The purpose of this study was to characterize the association between anti-prothrombin antibodies and the lupus anticoagulant (LA) in order to elucidate the antigenic site of the LA. Plasma from 8 patients with the LA had evidence of anti-prothrombin antibodies on prothrombin crossed immunoelectrophoresis as characterized by material moving slower in the first dimension of electrophoresis than normal prothrombin, i.e., a trailing shoulder. Four of 5 LA patients with a prolonged prothrombin time demonstrated the most pronounced evidence of anti-prothrombin antibodies. All patients were shown to have an essentially normal level of prothrombin antigen. Using an enzyme-linked immunosorbent assay (ELISA), six of 8 LA patients tested positive for anticardiolipin antibodies (aCL) of the IgG isotype while 7 of 8 LA patients tested positive for antiphosphatidylserine antibodies (aPS) of the IgG isotype.An anti-human Factor II (prothrombin) ELISA was developed to confirm the presence of anti-Factor II (aFII) activity in LA patients. Seven of 8 LA patients were positive for aFII activity. A strong parallel existed between the presence of aPS activity, anti-human Factor II activity, and the LA, i.e., 7 of 8 LA plasmas were aPS (+)/aFII (+). An antibovine Factor II ELISA was developed to determine if the aFII activity associated with LA patients is speciesspecific. Three of 5 LA patients positive for anti-human Factor II activity were also shown to be positive for antibovine Factor II activity. Antibodies with specificity for human prothrombin were purified from LA plasmas using a prothrombin affinity column. Three of 8 LA patient eluates were shown to be positive for aPS (IgG) while none were positive for aCL (IgG or IgM) or human aFII activity. Affinity-purified eluates were assayed for LA activity using the dilute Russell viper venom time (dRVVT). None of the LA patient eluates were shown to prolong the dRVVT when present with normal plasma in concentrations up to 100 micrograms/mL. / Department of Chemistry

Thesis (Ph.D.)--Cleveland State University, 2008. / Abstract. Title from PDF t.p. (viewed on Oct. 8, 2008). Includes bibliographical references. Available online via the OhioLINK ETD Center. Also available in print.

No description available.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior / CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / RESUMO As doenÃas trombÃticas constituem um sÃrio problema de saÃde pÃblica. Diversas desordens hereditÃrias e ambientais, que afetam o sistema fisiolÃgico anticoagulante, estÃo atualmente estabelecidas como fatores de risco para a ocorrÃncia do evento trombÃtico. Dentre os fatores hereditÃrios, as mutaÃÃes G1691A do gene do fator V e G20210A do gene da protrombina sÃo os mais freqÃentes. A associaÃÃo entre estas alteraÃÃes genÃticas e a ocorrÃncia de eventos trombÃticos desencadeou o desenvolvimento de diversas pesquisas. Neste estudo, 189 pacientes portadores de eventos trombÃticos, atendidos no ambulatÃrio de Hematologia do Centro de Hematologia e Hemoterapia do Cearà - HEMOCE/SESA/UFC, foram analisados para a detecÃÃo da presenÃa das mutaÃÃes G1691A do gene do fator V e G20210A do gene da protrombina. O grupo controle consistiu de 349 voluntÃrios. A freqÃÃncia encontrada na populaÃÃo controle foi de 2% (7/349) para a mutaÃÃo do fator V e 1,7% (6/349) para a mutaÃÃo da protrombina, enquanto que nos pacientes trombofÃlicos a freqÃÃncia destas mutaÃÃes foi de 9% (17/189) e 2,1% (4/349), respectivamente. Dentre os fatores hereditÃrios, apenas a mutaÃÃo do fator V foi significante (p<0,001). Considerando os fatores ambientais de risco, o tabaco, idade > 40 anos e sexo feminino apresentaram significÃncia estatÃstica (p<0,001). Os riscos foram estimados em anÃlises pareadas e nÃo pareadas para o fator V de Leiden (4,8; 5,3; 9,8), tabaco (17,6; 14,9; 33,3), idade idade > 40 anos (2) e sexo feminino (3,7 e 4,1). Os fatores de risco para eventos trombÃticos no Cearà foram tabagismo, idade > 40 anos, sexo feminino, e a mutaÃÃo G1691A do fator V.foram associados com o desenvolvimento de trombose no estado do CearÃ. / Thrombotic diseases are a serious problem for public health. Several hereditary and environmental factors, that affect physiological anticoagulant system, have been nowadays well established as risk factors for thrombosis. Among hereditary factor V Leiden and prothrombin G20210A mutation are the most frequents. The association between several modifications on factor V gene and prothrombin gene in the development of thrombotic events has brought about future searches. In this study, 189 patients with thrombosis attended at the Hematology and Hemoterapy Center of Cearà state âHEMOCE/ Brazil, were analyzed to find out the presence of factor V Leiden and prothrombin G20210A mutation The control group was made up 349 healthy volunteers. In this study, the frequency found of factor V Leiden the control population was of 2% (7/349) and in the patients was 9% (17/189) while the frequency found of prothrombin G20210A mutation the control population was of 1,7% (6/349) and in the patients was 2,1% (4/189). Among hereditary factors only factor V Leiden was significant statistic (p<0,001). Among environmental factors studied, tabagism, age > 40 anos and femele were significant statistic (p<0,001). The ODDS RATIO of the risk factors with significant statistic were factor V de Leiden (4,8; 5,3; 9,8), tabaco (17,6; 14,9; 33,3), age > 40 years old (2) and female (3,7 e 4,1). Our results demonstrate that factor V Leiden, tabagism, age > 40 years old and female were associated with development trombosis in CearÃ.

It was looked to stand out the importance of discover laboratorial parameters that are auxiliary to the diagnostic of acute infarction of myocardial (AMI). The AMI is one of the biggest problems of public health in the world. Due to this fact, it is of major importance to find laboratorial parameters with quality and reasonable costs. In the present study fibrinogen concentrations measured during acute phase of AMI were related to cardiovascular death or a new AMI event. This incidence was higher in the etary range of 44 to 75 years in men; and 56 to 90 in women. Approximately 73% of patients presented familiar history of Coronary Heart Disease (CHD), 66% smoked, 63% presented hypertension and 81% sedentary. It was also observed elevated cases of AMI in extreme temperature days. For the fibrinogen concentrations (FBR), results demonstrated significant difference (p<0.05) between the control and infarction group patients. For protrombin time, troponine (TROP), creatinokinase, CK-MB and leukocytes count, results showed statically difference between groups. However, TTPa, total cholesterol, HDL, LDL, triglycerides levels presented no significant difference between studied groups. In conclusion, this work demonstrated a increasing fibrinogen concentration in patients with AMI, revealing that it may be adequate as a cardiac marker for AMI. / Procurou-se ressaltar a importância de determinados parâmetros laboratoriais que auxiliem o diagnóstico do infarto agudo do miocárdio (IAM). O IAM é um dos maiores problemas de saúde pública no mundo. Devido a isso, torna-se importante encontrar parâmetros laboratoriais de qualidade e baixo custo, para a caracterização do IAM. Concentrações altas de fibrinogênio determinados durante a fase aguda do IAM, foram associadas com morte cardiovascular ou um novo evento de IAM. A incidência de IAM é maior em homens na faixa etária de 44 a 75 anos; e nas mulheres entre 56 a 90 anos. Dos pacientes avaliados neste estudo, 73% apresentavam história familiar de doença arterial coronariana (DAC); 66% fumavam, 63% apresentavam hipertensão e 81% era sedentária. Foi observado que nos dias frios ou com temperaturas extremas aumentou o número de IAM. Para as concentrações de fibrinogênio (fbr), tempo de protrombina (TP), tempo de tromboplastina ativada (TTPa), troponina (TROP), creatinoquinase (CK), creatinoquinase fração MB, CK-MB, contagem de leucócitos, a média dos resultados obtidos apresentou diferença significativa entre os grupos controle e infartados. No entanto para o TTPa, colesterol total, HDL, LDL, triglicerídeos as médias observadas não apresentaram diferença significativa. Neste trabalho foi possível observar o aumento da concentração de fibrinogênio e no tempo de protrombina dos pacientes com IAM.

Class of 2015 Abstract / Objectives: To determine if using three-factor prothrombin complex concentrate (PCC) immediately prior to heart transplantation reduces blood product transfusions in patients bridged to heart transplantation by mechanical circulatory support (MCS) devices who are treated with warfarin. Methods: This study retrospectively reviewed patients that either received PCC or received usual care (i.e. fresh frozen plasma – FFP) prior to heart transplantation. Outcomes that were evaluated included packed red blood cell (RBC), FFP, platelet and cryoprecipitate transfusions intra and five days post-operatively, Cell Saver autologous blood volume administered intra-operatively, chest tube output for the five days post-operatively, and thromboembolic events post-operatively. Results: There were 24 patients included in the study, 12 from each group. The PCC group showed significantly less intra-operative RBC transfusion (2.60 ± 1.49 units vs. 5.09 ± 2.42 units, p=0.018), Cell Saver autologous blood usage (2.60 ± 1.49 units vs. 4.02 ± 1.55 units, p=0.032), and FFP transfusion (2.14 ± 2.30 units vs. 10.94 ± 5.96 units, p=0.0005) than the usual care group. There was no difference in amount of vitamin K given, change in INR, platelets administered, cryoprecipitate administered, chest tube output, or thromboembolic events between the groups. The average dose of PCC was 31 units/kg IV; repeat doses were given to 2 patients. Conclusions: We propose that the use of PCC prior to heart transplant surgery for patients on MCS devices anticoagulated with warfarin may result in the reduction for the need of RBC’s, autologous blood use and FFP during surgery.

Warfarin är ett läkemedel som används för att förhindra att högriskpatienter såsom de med förmaksflimmer får tromboembolism. Denna verkan uppnås genom att hämma de K-vitaminberoende faktorerna VII, X och protrombin och på så sätt minska blodets förmåga att koagulera. Att hitta rätt dosering av läkemedlet för warfarinbehandlade patienter har visat sig vara svårt eftersom det kräver regelbunden provtagning och påverkas av mat- och levnadsvanor. Det vanligaste sättet att mäta protrombinkomplexhalten är med venös plasma men det är även möjligt att använda sig av kapillär plasma. Helblod kan användas för mekaniska metoder som inte använder sig av optisk detektion. Fördelen är att helblod inte kräver centrifugering. Studiens syfte var att undersöka om det fanns en signifikant skillnad (p≤0,05) mellan helblod och plasma som används i den nuvarande metoden för kapillära prover och om det finns en skillnad i stabiliteten av dessa prov. Dubbla prover togs från 30 warfarinbehandlade patienter och 5 icke warfarinbehandlade individer. Ett av proven centrifugerades och analyserades på plasma, det andra analyserades på helblod. Resultaten visade att det fanns en signifikant skillnad (p≤0,05) mellan metoderna. Bland-Altman diagrammet visade att 95 % av helblodsproverna inte var högre än 0,25 INR och lägre än 0,14 INR. Detta har en låg klinisk inverkan. 4 Proverna förvarades i rumstemperatur i upp till 24 timmar och analyserades sedan om. Ingen förändring över 10 % kunde observeras i hållbarheten. Studien visade att trots att det finns en signifikant skillnad är det möjligt att ersätta den nuvarande metoden med plasma och använda helblod istället. / Warfarin is a drug used to prevent high-risk patients such as those with atrial fibrillation from thromboembolisms. This effect is achieved by suppressing vitamin-K dependent factors VII, X and prothrombin and therefore decreasing the bloods ability to clot. Finding the right dosage of the drug for warfarin treated patients has proven difficult, as it demands regular blood draws to monitor their prothrombin complex level, which is affected by dietary and living habits. The most common way to measure prothrombin complex levels is by using venous plasma but it is also possible to use capillary plasma. Whole blood can be used for mechanical methods, which don’t use optical detection. The benefit is that whole blood doesn’t require centrifugation. The aim of this study was to investigate if there was a significant difference (p≤0,05) between using whole blood and plasma which is the existing method for capillary sample and also if there is any differences between the stability of these samples. Double samples from 30 warfarin treated patients and 5 non-treated persons were taken. One of the samples were centrifuged and analyzed on plasma and the other analyzed on whole blood. The results showed that there was a significant difference (p≤0,05) between the methods. Bland-Altman plot comparison showed that 95 % of the whole blood samples would not be higher than 0,25 INR and lower than 0,14 INR. This has low clinical impact. The samples were stored at room temperature for up to 24 hours and reanalyzed. No changes over 10 % in INR values were observed. This study showed that even though there is a significant difference, it is possible to replace the existing method which using plasma with the whole blood instead.

Abstract Intracerebral hemorrhage (ICH) is the most pernicious form of stroke, with high mortality. Warfarin-associated ICH (WA-ICH) carries an even higher mortality rate. The major reason for the high mortality is explained by early hematoma growth. Warfarin use has rapidly increased with the aging of the population. We investigated temporal trends in the incidence and outcome of WA-ICHs. We found that although the proportion of warfarin users almost quadrupled in our population, the annual incidence and case fatality of WA-ICHs decreased. Management of ICH is mostly supportive. Prevention of associated complications is the issue in improving outcome. Hypertension is the most important modifiable risk factor for primary ICH, but little is known of the effect of preceding hypertension on outcome. Aggressive lowering of blood pressure is suggested to be a feasible treatment option. Reversal of warfarin anticoagulation with prothrombin complex concentrate (PCC) has been implemented as an acute treatment option for patients with WA-ICH. We found that the survival of WA-ICH subjects among our population improved after implementation of reversal of warfarin anticoagulation with PCC, likely because of the introduction of PCC. Because high mean arterial blood pressure (BP) at admission is an independent predictor of early death in patients with ICH, we explored its role in survival and poor outcome separately in normotensive subjects and subjects with treated and untreated hypertension. We found that despite their higher BP values at admission, subjects with untreated hypertension showed better survival and more often a favorable outcome after BP-lowering therapy than other patients. Studies on recurrent ICH are scarce. Underlying comorbidities, prior strokes, and drug-induced impaired platelet function may increase the risk for primary ICH (PICH). A lobar location of primary ICH may predict recurrent ICH. We investigated whether these factors predicted recurrence of PICH. In our study the annual incidence of recurrent ICH was 1.67%. Cumulative 5- and 10-year incidences were 9.6% and 14.2%. In multivariable analyses, prior ischemic stroke and diabetes proved to be independent predictors for recurrence. Moreover, diabetes was an independent risk factor for fatal recurrent PICH. Use of aspirin and serotonergic drugs did not significantly contribute to the risk. / Tiivistelmä Aivoverenvuoto (ICH) on aivoverenkiertohäiriöistä vakavin. Sille on tyypillistä korkea kuolleisuus erityisesti varfariinihoitoon liittyen, ja eloonjääneetkin vammautuvat usein vakavasti. Verenvuodon koon kasvu alkuvaiheessa selittänee korkean kuolleisuuden. Väestön ikääntymisen myötä varfariinin käyttö on lisääntynyt nopeasti. Aivoverenvuodon hoito perustuu pitkälti ennusteen parantamiseen komplikaatioita estämällä. Verenpaine on tärkein hoidettavissa oleva riskitekijä, mutta tutkimustieto akuutin vaiheen verenpainetason merkityksestä ennusteeseen on vähäistä. Tehokasta verenpaineen alentamista alkuvaiheessa pidetään lupaavana hoitomenetelmänä. Vuodon koon kasvua pyritään rajoittamaan kumoamalla varfariinin antikoaguloiva vaikutus protrombiinikompleksi-konsentraatilla (PCC). Väitöstyössäni selvitän varfariinin käyttöön liittyvien aivoverenvuotojen (WA-ICH) esiintymistiheyttä ja ennustetta ajan myötä. Tutkin myös vuodon koon kasvun rajoittamista ja alkuvaiheen korkean verenpaineen alentamista hoitomenetelminä sekä selvitän, mitkä tekijät johtavat ICH:n uusiutumiseen. Totesimme WA-ICH:n ilmaantuvuuden ja tapauskuolleisuuden pienentyneen, vaikka varfariinin käyttö miltei nelinkertaistui väestössämme. Toisaalta WA-ICH -potilaiden kuolleisuus pieneni PCC-hoidon aloittamisen jälkeen, mahdollisesti sen ansiosta. Tutkiessamme riippumattomasti varhaista kuolemaa ennustavan korkean tulovaiheen verenpaineen roolia normaaliverenpaineisilla, hoidettua ja hoitamatonta verenpainetautia sairastavilla totesimme hoitamattomien hypertonia-potilaiden selvinneen akuutin vaiheen lääkehoidon myötä muita useammin hengissä ja hyväkuntoisina korkeista tulovaiheen verenpainearvoista huolimatta. Aivoverenvuodon uusiutumiseen vaikuttavista tekijöistä on vähän tutkimustietoa. Muu sairastavuus, aiemmat aivoverenkiertohäiriöt ja trombosyyttien toimintaan vaikuttavat lääkkeet saattavat lisätä ICH:n uusiutumisriskiä. Totesimme vuosittaisen uuden ICH:n esiintymistiheyden olevan 1,67&#160;%. Aikaisempi aivoinfarkti ja diabetes osoittautuivat riippumattomiksi uusiutumista ennustaviksi riskitekijöiksi, minkä lisäksi diabetes ennusti kuolemaan johtavaa uutta ICH:a. Asetyylisalisyylihapon ja selektiivisten serotoniinin takaisinoton estäjien käyttäminen ei vaikuttanut merkittävästi uusiutumisriskiin.

Introdução: Pacientes com cirrose possuem altos níveis de fator VIII e preservação da trombomodulina (TM) (ativador da proteína C) apesar da redução global nas concentrações dos procoagulantes e anticoagulantes naturais. Isto não é levado em conta no teste de TP/INR, o qual não requer a adição de trombomodulina. Deste modo, o TP/INR não é capaz de demonstrar a magnitude da geração de trombina, em condições similares à que ocorre in vivo. De fato, o teste de TP/INR mede o lado procoagulante e se correlaciona com somente 5% do total de trombina gerada. Nossa hipótese é que a geração de trombina está bem preservada na cirrose, ainda que avançada, apesar dos resultados anormais do TP/INR, os quais indicariam coagulopatia. Objetivo: correlacionar os resultados do teste TP/INR com a geração de trombina nos pacientes com cirrose após procedimento invasivo (ligadura elástica de varizes esofagianas - LEVE). Pacientes e métodos: 97 pacientes foram consecutivamente incluídos no estudo (58 homens; 54±10 anos) e divididos em dois grupos INR < 1,5 e INR >= 1,5. Todos os pacientes passaram por uma criteriosa análise clínica e laboratorial, que incluiu revisão dos prontuários, determinação do TP/INR e da geração de trombina (ETP) com e sem adição de trombomodulina e cálculo do rETP (razão dos resultados com e sem adição de trombomodulina). Resultados: Não houve diferença significante na média dos valores de ETP sem trombomodulina no grupo INR < 1,5 (n=72), que foi 1.250±315,7 nmol/min quando comparada ao grupo INR >= 1,5 (n=25), cujos valores foram 1.186±238 nmol/min, p=0,3572. Após adição de trombomodulina, os valores mudaram para 893,0±368,6 e 965,9±232,3 nmol/min, respectivamente (p=0,6265). Ambos os grupos apresentaram preservação da geração de trombina, com valores mais elevados no grupo INR >= 1,5 do que no grupo de pacientes com INR < 1,5 (rETP 0,81±0,1 versus 0,69±0,2; p=0,0042). Evidência de hipercoagulabilidade (valores altos de rETP) foi demonstrada em 80% dos pacientes. Mesmo pacientes com INR >= 1,5 apresentam geração de trombina preservada, o que justificaria a baixa prevalência de sangramento após ligadura elástica de varizes esofagianas (5,2%; 3 pacientes no grupo INR < 1,5 e 2 pacientes no grupo INR >= 1,5). Conclusões: a geração de trombina se encontrou preservada nos pacientes com cirrose e os valores anormais de INR não refletiram a ocorrência de sangramento. A maioria dos pacientes mostrou evidência de hipercoagulabilidade, apesar do INR alargado. Sangramento após LEVE ocorreu em pequena parcela dos pacientes e não foi relacionado ao status da coagulação / Introduction: Patients with cirrhosis have higher levels of factor VIII and preservation of endothelial thrombomodulin (protein C activator) in spite of the global reduction in procoagulant and natural anticoagulant concentrations. This is not taken into account in the laboratory test of INR/PT, which does not require the addition of thrombomodulin and, thus, is not able to emulate the generation of thrombin that happens in vivo. In fact, INR/PT is a measure of procoagulant status and correlates with only 5% of the total amount of generate thrombin. We hypothesized that thrombin generation is well preserved in cirrhosis, even in advanced stages, despite the abnormal result of INR/PT, which would indicate coagulopathy. Aims: to correlated INR/PT with thrombin generation in patients with cirrhosis in the elective setting of an invasive procedure (endoscopic variceal ligation- EVL). Patients and Methods: 97 consecutive patients were prospectively included in this study (58 men; 54±10 years old) and divided into two groups INR < 1.5 and INR >= 1.5. All patients underwent a stringent clinical and laboratory assessment which included review of the clinical chart, INR/PT determinations and assessment of endogenous thrombin potencial (ETP) without and with the addition of thrombomodulin and calculation of the ETP ratio (rETP= without/with thrombomodulin). Results: There was no significant difference in the mean value of ETP without thrombomodulin that was 1,250±315.7nmol/min for patients with INR < 1.5 (n=72) and 1,186±238 in those with INR >= 1.5 (n=25); p= 0.3572. After the addition of thrombomodulin, values changed to 893.0±368.6 and 965.9±232.3, respectively (p= 0.6265). Both groups had preserved thrombin generation, which was higher in patients with INR >=1.5 than in patients with INR < 1.5 (rETP 0.81±0.1 versus 0.69±0.2; p=0.0042). Evidence of hypercoagulability (high rETP) was demonstrated in 80% of patients. Even patients with INR >= 1.5 had preserved thrombin generation, which is likely to account for the low prevalence of post-EVL bleeding (5.2%; n=3 with INR < 1.5 and n=2 with INR >= 1.5). Conclusions: thrombin generation was well preserved in patients with cirrhosis and was not reflected by abnormal results of INR. Most of the patients had evidence of hypercoagulability, despite enlarged INR. Post-procedure bleeding occurred in a small subset of the patients and was not related to the coagulation status