OBJECTIVES: Degradation of coagulation proteins in frozen plasma may influence assay results. The aims of this study were to explore the changes in coagulation parameters in patient plasma and internal quality control (IQC) after different freezing and storage conditions during the short-term and long-term periods. METHODS: Platelet poor plasma was prepared from citrated peripheral blood collected from a group of healthy donors. The plasma was pooled, frozen and stored in a variety of freezing and storage conditions. The changes were monitored using routine coagulation assays, as well as factor VIII (FVIII) and protein S (PS) assays. RESULTS: Plasma stored in liquid nitrogen (LN 2 ) or in -80°C showed long-term stable values for routine tests for a period of over 12 months, and 6 months for FVIII. Interestingly, the activated partial thromboplastin time (aPTT) showed a temporary significant prolongation over the first two weeks. Plasma frozen and stored in -40°C is not viable for aPTT and FVIII testing, otherwise it can be used for other parameters for up to 4 months. PS showed a significant increase in all frozen samples. Freezing rate has a significant impact on plasma quality and the final storage temperature influences the long-term stability. CONCLUSION: The optimal storage conditions are ultra-low temperatures (LN 2 or -80°C) and the highest freezing rate possible. However, frozen plasma is not viable for IQC of aPTT during a period of two weeks after freezing. This study is unique in its conception as a practical guide for the handling of frozen plasma samples in modern laboratory settings.

• MeSH
• hemokoagulace MeSH
• hemostatika * MeSH
• krevní plazma * MeSH
• lidé MeSH
• parciální tromboplastinový čas MeSH
• vyšetření krevní srážlivosti MeSH
• zmrazování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Inherited protein S deficiency is a thrombophilic risk factor associated with venous thromboembolism. However, there is not much data on the impact of mutation position on thrombotic risk. OBJECTIVES: The aim of this study was to evaluate the risk of thrombosis due to mutations located in the sex hormone-binding globulin (SHBG)-like region as opposed to the rest of the protein. METHODS: Genetic analysis of PROS1 was performed in 76 patients with suspected inherited protein S deficiency, and the effect of missense mutations present in the SHBG region on thrombosis risk was analyzed by statistical methods. RESULTS: We found 30 unique mutations (13 of them novel), of which 17 were missense mutations, in 70 patients. Patients with missense mutations were then divided into 2 groups: the "SHBG-region" mutation group (27 patients) and the "non-SHBG" group (24 patients). The multivariable binary logistic regression analysis showed that mutation position in the SHBG region of protein S is an independent risk factor for thrombosis in deficient patients (OR, 5.17; 95% CI, 1.29-20.65; P = .02). The patients with a mutation in the SHBG-like region also developed a thrombotic event at a younger age compared to the "non-SHBG" group in the Kaplan-Meier analysis (median thrombosis-free survival of 33 vs 47 years, respectively; P = .018). CONCLUSION: Our findings show that a missense mutation located in the SHBG-like region may contribute to higher thrombotic risk rather than a missense mutation located elsewhere in the protein. However, as our cohort was relatively small, these findings should be taken with this limitation.

• Publikační typ
• časopisecké články MeSH

BACKGROUND: Extracorporeal membrane oxygenation (ECMO) support is often associated with bleeding complications caused by secondary or primary hemostasis pathology. However, there are limited data investigating primary hemostasis using Multiplate aggregometry with specific diagnostics tests for vWF (von Willebrand factor) deficiency. AIMS: The aim of this study was to find out whether short-term ECMO produces the pathology of primary hemostasis that is detected by Multiplate aggregometry and to investigate the pathology of vWF. METHODS: In this study, blood samples of 20 patients undergoing lung transplantations with short-term perioperative ECMO support were analyzed. The multimeric structure, the levels of von Willebrand factor antigen (vWF), ristocetin cofactor (RCo), collagen-binding protein (CB), and the results of multiple electrode aggregometry RISTO (ristocetin), ADP (adenosine diphosphate), ASPI (Aspirin®; arachidonic acid), and TRAP (thrombin receptor activating peptide) tests were compared to the samples obtained before and after ECMO support. RESULTS: The Multiplate ADP and RISTO tests showed the presence of significant pathology in primary hemostasis after surgery (p < 0.05), suggesting the presence of acquired platelet dysfunction. Although the RISTO tests suggest the presence of acquired vWF deficiency, laboratory tests for vWF antigen and RCo and CB tests showed an increase in this case. The multimeric structure of vWF did not show clinically significant deterioration. CONCLUSIONS: Multiple aggregometry ADP, ASPI, and TRAP tests seem to be able to detect primary hemostasis pathology (platelets aggregation and adhesion pathology) that is present during short-term perioperative ECMO support in lung transplantation procedures. Interestingly, RISTO tests seem to be more suitable for the diagnosis of platelet dysfunction than the diagnosis of acquired vWF deficiency in this situation.

• MeSH
• adenosindifosfát MeSH
• hemostáza MeSH
• lidé MeSH
• mimotělní membránová oxygenace * škodlivé účinky   metody   MeSH
• retrospektivní studie MeSH
• trombocytopatie * MeSH
• von Willebrandova nemoc * MeSH
• von Willebrandův faktor metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

BACKGROUND: Contemporary continuous-flow left ventricular assist devices (CF-LVADs) are associated with degradation of von Willebrand factor (vWF) high-molecular-weight multimers (HMWMs), a critical factor supporting platelet function. We hypothesized that the HeartMate 3 fully magnetically levitated LVAD, designed to reduce circulatory shear stress, favorably influences these hemostatic parameters. METHODS: Fifteen consecutive HeartMate 3 LVAD patients were compared with 11 consecutive HeartMate II controls. Serial plasma samples were collected pre-implant and on Days 2, 7, 30 and 45 post-operatively. Changes in vWF HMWMs were evaluated by 2 independent, study-blind hematologists and confirmed using densitometry-based computerized software. Ristocetin cofactor (RiCO) and vWF antigen (vWF Ag) were measured using standard protocols with enzyme-linked immunosorbent assay. RESULTS: HeartMate 3 patients and HeartMate II controls had a mean age of 67.3 ± 1.4 and 52.8 ± 2.5 years, respectively (INTERMACS Profiles 2 to 4 in 93.3% and 91%, respectively). HeartMate 3 group demonstrated a significantly greater preservation of HMWMs compared with the HeartMate II group, with the most prominent decrease occurring by Day 2 post-operatively and sustained through 45 days (71.94% vs 31.16%, p = 0.001). Laboratory values (normalized to baseline) for RiCO activity, vWF Ag and RiCO:vWF Ag ratio remained in the functional range with no statistically significant differences observed between groups. CONCLUSION: The HeartMate 3 LVAD is associated with enhanced hemocompatibility compared with the HeartMate II LVAD, as demonstrated by the improved preservation of vWF HMWMs, In contrast, effects on HMWM degradation appeared to be dissociated from functional attributes. Further confirmation of these findings in randomized clinical trials is warranted.

• MeSH
• ELISA MeSH
• lidé středního věku MeSH
• lidé MeSH
• mechanický stres MeSH
• podpůrné srdeční systémy MeSH
• srdeční selhání * MeSH
• von Willebrandova nemoc MeSH
• von Willebrandův faktor MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Práce ověřuje odolnost imunochemických souprav pro stanovení D-dimeru k působení interferujících složek lidské plazmy. Testované turbidimetrické soupravy Diagnostica Stago STA LIATEST DDi a její nová verze STA LIATEST DDi Plus byly použity pro stanovení koncentrace D-dimeru ve vzorcích s potvrzenou přítomností interferujících složek. Jako referenční metoda sloužila imunoanalýza na pevné fázi s promývacím krokem a fluorimetrickou detekcí VIDAS? D-Dimer ExclusionTM. Ve skupině 12 vzorků s interferujícími složkami testovaných soupravou STA LIATEST DDi se falešná pozitivita vyskytla 12krát, v paralelní skupině testované soupravou STA LIATEST DDi Plus se falešná pozitivita vyskytla 4krát. Byla stanovena korelace obou turbidimetrických LIA metod. V intervalu koncentrací D-dimeru 70–12 000 μg/l – FEU bylo dosaženo korelačního koeficientu R2 = 0,9944. Pomocí kontrolních vzorků byla stanovena preciznost měření soupravy STA LIATEST DDi Plus v sérii i v čase. V sérii bylo dosaženo variačního koeficientu na normální hladině 7,38 % a na patologické hladině 1,61 %. Při testováním mezi sériemi bylo dosaženo variačního koeficientu na normální hladině 7,78 % a na patologické hladině 2,16 %. Soupravu je možno použít pro detekci D-dimeru až do 20 000 μg/l – FEU. STA LIATEST DDi Plus má prokazatelně vyšší odolnost proti vlivu interferujících složek než stávající souprava STA LIATEST DDi.

Two immunoturbidimetric D-dimer assays were tested in terms of their resistance to plasma interferents. The Diagnostica Stago products STA LIATEST DDi and its new version STA LIATEST D-Di Plus were used to detect D-dimer concentrations in samples with confirmed presence of interferents. A solid phase immunoassay with fluorimetric detection VIDAS? D-Dimer ExclusionTM was used as the reference method. In the group of 12 samples containing interferents false positivity was detected in all 12 cases during testing with the STA LIATEST DDi kit. Results using the STA LIATEST DDi Plus kit were falsely positive in 4 cases. A correlation study on plasma samples with a range of 70–12000 μg/l – FEU was carried out. The coefficient of correlation between both assays was R2 = 0.9944. Precision studies of STA LIATEST DDi Plus were performed with the control samples on normal and pathological level. Intra-assay precision (expressed as coefficient of variation) was 7.38% on normal and 1.61% on pathological D-dimer levels. Inter-assay coefficient of variation values were 7.78% on the normal and 2.16% on the pathological D-dimer levels. The working range of the D-dimer concentration of STA LIATEST DDi Plus is up to 20000 μg/l–FEU. An increased resistance of the STA LIATEST DDi Plus to interferents was demonstrated.

• Klíčová slova
• matricový efekt, imunochemická diagnostika, interferent,
• MeSH
• dospělí MeSH
• falešně pozitivní reakce MeSH
• fibrin-fibrinogen - produkty degradace * analýza   MeSH
• imunoanalýza metody   MeSH
• krevní plazma * chemie   imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• nefelometrie a turbidimetrie MeSH
• plicní embolie krev   MeSH
• reagenční diagnostické soupravy * MeSH
• referenční standardy MeSH
• senioři MeSH
• statistika jako téma MeSH
• žilní trombóza krev   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• hodnotící studie MeSH
• srovnávací studie MeSH

The aim of the present work is the laboratory confirmation (or exclusion) of the von Willebrand disease (VWD) diagnosis in patients with a suspect bleeding disorder and determination of the disease type We used electrophoretic separation followed by an immunochemical chemiluminescent detection of the von Willebrand factor multimers. In the tested sample of 260 patients (in 2000– 2010), all the three primary categories of the VWD were detected. The VWD categories were of type 1 (71 %), of type 2A/B/E (24.7 %) and of type 3 (3.6 %). Our findings in the Czech population are close to the data of other research groups.

• MeSH
• adhezivita trombocytů MeSH
• dospělí MeSH
• faktor VIII MeSH
• hemofilie A * MeSH
• hemoragické poruchy * MeSH
• hemostáza MeSH
• koagulopatie MeSH
• lidé MeSH
• von Willebrandova nemoc MeSH
• von Willebrandův faktor * MeSH
• vyšetření krevní srážlivosti * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

Cíl studie: Alfa-2 sympatomimetika se stále častěji používají v premedikaci ke zlepšení perioperačního průběhu. Cílem práce bylo zhodnotit účinnost dexmedetomidinu v premedikaci před laparoskopickou cholecystektomií v porovnání s dosavadním postupem. Typ studie: Prospektivní, randomizovaná, dvojitě slepá klinická studie. Typ pracoviště: Fakultní nemocnice. Materiál a metoda: Po souhlasu etické komise a pacientů byl zaslepeně a randomizovaně podán 15 minut před úvodem do anestezie do m. deltoideus atropin 0,5 mg + jedna ze 4 premedikací: dexmedetomidin 1,0 µg . kg-1 + ketamin 0,5 mg . kg-1 + fentanyl 1,0 µg . kg-1 (skupina DexKeFNT), petidin 1,5 mg . kg-1 (skupina Dolsin), dexmedetomidin 1,0 µg . kg-1 + morfin 0,05 mg . kg-1 (DexMor) a morfin 0,05 mg . kg-1 (skupina Morfin). Úvod a vedení anestezie byly standardní. Byly zaznamenávány hodnoty vitálních funkcí, nežádoucí reakce pacienta a doba do podání prvního analgetika po operaci. Výsledky byly zhodnoceny Kruskal-Wallisovým, chí-kvadrát testem a Fisherovým testem. Výsledky: Do studie bylo zařazeno 61 pacientů indikovaných k laparoskopické cholecystektomii. Hypertenzní reakce na kapnoperitoneum byla nejčastější (68,7 %) ve skupině Dolsin (p < 0,001 oproti ostatním), ve skupině Dolsin byla i nejvyšší peroperační spotřeba fentanylu (165 ± 111 µg, p < 0,001 vs 13 ± 15 µg u DexMor, 25 ± 42 µg u Morfin i 31,5 ± 53,8 µg u DexKeFNT). Amnézii mělo 92,9 % v DexKeFNT (p < 0,001 oproti ostatním) a u obou skupin s dexmedetomidinem byla delší pooperační analgezie, p < 0,01 (DexMor 3,7 ± 1,0 h, Morfin 0,7 ± 1,1 h, Dolsin 0,9 ± 1,1 h, DexKeFNT 2,3 ± 1,0 h). Závěr: Dexmedetomidin v kombinaci s fentanylem, ketaminem a atropinem potlačí nežádoucí oběhové účinky kapnoperitonea, snižuje peroperační spotřebu fentanylu, prodlužuje dobu pooperační analgezie a nemá významné vedlejší účinky.

Objective: Alpha-2 sympatho-mimetic drugs are used to prevent adverse peri-operative reactions. The aim of this study was to compare premedication with dexmedetomidine against other currently used agents. Design: Prospective, randomised, double-blinded clinical study. Setting: University Hospital. Materials and methods. After ethic committee approval and patients' consent, premedication with atropine 0.5 mg and one of the following agents was administered in a randomised and blind fashion to the deltoid muscle 15 min before induction to anaesthesia: dexmedetomidine 1.0 µg . kg-1 + ketamine 0.5 mg . kg-1 + fentanyl 1.0 µg . kg-1 (group DexKeFNT), pethidine 1.5 mg . kg-1 (group Dolsin), dexmedetomidine 1.0 µg . kg-1 + morphine 0.05 mg . kg-1 (group DexMor) and morphine 0.05 mg . kg1 (group Morfin). Induction and maintenance of anaesthesia were standardised. Parameters of the vital functions, side-effects and time to request for the first dose of analgesics after surgery were examined and processed by Kruskal-Wallis, chi-squared and Fisher tests. Results: Total 61 patients scheduled for laparoscopic cholecystectomy were enrolled in the trial. Capnoperitoneum-induced hypertension and peri-operative fentanyl consumption were the most frequent in Dolsin group (68.7 %, p < 0.001 vs. all others; and 165 ± 111 µg vs. 13 ± 15 µg in DexMor, 25 ± 42 µg in Morfin and 31.5 ± 53.8 µg in DexKeFNT group, respectively). Amnesia was present in 92.9 % in DexKeFNT (p < 0.001 vs. all others) and longer postoperative analgesia was found in both the dexmedetomidine groups, p < 0.01 (DexMor 3.7 ± 1.0 h, Morfin 0.7 ± 1.1 h, Dolsin 0.9 ± 1.1 h, DexKeFNT 2.3 ± 1.0 h). Conclusion: The combination of dexmedetomidine, ketamine, fentanyl and atropine suppresses the adverse haemodynamic effects of the capnoperitoneum, decreases fentanyl consumption and prolongs postoperative analgesia without side-effects.

• Klíčová slova
• opioidy, alfa-2 sympatomimetika, operace,
• MeSH
• analgetika terapeutické užití   MeSH
• anestetika klasifikace   terapeutické užití   MeSH
• atropin terapeutické užití   MeSH
• cholecystektomie laparoskopická metody   využití   MeSH
• dexmedetomidin aplikace a dávkování   škodlivé účinky   terapeutické užití   MeSH
• financování organizované MeSH
• hemodynamika MeSH
• ketamin terapeutické užití   MeSH
• lidé MeSH
• opioidní analgetika terapeutické užití   MeSH
• premedikace anestezie metody   využití   MeSH
• prospektivní studie MeSH
• statistika jako téma MeSH
• sympatomimetika aplikace a dávkování   škodlivé účinky   terapeutické užití   MeSH
• výsledky a postupy - zhodnocení (zdravotní péče) MeSH
• Check Tag
• lidé MeSH