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1.
Sci Rep ; 11(1): 16841, 2021 08 26.
Artigo em Inglês | MEDLINE | ID: mdl-34446727

RESUMO

The pathogenesis of kidney stone formation includes multi-step processes involving complex interactions between mineral components and protein matrix. Calcium-binding proteins in kidney stones have great influences on the stone formation. The spatial distributions of these proteins in kidney stones are essential for evaluating the in vivo effects of proteins on the stone formation, although the actual distribution of these proteins is still unclear. We reveal micro-scale distributions of three different proteins, namely osteopontin (OPN), renal prothrombin fragment 1 (RPTF-1), and calgranulin A (Cal-A), in human kidney stones retaining original mineral phases and textures: calcium oxalate monohydrate (COM) and calcium oxalate dihydrate (COD). OPN and RPTF-1 were distributed inside of both COM and COD crystals, whereas Cal-A was distributed outside of crystals. OPN and RPTF-1 showed homogeneous distributions in COM crystals with mosaic texture, and periodically distributions parallel to specific crystal faces in COD crystals. The unique distributions of these proteins enable us to interpret the different in vivo effects of each protein on CaOx crystal growth based on their physico-chemical properties and the complex physical environment changes of each protein. This method will further allow us to elucidate in vivo effects of different proteins on kidney stone formation.


Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Cálculos Renais/diagnóstico por imagem , Rim/patologia , Osteopontina/metabolismo , Fragmentos de Peptídeos/metabolismo , Precursores de Proteínas/metabolismo , Protrombina/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Oxalato de Cálcio/química , Oxalato de Cálcio/metabolismo , Cristalização/métodos , Feminino , Humanos , Rim/metabolismo , Masculino , Microscopia Eletrônica de Varredura/métodos , Pessoa de Meia-Idade
2.
Biosci Rep ; 41(8)2021 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-34328172

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2)-induced infection, the cause of coronavirus disease 2019 (COVID-19), is characterized by unprecedented clinical pathologies. One of the most important pathologies, is hypercoagulation and microclots in the lungs of patients. Here we study the effect of isolated SARS-CoV-2 spike protein S1 subunit as potential inflammagen sui generis. Using scanning electron and fluorescence microscopy as well as mass spectrometry, we investigate the potential of this inflammagen to interact with platelets and fibrin(ogen) directly to cause blood hypercoagulation. Using platelet-poor plasma (PPP), we show that spike protein may interfere with blood flow. Mass spectrometry also showed that when spike protein S1 is added to healthy PPP, it results in structural changes to ß and γ fibrin(ogen), complement 3, and prothrombin. These proteins were substantially resistant to trypsinization, in the presence of spike protein S1. Here we suggest that, in part, the presence of spike protein in circulation may contribute to the hypercoagulation in COVID-19 positive patients and may cause substantial impairment of fibrinolysis. Such lytic impairment may result in the persistent large microclots we have noted here and previously in plasma samples of COVID-19 patients. This observation may have important clinical relevance in the treatment of hypercoagulability in COVID-19 patients.


Assuntos
COVID-19/patologia , Fibrina/metabolismo , Fibrinólise/fisiologia , Glicoproteína da Espícula de Coronavírus/metabolismo , Trombose/patologia , Adulto , Idoso , Amiloide/metabolismo , Plaquetas/metabolismo , Complemento C3/metabolismo , Feminino , Fibrinogênio/metabolismo , Humanos , Pulmão/patologia , Masculino , Técnicas Analíticas Microfluídicas , Pessoa de Meia-Idade , Protrombina/metabolismo , SARS-CoV-2/metabolismo , Trombose/virologia , Tripsina/metabolismo
3.
Sci Rep ; 11(1): 15572, 2021 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-34330995

RESUMO

Factor (F) VIII deficiency causes bleeding in haemophilia A patients because of the reduced formation of procoagulant enzyme thrombin, which is needed to make the blood clot. We measured the dynamics of coagulation in haemophilia A patients by measuring thrombin generation (TG). Additionally, we quantified the procoagulant process of prothrombin conversion and anticoagulant process of thrombin inhibitor complex formation. In haemophilia A, prothrombin conversion is severely reduced, causing TG to be low. Nevertheless, the thrombin inactivation capacity of these patients is comparable to that in healthy subjects, leading to a severe imbalance between procoagulant and anticoagulant processes and a subsequent increased bleeding risk. A novel therapy in haemophilia A is the targeting of anticoagulant pathway, e.g. thrombin inhibitor antithrombin (AT), to restore the haemostatic balance. We simulated the effect of AT reduction on TG in silico. Lowering AT levels restored TG dose-dependently and an AT reduction of 90-95% led to almost normal TG in most patients . However, the variation in response to AT reduction was large between patients, indicating that this approach should be tailored to each individual patients. Ideally, TG and thrombin dynamics simulation could in the future contribute to the management of patients undergoing AT targeting therapy.


Assuntos
Antitrombinas/farmacologia , Hemofilia A/tratamento farmacológico , Adulto , Coagulação Sanguínea/efeitos dos fármacos , Hemofilia A/metabolismo , Hemofilia B/tratamento farmacológico , Hemofilia B/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Protrombina/metabolismo , Trombina/metabolismo
4.
BMC Cancer ; 21(1): 775, 2021 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-34218801

RESUMO

BACKGROUND: Few biomarkers can predict the efficiency of PD-1 blockade in patients with hepatocellular carcinoma (HCC). This study aimed to investigate the prognostic role of AFP and PIVKA-II in HCC patients receiving anti-PD-1 immunotherapy. METHODS: A total of 235 HCC patients treated with PD-1 blockade were enrolled. Serum AFP and PIVKA-II levels were collected before and after treatments. The patients were divided into groups based on the reduction in AFP and PIVKA-II: AFP reduction ≤50% vs AFP reduction > 50% and PIVKA-II reduction ≤50% vs PIVKA-II reduction > 50%. The primary endpoints included objective response rate (ORR), progression-free survival (PFS) and overall survival (OS). Binary logistic regression analyses were used to explore the related factors of ORR. A Cox proportional hazards model was employed to identify the potential prognostic factors of PFS and OS. RESULTS: Among all the patients, 34.9% (82/235) achieved a complete or partial response. There was a positive correlation between AFP reduction > 50% or PIVKA-II reduction> 50% and the ORR of PD-1 blockade (P < 0.001 and = 0.003). PFS was significantly improved in patients with AFP reduction > 50% and PIVKA-II reduction > 50% (p < 0.001 and = 0.021). In addition, AFP reduction > 50% and PIVKA-II reduction> 50% were positively correlated with longer OS (p = 0.003 and 0.006). CONCLUSION: Early reductions in AFP and PIVKA-II can be predictors of the efficacy of PD-1 blockade in HCC patients.


Assuntos
Biomarcadores Tumorais/sangue , Biomarcadores/metabolismo , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/tratamento farmacológico , Inibidores de Checkpoint Imunológico/uso terapêutico , Imunoterapia/métodos , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/tratamento farmacológico , Precursores de Proteínas/metabolismo , Protrombina/metabolismo , alfa-Fetoproteínas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Inibidores de Checkpoint Imunológico/farmacologia , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Adulto Jovem
5.
PLoS One ; 16(5): e0251656, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34015010

RESUMO

BACKGROUND: Pancreatic adenocarcinoma (PDAC) is an incurable cancer without adequate tumor markers. Our previous study has showed a better diagnostic performance of Protein Induced by Vitamin K Absence II (PIVKA-II) compared to currently used PDAC biomarkers. To corroborate our previous data with a larger sample size and to assess a possible role of PIVKA-II in predicting surgical success. Additionally, to further evaluate the hypothesis of a direct PIVKA-II production by PDAC cells, we examined PIVKA-II tissue expression in a case of PDAC using immunofluorescence. METHODS: We enrolled 76 newly diagnosed PDAC patients and selected 11 patients to determine PIVKA-II levels also after surgical resection. An immunofluorescence (IF) study of PIVKA-II tissue expression was carried out in one of them. PIVKA-II serum values were measured by chemiluminescent enzyme immunoassay method (CLEIA) on LUMIPULSE G1200 (Fujirebio-Europe, Belgium). RESULTS: PIVKA-II serum levels were above the cut-off at baseline in 71 patients (94%) with a median value of 464 mAU/Ml (range 27-40783 mAU/mL); the sensitivity and specificity were 78.67% and 90.67% respectively. Patients with pre-operative PIVKA-II positivity showed a significant decrease (P < 0.015) of median PIVKA-II serum concentrations after surgery: 820 (91-40783) mAU/mL at diagnosis vs 123 (31-4666) mAU/mL post-operatively. IF assay on PDAC sections demonstrated PIVKA-II expression in cancer cells. CONCLUSION: These data are the first showing a decreased PIVKA-II serum levels after surgery in PDAC patients and reporting PIVKA-II expression in PDAC tissue. Further studies are needed to confirm these findings and to determine PIVKA-II usefulness in diagnosing and monitoring PDAC patients.


Assuntos
Biomarcadores Tumorais/metabolismo , Biomarcadores/metabolismo , Neoplasias Pancreáticas , Precursores de Proteínas/metabolismo , Protrombina/metabolismo , Adenocarcinoma/diagnóstico , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Adenocarcinoma/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/cirurgia
6.
Sci Rep ; 11(1): 10738, 2021 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-34031483

RESUMO

Physicians taking care of patients with COVID-19 have described different changes in routine blood parameters. However, these changes hinder them from performing COVID-19 diagnoses. We constructed a machine learning model for COVID-19 diagnosis that was based and cross-validated on the routine blood tests of 5333 patients with various bacterial and viral infections, and 160 COVID-19-positive patients. We selected the operational ROC point at a sensitivity of 81.9% and a specificity of 97.9%. The cross-validated AUC was 0.97. The five most useful routine blood parameters for COVID-19 diagnosis according to the feature importance scoring of the XGBoost algorithm were: MCHC, eosinophil count, albumin, INR, and prothrombin activity percentage. t-SNE visualization showed that the blood parameters of the patients with a severe COVID-19 course are more like the parameters of a bacterial than a viral infection. The reported diagnostic accuracy is at least comparable and probably complementary to RT-PCR and chest CT studies. Patients with fever, cough, myalgia, and other symptoms can now have initial routine blood tests assessed by our diagnostic tool. All patients with a positive COVID-19 prediction would then undergo standard RT-PCR studies to confirm the diagnosis. We believe that our results represent a significant contribution to improvements in COVID-19 diagnosis.


Assuntos
COVID-19/diagnóstico , Aprendizado de Máquina , Idoso , Área Sob a Curva , Biomarcadores/sangue , COVID-19/patologia , COVID-19/virologia , Eosinófilos/citologia , Feminino , Testes Hematológicos , Humanos , Masculino , Protrombina/metabolismo , Curva ROC , SARS-CoV-2/isolamento & purificação , Sensibilidade e Especificidade , Albumina Sérica/análise , Índice de Gravidade de Doença , Tórax/diagnóstico por imagem , Tomografia Computadorizada por Raios X
7.
Theranostics ; 11(9): 4251-4261, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33754059

RESUMO

Rationale: The interaction between coagulation and inflammation resolution remains elusive. We recently highlighted a link between fibrinogen-like protein 2 (Fgl2) and a specialized pro-resolving mediator (SPM)-n-3 docosapentaenoic acid-derived resolvin D5 (RvD5n-3 DPA) in sepsis. This study aimed to investigate the functions of commonly used anticoagulants warfarin, dabigatran and heparin in regulating inflammation resolution. Methods: Peripheral blood was collected from clinical sepsis patients and healthy control for the determination of indicated indexes. Mouse sepsis models of zymosan-induced peritonitis and cecal ligation and puncture (CLP) were employed for the measurement of inflammation- and coagulation-related indexes. Western-blotting, ELISA and flow cytometry were applied to assess proteins. UPLC-MS/MS was used to evaluate lipid metabolites. Results: Here we report that the transmembrane Fgl2 (mFgl2) was positively associated with coagulation, while soluble Fgl2 (sFgl2) level correlated with the enhanced number of peripheral blood mononuclear cells in the sepsis patients. The anticoagulants dabigatran and warfarin attenuated zymosan-induced peritonitis, which was not shared by heparin, while only dabigatran significantly improved sepsis survival in the CLP sepsis mouse model. Although these anticoagulants consistently inhibited pro-inflammatory mediators including prostaglandin E2 and leukotriene B4, only dabigatran increased sFgl2 at both the initiation and resolution phases of inflammation. Mechanistically, dabigatran elicited the shedding of sFgl2 via prothrombin-related metalloproteases, thereby enhanced the subsequent biosynthesis of RvD5n-3 DPA via STAT6-ALOX15 axis. Blocking metalloproteases or ALOX15 significantly impaired dabigatran-enhanced macrophage efferocytosis in vitro, as well as delayed the dabigatran-accelerated inflammation resolution in vivo. Conclusions: Our findings identify the dual anti-inflammatory and pro-resolving actions of dabigatran, through promoting sFgl2-triggered RvD5n-3 DPA production, which has important implications for promoting tissue homeostasis of sepsis.


Assuntos
Dabigatrana/farmacologia , Ácidos Docosa-Hexaenoicos/metabolismo , Ácidos Graxos Insaturados/metabolismo , Fibrinogênio/metabolismo , Inflamação/metabolismo , Animais , Anticoagulantes/farmacologia , Cromatografia Líquida/métodos , Modelos Animais de Doenças , Humanos , Inflamação/tratamento farmacológico , Mediadores da Inflamação/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Metaloproteases/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Protrombina/metabolismo , Sepse/tratamento farmacológico , Sepse/metabolismo , Espectrometria de Massas em Tandem/métodos , Zimosan/farmacologia
8.
Clin Toxicol (Phila) ; 59(10): 905-912, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33641566

RESUMO

OBJECTIVE: This retrospective study investigated the nature and severity of venom-induced consumption coagulopathy (VICC) and determined the clotting factors involved in VICC in patients after envenomation by South Korea's snakes. Additionally, we studied the effectiveness of antivenom for the treatment of VICC after envenomation. METHODS: Included patients were divided into three groups according to the severity of VICC (no VICC, partial VICC, and complete VICC). Data, including changes in coagulation parameters during hospitalization and clotting factors at presentation, were collected and analyzed. RESULTS: One hundred nineteen patients who presented at our emergency department within 3 h after snake envenomation were included. VICC developed in 34 patients (27 patients with partial VICC and 7 patients with complete VICC). Two of 34 patients with VICC required blood transfusions. Five patients with complete VICC had an undetectable fibrinogen concentration at presentation. Three patients with complete VICC had an unmeasurable INR and aPTT within 24 h. The median times of the most extreme values were 10 h for INR, 12 h for aPTT, and 16 h for fibrinogen after presentation in the VICC group. The D-dimer concentration peaked at a median of 63.5 h after presentation. The activities of factors II and X were significantly reduced in the complete VICC group (factor II: 88 (84-99.3)% in the non-VICC group vs. 69 (49.5-83.5)% in the complete VICC group; factor X:94 (83-102) in the non-VICC group vs. 70 (66.5-79.8)% in the complete VICC group), while there was no difference in factor V activity at presentation. The time from bite to first antivenom administration did not correlate with the time course and most extreme concentrations for fibrinogen and D-dimer within the VICC groups. DISCUSSION AND CONCLUSION: VICC occurs in approximately one-quarter of snakebite patients in South Korea; however, VICC itself does not appear to lead to clinical deterioration. Fibrinogen is an early diagnostic maker for complete VICC. Clotting factors II and X are involved in VICC. Future investigations should explore the mechanism of VICC from Korean snakebites and the effect of antivenom on VICC.


Assuntos
Coagulação Sanguínea , Coagulação Intravascular Disseminada/etiologia , Mordeduras de Serpentes/complicações , Venenos de Serpentes/antagonistas & inibidores , Serpentes , Idoso , Animais , Antivenenos/uso terapêutico , Biomarcadores/sangue , Coagulação Sanguínea/efeitos dos fármacos , Testes de Coagulação Sanguínea , Coagulação Intravascular Disseminada/sangue , Coagulação Intravascular Disseminada/diagnóstico , Coagulação Intravascular Disseminada/tratamento farmacológico , Fator X/metabolismo , Feminino , Fibrinogênio/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Protrombina/metabolismo , República da Coreia , Estudos Retrospectivos , Índice de Gravidade de Doença , Mordeduras de Serpentes/sangue , Mordeduras de Serpentes/diagnóstico , Mordeduras de Serpentes/tratamento farmacológico , Venenos de Serpentes/metabolismo , Fatores de Tempo , Resultado do Tratamento
9.
Int J Mol Sci ; 22(5)2021 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-33668986

RESUMO

The outcome of congenital fibrinogen defects (CFD) is often unpredictable. Standard coagulation assays fail to predict the clinical phenotype. We aimed to assess the pheno- and genotypic associations of thrombin generation (TG) and ROTEM in CFD. We measured fibrinogen (Fg) activity and antigen, prothrombin fragments F1+2, and TG by ST Genesia® with both Bleed- and ThromboScreen in 22 patients. ROTEM was available for 11 patients. All patients were genotyped for fibrinogen mutations. Ten patients were diagnosed with hypofibrinogenemia, nine with dysfibrinogenemia, and three with hypodysfibrinogenemia. Among the 17 mutations, eight were affecting the Fg γ chain, four the Fg Bß chain, and five the Fg Aα chain. No statistical difference according to the clinical phenotypes was observed among FGG and FGA mutations. Median F1+2 and TG levels were normal among the different groups. Fg levels correlated negatively with F1+2 and peak height, and positively with lag time and time to peak. The pheno- and genotypes of the patients did not associate with TG. FIBTEM by ROTEM detected hypofibrinogenemia. Our study suggests an inverse link between low fibrinogen activity levels and enhanced TG, which could modify the structure-function relationship of fibrin to support hemostasis.


Assuntos
Afibrinogenemia/sangue , Fibrinogênio/metabolismo , Tromboelastografia/métodos , Trombina/metabolismo , Adulto , Afibrinogenemia/enzimologia , Afibrinogenemia/genética , Idoso , Idoso de 80 Anos ou mais , Transtornos da Coagulação Sanguínea/sangue , Transtornos da Coagulação Sanguínea/genética , Testes de Coagulação Sanguínea , Feminino , Fibrinogênio/genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Fenótipo , Protrombina/metabolismo , Relação Estrutura-Atividade
10.
J Am Heart Assoc ; 10(5): e018243, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33624506

RESUMO

Background White blood cell count, which is inexpensive and widely available in clinical practice, has been proposed to provide prognostic information in coronary artery disease (CAD). Elevated levels of white blood cell subtypes may play different roles in atherothrombosis and predict cardiovascular outcomes. Methods and Results The association between white blood cell counts and mortality was evaluated in 823 subjects with angiographically demonstrated and clinically stable CAD in an observational-longitudinal study. The correlation among white blood cell counts and factor II plasma coagulant activity was analyzed in 750 subjects (554 CAD and 196 CAD-free) not taking anticoagulant drugs. Subjects with overt leukocytosis or leukopenia were excluded. In the longitudinal study after a median follow-up of 61 months, 160 (19.4%) subjects died, 107 (13.0%) of whom from cardiovascular causes. High levels of neutrophils, monocytes, eosinophils, and basophils were associated with an increased mortality rate. In multiadjusted Cox regression models, only neutrophils and basophils remained predictors of total and cardiovascular mortality. The associations remained significant after adjustment for traditional cardiovascular risk factors and by including D-dimer and the chemokine CXCL12 in the regression models. Neutrophils and basophils were also significant predictors of factor II plasma coagulant activity variability after adjustment for blood cell counts, age, sex, inflammatory markers, CAD diagnosis, and prothrombin G20210A polymorphism. Factor II plasma coagulant activity was similarly increased in subjects with high neutrophil and basophil counts and in carriers of the prothrombin 20210A allele. Conclusions Both high neutrophil and basophil blood counts may predict mortality in patients with clinically stable CAD and are associated with enhanced factor II plasma coagulant activity, thereby suggesting underlying prothrombotic mechanisms.


Assuntos
Basófilos/patologia , Doença da Artéria Coronariana/sangue , Isquemia Miocárdica/sangue , Neutrófilos/patologia , Protrombina/metabolismo , Medição de Risco/métodos , Biomarcadores/sangue , Doença da Artéria Coronariana/complicações , Doença da Artéria Coronariana/mortalidade , Feminino , Seguimentos , Humanos , Itália/epidemiologia , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Isquemia Miocárdica/etiologia , Isquemia Miocárdica/mortalidade , Prognóstico , Estudos Retrospectivos , Fatores de Risco , Taxa de Sobrevida/tendências
11.
Biosci Biotechnol Biochem ; 85(4): 824-833, 2021 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-33589932

RESUMO

At the last stage of the blood coagulation cascade, thrombin plays a central role in the processing of fibrinogen for the polymerization and in the additional activation of Factor XIII for the stable cross-linking of fibrin. In addition, thrombin carries out possible multiple roles via processing or interaction with various functional proteins. Several studies conducted in order to elucidate additional physiological significance are ongoing. To clarify further significance of thrombin and to establish an associated disease model, we characterized the orthologue gene for medaka (Oryzias latipes), a research model fish. Tissue distribution of medaka prothrombin has been immunotechnically analyzed. Furthermore, thrombin-deficient medaka mutants were viably established by utilizing a genome-editing method. The established gene-deficient mutants exhibited retarded blood coagulation even in the heterozygous fish. Taking advantage of their ease of handling, this specific model is useful for further investigation in medical research areas on human coagulation diseases.


Assuntos
Transtornos da Coagulação Sanguínea/genética , Trombina/genética , Animais , Edição de Genes , Modelos Animais , Oryzias , Protrombina/metabolismo , Distribuição Tecidual
12.
J Fluoresc ; 31(2): 385-392, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33403518

RESUMO

A new three-dimensional lanthanide metal-organic framework (Ln-MOF), [Eu4(L)4(H2O)8]·10H2O (1, H3L = biphenyl-3'-nitro-3,4',5-tricarboxylic acid), has been constructed via solvothermal technology and its framework has been detected by the single-crystal X-ray diffraction and elemental analyses. Complex 1 with typical emission of Eu3+ ion represents dramatic luminescence quenching affect for picric acid (PA) and the linear Stern-Volmer plot was surveyed in the consistence, ranging from 0.05 to 0.15 mM (Ksv = 98,074 M- 1). Its therapeutic effect of the compound on the cerebral edema caused by cerebral hemorrhage was estimated and the mechanism was explored. Possible binding interactions have been investigated by molecular docking simulations, from which the binding interactions are identified and the carboxyl oxygens are responsible for those identified interactions.


Assuntos
Edema Encefálico/diagnóstico , Hemorragia Cerebral/diagnóstico , Complexos de Coordenação/química , Európio/química , Estruturas Metalorgânicas/química , Picratos/análise , Animais , Edema Encefálico/metabolismo , Hemorragia Cerebral/metabolismo , Complexos de Coordenação/síntese química , Ensaio de Imunoadsorção Enzimática , Estruturas Metalorgânicas/síntese química , Modelos Moleculares , Oxirredução , Protrombina/metabolismo , Ratos , Ratos Sprague-Dawley
13.
Proc Natl Acad Sci U S A ; 118(1)2021 01 05.
Artigo em Inglês | MEDLINE | ID: mdl-33443167

RESUMO

The blood-clotting protein fibrinogen has been implicated in host defense following Staphylococcus aureus infection, but precise mechanisms of host protection and pathogen clearance remain undefined. Peritonitis caused by staphylococci species is a complication for patients with cirrhosis, indwelling catheters, or undergoing peritoneal dialysis. Here, we sought to characterize possible mechanisms of fibrin(ogen)-mediated antimicrobial responses. Wild-type (WT) (Fib+) mice rapidly cleared S. aureus following intraperitoneal infection with elimination of ∼99% of an initial inoculum within 15 min. In contrast, fibrinogen-deficient (Fib-) mice failed to clear the microbe. The genotype-dependent disparity in early clearance resulted in a significant difference in host mortality whereby Fib+ mice uniformly survived whereas Fib- mice exhibited high mortality rates within 24 h. Fibrin(ogen)-mediated bacterial clearance was dependent on (pro)thrombin procoagulant function, supporting a suspected role for fibrin polymerization in this mechanism. Unexpectedly, the primary host initiator of coagulation, tissue factor, was found to be dispensable for this antimicrobial activity. Rather, the bacteria-derived prothrombin activator vWbp was identified as the source of the thrombin-generating potential underlying fibrin(ogen)-dependent bacterial clearance. Mice failed to eliminate S. aureus deficient in vWbp, but clearance of these same microbes in WT mice was restored if active thrombin was administered to the peritoneal cavity. These studies establish that the thrombin/fibrinogen axis is fundamental to host antimicrobial defense, offer a possible explanation for the clinical observation that coagulase-negative staphylococci are a highly prominent infectious agent in peritonitis, and suggest caution against anticoagulants in individuals susceptible to peritoneal infections.


Assuntos
Fibrinogênio/metabolismo , Peritonite/metabolismo , Protrombina/metabolismo , Animais , Antibacterianos/metabolismo , Anti-Infecciosos/metabolismo , Coagulação Sanguínea , Coagulase/metabolismo , Feminino , Fibrina/metabolismo , Fibrinogênio/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/metabolismo , Staphylococcus aureus/patogenicidade , Tromboplastina
14.
J Thromb Thrombolysis ; 51(3): 577-583, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33389608

RESUMO

In recent years a variety of metals (cadmium, chromium, copper, iron) have been demonstrated to modulate coagulation in vitro and in vivo. One group of metals, the platinoids, have not been assessed, and such investigation is justified given the thromboembolic phenomena associated with platinum-based chemotherapy. Thus, the goal of the present investigation was to assess the effects of carboplatin, cisplatin (platinum compounds), NAMI-A, and ruthenium chloride (ruthenium compounds) on human plasmatic coagulation. Human plasma was exposed to clinically relevant, equimolar concentrations of the aforementioned platinum and ruthenium compounds, with changes in plasmatic coagulation assessed via thrombelastography. The first series of experiments demonstrated no significant modulation of coagulation by the platinum compounds, while NAMI-A demonstrated mild hypercoagulability and ruthenium chloride exerted marked hypercoagulability. A second series of experiments utilizing a variety of specialized modifications of thrombelastography focused on ruthenium chloride revealed that this compound enhances prothrombin activation. While the hypercoagulability associated with platinum compounds in vivo do not appear to have a basis in plasmatic biochemistry, it appears that ruthenium compounds can exert procoagulant properties by enhancing the common pathway of human plasmatic coagulation. Future investigation of Ru based chemotherapeutic agents in development to assess procoagulant activity as part of evaluating their potential clinical safety is warranted.


Assuntos
Coagulação Sanguínea , Carboplatina/farmacologia , Cisplatino/farmacologia , Dimetil Sulfóxido/análogos & derivados , Compostos Organometálicos/farmacologia , Protrombina/metabolismo , Compostos de Rutênio/farmacologia , Tromboelastografia/métodos , Antineoplásicos/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Coagulação Sanguínea/fisiologia , Testes de Coagulação Sanguínea/métodos , Dimetil Sulfóxido/farmacologia , Humanos , Compostos de Platina/farmacologia , Trombofilia/sangue , Trombofilia/induzido quimicamente
15.
Oncology ; 99(2): 114-123, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32998139

RESUMO

INTRODUCTION: It remains unclear whether TERT promoter mutation (TERT C228T) in serum cell-free DNA (cfDNA) is useful for the diagnosis of hepatocellular carcinoma (HCC) in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: In this retrospective study, we analyzed the relationships between TERT C228T in serum cfDNA and levels of AFP and PIVKAII in 57 Japanese patients with histopathologically confirmed NAFLD background, consisting of 36 patients with HCC and 21 without HCC. We also examined the liver-related survival rate and HCC recurrence rate after the initial treatment for HCC. TERT C228T was detected using a highly sensitive method based on wild-type blocking PCR (detection limit in excess of 0.7% mutant-type DNA). RESULTS: In all of the 57 patients, multivariate analysis identified TERT C228T positive as significant determinant of primary HCC. In the 36 patients with HCC, the percentage of patients positive for TERT C228T was 63.9%. The percentage of patients positive for TERT C228T with normal AFP and PIVKAII was 35.3%. The positive predictive value and specificity for prediction of BCLC stage 0 or A were both high. In 6 patients, TERT C228T was repeatedly negative during follow-up but became positive at the time of HCC diagnosis. Four patients who underwent HCC surgical resection had well-differentiated solitary HCC measuring <30 mm, and all were TERT C228T positive with normal AFP and PIVKAII. TERT C228T status had no influence on the cumulative liver-related survival rate and HCC recurrence rate. CONCLUSIONS: Our results highlight the superiority of TERT C228T in serum cfDNA compared with AFP and PIVKAII in the early diagnosis of primary HCC in NAFLD patients.


Assuntos
Carcinoma Hepatocelular/diagnóstico , Ácidos Nucleicos Livres/sangue , Neoplasias Hepáticas/diagnóstico , Mutação , Hepatopatia Gordurosa não Alcoólica/genética , Telomerase/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/metabolismo , Biomarcadores Tumorais/genética , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/cirurgia , Feminino , Humanos , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/cirurgia , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/sangue , Prognóstico , Regiões Promotoras Genéticas , Precursores de Proteínas/metabolismo , Protrombina/metabolismo , Estudos Retrospectivos , Análise de Sobrevida , alfa-Fetoproteínas/metabolismo
16.
Pediatr Cardiol ; 42(1): 47-58, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32886153

RESUMO

To develop and internally validate nomogram predicting postoperative blood loss risk among pediatric patients with pulmonary atresia (PA) undergoing cardiopulmonary bypass (CPB). All patients aged from 6 months to 6 years with PA who underwent surgery at Fuwai Hospital from June 2015 to December 2019 were selected. And the prediction nomogram model was developed in the training set based on the selected patients. The demographic characteristics and laboratory data from each enrolled patient were gathered. Postoperative blood loss was defined as a blood loss exceeding 20.0 ml/kg within the first 24 postoperative hours. The least absolute shrinkage and selection operator (LASSO) method was used to optimize feature selection for multivariate logistic regression analysis that was applied to build a nomogram composed of all the features selected in the LASSO algorithm. The concordance index (C-index), calibration plot, and decision curve analysis (DCA) were used to evaluate the discrimination, calibration, and clinical net benefit of the nomogarm, respectively. Finally, internal validation was performed using the bootstrap technique. Of the 66 pediatric patients in the training set, 21 (31.82%) and 45 (68.18%) patients were assigned into bleeding group and non-bleeding group, respectively. The first postoperative 24-h blood loss in the bleeding group was significantly higher than that in the non-bleeding group during ICU stay (P = 0.000). Multivariate logistic regression analysis showed that, the immediate postoperative prothrombin time (odds ratio = 1.419, 95% confidence interval: 1.094-1.841, P = 0.008), the immediate postoperative platelet count (odds ratio = 0.985, 95% confidence interval: 0.973-0.997, P = 0.015) and the immediate postoperative red blood cell (RBC) count (odds ratio = 0.335, 95% confidence interval: 0.166-0.667, P = 0.002) were independent predictors of postoperative blood loss risk. The model presented favorable calibration and good discrimination with satisfactory calibration curve and a C-index of 0.858 (95% confidence interval: 0.758-0.958). High C-index value of 0.837 was achieved in the internal validation. The DCA revealed that the nomogram was great clinical effect when intervention was decided among nearly the entire range of threshold probabilities. We developed and internally validated an accurate nomogram to assist in the clinical decision-making concerning the presence of postoperative blood loss in pediatric patients with PA undergoing CPB. However, the nomogram should be endorsed by external validation before it can be recommended for routine practice.


Assuntos
Ponte Cardiopulmonar/efeitos adversos , Nomogramas , Hemorragia Pós-Operatória/diagnóstico , Atresia Pulmonar/cirurgia , Ponte Cardiopulmonar/métodos , Criança , Pré-Escolar , Tomada de Decisão Clínica , Contagem de Eritrócitos/métodos , Feminino , Humanos , Lactente , Modelos Logísticos , Masculino , Contagem de Plaquetas/métodos , Hemorragia Pós-Operatória/etiologia , Protrombina/metabolismo , Atresia Pulmonar/complicações , Fatores de Risco
17.
Eur Rev Med Pharmacol Sci ; 24(24): 12675-12685, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33378014

RESUMO

OBJECTIVE: Hepatocellular carcinoma (HCC) is a primary liver tumor derived from metabolic or viral chronic hepatitis, with few treatment options in advanced cases. New biomarkers that allow improving diagnosis and staging are widely desired. Here, we aim to evaluate the performance of Protein Induced by Vitamin K Absence or Antagonist-II (PIVKA-II) in combination with α-fetoprotein (AFP), in the diagnosis of HCC in patients with metabolic or viral hepatitis. PATIENTS AND METHODS: We enrolled 60 HCC patients (20 metabolic and 40 viral) and 20 healthy subjects (HS) as negative controls. PIVKA-II, AFP, Matrix metalloproteinase-9 (MMP-9) and Fibroblast growth factor (FGF) serum levels were assessed by immunoassays. RESULTS: AFP and PIVKA-II levels were obviously higher in patients than in HS. AFP displayed a better diagnostic performance than PIVKA-II for viral HCC while PIVKA-II was better for metabolic HCC. The combination of the two biomarkers did not improve the discriminating ability. CONCLUSIONS: PIVKA-II may be considered an independent predictor of macrovascular invasion from HCC cells and it can be used to better stratify HCC patients and should be evaluated in prospective studies for early detection of advanced HCC in metabolic subjects.


Assuntos
Biomarcadores Tumorais/sangue , Biomarcadores/sangue , Carcinoma Hepatocelular/sangue , Neoplasias Hepáticas/sangue , Precursores de Proteínas/sangue , Biomarcadores/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/virologia , Humanos , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/virologia , Projetos Piloto , Precursores de Proteínas/metabolismo , Protrombina/metabolismo , alfa-Fetoproteínas/análise
18.
Mol Reprod Dev ; 87(12): 1206-1218, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33216420

RESUMO

Although the epididymal environment promotes the maturation and survival of spermatozoa, not all spermatozoa remain viable during passage through the epididymis. Does the epididymis has a protective mechanism(s) to segregate the viable sperm from defective spermatozoa? Previously, we identified 260/280 kDa oligomers (termed eFGL-Epididymal Fibrinogen-Like oligomer) are composed of two disulfide-linked subunits: a 64 kDa polypeptide identified as fibrinogen-like protein-2 (FGL2) and a 33 kDa polypeptide identified as fibrinogen-like protein-1 (FGL1). Our morphological studies demonstrated that the eFGL, secreted from the principal cells of the cauda epididymis, is polymerized into a death cocoon-like complex (DCF), masking defective luminal spermatozoa but, not the viable sperm population. In the present study, we purified FGL2 from hamster cauda epididymal fluid toward homogeneity and its prothrombinase catalytic activity was examined. Time-course conversion studies revealed that all prothrombin was converted to thrombin by purified hamster FGL2. Our biochemical studies demonstrate that FGL2 is a lipid-activated serine protease and functions as a lectin by binding specific carbohydrate residues. Co-immunoprecipitation analysis demonstrated that FGL2 of cauda epididymal fluid is ubiquitinated but not the FGL1. We propose that FGL2/FGL1 oligomers represent a novel and unique mechanism to shield the viable sperm population from degenerating spermatozoa contained within the tubule lumen.


Assuntos
Epididimo/metabolismo , Fibrinogênio/metabolismo , Peptídeos/metabolismo , Espermatozoides/metabolismo , Tromboplastina/metabolismo , Animais , Cricetinae , Fibrinogênio/isolamento & purificação , Lectinas/metabolismo , Masculino , Protrombina/metabolismo , Serina Proteases/metabolismo , Trombina/metabolismo
19.
Biomed Res Int ; 2020: 5087643, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33015170

RESUMO

The role of α-fetoprotein (AFP) in the surveillance and diagnosis of hepatocellular carcinoma (HCC) has been questioned in recent years due to its low sensitivity and specificity. In addition to AFP, several new serum biomarkers, such as lens culinaris agglutinin-reactive fraction of AFP (AFP-L3) and des-gamma-carboxy prothrombin (DCP), have also been identified as useful HCC serological markers. However, the exact diagnostic value of the combinations of these biomarkers for detecting HCC in patients with liver disease remains unclear. Thus, we performed the current meta-analysis to assess performance of AFP+AFP-L3%+DCP for diagnosing HCC. Studies were systematically searched in PubMed, Embase, the Cochrane Library, CNKI, and WanFang Data databases. After full-text evaluation, 13 studies from 11 articles focusing on the combination of the three serum biomarkers for HCC detection were enrolled. Random-effects models were used due to the presence of heterogeneity. The pooled sensitivity and specificity for AFP+AFP-L3%+DCP were 88% and 79%, respectively. The area under the summary receiver operating characteristic (sROC) curve was 0.91, and the diagnostic odds ratio (DOR) was 28.33 (95% CI 16.78-47.83). Subgroup analysis showed that the pooled sensitivity and specificity of AFP+AFP-L3%+DCP in the diagnosis of HCC versus cirrhosis patients were 0.81 and 0.82, respectively. In conclusion, the combination of AFP, AFP-L3%, and DCP may prove to be useful in the diagnosis and screening of HCC.


Assuntos
Carcinoma Hepatocelular/diagnóstico , Neoplasias Hepáticas/diagnóstico , Protrombina/metabolismo , alfa-Fetoproteínas/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/metabolismo , Humanos , Neoplasias Hepáticas/metabolismo
20.
Scand J Clin Lab Invest ; 80(8): 694-698, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33026843

RESUMO

Portal vein thrombosis (PVT) is a rare but severe condition. Several risk factors predispose to PVT. However, it remains unclear to which degree thrombophilia contributes to the risk of PVT and whether PVT patients should be routinely referred for thrombophilia testing. The aim of the present study was to investigate the prevalence of thrombophilia in PVT patients to clarify the relevance of thrombophilia testing in PVT patients. Clinical data and results from thrombophilia investigations were systematically obtained from all PVT patients referred to Centre for Hemophilia and Thrombosis, Aarhus University Hospital, Denmark for thrombophilia testing between 1st of January 2010 and 31st of December 2018 (n = 93). The investigated thrombophilias included factor V Leiden and prothrombin G20210A mutations, deficiency of protein S, protein C and antithrombin, antiphospholipid syndrome, and increased levels of factor VIII. The prevalence of thrombophilia was compared to healthy controls obtained from previously published data on thrombophilia distribution in cohorts of the Western European adult general population. Comparing PVT patients with healthy controls, significantly increased odds of presence of lupus anticoagulant (crude odds ratio (OR) 6.2, 95% confidence interval (CI) 1.8-20.6) were found, whereas no significantly increased odds of inherited thrombophilia were demonstrated. In conclusion, routine testing for inherited thrombophilia in PVT patients does not seem indicated. However, PVT patients should still be tested for antiphospholipid antibodies because patients meeting the criteria for antiphospholipid syndrome preferentially should receive vitamin K antagonists as anticoagulant therapy.


Assuntos
Síndrome Antifosfolipídica/diagnóstico , Fator V/metabolismo , Deficiência de Proteína C/diagnóstico , Protrombina/metabolismo , Trombofilia/diagnóstico , Trombose Venosa/diagnóstico , Adulto , Anticoagulantes/uso terapêutico , Síndrome Antifosfolipídica/sangue , Síndrome Antifosfolipídica/epidemiologia , Síndrome Antifosfolipídica/patologia , Antitrombinas/sangue , Estudos de Casos e Controles , Dinamarca/epidemiologia , Fator V/genética , Fator VIII/metabolismo , Feminino , Humanos , Inibidor de Coagulação do Lúpus/sangue , Masculino , Pessoa de Meia-Idade , Mutação , Veia Porta/metabolismo , Veia Porta/patologia , Prevalência , Proteína C/metabolismo , Deficiência de Proteína C/sangue , Deficiência de Proteína C/epidemiologia , Deficiência de Proteína C/patologia , Proteína S/metabolismo , Protrombina/genética , Trombofilia/sangue , Trombofilia/epidemiologia , Trombofilia/patologia , Trombose Venosa/sangue , Trombose Venosa/epidemiologia , Trombose Venosa/patologia
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