Diagnostic value of clot formation parameters determined by rotational thromboelastometry in 63 patients with congenital dysfibrinogenemia.

Rotational thromboelastometry (ROTEM) is a global hemostasis assay. The diagnosis added value of ROTEM in congenital dysfibrinogenemia remains to be established. The aim of this study was to analyze clot formation by ROTEM in a cohort of dysfibrinogenemic patients and to establish correlations with genotype, clinical features, and coagulation parameters. The study included genetically confirmed congenital dysfibrinogenemia cases (n = 63) and healthy controls ( n  = 50). EXTEM, INTEM, FIBTEM tests were used to measure ROTEM parameters, that is, clotting time (CT), clot formation time (CFT), maximal clot firmness (MCF) and amplitude 10 min after CT (A10). The ISTH bleeding assessment tool was used to determine bleeding episodes. CT (INTEM) was statistically significantly shorter in congenital dysfibrinogenemia patients compared to controls while CFT (EXTEM) was prolonged. Patients's MCF in EXTEM, INTEM, and FIBTEM were similar to controls while A10 (FIBTEM) was statistically significantly lower. Fibrinogen activity was positively correlated with fibrinogen antigen, A10 and MCF in all three assays. Bleeding phenotypes were observed in 23 (36.5%) patients. Only CFT in EXTEM and CT in INTEM were statistically different in patients with bleeding phenotype versus controls. Carriers of the FGA mutation p.Arg35His had a CT (EXTEM) slightly prolonged and a reduced A10 (FIBTEM) compared to controls. Some ROTEM parameters were able to distinguish congenital dysfibrinogenemia patients from controls, and patients with a bleeding phenotype. Prolonged CFT in EXTEM were associated with congenital dysfibrinogenemia and bleeding phenotype. Bleeding episodes in most patients were generally mild and prevalence of thrombosis was very low.

QOLEC2: a randomized controlled trial on nutritional and respiratory counseling after esophagectomy for cancer.

BACKGROUND: Esophagectomy for cancer strongly impairs quality of life. The aim of this trial was to evaluate the effect of the nutritional and respiratory counseling on postoperative quality of life. METHODS: At hospital discharge, patients were randomized into four groups receiving respectively: nutritional and respiratory counseling, nutritional counseling alone, respiratory counseling alone, or standard care. The main endpoint was the impairment in quality of life in the first month after surgery. Linear mixed effect models were estimated to assess mean score differences (MDs) in quality of life scores. RESULTS: Patients receiving nutritional counseling reported less appetite loss (MD - 17.7, 95% CI - 32.2 to -3.3) than those not receiving nutritional counseling at 1 month after surgery. Dyspnea was similar between patients receiving vs. those not receiving respiratory counseling (MD - 3.1, 95% CI - 10.8 to 4.6). Global quality of life was clinically similar between patients receiving vs. those not receiving nutritional counseling over time (MD 0.9, 95% CI - 5.5 to 7.3), as well as in patients receiving vs. those not receiving respiratory counseling over time (MD 0.7, 95% CI - 5.9 to 7.2). CONCLUSIONS: Intensive postoperative care does not affect global quality of life even if nutritional counseling reduced appetite loss.

Purificación del fibrinógeno gamma A/gamma prima (үA/ү') por cromatografía líquida rápida de proteínas / Purification of fibrinogen gamma A/gamma prima (үA/ү') by fast protein liquid chromatography / Purificação do fibrinogênio gamma A/gamma prima (үA/ү') por cromatografía liquida rápida de proteínas

Resumen Una fracción del fibrinógeno circulante contiene una variante de la cadena ү que se origina por empalme alternativo del ARNm, denominada ү' cuya concentración en plasma se ha relacionado con un incremento en el riesgo de padecer enfermedades cardiovasculares. Por tanto, el objetivo de este trabajo fue diseñar un método de purificación del fibrinógeno үA/ү' más eficiente en relación a los descritos en la literatura, a partir de plasma humano. Se purificó el fibrinógeno үA/ү' a partir del fibrinógeno total obtenido por precipitación con β-alanina, mediante la separación por cromatografía líquida rápida de proteínas. Se confirmó la presencia de fibrinógeno үA/ү' mediante Western blot; su concentración fue determinada por ELISA. El método mostró ventajas en comparación con los métodos clásicos de separación, por ejemplo, que cantidades menores de muestra pudieron ser fraccionadas cuantitativamente en componentes puros en menor tiempo (30 min). Por tanto, se puede concluir que la técnica utilizada para la purificación de las variantes del fibrinógeno, correspondiente al Fg үA/үA y Fg үA/ү', es un método de separación eficiente que permite purificar el Fg үA/ү' libre de contaminantes principales, como lo confirma la inmunoelectroforesis.

Abstract A fraction of the circulating fibrinogen contains a variant of the y chain that is originated by mRNA alternative splicing denominated ү' whose concentration in plasma has been related to an increase in the risk of cardiovascular diseases. Thus, the objective of this work was to design a more efficient үA/ү' fibrinogen purification method in relation to those described in the literature from human plasma. The үA/ү' fibrinogen was purified from the total fibrinogen obtained by precipitation with β-alanine, using separation by fast protein liquid chromatography. Fibrinogen үA/ү', was confirmed by Western blot and its concentration was determined by ELISA. The method showed advantages compared to classical separation methods, for example, smaller amounts of sample could be fractionated quantitatively into pure components in less time (30 min). Therefore, it can be concluded that the technique used for the purification of fibrinogen variants, corresponding to Fg үA/үA and Fg үA/ү', is an efficient separation method that allows purifying the Fg үA/ү' free of main contaminants, as confirmed by immunoelectrophoresis.

Resumo Uma fracção do fibrinogênio circulante contém uma variante da cadeia y que origina-se por junção alternativa do ARNm, chamada ү' cuja concentração em plasma está asociada com um incremento no resgo de sofrer doenças cardiovasculares. Asim, o objetivo em este trabalho foi desenhar um método de purificação do fibrinogênio үA/ү' mais eficiente em relação aos que estão descritos na literatura, a partir do plasma humano. Purificou-se o fibrinogênio үA/ү' a partir do fibrinogênio total conseguido pela precipitação com β-alanina, através da separação por cromatografia líquida rápida de proteínas. Se confirmou por Western blot o fibrinogênio үA/ү'. A sua concentração foi determinada por ELISA. O método apresenta vantagens em comparação com os métodos clássicos de separação, por exemplo, quantidades menores de mostra podem ser fraccionadas quantitativamente em componentes puros, em tempos mais curtos (30 min). Entao, pode-se concluir que a técnica utilizada, para a purificação das variantes do fibrinogênio correspondente ao Fg үA/үA e Fg үA/ү', é um método de separação eficiente que permite purificar o Fg үA/ү' livre de contaminantes principais, como o confirma a imunoeletroforese.

Efecto de los anticuerpos antifosfolipídicos en la formación y degradación de la malla de fibrina en pacientes con aborto recurrente / Effect of antiphospholipid antibodies on the formation and lysis of fibrin network in patients with recurrent miscarriage

En el presente trabajo se estudió el proceso de formación y disolución de la malla de fibrina y la generación de plasmina en un grupo de pacientes con aborto recurrente (AR) debido a la presencia de anticuerpos antifosfolipídicos (N= 10), mujeres con AR sin el síndrome antifosfolipídico (SAF) (N= 6) y se comparó con un grupo de mujeres sanas (N= 8). Del grupo de pacientes estudiadas con SAF, nueve fueron positivas para anticuerpos anticardiolipina (aCL), cinco para la anti-b2-glicoproteína I (anti-b2GPI), cuatro para ambos anticuerpos, una para anticuerpos antiprotrombina (aPT) y anticoagulante lúpico (AL). El proceso de formación de la fibrina y su disolución fue estudiado por turbidimetría y la generación de plasmina mediante sustrato cromogénico S2251. Las curvas de polimerización de la(s) paciente(s) con AR sin SAF y AL presentaron un incremento en la pendiente y turbidez final, comparado con las del grupo control de mujeres sanas. La velocidad de disolución del coágulo fue mayor en la paciente con AL (21 ± 0) 10-4 DDO/seg y en las AR sin SAF (19,6 ± 5,7) 10-4 DDO/seg, comparado con el grupo control (14,5 ± 2,8) 10-4 DDO/seg. La generación de plasmina estuvo incrementada solamente en las AR sin SAF (85 ± 24%) comparado con 52 ± 3% en el grupo control, p= 0,005. Los cambios observados en el proceso de polimerización y fibrinólisis de la(s) paciente(s) con AR sin SAF y AL pudieran estar relacionados con el incremento en los niveles de fibrinógeno, mientras que los de la generación de plasmina con la entidad mórbida.

The present work was intended to study the process of fibrin formation and lysis and plasmin generation in a group of patients with recurrent miscarriage (RM), due to the presence of antiphospholipid antibodies (N= 10); as well as in women with RM without the antiphospholipid syndrome (APS) (N= 6), compared with those of a group of healthy women (N= 8). In the group of patients with APS, nine were positive for antibodies against cardiolipin (aCL), five for anti-b2-glycoprotein I (anti-b2GPI), four for both antibodies, and one for antibodies against prothrombin (aPT) and lupus anticoagulant (LA). Fibrin formation and lysis was followed by turbidity and plasmin generation using chromogenic substrate S2251. The polymerization curves from RM patients without APS and the LA patient showed an increased slope and maximum turbidity compared to those of the control group. The speed of lysis was higher in the LA patient (21 ± 0) 10-4 DOD/seg and the RM patients without APS (19.6 ± 5.7) 10-4 DDO/seg, compared to that of the control group (14.5 ± 2.8) 10-4 DDO/seg. Plasmin generation increased only in RM patients without APS (85 ± 24%) against the control group (52 ± 3%), p= 0.005. The changes observed in the fibrin polymerization and lysis process of women with RM without APS and LA seem to be related to their higher fibrinogen levels, while the increased plasmin generation was related to the patients´ morbidity.