The relationship between homocysteine, coagulation dysfunction and breast cancer risk / 中华肿瘤杂志

Objective: To investigate the correlation of homocysteine (HCY) and coagulation function index with the risk of breast cancer and its clinicopathological characteristics. Methods: The HCY, coagulation function test index, and clinicopathological information of female breast cancer patients (333 cases) treated in Tianjin Medical University Cancer Hospital from January 2018 to December 2018 were collected, and female patients with benign breast (225 cases) were selected during the same period for the control group. The t-test was used to compare measurement data with normal distribution, D-Dimer data were distributed discreetly and described by median, non-parametric Mann-Whitney U test was used to compare the two groups. The chi-square test was used to compare enumeration data, and the Logistic regression analysis was used for the risk analysis. Results: The levels of HCY, fibrinogen (Fbg), protein C (PC), and median D-Dimer (D-D) in peripheral blood of breast cancer patients group [(13.26±5.24) μmol/L, (2.61±0.83) g/L, (117.55±19.67)%, and 269.68 ng/ml, respectively] were higher than those in the control group [(11.58±0.69) μmol/L, (2.49±0.49) g/L, (113.42±19.82)% and 246.98 ng/ml, respectively, P<0.05]. The prothrombin time (PT), PT(INR), α2-antiplasmin (α2-AP) levels [(10.19±0.63) s, 0.91±0.07 and (110.64±13.93)%, respectively] were lower than those in the control group [(10.58±0.65) s, 0.93±0.01 and (123.81±14.77) %, P<0.05]. The serum levels of PC and median D-D in premenopausal breast cancer patients [(112.57±17.86)% and 242.01 ng/ml, respectively] were higher than those in the control group [(105.31±22.31)% and 214.75 ng/ml, respectively, P<0.05]. The levels of PT(INR), α2-AP [0.91±0.07 and (111.29±12.54)%, respectively] were lower than those of the control group[0.98±0.15 and (120.17±16.35)%, respectively, P<0.05]. The levels of HCY and median D-D in postmenopausal breast cancer patients [(14.25±5.76) μmol/L and 347.53 ng/ml, respectively] were higher than those in the control group [(11.67±2.38) μmol/L and 328.28 ng/ml, P<0.05]. The levels of PT, PT(INR), antithrombin Ⅲ (AT-Ⅲ), α2-AP levels [(10.18±0.66) s, 0.87±0.09, (97.30±12.84)% and (110.13±14.96)%] were lower than those in the control group [(10.38±0.61) s, 0.90±0.08, (102.89±9.12)%, and (127.05±12.38)%, respectively, P<0.05]. The levels of α2-AP and median D-D in T2-4 stage breast cancer patients [(111.69±14.41)% and 289.25 ng/ml, respectively] were higher than those in Tis-1 stage patients [(108.05±12.37)% and 253.49 ng/ml, respectively, P<0.05]. The levels of PT, PT (INR), Fbg, AT-Ⅲ, α2-AP, median D-D [(10.62±0.63) s, 0.95±0.06, (3.04±1.52) g/L, (103.21±9.45)%, (118.72±14.77)% and 331.33 ng/ml, respectively] in breast cancer patients with lymph node metastasis were higher than those of patients without lymph node metastasis [(10.42±0.58) s, 0.93±0.06, (2.52±0.54) g/L, (95.20±13.63)%, (106.91±13.13)% and 263.38 ng/ml, respectively, P<0.05]. In non-menopausal breast cancer patients, the level of HCY [(12.63±4.41) μmol/L] in patients with T2-4 stage was higher than that of patients with Tis-1 stage [(10.70±3.49) μmol/L, P=0.010], and the level of thrombin time [(19.35±0.90) s] of patients with T2-4 stage was lower than that of patients with Tis-1 stage [(19.79±1.23) s, P=0.015]. The levels of PT(INR), Fbg, AT-Ⅲ, α2-AP [0.97±0.56, (3.37±2.34) g/L, (102.38±8.77)% and (120.95±14.06)%] in patients with lymph node metastasis were higher than those of patients without lymph node metastasis [0.94±0.05, (2.36±0.48) g/L, (94.56±14.37)% and (109.51±11.46)%, respectively, P<0.05]. Among postmenopausal breast cancer patients, the levels of AT-Ⅲ and α2-AP in T2-4 stage patients [(98.48±11.80)% and (111.84±15.35)%, respectively] were higher than those in patients with the Tis-1 stage [(94.12±14.98)% and (105.49±12.89)%, respectively, P<0.05]. The levels of AT-Ⅲ and α2-AP in N1-3 stage patients [(103.74±9.94)% and (117.29±15.23)%] were higher than those in N0 stage patients [(95.75±13.01)% and (108.39±14.42)%, P<0.05]. Conclusions: HCY and abnormal coagulation function are related to the risk of breast cancer, T stage and lymph node metastasis in breast cancer patients.

A novel prognostic index in patients with resectable esophageal squamous cell carcinoma: fibrinogen/prealbumin ratio

ABSTRACT Background: Fibrinogen (Fib) to albumin (ALB) fibrinogen-to-albumin ratio as a prognostic index for esophageal cancer has been confirmed. A novel prognostic index was initially proposed with fibrinogen to prealbumin ratio (FPR) in patients with resectable esophageal squamous cell carcinoma (ESCC). Objective: The objective of the study was to study the prognostic role of the novel prognostic index (FPR) in patients with resectable ESCC without any neoadjuvant treatment. Methods: In this retrospective study, a total of 372 resectable ESCC patients without any neoadjuvant treatment were included. The best cutoff values were selected by the receiver operating characteristic curves. Two Cox regression analyses with forward stepwise (one for categorical variables and the other for continuous variables) were used to evaluate the overall survival (OS) and cancer-specific survival (CSS). Results: The best cutoff point was 0.014 for FPR. Patients with lower levels of FPR (≤0.014) had better CSS (50.7% vs. 18.0%, p < 0.001) and OS (48.0% vs. 17.6%, p < 0.001) than patients with higher levels of FPR (> 0.014). Multivariate Cox analyses (categorical and continuous) demonstrated that FPR was an independent prognostic factor in CSS (categorical: hazard ratio [HR]: 2.014, 95% confidence interval [CI]: 1.504-2.697, p < 0.001; continuous per 0.01: HR: 1.438, 95% CI: 1.154-1.793, p = 0.001) and OS (categorical: HR: 1.964, 95% CI: 1.475-2.617, p < 0.001; continuous per 0.01: HR: 1.429, 95% CI: 1.146-1.781, p = 0.002). Conclusions: Our study indicated that FPR served as an independent prognostic factor in patients with resectable ESCC.

Pretreatment plasma fibrinogen level as a prognostic biomarker for patients with lung cancer

Many researchers have shown that pretreatment plasma fibrinogen levels are closely correlated with the prognosis of patients with lung cancer (LC). In this study, we thus performed a meta-analysis to systematically assess the prognostic value of pretreatment plasma fibrinogen levels in LC patients. A computerized systematic search in PubMed, EMBASE, Web of Science and China National Knowledge Infrastructure (CNKI) was performed up to March 15, 2018. Studies with available data on the prognostic value of plasma fibrinogen in LC patients were eligible for inclusion. The pooled hazard ratios (HRs) and odd ratios (ORs) with 95% confidence intervals (CIs) were used to evaluate the correlation between pretreatment plasma fibrinogen levels and prognosis as well as clinicopathological characteristics. A total of 17 studies with 6,460 LC patients were included in this meta-analysis. A higher pretreatment plasma fibrinogen level was significantly associated with worse overall survival (OS) (HR: 1.57; 95% CI: 1.39-1.77; p=0.001), disease-free survival (DFS) (HR: 1.53; 95% CI: 1.33-1.76; p=0.003), and progression-free survival (PFS) (HR: 3.14; 95% CI: 2.15-4.59; p<0.001). Furthermore, our subgroup and sensitivity analyses demonstrated that the pooled HR for OS was robust and reliable. In addition, we also found that a higher fibrinogen level predicted advanced TNM stage (III-IV) (OR=2.18, 95% CI: 1.79-2.66; p<0.001) and a higher incidence of lymph node metastasis (OR=1.74, 95% CI: 1.44-2.10; p=0.02). Our study suggested that higher pretreatment plasma fibrinogen levels predict worse prognoses in LC patients.

Efeito compensatório do fibrinogênio em paciente com aplasia de medula óssea, choque séptico e trombocitopenia grave guiado por tromboelastometria: relato de caso / Compensatory effect of fibrinogen in a patient with bone marrow aplasia, septic shock, and severe thrombocytopenia guided by thromboelastometry: a case report

RESUMO A transfusão de concentrado de plaquetas é prática comum para prevenção de sangramento espontâneo ou decorrente de procedimentos invasivos; sabe-se que a transfusão de componentes alogênicos do sangue se associa a aumento da mortalidade e piora do desfecho clínico. A força do coágulo é avaliada por meio da tromboelastometria rotacional e determinada pela interação entre plaquetas e fibrinogênio. O efeito compensatório do incremento na concentração sérica de fibrinogênio na força do coágulo, em pacientes com trombocitopenia, tem sido demonstrado em diferentes contextos clínicos, incluindo sepse. Relatamos o caso de uma paciente com trombocitopenia grave, cujo resultado da tromboelastometria rotacional demonstrou efeito compensatório na força do coágulo determinada pelos níveis plasmáticos aumentados de fibrinogênio como reagente de fase aguda em pacientes sépticos. Relatamos o caso de uma paciente de 62 anos com diagnóstico de aplasia de medula óssea admitida a uma unidade de terapia intensiva com choque séptico e trombocitopenia grave. Nas primeiras 24 horas na unidade de terapia intensiva, ela apresentou quadro clínico de insuficiência respiratória aguda e choque. Foi necessário utilizar ventilação mecânica invasiva e fármaco vasoativo. A radiografia de tórax mostrou padrão de lesão pulmonar bilateral. Desta forma, foi solicitada broncoscopia com lavagem broncoalveolar para investigação diagnóstica. Conduziu-se uma tromboelastometria rotacional, e seu resultado mostrou perfil de coagulação normal. Apesar da trombocitopenia grave (1.000/mm3), os níveis de fibrinogênio aumentaram (1.050mg/dL) devido ao choque séptico. A broncoscopia foi realizada sem que subsequentemente ocorresse sangramento ativo. Este caso relata o uso da tromboelastometria como ferramenta diagnóstica em distúrbios da coagulação de pacientes graves, permitindo prevenir o uso desnecessário de transfusões profiláticas de concentrado de plaquetas.

ABSTRACT Platelet transfusion is a common practice to prevent spontaneous bleeding or bleeding due to invasive procedures. Transfusion of allogeneic blood components is associated with increased mortality and a worse clinical outcome. The clot strength is assessed by thromboelastometry and determined by the interaction between platelets and fibrinogen. The compensatory effect of high levels of fibrinogen on clot strength in patients with thrombocytopenia has been demonstrated in different clinical settings including sepsis. We report the case of a patient with severe thrombocytopenia whose thromboelastometry showed clot strength that was compensated for by the increase in plasma fibrinogen levels as an acute phase reactant of septic patients. Here, we report a case of a 62-year-old female diagnosed with bone marrow aplasia admitted in the intensive care unit with septic shock and severe thrombocytopenia. During the first 24 hours in the intensive care unit, she presented acute respiratory insufficiency and circulatory shock. The use of invasive mechanical ventilation and norepinephrine was required. Her chest X-ray showed bilateral lung injury. Thus, bronchoscopy with bronchoalveolar lavage was requested. Thromboelastometry was performed and resulted in a normal coagulable profile. Despite severe thrombocytopenia (1,000/mm3), fibrinogen levels were increased (1,050mg/dL) due to septic shock. Bronchoscopy was performed without any active or further bleeding. Here, we report the use of thromboelastometry in the diagnosis of coagulation disorders, preventing unnecessary prophylactic platelet transfusion.

Evaluación de parámetros inflamatorios locales y sistémicos de quemadura periférica en un modelo animal / Assessment of local and systemic inflammatory parameters of peripheral burn in an animal model

RESUMEN Evaluar el volumen de edema y el recuento leucocitario, plaquetario y de fibrinógenos de quemadura periférica en un modelo animal. Se introdujo en agua a 60 °C y durante 60 s, la pata posterior izquierda de Rattus rovergicus (grupo experimental) o a temperatura ambiente (grupo control). Se realizó el análisis antes y después de la quemadura inducida (a las 4, 8, 12 y 24 h). El volumen del edema se determinó por fotografía ortogonal, los leucocitos y el recuento plaquetario, en un equipo automatizado, y el fibrinógeno por el método gravimétrico. El valor máximo del edema se obtuvo a las 4 h, y de leucocitos a las 24 h. El recuento plaquetario no varió a los diferentes intervalos de tiempo posinflamación. El fibrinógeno se incrementó a las 4 h y 24 h. Este modelo animal induce inflamación sistémica caracterizada por leucocitosis, nivel elevado de fibrinógeno y edema localizado en la zona de inducción.

ABSTRACT To evaluate the edema volume and leukocyte, platelet, and fibrinogen count of peripheral burn in an animal model. The back left leg of Rattus norvegicus (experimental group) was placed in water at 60 °C for 60 seconds or at room temperature (control group). An analysis was carried out before and after the induced burn (at 4, 8, 12, and 24 h). The edema volume was determined by an orthogonal photo, the leukocyte and platelet counts were determined using automated equipment, and the fibrinogen count was determined using the gravimetric method. The maximum value of the edema was recorded at 4 h and leukocytes at 24 h. The platelet count did not vary at different post-edema time intervals. The fibrinogen level increased at 4 h and 24 h. In this animal model we induced systemic inflammation characterized by leukocytosis and elevated fibrinogen levels, combined with edema located at the induction area.

Thrombin activatable fibrinolysis inhibitor como preditor de sangramento no transplante hepático: estudo piloto observacional / Thrombin activatable fibrinolysis inhibitor as a bleeding predictor in liver transplantation: a pilot observational study

RESUMO Objetivo: Correlacionar os níveis de thrombin activatable fibrinolysis inhibitor no pós-operatório imediato e com 24 horas de pós-operatório com o volume de sangramento tansoperatório. Métodos: Foram analisados vinte e um pacientes alocados imediatamente antes do transplante hepático (eletivo ou de urgência), com coleta de amostras sanguíneas para análise de thrombin activatable fibrinolysis inhibitor em três diferentes momentos: imediatamente antes do transplante hepático (thrombin activatable fibrinolysis inhibitor pré-operatório), imediatamente após o procedimento cirúrgico (thrombin activatable fibrinolysis inhibitor pós-operatório imediato) e após 24 horas do final da cirurgia (thrombin activatable fibrinolysis inhibitor 24 horas pós-operatório). O principal desfecho do estudo foi correlacionar os níveis de thrombin activatable fibrinolysis inhibitor pré-operatório e de thrombin activatable fibrinolysis inhibitor pós-operatório imediato com perda sanguínea no transoperatório. Resultados: Houve correlação entre thrombin activatable fibrinolysis inhibitor pré-operatório e o volume de sangramento (ρ = -0,469; p = 0,05), mas não de thrombin activatable fibrinolysis inhibitor pós-operatório imediato (ρ = -0,062; p = 0,79). Em análise de regressão linear, nenhuma das variáveis incluídas (hemoglobina pré, fibrinogênio pré e thrombin activatable fibrinolysis inhibitor pré-operatório) se mostrou preditor de sangramento. Houve tendência semelhante na variação entre os níveis de thrombin activatable fibrinolysis inhibitor durante os três diferentes momentos e os níveis de fibrinogênio. Pacientes que evoluíram a óbito em até 6 meses (14,3%) apresentaram níveis diminuídos de thrombin activatable fibrinolysis inhibitor pré-operatório e de thrombin activatable fibrinolysis inhibitor pós-operatório imediato, comparando-se aos sobreviventes (pré-operatório: 1,3 ± 0,15 versus 2,55 ± 0,53; p = 0,06; e pós-operatório imediato: 1,2 ± 0,15 versus 2,5 ± 0,42; p = 0,007). Conclusão: Houve correlação moderada entre thrombin activatable fibrinolysis inhibitor pré-operatório e o sangramento transoperatório em transplante hepático, porém seu papel preditivo independente de outras variáveis ainda permaneceu incerto. Thrombin activatable fibrinolysis inhibitor pré-operatório e pós-operatório imediato podem ter um papel na avaliação da sobrevida dessa população, necessitando-se confirmar em novos estudos, de maior tamanho amostral.

ABSTRACT Objective: To correlate the levels of thrombin activatable fibrinolysis inhibitor in the immediate postoperative period and at 24 hours postoperatively with the volume of intraoperative bleeding. Methods: Twenty-one patients allocated immediately before (elective or emergency) liver transplantation were analyzed. Blood samples were collected for thrombin activatable fibrinolysis inhibitor analysis at three different time points: immediately before liver transplantation (preoperative thrombin activatable fibrinolysis inhibitor), immediately after the surgical procedure (immediate postoperative thrombin activatable fibrinolysis inhibitor), and 24 hours after surgery (thrombin activatable fibrinolysis inhibitor 24 hours after surgery). The primary outcome of the study was to correlate the preoperative and immediate postoperative levels of thrombin activatable fibrinolysis inhibitor with intraoperative blood loss. Results: There was a correlation between the preoperative thrombin activatable fibrinolysis inhibitor levels and bleeding volume (ρ = -0.469; p = 0.05) but no correlation between the immediate postoperative thrombin activatable fibrinolysis inhibitor and bleeding volume (ρ = -0.062; p = 0.79). No variable included in the linear regression analysis (prehemoglobin, prefibrinogen and preoperative thrombin activatable fibrinolysis inhibitor) was a bleeding predictor. There was a similar trend in the variation between the levels of thrombin activatable fibrinolysis inhibitor at the three different time points and fibrinogen levels. Patients who died within 6 months (14.3%) showed decreased preoperative and immediate postoperative levels of thrombin activatable fibrinolysis compared with survivors (preoperative: 1.3 ± 0.15 versus 2.55 ± 0.53, p = 0.06; immediate postoperative: 1.2 ± 0.15 versus 2.5 ± 0.42, p = 0.007). Conclusion: There was a moderate correlation between preoperative thrombin activatable fibrinolysis inhibitor and intraoperative bleeding in liver transplantation patients, although the predictive role of this variable independent of other variables remains uncertain. Preoperative and immediate postoperative thrombin activatable fibrinolysis inhibitor levels may have a role in the survival prognosis of this population; however, this possibility requires confirmation in further studies with larger sample sizes.

Differential expression of CC chemokines (CCLs) and receptors (CCRs) by human T lymphocytes in response to different Aggregatibacter actinomycetemcomitans serotypes

ABSTRACT In Aggregatibacter actinomycetemcomitans, different serotypes have been described based on LPS antigenicity. Recently, our research group has reported a differential immunogenicity when T lymphocytes were stimulated with these different serotypes. In particular, it was demonstrated that the serotype b of A. actinomycetemcomitans has a stronger capacity to trigger Th1- and Th17-type cytokine production. Objective This study aimed to quantify the expression of different CC chemokines (CCLs) and receptors (CCRs) in T lymphocytes stimulated with the different A. actinomycetemcomitans serotypes. In addition, the expression of the transcription factors T-bet, GATA-3, RORC2, and Foxp3, master-switch genes implied in the Th1, Th2, Th17, and T-regulatory differentiation, respectively, was analyzed in order to determine T-cell phenotype-specific patterns of CCL and CCR expression upon A. actinomycetemcomitans stimulation. Material and Methods Human naïve CD4+ T lymphocytes were obtained from healthy subjects and stimulated with autologous dendritic cells primed with the different A. actinomycetemcomitans serotypes. The expression levels for the chemokines CCL1, CCL2, CCL3, CCL5, CCL11, CCL17, CCL20, CCL21, CCL25, and CCL28, as well as the chemokine receptors CCR1, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, and CCR10 were quantified by qPCR. Similarly, the expression levels for the transcription factors T-bet, GATA-3, RORC2, and Foxp3 were quantified and correlated with the CCL and CCR expression levels. Results Higher expression levels of CCL2, CCL3, CCL5, CCL20, CCL21, CCL28, CCR1, CCR2, CCR5, CCR6, CCR7, and CCR9 were detected in T lymphocytes stimulated with the serotype b of A. actinomycetemcomitans compared with the other serotypes. In addition, these higher expression levels of CCLs and CCRs positively correlated with the increased levels of T-bet and RORC2 when T lymphocytes were stimulated with the serotype b. Conclusion A T-lymphocyte response biased towards a Th1- and Th17-pattern of CCL and CCR expression was detected under stimulation with the serotype b of A. actinomycetemcomitans.

Fibrinogen, bFGF and VEGF levels during antibiotic therapy in gynecologic cancer: A preliminary report.

The role of angiogenesis in the development of neoplasia has been identified and characterized. However, anti-angiogenic therapeutic intervention still requires more evidence to become recognized and successful. The aim of this study was to evaluate levels of selected proangiogenic factors, such as fibrinogen, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in serum of patients with the gynecologic cancer on the first, third and sixth day of antibiotic therapy, routinely administered as a perioperative treatment. In addition, serum concentrations of γ-γ dimers and α-polymers of cross-linked fibrin structure and the degree of bFGF binding with the fibrin network were investigated. Immunohistochemistry staining of the excised tumor tissue was also performed. We observed higher levels of bFGF, VEGF, as well as fibrinogen in patients with gynecologic malignancy, as compared to healthy women. In cancer patients, the concentration of α-polymers and γ-γ dimers of fibrin network increased. Further only γ-γ dimers fraction of fibrin was found to bind to bFGF. Immunohistochemical analysis indicated the presence of bFGF in an excised tumor tissue. In conclusion, the decrease of proangiogenic bFGF and fibrinogen levels in a clinical trial of gynecologic patients may confirm anti-angiogenic properties of selected antibiotic therapy.

Training-induced changes in clotting parameters of athletic horses

The purpose of this study was to investigate the effects of training on prothrombin time, activated partial thromboplastin time, and fibrinogen (Fb) concentrations in horses to assess potential adaptive response to training. Fifteen clinically healthy horses were enrolled in the present study and equally divided into three groups. Group A completed an intense training program, group B participated in a light training program, and group C included sedentary horses. After 5 weeks, group B was subjected to the same training program completed by group A and renamed group B1. Blood samples were collected by jugular venipuncture from each animal at rest and analyzed within 2 h after sampling. A two-way ANOVA for repeated measures showed a significant effect of training (p < 0.05) on Fb concentrations in group B1 alone during the first week after changing the training program. Our findings demonstrated that Fb is a parameter susceptible to training. Fb plasma levels increase with a more intense training program. However, Fb plasma levels decreased after the first week and returned to basel levels, suggesting that the horses had adapted to the new training program.

Proteinograma sérico, com ênfase em proteínas de fase aguda, de bovinos sadios e bovinos portadores de enfermidade aguda de ocorrência natural / Serum proteinogram emphasizing acute phase proteins from healthy and acute naturally occurring diseased cattle

Nas últimas décadas, as proteínas de fase aguda (PFAs) tornaram-se biomarcadores de escolha em medicina humana para identificação e monitoração de doenças. Não há razão para imaginar que tais pesquisas clínicas não sejam igualmente úteis na medicina veterinária. Com o objetivo de verificar a importância das PFAs como biomarcadores de doenças inflamatórias em bovinos, determinou-se o proteinograma sérico, por meio da técnica de eletroforese SDS-PAGE, com interesse especial nas PFAs. Foram utilizados 30 animais, distribuídos em dois grupos: 15 bovinos sadios e 15 bovinos doentes (cinco com mastite estafilocócica, cinco com fotossensibilização e cinco com onfaloflebite). Os animais foram submetidos a colheitas diárias de sangue durante sete dias, enquanto internados no Hospital Veterinário da Unesp, Campus de Jaboticabal. Ceruloplasmina e haptoglobina apresentaram elevação significativa em animais acometidos por mastite, fotossensibilização e onfaloflebite (275,17% e 343,71%; 175,17% e 230,19%; 114,47% e 144,47%, respectivamente). A α1-glicoproteína ácida foi um bom biomarcador apenas em animais com mastite e fotossensibilização, elevando, respectivamente, suas concentrações séricas em 198,14% e 145,89%. Fibrinogênio mostrou-se um indicador confiável apenas em bovinos com mastite, com elevação de 146,5% em relação ao grupo sadio. Ficou clara a diferença na responsividade de distintas PFAs frente a diferentes estímulos inflamatórios. Ceruloplasmina e haptoglobina foram biomarcadores mais sensíveis e, portanto, mais confiáveis entre as PFAs estudadas nessa espécie.

Over the last few decades acute phase proteins (APP) have become the biomarkers of choice in human medicine to identify and monitor inflammation and infection. There is no reason to suppose that clinical investigations in veterinary medicine would not be equally assisted by APP assays. Aiming to verify the importance of APPs as biomarkers of inflammatory diseases in domestic cattle, serum protein profiles, especially APPs, were determined through the SDS-PAGE electrophoresis technique. Thirty animals were allotted in two groups: 15 healthy cattle and 15 clinically ill cattle (5 with staphylococcal mastitis, 5 with photosensitization and 5 with onphalophlebitis). All animals were submitted to daily blood sampling during 7 days, while interned in the Veterinarian Hospital from UNESP, Jaboticabal campus. Ceruloplasmin and haptoglobin were significantly elevated in animals with mastitis, photosensitization and onphalophlebitis (275.17% and 343.71%; 175.17% and 230.19%; 114.47% and 144.47%, respectively). α1-acid glycoprotein behaved as a good biomarker only in animals with mastitis and photosensitization, elevating respectively 198.14% and 145.89% of their serum levels. Fibrinogen was a reliable indicator only in animals undergoing mastitis, with a raise of 146.5%. The diverse responsiveness of different APP under distinct inflammatory stimuli was clear. Ceruloplasmin and haptoglobin were more sensible and, therefore, reliable biomarkers to the diseases studied in this species.

Interaction between Paracoccidioides brasiliensis conidia and the coagulation system: involvement of fibrinogen

The infectious process starts with an initial contact between pathogen and host. We have previously demonstrated that Paracoccidioides brasiliensis conidia interact with plasma proteins including fibrinogen, which is considered the major component of the coagulation system. In this study, we evaluated the in vitro capacity of P. brasiliensis conidia to aggregate with plasma proteins and compounds involved in the coagulation system. We assessed the aggregation of P. brasiliensis conidia after incubation with human serum or plasma in the presence or absence of anticoagulants, extracellular matrix (ECM) proteins, metabolic and protein inhibitors, monosaccharides and other compounds. Additionally, prothrombin and partial thromboplastin times were determined after the interaction of P. brasiliensis conidia with human plasma. ECM proteins, monosaccharides and human plasma significantly induced P. brasiliensis conidial aggregation; however, anticoagulants and metabolic and protein inhibitors diminished the aggregation process. The extrinsic coagulation pathway was not affected by the interaction between P. brasiliensis conidia and plasma proteins, while the intrinsic pathway was markedly altered. These results indicate that P. brasiliensis conidia interact with proteins involved in the coagulation system. This interaction may play an important role in the initial inflammatory response, as well as fungal disease progression caused by P. brasiliensis dissemination.

Fresh Frozen Plasma in Pump Priming for Congenital Heart Surgery: Evaluation of Effects on Postoperative Coagulation Profiles Using a Fibrinogen Assay and Rotational Thromboelastometry

PURPOSE: In this prospective study, the effects of fresh frozen plasma (FFP) included in pump priming for congenital heart surgery in infants and children on post-bypass coagulation profiles were evaluated. MATERIALS AND METHODS: Either 20% albumin (50-100 mL) or FFP (1-2 units) was added to pump priming for patients randomly allocated into control or treatment groups, respectively. Hematologic assays, including functional fibrinogen level, and rotational thromboelastometry (ROTEM(R)) were measured before skin incision (baseline), after weaning from cardiopulmonary bypass (CPB) and heparin reversal, and at 24 hours (h) in the intensive care unit (ICU). RESULTS: All the baseline measurements were comparable between the control and treatment groups of infants and children. After heparin reversal, however, significantly higher fibrinogen levels and less reduced ROTEM parameters, which reflect clot formation and firmness, were demonstrated in the treatment groups of infants and children. At 24 h in the ICU, hematologic assays and ROTEM measurements were comparable between the control and treatment groups of infants and children. Transfusion requirements, excluding FFP in pump prime, and postoperative bleeding were comparable between the control and treatment groups of infants and children. CONCLUSION: Although clinical benefits were not clearly found, the inclusion of FFP in pump priming for congenital heart surgery in infants and children was shown to improve the hemodilution-related hemostatic dysfunction immediately after weaning from CPB and heparin reversal.

Porcine uterine and placental extracellular matrix molecules throughout pregnancy / Moléculas de la matríz extracelular placentaria y uterina durante la preñez porcina

The molecules that constitute the extracellular matrix are important in several functions related to tissue support and cell-cell, cell-extracellular matrix interaction. Among the macromolecules that constitute the mentioned matrix we find osteopontin, fibrinogen and collagen. The present study was undertaken to analyze the rol of osteopontin, fibrinogen and collagen in uterine-placental interface during normal porcine gestation. Uterine and placental tissues from crossbred gilts of 30 (n=5), 60 (n=5), 70 (n=5) and 114 (at term, n=5) days of gestation were used. Macroscopic analysis of the embryos/fetuses allowed us to determine their gestational age by means of the crown-rump lenght. Haematoxylin-Eosin and Masson's Trichrome dyes along with light microscopy were used to structure analysis of every selected period of gestation. A spacial and temporal study of osteopontin and fibrinogen was performed through immunohistochemical technique. Determination of collagen fibers was carried out through Picrosirius red technique and polarizing microscopy. Results were expressed as semi-quantitative. Higher expression of osteopontin was observed at early periods of gestation, mainly in uterine and placental villi, endometrial gland epithelium and histotroph. Fibrinogen expressed abundantly in fetal mesenchyme in every period analyzed and in fetal and maternal vessels at Day 70. Negative expression of collagen fibers was observed in villi, however increasing expression of thick fibers throughout pregnancy was detected in uterine stroma and myometrium. These results confirm the importance of osteopontin, fibrinogen and collagen in the support of uterine and placental structures and in the suitable maintenance of pregnancy.

Las moléculas que constituyen la matriz extracelular son importantes en varias funciones relacionadas con el soporte del tejido y la interacción célula-célula, célula-matriz extracelular. Entre las macromoléculas que constituyen la matriz mencionada se encuentra la osteopontina, el fibrinógeno y el colágeno. Este estudio se realizó para analizar el rol de la osteopontina, el fibrinógeno y el colágeno en la interface útero-placentaria durante la gestación porcina normal. Tejidos uterinos y la placentarios de hembras porcinas cruzadas de 30 (n=5), 60 (n=5), 70 (n=5) y 114 (a término, n=5) días de gestación fueron utilizados. El análisis macroscópico de los embriones/fetos nos permitió determinar la edad gestacional por medio de la longitud cráneo-rabadilla. Tinciones de Hematoxilina-Eosina y Tricrómico de Masson con microscopía de luz se utilizó para estructurar el análisis de cada periodo de tiempo seleccionado de la gestación. Un estudio espacial y temporal de la osteopontina y el fibrinógeno se realizó mediante técnicas de inmunohistoquímica. La determinación de fibras colágenas se llevó a cabo a través de la técnica Picrosirius rojo por microscopia de polarización. Los resultados se expresaron como semi-cuantitativos. La expresión de osteopontina se observó en los primeros períodos de gestación, principalmente en las vellosidades del útero y la placenta, epitelio de las glándulas endometriales e histotrofos. El fibrinógeno se expresa abundantemente en mesénquima fetal en todos los períodos analizados y en los vasos fetales y maternos el día 70. Una expresión negativa de fibras colágenas se observó en las vellosidades, sin embargo, un aumento de expresión de las fibras gruesas durante la gestación se detectó en el estroma uterino y el miometrio. Estos resultados confirman la importancia de la osteopontina, fibrinógeno y colágeno en el soporte de las estructuras del útero y placenta, así como el mantenimiento adecuado durante la gestación.

Effect of natural honey on human platelets and blood coagulation proteins

Present study was conducted to determine the effects of honey on blood hemostasis, in-vitro effect of honey was observed on platelet aggregation and blood coagulation employing, activated partial prothrombin time [aPTT], prothrombin time [PT], thrombin time [TT] and fibrinogen levels in blood. Honey samples showed moderate inhibition of platelet aggregation with IC[50] 5-7.5%. The coagulation assays showed that at higher concentrations [>/= 15%] honey samples increased whole blood clotting time. When assayed in platelet poor plasma [PPP], honey samples significantly [P >/= 0.005] prolonged aPTT, PT, and TT. The honey samples [at 3.75% and 7.5% concentrations] cause mean increment of aPTT = 19 +/- 10% and 62 +/- 10%; PT 6 +/- 5% and 40 +/- 5%; TT 35 +/- 15% and 112 +/- 30% respectively. Moreover, PPP isolated from whole blood pre-incubated with honey samples [9.0% for 10 minutes] showed mean prolongation of aPTT, PT and TT of 45 +/- 21%, 26 +/- 9% and 105 +/- 24% respectively. Interestingly, incubation of honey at 6.25% and 11.75% concentrations in PPP considerably [P >/= 0.005] reduced fibrinogen levels i.e. 13 +/- 4% and 86 +/- 30% respectively. The present study outlines the inhibitory effect of natural honey on platelet aggregation and blood coagulation. These observations provide first line data for modulatory role [s] of honey on process of hemostasis

Hemophilia a and congenital Hypofibrinogenemia: A rare association in same family.

Hemophilia A is the commonest inherited coagulation defect in human beings, whereas congenital hypofibrinogenemia is a much rarer disease. Occurrence of these two inherited diseases in the same family has not been reported so far. Younger sibling of a known case of Hemophilia A presented with recurrent, spontaneously occurring echymotic spots having prolonged PT, APTT, TT and very low absolute fibrinogen level with normal factor VIII level ultimately diagnosed as a case of congenital hypofibrinogenemia.

Inhibition of local effects of Indian Daboia/Vipera russelli venom by the methanolic extract of grape (Vitis vinifera L.) seeds.

Although anti-venom therapy is available for the treatment of fatal bite by snakes, it offers less or no protection against the local effects such as dermo- and myonecrosis, edema, hemorrhage and inflammation at the bitten region. The viper species are known for their violent local effects and such effects have been commonly treated with plant extracts without any scientific validation in rural India. In this investigation, the methanolic extract of grapes (Vitis vinifera L.) seed was studied against the Indian Daboia/Vipera russelli venom-induced local effects. The extract abolished the proteolytic and hyaluronidase activities and also efficiently neutralized the hemorrhage, edema-inducing and myonecrotic properties of the venom. In addition, the extract also inhibited partially the pro-coagulant activity of the venom and abolished the degradation of Aα and Bβ chains of human fibrinogen. Thus, the extract possesses potent anti-snake venom property, especially against the local effects of viper bites.

Short-term treatment with metformin improves the cardiovascular risk profile in first-degree relatives of subjects with type 2 diabetes mellitus who have a metabolic syndrome and normal glucose tolerance without changes in C-reactive protein or fibrinogen

OBJECTIVE: To study if metformin, when administered to first-degree relatives of type 2 diabetes mellitus subjects who have metabolic syndrome and normal glucose tolerance, could improve the cardiovascular risk profile and reduce the levels of both C-reactive protein and fibrinogen. INTRODUCTION: Metabolic syndrome is associated with higher cardiovascular morbidity and mortality. Metformin has vasculo-protective effects even in normoglycemic subjects, and C-reactive protein and fibrinogen are considered markers of endothelial injury and inflammation. METHODS: Thirty-one non-diabetic first-degree relatives of type 2 diabetes mellitus subjects with metabolic syndrome were randomized (1:1) and double-blinded for placement in the placebo and metformin groups (850mg bid/±90days); 16 subjects were administered metformin (mean age 40.0 [33.5-50] years; 13 females) and 15 subjects were in the placebo group (mean age 37.0 [32-42] years; 9 females). Blood samples were collected at baseline and at the end of treatment for biochemical analyses, including an assessment of C-reactive protein and fibrinogen levels. RESULTS: Metformin improved the lipid profile and decreased fasting plasma glucose, systolic blood pressure, weight and body mass index without changing body composition. For those in the placebo we identified no changes in fibrinogen (282.2 [220.4-323.7] mg/L vs. 286.7 [249.6-295.1] mg/L; NS) or in C-reactive protein levels (0.68 [0.3-1.2] vs. 0.64 [0.3-1.0] mg/L; NS). The same was also observed for the levels of fibrinogen (303.9 [217.6-347.6] mg/L vs. 290.9 [251.5-301.9] mg/L; NS) and C-reactive proteins (0.78 [0.3-1.1] vs. 0.80 [0.4-0.9] mg/L; NS) in the metformin group. CONCLUSIONS: Metformin treatment in first-degree relatives of type 2 diabetes mellitus sufferers who have metabolic syndrome and normal glucose tolerance improved the cardiovascular risk profile without changing the levels of C-reactive protein and fibrinogen.

Hypercoagulable state in idiopathic ulcerative colitis: role of hyperhomocysteinemia and hyperfibrinogenemia.

BACKGROUND: Previous reports on hypercoagulable factors in inflammatory bowel diseases involve heterogeneous populations and patients on various medications. AIMS: To determine the frequency of thrombotic complications in ulcerative colitis (UC); to evaluate for hyperhomocysteinemia and its relationship to vitamin B12 and folate levels and methylene tetrahydrofolate reductase (MTHFR) mutation; and to evaluate for hyperfibrinogenemia and factor V Leiden mutation. METHODS: Eighty-six adult patients with UC were seen during the study period; 28 of them underwent blood tests and constituted the study population. Patients who received medications that affect these factors were among the 58 excluded. Tests were obtained at baseline and after 2 months during remission. Patients received folic acid in addition to treatment for UC. RESULTS: Vascular thrombotic events were noted in 4 patients during follow up. Hyperhomocysteinemia was detected in 11 (39.3%) patients (controls 15/100, p=0.007). Heterozygous state for MTHFR C677T mutation was found in 5 (17.9%) patients (controls: 0.2% homozygous, 13.6% heterozygous, p>0.05). Plasma homocysteine did not correlate with extent, severity or duration of disease, or with MTHFR C677T heterozygous state, but correlated with serum folic acid level (p=0.003) and BMI (p=0.03). With folate supplementation, homocysteine decreased significantly in patients who had hyperhomocysteinemia at baseline. Hyperfibrinogenemia was detected in 3 patients (none in 100 controls). Plasma fibrinogen was not affected by duration, extent or severity of UC and did not decrease with remission of disease. Only one patient had heterozygous factor V Leiden mutation. CONCLUSION: Vascular thrombosis occurred in less than a fifth of the UC population studied. Hyperhomocysteinemia reversible by folate supplementation and hyperfibrinogenemia were observed, but their contribution and that of factor V Leiden mutation appear to be insignificant.

Capacidade dos biomarcadores inflamatórios em predizer a síndrome metabólica / Inflammation biomarkers capacity in predicting the metabolic syndrome

O processo inflamatório é o elo entre a síndrome metabólica e as doenças cardiovasculares. Para medir o grau da inflamação subclínica, vários biomarcadores inflamatórios têm sido propostos. Este trabalho tem como objetivo revisar as recentes pesquisas das associações entre os biomarcadores inflamatórios e a síndrome metabólica, bem como a capacidade daqueles em predizer a síndrome metabólica. Estes biomarcadores incluem as citocinas pró-inflamatórias, citocinas antiinflamatórias, adipocinas, chemocinas, marcadores de inflamação derivados de hepatócitos, marcadores de conseqüência da inflamação e enzimas. Com esta revisão pode-se integrar o novo conhecimento referente às interações possíveis de mediadores inflamatórios com a síndrome metabólica, visto que estes biomarcadores desempenham vários papéis e seguem diversos caminhos metabólicos.

The inflammatory process is the link between metabolic syndrome and cardiovascular diseases. To measure the degree of subclinical inflammation some inflammatory biomarkers have been considered. This work reviews the recent researches of the associations between inflammatory biomarkers and metabolic syndrome, as well as the capacity in predicting the metabolic syndrome. These biomarkers include pro-inflammatory cytokines, anti-inflammatory cytokines, adipokines, chemokines, inflammation markers derived from hepatocites, the consequence markers of inflammation and enzymes. This review integrates the new knowledge of inflammatory mediators interactions with metabolic syndrome, since these biomarkers play different roles and follow diverse metabolic ways.