar spine, BMD in hemodialysis patients was above average in persons without known CKD.

Examining an association between myocardial infarction (MI) and the Val/Ile polymorphism in the gene for CC chemokine receptor (CCR)2 at the position 64 (CCR2-V64I), 122 MI Czech patients and 277 unrelated control (C) subjects were genotyped by PCR-SSP. The frequency of the VI genotype of CCR2-V64I was increased in MI patients in comparison with the control population (P=0.03). Further analysis revealed that relationship between the VI genotype and MI is specific only for females and, strikingly, this genotype was associated to an early MI onset (before or at the age of 50 years). Females with the VI genotype were seven times more prone to suffer from MI before 50 years than those with the VV genotype (P

Aims. To examine biological materials (blood, urine, cerebrospinal fluid) of patients with suspected leptospirosis using real-time PCR for detecting the gene that codes the superficial LipL32 lipoprotein, and to evaluate the contribution of the real-time PCR method for the laboratory diagnosis of the acute form of leptospirosis. Material and Methods. During the monitored period of April 2010 – December 2011, a total of 340 biological materials samples were examined (177x blood plasma, 88x urine, 68x, cerebrospinal fluid, 6x bronchoalveolar lavage and 1x sputum) from 216 patients with suspected leptospirosis using real-time PCR LipL32 gene detection. Results. From the mentioned 216 patients suspected of leptospirosis, 8 patients were evaluated as being PCR LipL32 positive, from which 14 positive biological materials originated (9 x urine, 4x blood and 1x liquor). Conclusion. As demonstrated in the study, the real-time PCR method for detecting the gene for the superficial lipoprotein LipL32 is an appropriate, quick and reliable method for the diagnosis of the acute form of leptospirosis.

Summary Monocyte chemoattractant protein (MCP)-1 is the key chemokine in the process of atheroslerotic vascular inflammation. Examining already reported association between coronary artery disease (CAD) and the SNP A/G in the MCP-1 gene (position −2518), 139 Czech patients with CAD manifested as myocardial infarction (MI) and 359 unrelated healthy control (C) subjects were genotyped by PCR-SSP. Genotype and allele frequencies were not different in MI and C groups (allele G: MI, 20.5%; C, 23.8%, OR = 0.8, P > 0.05). No differences were detected when the patients were subdivided based on sex or the age of MI first occurrence. Further, no relationship was observed between circulating MCP-1 levels and carriage of the G allele. The data do not support a role for the MCP-1 −2518 single nucleotide polymorphism in susceptibility to CAD manifested by myocardial infarction.

Antiborrelia antibodies in Lyme borreliosis (LB) are mostly detected by enzyme immunoassay (EIA), confirmed by immunoblot (the "two-step system"). In indicated cases, direct evidence of Borrelia burgdorferi is obtained with the PCR method, electron microscopy and cultivation. The "one-step system" of testing for IgM and IgG antibodies in LB is economically preferably, but it requires an EIA kit with more than 90% sensitivity and specificity.90 blood samples were collected, 54 from patients with clinically defined LB and 36 samples from individuals free of LB. IgM and IgG antibodies against Borrelia burgdorferi were detected in parallel with five different EIA kits from various producers. The results were verified clinically in all cases, in disputable cases with additional immunoblot (BAG-Med), and analyzed statistically.Specificity and sensitivity were calculated from the measured values, and diagnostic efficiency was determined for each EIA kit. EIA kits for antiborrelia antibody assay with high specificity have low sensitivity and vice versa. In 9 samples from patients with clinical diagnoses (multiple sclerosis, Parkinson disease, epilepsy, rheumatoid arthritis) we found false positives in EIA and WB tests.The best results for a "one-step system" of examinations for antiborrelia antibodies were obtained with the Abbot and Euroimmun EIA kits in our set. A "two-step system" of serological examination could be composed from the basic IgM and IgG examination with a high sensitivity EIA kit (Viroimmun, Test-Line) followed with confirmation of positives by specific immunoblot.