RESUMO
OBJECTIVES: The primary objective of this study was to assess the pharmacokinetic profiles of acetylsalicylic acid (ASA) and salicylic acid (SA) after administration of two different formulations of aspirin under fasting and fed conditions. MATERIALS AND METHODS: The study was a randomized, open-label, parallel-group, 2-arm crossover study conducted at a single center. Healthy subjects were randomized to receive 300 mg of aspirin in either a 15-mL oral solution (pre-packaged vial containing powder and solvent that are combined at the time of administration) or a single solid tablet to be chewed and swallowed with 150 mL of water. Treatment visits were separated by a 10-day wash-out period. RESULTS: At 3 minutes, ASA concentrations for the oral solution fed state and fasting state arms exceeded those for the chewed tablet (fed 299 vs. 139 ng/mL; fasting 356 vs. 204 ng/mL). Compared to the chewed tablet, the mean plasma ASA concentration was 74% greater with the oral solution under fasting conditions, and 115% greater under fed conditions. Similarly, at 3 minutes, the mean SA plasma concentration with the oral solution under fed and fasting conditions exceeded those for the chewed tablet (fed 310 vs. 160 ng/mL; fasting 330 vs. 185 ng/mL). Under fasting conditions, the mean plasma ASA AUC0-last, with the oral solutions was 168,076.8 min.ng/mL compared to 163,726.3 min.ng/mL with the chewed tablet. Under fed conditions, the mean plasma ASA AUC0-last, with the oral solutions was 179,116.7 min.ng/mL compared to 164,704.3 min.ng/mL with the chewed tablet. CONCLUSION: This phase 1 study showed that use of an aspirin oral solution provided more rapid exposure to higher plasma concentration levels of ASA and SA than chewing a solid tablet.
Assuntos
Aspirina , Mastigação , Administração Oral , Área Sob a Curva , Estudos Cross-Over , Jejum , Humanos , Pós , Ácido Salicílico , Solventes , Comprimidos , Equivalência Terapêutica , ÁguaRESUMO
BACKGROUND: The purpose of this study was to investigate mediators of inflammation and haemostasis in patients with chronic necrotizing pulmonary aspergillosis (CNPA), a locally, destructive process of the lung due to invasion by Aspergillus species. METHODS: Measurements of selected biomarkers in 10 patients with CNPA and 19 healthy, matched controls were performed with enzyme-linked immunosorbent assay (ELISA) and multiplex methodology. The gene expressions of relevant biomarkers were analyzed with real-time quantitative RT-PCR. RESULTS: Increased concentrations of circulating mediators of inflammation interleukin (IL)-6, IL-8, RANTES, TNF-α, ICAM-1 and mediators involved in endothelial activation and thrombosis (vWF, TF and PAI-1) were observed in patients with CNPA. The concentration of the anti-inflammatory cytokine IL-10 was increased both in plasma and in PBMC in the patient population. The gene expression of CD40L was decreased in PBMC from the patient group, accompanied by decreased concentrations of soluble (s) CD40L in the circulation. CONCLUSIONS: The proinflammatory response against Aspergillus may be counteracted by reduced CD40L and sCD40L, as well as increased IL-10, which may compromise the immune response against Aspergillus in patients with CNPA.
Assuntos
Biomarcadores/sangue , Fatores de Coagulação Sanguínea/análise , Citocinas/sangue , Aspergilose Pulmonar Invasiva/patologia , Idoso , Estudos de Casos e Controles , Ensaio de Imunoadsorção Enzimática , Feminino , Perfilação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo RealRESUMO
OBJECTIVE: The purpose of this study was to measure the in vivo platelet activation and thrombin generation in arterial blood after passing a subintimal conduit. METHODS: Subintimal percutaneous transluminal angioplasty (SPTA) is a technique where a subintimal channel is created, allowing recanalization of long peripheral arterial occlusion. From 10 patients with intermittent claudication, undergoing successful SPTA for femoropopliteal occlusive disease, we collected antecubital venous blood samples immediately before treatment, preprocedural arterial blood samples taken at the entry level proximal to the vessel occlusion, and subsequently at the reentry level after successful recanalization. Venous follow-up blood samples were taken after 24 hours. Plasma concentrations of ß-thromboglobulin (ß-TG), RANTES, and Prothrombin fragment (F1 + 2), were determined by immunoassay. Fibrinogen binding to platelets, leukocyte-platelet adhesion, and P-selectin were determined by flow cytometry. RESULTS: We found a statistically significant transluminal increase in the plasma concentrations of RANTES, ß-TG and F1 + 2 (p = 0.002, 0.001 and 0.001 respectively), which all normalized within 24 hours. Platelet-leukocyte aggregates significantly decreased after 24 hours compared with preprocedural and preentry levels (3.26% versus 5.26 %, p = 0.017). P-selectin expression on circulating platelets was statistically significantly increased in the blood sample taken at the re-entry level compared with the pre-procedural and pre-entry level (p = 0.007). After 24 hours there was no statistically significant difference to pre-procedural levels. There was no significant change in platelet fibrinogen binding at any levels. CONCLUSION: When passing a subintimal conduit, in vivo sampled blood demonstrated an extremely rapid and substantial uniform platelet activation and thrombin generation.
Assuntos
Angioplastia , Aterosclerose/sangue , Ativação Plaquetária , Trombina/metabolismo , Idoso , Idoso de 80 Anos ou mais , Aterosclerose/terapia , Plaquetas/metabolismo , Plaquetas/patologia , Quimiocina CCL5/sangue , Feminino , Artéria Femoral/patologia , Humanos , Claudicação Intermitente/sangue , Claudicação Intermitente/terapia , Masculino , Pessoa de Meia-Idade , Monócitos/patologia , Selectina-P/sangue , Artéria Poplítea/patologia , beta-Tromboglobulina/metabolismoRESUMO
BACKGROUND: Fast platelet function tests can identify weak clopidogrel responders, but data on variability over time in clopidogrel responsiveness in several clinical settings are lacking. We wanted to explore long-term variability of multiple electrode aggregometry (MEA) measurements and the agreement between MEA and light transmission aggregometry (LTA) in patients with non-ST elevation myocardial infarction (NSTEMI) treated with aspirin and clopidogrel. METHODS: Parallel MEA and LTA were performed at baseline and after 6 and 12 weeks in 31 patients treated with percutaneous coronary intervention after NSTEMI. Adenosine diphosphate (ADP) concentrations 2 µM, 6.5 µM and 10 µM were used. Parallel testings in both arterial and venous blood were performed at baseline. MEA and LTA cut-off levels were applied to discriminate aggregation values suggesting presence or absence of high platelet reactivity (HPR). RESULTS: Arterial and venous MEA and LTA aggregation were similar. Within-subject variability in both MEA and LTA aggregation throughout the study was moderate. According to MEA, eight patients had HPR at baseline (MEA aggregation > 47 U). Defining > 47% as the LTA aggregation HPR cut-off level, the same number of patients (eight) had HPR according to LTA. Of the 93 MEA/LTA observations 81 (87.1%) gave the same HPR classification. MEA vs. LTA agreement at baseline was slightly inferior to that obtained after 12 weeks. CONCLUSIONS: MEA and LTA aggregation in arterial and venous blood seem similar. Within-subject variability over time was moderate, and the agreement between LTA and MEA was good, and stable in most patients.
Assuntos
Aspirina/uso terapêutico , Infarto do Miocárdio/tratamento farmacológico , Inibidores da Agregação Plaquetária/uso terapêutico , Agregação Plaquetária/efeitos dos fármacos , Ticlopidina/análogos & derivados , Idoso , Aspirina/farmacologia , Clopidogrel , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Inibidores da Agregação Plaquetária/farmacologia , Testes de Função Plaquetária , Reprodutibilidade dos Testes , Ticlopidina/farmacologia , Ticlopidina/uso terapêuticoRESUMO
The interaction between inflammatory cytokines and endothelial cells is a critical step in atherogenesis leading to endothelial dysfunction and inflammation. We have previously reported that the tumor necrosis factor superfamily member LIGHT could be involved in atherogenesis through its ability to promote vascular inflammation. In the present study we identified proteinase-activated receptor (PAR)-2 as an inflammatory mediator that was markedly enhanced by LIGHT in endothelial cells. We also found that LIGHT acted synergistically with PAR-2 activation to promote enhanced release of the proatherogenic chemokines interleukin-8 and monocyte chemoattractant protein-1, underscoring that the interaction between LIGHT and PAR-2 is biologically active, promoting potent inflammatory effects. We showed that the LIGHT-mediated upregulation of PAR-2 in endothelial cells is mediated through the HVEM receptor, involving Jun N-terminal kinase signaling pathways. A LIGHT-mediated upregulation of PAR-2 mRNA levels was also found in human monocytes when these cells were preactivated by tumor necrosis factor alpha. We have previously demonstrated increased plasma levels of LIGHT in unstable angina patients, and here we show a similar pattern for PAR-2 expression in peripheral blood monocytes. We also found that LIGHT, LIGHT receptors, and PAR-2 showed enhanced expression, and, to some degree, colocalization in endothelial cells and macrophages, in the atherosclerotic plaques of ApoE(-/-) mice, suggesting that the inflammatory interaction between LIGHT and PAR-2 also may be operating in vivo within an atherosclerotic lesion. Our findings suggest that LIGHT/PAR-2-driven inflammation could be a pathogenic loop in atherogenesis potentially representing a target for therapy in this disorder.
Assuntos
Aterosclerose/etiologia , Células Endoteliais/metabolismo , Endotélio Vascular/patologia , Receptor PAR-2/fisiologia , Membro 14 de Receptores do Fator de Necrose Tumoral/fisiologia , Membro 14 da Superfamília de Ligantes de Fatores de Necrose Tumoral/fisiologia , Vasculite/metabolismo , Idoso , Angina Pectoris/metabolismo , Angina Pectoris/patologia , Angina Instável/metabolismo , Angina Instável/patologia , Animais , Aterosclerose/metabolismo , Aterosclerose/patologia , Células Cultivadas/metabolismo , Quimiocina CCL2/metabolismo , Células Endoteliais/patologia , Feminino , Regulação da Expressão Gênica , Humanos , Interleucina-8/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Pessoa de Meia-Idade , Óxido Nítrico Sintase Tipo III/metabolismo , Receptor PAR-2/agonistas , Proteínas Recombinantes de Fusão/fisiologia , Transdução de Sinais/fisiologia , Membro 14 da Superfamília de Ligantes de Fatores de Necrose Tumoral/genética , Vasculite/complicações , Vasculite/patologiaRESUMO
BACKGROUND: We investigated whether the heparin-coated AN69 ST hemodialysis (HD) filter induced less hypercoagulability during HD than a conventional polysulfone filter (F×8). METHODS: In a crossover design, 11 patients were treated alternately with AN69 ST and F×8 filters (45 sessions). All filters were primed with unfractionated heparin (UFH) and unadsorbed UFH was removed by saline flushing. Half the conventional dalteparin dose was given as a bolus dose at the start of HD. Clotting was evaluated hourly in the venous air trap. Prothrombin fragments 1 and 2 (PF1 + 2), antithrombin (AT), ß-TG and anti-FXa activity were repeatedly measured. RESULTS: One patient treated with enalapril had two repeated adverse reactions to the AN69 ST filter and was excluded from the study. Use of the AN69 ST filter did not decrease the mean clot score or PF1 + 2, but decreased ß-TG compared to the F×8 filter. CONCLUSION: The heparin-coated AN69 ST filter did not induce less coagulation when compared to the F×8 filter.
Assuntos
Anticoagulantes/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Materiais Revestidos Biocompatíveis , Heparina/farmacologia , Membranas Artificiais , Polímeros , Diálise Renal/instrumentação , Sulfonas , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos Cross-Over , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Diálise Renal/métodosRESUMO
Common variable immunodeficiency (CVID) is a heterogeneous syndrome characterized by defective immunoglobulin production and high frequency of bacterial infections, autoimmunity and manifestations of chronic inflammation. The urokinase plasminogen activator (uPA), its cell bound and soluble receptor (uPAR, suPAR) have complex biological functions involving innate immune defense mechanisms and regulation of inflammation. Based on this dual role, we hypothesized that the uPA system could be affected in CVID, and examined expression of components of the uPA system in subgroups of CVID. All CVID-patients had increased plasma levels of suPAR with particularly high levels in those with splenomegaly and thrombocytopenia. Plasma uPA levels were also raised in these patients, and both suPAR and uPA levels correlated with the monocyte activation marker neopterin. Monocytes from CVID patients had increased expression of uPAR. We show an increased activation of the uPA system possibly contributing to the inflammatory phenotype seen in subgroups of CVID patients.
Assuntos
Imunodeficiência de Variável Comum/imunologia , Monócitos/imunologia , Receptores de Ativador de Plasminogênio Tipo Uroquinase/sangue , Ativador de Plasminogênio Tipo Uroquinase/sangue , Adulto , Imunodeficiência de Variável Comum/sangue , Imunodeficiência de Variável Comum/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismoRESUMO
BACKGROUND: Platelet microparticles (PMPs) possess proatherogenic and procoagulant properties which may play a role in atherogenesis and subsequent thromboembolic complications. The present study was conducted to investigate the possible relationship between carotid atherosclerosis and plasma concentrations of PMPs, and elucidate if plasma levels of PMPs were affected by postprandial hypertriglyceridemia. METHODS AND RESULTS: Subjects with ultrasound-assessed carotid atherosclerotic plaques (echogenic; n=20 and echolucent; n=20), assessed by ultrasonography, and subjects without carotid plaques (n=20) were recruited from a population-based study and underwent a standard fat tolerance test. Subjects with carotid plaques had significantly higher levels of large PMPs than subjects without carotid atherosclerotic plaques (96.7+/-50.4 microg/l versus 56.1+/-34.9 microg/l), after adjustments for traditional cardiovascular risk factors and use cardiovascular drugs (p=0.021). Plasma PMPs were not associated with plaque echogenicity. Postprandial hypertriglyceridemia induced a similar increase in plasma PMPs within all groups. Significant correlations were found between an increase in plasma triglycerides and percent elevation in total PMPs (r=0.29, p<0.05) and large PMPs (r=0.34, p<0.01) in the postprandial phase. CONCLUSIONS: Individuals with echogenic and echolucent carotid atherosclerotic plaques have statistically significant elevation of large plasma PMPs compared to age/sex-matched normal controls. Postprandial hypertriglyceridemia induces a significant, similar increase in plasma PMPs in individuals with and without carotid atherosclerotic plaques which could be of pathophysiological importance in atherogenesis.
Assuntos
Plaquetas/fisiologia , Plaquetas/ultraestrutura , Doenças das Artérias Carótidas/sangue , Período Pós-Prandial/fisiologia , Idoso , Doenças das Artérias Carótidas/diagnóstico por imagem , Doenças das Artérias Carótidas/etiologia , Estenose das Carótidas/sangue , Estenose das Carótidas/diagnóstico por imagem , Estenose das Carótidas/etiologia , Estudos de Casos e Controles , Gorduras na Dieta/administração & dosagem , Feminino , Humanos , Hipertrigliceridemia/sangue , Hipertrigliceridemia/etiologia , Masculino , Triglicerídeos/sangue , UltrassonografiaRESUMO
BACKGROUND: Evaluation of clopidogrel therapy by in vitro methods has limitations which may be of clinical importance. We wanted to explore the variability in aggregometry response in aspirin sensitive patients before and after initiation of clopidogrel therapy. METHODS: ADP 9.37 microM, AA 1.2mM and TRAP 25 mM stimulated light transmissions aggregometry (LTA) were performed twice before (Exams 1 and 2; 3 weeks apart)-and within one year after-initiation of clopidogrel therapy (Exam 3) in 79 patients treated with PCI. Repeated ADP aggregometry was also performed in 16 healthy volunteers in order to estimate LTA measurement error. RESULT: Inter-individual differences in ADP aggregation e.g. at Exam 1 were substantial (range 17-77%, SD 15.8%). Intra-individual changes between Exams 1 and 2 were significant (-27 to +36%, SD 14.6%, p<0.05). Inter-individual differences at Exam 3 (on clopidogrel treatment) were larger than expected from Exams 1 and 2 (p<0.01). AA aggregation was the same before and during clopidogrel treatment. In controls, inter-individual differences were smaller at ADP 10 than at ADP 5 microM. CONCLUSIONS: Inter-individual differences in ADP aggregation were significant both before and during clopidogrel therapy, and there were significant intra-individual variations over time. Therefore, prediction of aggregometry response before or during clopidogrel therapy based on single tests may be unreliable. Inter-individual differences in healthy controls are smaller at high concentrations of ADP, and comparisons of aggregometry response should be performed with caution unless ADP concentrations are standardized.
Assuntos
Inibidores da Agregação Plaquetária/farmacologia , Agregação Plaquetária/efeitos dos fármacos , Testes de Função Plaquetária/métodos , Ticlopidina/análogos & derivados , Difosfato de Adenosina/farmacologia , Ácido Araquidônico/farmacologia , Aspirina/farmacologia , Estudos de Casos e Controles , Clopidogrel , Demografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fragmentos de Peptídeos/farmacologia , Reprodutibilidade dos Testes , Ticlopidina/farmacologiaRESUMO
RATIONALE: Thrombus formation and inflammation are involved in the pathogenesis of pulmonary arterial hypertension (PAH), and LIGHT (Lymphotoxin-like Inducible protein that competes with Glycoprotein D for Herpesvirus entry mediator on T lymphocytes) has been shown to promote vascular inflammation. OBJECTIVES: We sought to investigate the role of the tumor necrosis factor superfamily ligand LIGHT in the pathogenesis of PAH. METHODS: We studied 73 patients with severe PAH and 10 control subjects. LIGHT and pro- and antithrombotic markers were assessed by enzyme immunoassays. MEASUREMENTS AND MAIN RESULTS: (1) Patients with idiopathic PAH (n = 21), patients with PAH related to risk factors or associated conditions (n = 31), and those with chronic thromboembolic PAH (n = 21) all had raised serum levels of LIGHT compared with control subjects (n = 10). (2) LIGHT levels in femoral artery were significantly related to mortality in the patients with PAH. (3) Immunostaining of LIGHT and its receptors was seen in alveolar macrophages, vascular smooth muscle cells, and endothelial cells in lungs from patients with PAH. (4) Thirteen patients received prostacyclin infusion (3 mo), and all showed hemodynamic improvement, accompanied by decreased LIGHT levels. (5) Prostacyclin abolished the release of LIGHT from activated platelets in vitro, suggesting that the decrease in LIGHT during prostacyclin therapy could involve direct effects on platelets. (6) LIGHT increased tissue factor and plasminogen activator inhibitor type 1 and decreased thrombomodulin levels in endothelial cells, inducing a prothrombotic state in these cells. CONCLUSIONS: Our findings suggest prothrombotic effects of LIGHT in PAH involving endothelium-related mechanisms, potentially contributing to the progression of this disorder.
Assuntos
Hipertensão Pulmonar/etiologia , Trombose/etiologia , Membro 14 da Superfamília de Ligantes de Fatores de Necrose Tumoral/fisiologia , Biomarcadores/sangue , Progressão da Doença , Humanos , Ligantes , Membro 14 da Superfamília de Ligantes de Fatores de Necrose Tumoral/análise , Membro 14 da Superfamília de Ligantes de Fatores de Necrose Tumoral/sangue , Veias Umbilicais/citologiaRESUMO
OBJECTIVES: Patients with various inflammatory rheumatic diseases (IRDs) have increased risk of atherothrombotic disease. Lipoprotein (a) (Lp(a)) is a risk factor for atherosclerosis but its role in IRD with accompanying coronary artery disease (CAD) is still unclear. We aimed to examine if serum Lp(a) levels differed between CAD patients with and without accompanying IRD. DESIGN: A cross-sectional observational, patient-based cohort study. SETTING: Referred centre for coronary artery bypass grafting in the South Eastern part of Norway. PARTICIPANTS: 67 CAD patients with IRD (CAD/IRD) and 52 CAD patients without IRD (CAD/non-IRD). All patients were Caucasians, aged >18 years, without any clinically significant infection or malignancy. METHODS: Lp(a) levels in serum were analysed by particle enhanced immunoturbidimetric assay, and Lp(a) levels were related to clinical and biochemical characteristics of the patient population. RESULTS: We found no differences in serum levels of Lp(a) between CAD patients with and without IRD. In general, we found that Lp(a) correlated poorly with clinical and biochemical parameters including C reactive protein with the same pattern in the CAD/non-IRD and CAD/IRD groups. CONCLUSIONS: Our data do not support a link between inflammation and Lp(a) levels in CAD and in general Lp(a) levels were not correlated with other risk factors for cardiovascular disease.
Assuntos
Doença da Artéria Coronariana/sangue , Lipoproteína(a)/sangue , Doenças Reumáticas/sangue , Adulto , Idoso , Estudos de Coortes , Doença da Artéria Coronariana/complicações , Estudos Transversais , Feminino , Humanos , Inflamação/sangue , Inflamação/complicações , Masculino , Pessoa de Meia-Idade , Doenças Reumáticas/complicações , Fatores de RiscoRESUMO
BACKGROUND AND PURPOSE: Although cerebral microemboli are often detected by transcranial Doppler ultrasonography in mechanical heart valve patients, the clinical significance of such microemboli is unclear. The aim of this study was to determine the frequency and composition of cerebral microemboli in a prosthetic heart valve population and to correlate these findings to cerebrovascular symptoms, blood inflammation, and coagulation parameters. METHODS: Seventy-six consecutive patients with a total of 81 prosthetic (54 mechanical, 27 biologic) heart valves were monitored for cerebral microemboli by multifrequency transcranial Doppler ultrasonography 1 year after valve replacement. Cerebrovascular events in the first year were recorded by a neurologist. Inflammation and coagulation markers were measured by immunoassays. RESULTS: Microemboli were detected in mechanical heart valve patients only (28 patients, 56%). Twelve percent were solid, occurring in 17 (34%) of the mechanical heart valve population. The presence of solid cerebral microemboli was the only variable that was associated with cerebrovascular symptoms after a final regression analysis (P=0.026). The plasma monocyte chemotactic protein-1 level was raised in patients with solid microemboli (P=0.014). CONCLUSIONS: Solid cerebral microemboli were detected by multifrequency transcranial Doppler ultrasonography in 35% of a mechanical heart valve population, and the frequency was higher in patients who experienced cerebrovascular events during the first year after valve replacement. The results suggest that the detection of solid cerebral microemboli may be helpful in predicting the risk of ischemic stroke in mechanical heart valve patients.
Assuntos
Próteses Valvulares Cardíacas/efeitos adversos , Embolia Intracraniana/diagnóstico por imagem , Embolia Intracraniana/epidemiologia , Acidente Vascular Cerebral/diagnóstico por imagem , Acidente Vascular Cerebral/epidemiologia , Ultrassonografia Doppler Transcraniana , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Amaurose Fugaz/diagnóstico por imagem , Amaurose Fugaz/epidemiologia , Coagulação Sanguínea , Embolia Aérea/diagnóstico por imagem , Embolia Aérea/epidemiologia , Feminino , Humanos , Incidência , Inflamação/epidemiologia , Ataque Isquêmico Transitório/diagnóstico por imagem , Ataque Isquêmico Transitório/epidemiologia , Masculino , Pessoa de Meia-Idade , Ativação Plaquetária , Valor Preditivo dos Testes , Fatores de RiscoRESUMO
OBJECTIVE: Both animal and human studies demonstrate activation of coagulation during cardiac arrest. Prearrest anticoagulation is used routinely in many experimental studies. We studied the hemodynamic effects of prearrest anticoagulation with a low-molecular-weight heparin suitable for clinical use during cardiopulmonary resuscitation in pigs. DESIGN: Randomized and blinded experimental animal study. SETTING: University hospital-affiliated research laboratory. SUBJECTS: Sixteen female domestic pigs. INTERVENTIONS: Three minutes before electrically induced ventricular fibrillation, enoxaparin 1 mg/kg or physiologic saline was blinded and administered intravenously. After 10 mins of untreated ventricular fibrillation, advanced cardiac life support was initiated with continuous mechanical chest compressions and interposed manual ventilation with 100% oxygen. Epinephrine was administered after 2 mins of advanced cardiac life support followed by attempted defibrillation 1 min thereafter. Advanced cardiac life support was continued for 10 mins following international guidelines. Electrocardiogram was recorded continuously and ventricular fibrillation waveform was analyzed (median slope). Animals with return of spontaneous circulation were observed for ten more minutes. Blood specimens were drawn for analysis of coagulation activation (thrombin-antithrombin complex) and drug effect (anti-factor Xa activity). MEASUREMENTS AND MAIN RESULTS: Six of eight (75%) pigs in each group achieved return of spontaneous circulation. Thrombin-antithrombin complex levels were significantly lower in pigs that received enoxaparin. There was no significant difference either in measured hemodynamics between the groups during advanced cardiac life support and after return of spontaneous circulation or in median slope values during ventricular fibrillation. Epinephrine caused a significant decrease in femoral and increase in cerebral cortical blood flow with no difference between the groups. CONCLUSIONS: Prearrest anticoagulation with enoxaparin did not influence either hemodynamics during advanced cardiac life support and after return of spontaneous circulation or the frequency of return of spontaneous circulation in porcine cardiac arrest.
Assuntos
Reanimação Cardiopulmonar , Fibrinolíticos/farmacologia , Parada Cardíaca/terapia , Hemodinâmica/efeitos dos fármacos , Heparina de Baixo Peso Molecular/farmacologia , Animais , Feminino , SuínosRESUMO
Platelet-derived microparticles (PMPs) are considered a marker of platelet activation. They vary considerably in size, and flow cytometry, the predominant method used to assay PMPs, is only detecting larger PMPs (>0.1 microm). We describe here a method that quantifies the amount of PMP-located GPIIb antigen in detergent-treated platelet-free plasma (PPP) by means of a one-step time-resolved immunofluorometric assay (TR-IFMA). This assay uses a streptavidin-coated microwell plate and two different monoclonal antibodies to GPIIb (CD41), one conjugated to biotin and the other labeled with europium ion. A wide linear range standard curve with low background and a high sensitivity was obtained. Pre-assay ultracentrifugation or filtration of PPP extensively reduced the fluorometric signal, indicating that the GPIIb antigen is mainly particle-located. A strong correlation between the amount of GPIIb and PMP as detected by flow cytometry was found. Consequently, the assay can be used to study PMP-related phenomena and, in contrast to flow cytometry, can be used on frozen samples and is independent of PMP size.
Assuntos
Plaquetas/química , Fluorimunoensaio , Ativação Plaquetária/fisiologia , Glicoproteína IIb da Membrana de Plaquetas/análise , Anticorpos Monoclonais/metabolismo , Biomarcadores/sangue , Biotina/metabolismo , Biotinilação , Calcimicina/farmacologia , Detergentes/farmacologia , Európio/metabolismo , Citometria de Fluxo , Humanos , Ionóforos/farmacologia , Isótopos , Tamanho da Partícula , Fragmentos de Peptídeos/farmacologia , Ativação Plaquetária/efeitos dos fármacos , Sensibilidade e Especificidade , Estreptavidina/metabolismo , beta-Tromboglobulina/análiseRESUMO
BACKGROUND: Detailed studies of a family with a strong clustering of fibromyalgia revealed that the affected members had eight or more urinations per day and at least one per night. As this feature was not known to be an important symptom in this ailment, we carried out a questionnaire-based study. MATERIAL AND METHODS: Questionnaires were mailed to a random group of 285 subjects drawn from the member list of the Norwegian Fibromyalgia Association, and to 160 healthy females, free of urinary tract pains. Response rates were 70% and 74%. RESULTS: The fibromyalgia group reported 8 or more daily and 1 permanent nocturnal urinations; the corresponding figures for healthy females were 6 and 0.2. This corresponds to a sensitivity and specificity for fibromyalgia of 78 and 91% for daytime urinations, and to 89% and 92% for one or more permanent nocturia. INTERPRETATION: The study indicates that an abnormally high frequency of urinations is a characteristic feature in fibromyalgia and a useful diagnostic variable. It is also objectively verifiable by urodynamic investigation. The pattern is that of urge, sometimes with incontinence.
Assuntos
Fibromialgia/diagnóstico , Micção , Adulto , Feminino , Fibromialgia/complicações , Fibromialgia/genética , Fibromialgia/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Inquéritos e Questionários , Micção/fisiologia , Transtornos Urinários/diagnóstico , Transtornos Urinários/etiologiaRESUMO
Homozygosity for a novel D180G mutation in the protease domain of protein C, associated with plasma protein C activity and antigen levels of 8% of normal was identified in a thrombosis prone family. Transient expression of protein C in HK-293 cells and analysis of protein C antigen in culture media and cell lysates showed that the secretion of mutant protein as compared with wild-type protein was reduced by 79% while the intracellular contents were similar. Computer analysis of the X-ray structure of activated protein C and of a theoretical model of the zymogen predicts that the mutation destabilises the molecule locally. Our results are compatible with a relatively unstable mutant molecule that could be trapped inside the cell and degraded. However, if secreted the mutant molecule could have a relatively normal catalytic activity and structure consistent with the plasma levels of protein C activity and the late onset of thrombosis.
Assuntos
Mutação Puntual/genética , Deficiência de Proteína C/genética , Proteína C/genética , Adulto , Saúde da Família , Feminino , Genótipo , Homozigoto , Humanos , Pessoa de Meia-Idade , Modelos Moleculares , Linhagem , Proteína C/química , Proteína C/metabolismo , Deficiência de Proteína C/etiologia , Conformação Proteica , Trombose/etiologia , Trombose/genética , TransfecçãoRESUMO
BACKGROUND: Intraoperative autologous blood withdrawal protects the pooled blood from the deleterious effects of cardiopulmonary bypass. Following reinfusion after cardiopulmonary bypass, the fresh autologous blood contributes to less coagulation abnormalities and reduces postoperative bleeding and the need for allogeneic blood products. However, few data have been available concerning the quality and potential activation of fresh blood stored at room temperature in the operating room. METHODS: Forty coronary artery bypass grafting patients undergoing a consistent intraoperative and postoperative autotransfusion protocol had a median of 1,000 mL of autologous blood withdrawn before cardiopulmonary bypass. After heparinization the blood was drained from the venous catheter via venous cannula into standard blood bags and stored in the operating room until termination of cardiopulmonary bypass. Samples for hemostatic and inflammatory markers were taken from the pooled blood immediately before it was returned to the patient. RESULTS: There was some activation of platelets in the stored autologous blood, as measured by an increase of beta-thromboglobulin. Indications of thrombin formation, as assessed by plasma levels of thrombin-antithrombin complex and prothrombin fragment 1.2 were not seen, and there was no fibrinolytic activity. The red blood cells remained intact, indicated by the absence of plasma free hemoglobin. As for the inflammatory response, the levels of the terminal complement complex remained stable, and the cytokines tumor necrosis factor-alpha and interleukin 6 levels were not increased during storage. The complement activation products increased minimally, but remained within normal ranges. CONCLUSIONS: Except for slight activation of platelets, there was no indication of coagulation, hemolysis, fibrinolysis, or immunologic activity in the autologous blood after approximately 1 hour of operating room storage. The autologous blood was preserved in a condition of high quality, and retransfusion after cardiopulmonary bypass represents an uncomplicated and almost costless procedure for blood conservation.
Assuntos
Transfusão de Sangue Autóloga , Ponte Cardiopulmonar , Testes Hematológicos , Cuidados Intraoperatórios , Cuidados Pós-Operatórios , Adulto , Idoso , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Transfusions with incompatible blood products including plasma are a well-known problem. The present study was performed to test the effects of Uniplas, universal plasma that can be transfused regardless of a patient's blood group, with respect to bleeding and hemostatic activity. The study comprised 84 adult patients scheduled for elective openheart surgery. A total of 55 patients received plasma transfusions, while 29 patients not requiring plasma served as controls. If plasma transfusion was indicated during operation or over the two following days, patients were randomised 2:1 to receive Uniplas or Octaplas of blood group AB. Relevant clinical observations were recorded and blood tests taken repeatedly. The transfused patient groups were comparable, and no significant differences were observed with respect to activated clotting time (ACT), activated partial thromboplastin time (APTT) or postoperative bleeding into chest drains. The median number of transfused units of Uniplas was 3, with a range of 1-23, while the median for Octaplas was 2 with a range of 1-11. These differences were not significant.Thus, Uniplas has a similar effect as Octaplas in the treatment of bleeding in patients undergoing openheart surgery. Uniplas can therefore substitute for Octaplas and eliminate the risk of ABO-incompatible transfusions.
Assuntos
Transfusão de Componentes Sanguíneos/normas , Incompatibilidade de Grupos Sanguíneos , Plasma/imunologia , Idoso , Idoso de 80 Anos ou mais , Testes de Coagulação Sanguínea , Transfusão de Componentes Sanguíneos/efeitos adversos , Procedimentos Cirúrgicos Cardíacos , Detergentes , Feminino , Hemorragia , Humanos , Cuidados Intraoperatórios , Masculino , Pessoa de Meia-Idade , Plasma/microbiologia , Solventes , Esterilização/métodos , Resultado do TratamentoRESUMO
This study describes a purification protocol of salmon fibrinogen that gives a consumable and highly clottable fibrinogen. Some characteristics of salmon and human fibrinogen are compared. Fibrinogen was purified from barium sulphate adsorbed plasma of Atlantic salmon, using two steps of 25% ammonium sulphate precipitation followed by ultrafiltration. The clottability of the purified salmon fibrinogen was 91%. The Aalpha chains of salmon fibrinogen were heterogeneous with a molecular mass of 90-110 kDa, compared to approximately 67 kDa of human fibrinogen Aalpha chains. The Bbeta and gamma chains of salmon and human fibrinogen had molecular masses of approximately 55 and 50 kDa, respectively. Western blotting revealed that polyclonal rabbit anti-human fibrinogen antibodies had affinity for the gamma chains of salmon fibrinogen, making it possible to study factor XIII activity in purified salmon fibrinogen. Cross-linking of either gamma-gamma or gamma-alpha chains was not detected upon incubation of the purified fibrinogen with thrombin and calcium alone, but was detected when clotting was performed in plasma indicating absence of factor XIII activity in the purified product.