Autoantibodies against the fibrinolytic receptor, annexin A2, in cerebral venous thrombosis.

BACKGROUND AND PURPOSE: Cerebral venous thrombosis (CVT) may be a manifestation of underlying autoimmune disease. Antibodies against annexin A2 (anti-A2Ab) coincide with antiphospholipid syndrome, in which antiphospholipid antibodies (aPLA) are associated with thrombosis in any vascular bed. Annexin A2, a profibrinolytic receptor and binding site for ß2-glycoprotein-I, the main target for aPLA, is highly expressed on cerebral endothelium. Here we evaluate the prevalence of anti-A2Ab in CVT. METHODS: Forty individuals with objectively documented CVT (33 women and 7 men) and 145 healthy controls were prospectively studied for hereditary and acquired prothrombotic risk factors, classical aPLA, and anti-A2Ab. RESULTS: One or more prothrombotic risk factors were found in 85% of CVT subjects, (pregnancy/puerperium in 57.5%, classical aPLA in 22.5%, and hereditary procoagulant risk factors in 17.5%). Anti-A2Ab (titer >3 SD) were significantly more prevalent in patients with CVT (12.5%) than in healthy individuals (2.1%, P<0.01, OR, 5.9). CONCLUSIONS: Anti-A2Ab are significantly associated with CVT and may define a subset of individuals with immune-mediated cerebral thrombosis.

Tratamiento de la cervicitis aguda con Aloe Vera L: estrategia para su prevención / Treatment for acute cervicitis, with Aloe vera L: Strategy for its prevention

Se realizó un ensayo clínico en fase II b en grupos paralelos a ciegas por terceros, controlado y aleatorizado, con el objetivo de evaluar la eficacia y seguridad del Aloe vera en pacientes diagnosticadas con cervicitis aguda, además se diseñó una estrategia preventiva sobre la base de los factores de riesgo más influyentes en la aparición de la cervicitis aguda. El universo estuvo constituido por 267 mujeres con cervicitis aguda en la consulta de ginecobstetricia del Hospital 4 de Abril. La muestra quedó conformada por 120 pacientes, 60 pacientes cada grupo. Se logró una evolución favorable de las pacientes con respecto al comportamiento de síntomas, signos y la presencia de microorganismos. Resultó eficaz la terapia con el gel de Aloe vera en la cervicitis aguda, sin la aparición de efectos adversos. La estrategia preventiva diseñada es adecuada para ser aplicada para la profilaxis de la cervicitis aguda(AU)

A clinical blind assay with parallel, controlled and aleatorized groups at stage II b was carried out in order to evaluate the effectiveness and security of Aloe vera in patients diagnosed with acute cervicitis. A preventive strategy was also designed taking into account relevant risk factors at early stages aiming to decrease this disease. The universe was composed by 267 female patients from the gynecological consult at “4 de Abril” Hospital. The sample was made by 120 patients; both groups were formed by 60 patients using the simple aleatory sample. The outcome was favorable concerning signs and symptoms behaviour and the presence of microorganisms. The therapy with aloe vera gel was effective in acute cervicitis without consequences. The preventive strategy is proper to be used in the prophylaxis of acute cervicitis(EU)

Dehydroepiandrosterone inhibits the TNF-alpha-induced inflammatory response in human umbilical vein endothelial cells.

Dehydroepiandrosterone (DHEA) has a protective role against atherosclerosis. We determined the effect of pharmacological doses of DHEA upon the adhesion of monocytic U937 cells to human umbilical vein endothelial cells (HUVEC), as well as the expression of adhesion and chemoattractant molecules, the translocation of NF-kappaB, the degradation of IkappaB-alpha and the production of reactive oxygen species (ROS) in HUVEC. Adhesion of U937 cells to DHEA-treated HUVEC was evaluated by co-culture experiments using [(3)H]-thymidine-labeled U937 cells. The expression of adhesion and chemoattractant molecules was evaluated by flow cytometry and RT-PCR, respectively; NF-kappaB translocation was determined by Electrophoretic Mobility Shift Assay (EMSA) and IkappaB-alpha degradation by Western blot. ROS production was determined by the reduction of fluorescent DCFDA. TNF-alpha was used to induce inflammatory responses in HUVEC. One hundred micromolar of DHEA-treatment inhibited the TNF-alpha-induced expression of ICAM-1, E-selectin, ROS production and U937 cells adhesion to HUVEC, and interfered with NF-kappaB translocation and IkappaB-alpha degradation. DHEA at the above mention concentration also inhibited the mRNA expression of MCP-1 and IL-8 in basal conditions but not in TNF-alpha-stimulated conditions. Our results suggest that DHEA inhibits the expression of molecules involved in the inflammatory process, therefore it could be used as an alternative in the treatment of chronic inflammatory diseases such as atherosclerosis.

Hypolipidaemic and antiplatelet activity of phenoxyacetic acid derivatives related to alpha-asarone.

The phenoxyacetic acid derivatives 1-6 [2-methoxy-4-(2-propenyl)phenoxyacetic acid (1); 2-methoxy-5-nitro-4-(2-propenyl)phenoxyacetic acid (2); methyl 2-methoxy-4-(2-propenyl)phenoxyacetate (3); ethyl 2-methoxy-4-(2-propenyl)phenoxyacetate (4); methyl 2-methoxy-5-nitro-4-(2-propenyl)phenoxyacetate (5); ethyl 2-methoxy-5-nitro-4-(2-propenyl)phenoxyacetate (6)] related to alpha-asarone have been reported previously as hypolipidaemic agents in diet-induced hyperlipidaemic mice. We have aimed to expand the pharmacological profile of these derivatives by investigating their hypolipidaemic activity in rats and mice under different experimental conditions. The antiplatelet activity was tested also in-vitro from blood derived from consenting healthy volunteers. In normolipidaemic rats, compounds 2, 3 and 5 at oral doses of 40 and 80 mg kg(-1) significantly decreased total cholesterol and LDL-cholesterol levels. Moreover, analogues 3 and 5 administered to hypercholesterolaemic rats at the same doses for seven days also produced a reduction in the content of these same lipoproteins. In neither case were the high-density lipoprotein cholesterol and triglyceride concentrations affected. However, practically all tested compounds were found to be hypocholesterolaemic agents, and were shown to effectively lower low-density lipoprotein cholesterol and triglyceride levels in Triton-induced hyperlipidaemic mice at oral doses of 50 and 100 mg kg(-1). In all tests, all animals appeared to be healthy throughout the experimental period in their therapeutic ranges. Triton-induced hypercholesterolaemic mice appeared to be a desirable model for this class of hypolipidaemic drugs. On the other hand, compounds 1, 2, 4 and 5 significantly inhibited ADP-induced aggregation in-vitro. These findings indicated that all of these compounds appeared to be promising for the treatment of human hyperlipidaemia and thrombotic diseases.

In-vitro model for the ultrastructural study of the formation of thrombi in human platelets.

Platelets are cell fragments with dynamic properties involved in clot formation after tissue damage. Platelet activation causes a change in shape, secretion of intracellular granules and aggregation with each other through the cytoskeleton components and biochemical changes. Platelet adhesion, considered as the major event in haemostasis, has been studied in several in-vitro and in-vivo models to evaluate the feasible thrombogenicity of some materials, the dynamics of specific receptors, as well as the effect of different buffers and inhibitors in this process. In spite of the numerous reports about platelet activation, to date there is no information available about the fine structure of the platelet-platelet and platelet-substrate interactions. In the present report we describe an in-vitro system that allows the visualization of these interactions: platelets are adhered to an inert substrate, and interactions with suspended platelets as a process to initiate the formation of thrombi was followed by ultramicrotomy and transmission electron microscopy.

Effect of pulsed estrogen therapy on hemostatic markers in comparison with oral estrogen regimen in postmenopausal women.

BACKGROUND/AIMS: Hormone replacement therapy (HRT) is associated with an increased risk of thromboembolism dependent on the type of HRT; therefore, we compared therapy effects of intranasal with oral estrogens on coagulation and fibrinolysis markers in postmenopausal women. METHODS: A randomized study in which 29 healthy hysterectomized women received intranasal 17beta-estradiol or oral estrogens for 3 months. RESULTS: There were no significant differences in the baseline characteristics between groups. Those women receiving intranasal estradiol showed a mild increment in plasminogen activator inhibitor-1 (PAI-I) (from 6.8 +/- 3.5 to 9.6 +/- 3.9 U/ml, p < 0.01); however, fibrinogen, factor VII-tissue factor complex (VIIa-rTF), antithrombin III (ATIII), protein C (PC) activity, protein S (PS) activity, plasminogen (PLG), and tissue-type plasminogen activator antigen (t-PA) were unchanged. In contrast, oral unopposed estrogens elevated t-PA (from 4.9 +/- 2.9 to 9.6 +/- 5.1 ng/ml, p < 0.01) in parallel with a decrement in PAI-I (from 5.2 +/- 4.0 to 2.7 +/- 1.7 U/ml, p < 0.05) and VIIa-rTF (from 201.2 +/- 181.0 to 140.6 +/- 108.7 mU/ml, p < 0.05). Fibrinogen, ATIII, PC, PS, and PLG were unchanged. CONCLUSIONS: Nasal 17beta-estradiol had no effect on the coagulation markers, except a moderate increment in PAI-1. In contrast, oral estrogens elicited a decrement in both VIIa-rTF and PAI-1; however, those changes did not surpass normal limits.

Activated protein C resistance and lupus anticoagulant activity induced by plasma and purified monospecific human IgG anti-beta2-glycoprotein-I antibodies.

INTRODUCTION: We investigated the activated protein C resistance (APCR) phenotype and the lupus anticoagulant (LA), activity induced by anti-beta2-glycoprotein-I (anti-beta2GP-I) antibodies. PATIENTS AND METHODS: We studied plasma and sera samples from 29 patients with persistently positive anti-beta2GP-I: 22 with thrombosis (12 with primary APS, 10 with APS secondary to SLE) and seven without thrombosis (all with SLE); 25 healthy subjects were studied as controls. We detected anticardiolipin antibodies (ACA); IgG (and its subclasses) and IgM anti-beta2GP-I, on irradiated and non-irradiated plates by ELISA. APCR was assessed by the activated partial thromboplastin time (APTT)-based assay and by the modified test. The FV Leiden mutation was studied by PCR. LA determination included screening and confirmatory dRVVT. Serum anti-2 GP-I were affinity purified on sepharose columns and their isotype, subclass, and reactivity against various antigens were studied by ELISA. RESULTS: We found that titers of IgG anti-beta2GP-I on irradiated plates were higher than on non-irradiated plates (p = 0.002), IgG2 was the predominant subclass. Fifteen patients (13 with thrombosis) had LA and 15 (also 13 with thrombosis) induced the APCR phenotype. Eleven (all with thrombosis) had both. Two patients were heterozygous for the Leiden mutation. Two purified antibodies, monospecific for beta2GP-I, induced an in vitro APCR phenotype and LA activity. CONCLUSIONS: Our results seem to indicate that the inhibition of the APC anticoagulant function by IgG2 anti-beta2GP-I with LA activity may be one of the responsible mechanisms of thrombophilia in patients with APS.

Activated protein C resistance and lupus anticoagulant activity induced by plasma and purified monospecific human IgG anti-β2-glycoprotein-I antibodies / Resistencia a la proteína C activada y actividad de anticoagulante lúpico inducidas por plasma y por anticuerpos purificados humanos del IgG anti β2-glicoproteína I

ABSTRACT Introduction. We investigated the activated protein C resistance (APCR) phenotype and the lupus anticoagulant (LA), activity induced by anti-β2-glycoprotein-I (anti-β2GP-I) antibodies. Patients and methods. We studied plasma and sera samples from 29 patients with persistently positive anti-β2GP-I: 22 with thrombosis (12 with primary APS, 10 with APS secondary to SLE) and seven without thrombosis (all with SLE); 25 healthy subjects were studied as controls. We detected anticardiolipin antibodies (ACA); IgG (and its subclasses) and IgM anti-β2GP-I, on irradiated and non-irradiated plates by ELISA. APCR was assessed by the activated partial thromboplastin time (APTT)-based assay and by the modified test. The FV Leiden mutation was studied by PCR. LA determination included screening and confirmatory dRVVT. Serum anti-β2GP-I were affinity purified on sepharose columns and their isotype, subclass, and reactivity against various antigens were studied by ELISA. Results. We found that titers of IgG anti-β2GP-I on irradiated plates were higher than on non-irradiated plates (p = 0.002), IgG2 was the predominant subclass. Fifteen patients (13 with thrombosis) had LA and 15 (also 13 with thrombosis) induced the APCR phenotype. Eleven (all with thrombosis) had both. Two patients were heterozygous for the Leiden mutation. Two purified antibodies, monospecific for β2GP-I, induced an in vitro APCR phenotype and LA activity. Conclusions. Our results seem to indicate that the inhibition of the APC anticoagulant function by IgG2 anti-β2GP-I with LA activity may be one of the responsible mechanisms of thrombophilia in patients with APS.

Introducción. Investigamos la resistencia a la proteína C activada (RPCA) y la actividad de anticoagulante lápico (AL), inducidas por anticuerpos anti-β2-glicoproteína-I (anti-β2GP-I). Pacientes y métodos. Estudiamos los plasmas y sueros persistentemente positivos para anti-β2GP-I de 29 pacientes: 22 tuvieron trombosis (12 con síndrome de antifosfolípidos (SAF) primario y 10 con SAF secundario a lupus erítematoso generalizado (LEG)) y siete sin trombosis (todos con LEG). Como controles estudiamos 25 sueros de personas clínicamente sanas. Detectamos anticuerpos anticardiolipina, anti-β2GP-I IgG (y sus subclases) e IgM por ELISA en placas irradiadas y no irradiadas. Evaluamos la RPCA por medio del tiempo parcial de tromboplastina activada y por la prueba modificada. Estudiamos la mutación FV de Leiden por PCR y el anticoagulante lápico con el método de dRVVT screening y confirmatorio. Después de purificar los anti-β2GP-I séricos con una columna de antígeno unido a sefarosa, analizamos por ELISA sus isotipos, subclases y reactividad contra β2GP-I y algunos fosfolípidos. Resultados. Los títulos de anti-β2GP-I IgG fueron más altos en placas irradiadas que en no irradiadas (p = 0.002), predominó la subclase IgG2. Quince plasmas (13 de pacientes con trombosis) tuvieron AL y 15 (13 también de pacientes con trombosis) indujeron el fenotipo de RPCA. Once plasmas (todos de pacientes con trombosis) indujeron ambas actividades. Dos pacientes fueron heterocigotos para la mutación de Leiden. Dos anticuerpos purificados monoespecíficos para β2GP-I indujeron el fenotipo de la RPCA y la actividad de AL in vitro. Conclusiones. Nuestros resultados sugieren que la RPCA, inducida por los anti-β2GP-I que concomitantemente tienen actividad de AL, puede tener implicaciones patogénicas en la trombofílía del SAF.

Neocarzinostatin induces an effective p53-dependent response in human papillomavirus-positive cervical cancer cells.

Human papillomavirus (HPV) E6 viral oncoprotein plays an important role during cervical carcinogenesis. This oncoprotein binds the tumor suppressor protein p53, leading to its degradation via the ubiquitin-proteasome pathway. Therefore, it is generally assumed that in HPV-positive cancer cells p53 function is completely abolished. Nevertheless, recent findings suggest that p53 activity can be recovered in cells expressing endogenous E6 protein. To investigate whether p53-dependent functions controlling genome integrity, cell proliferation, and apoptosis can be reactivated in cervical cancer cells, we examined the capacity of HeLa, INBL, CaSki, C33A, and ViBo cell lines to respond to neocarzinostatin (NCS), a natural product which induces single- and double-strand breaks in DNA. We found that NCS treatment inhibits cellular proliferation through G2 cell cycle arrest and apoptosis induction. This effect was preceded by nuclear accumulation of p53 protein and by an increase of p21 transcripts. Although apoptosis was blocked in ViBo cells (HPV-negative), nuclear accumulation of transcriptionally active p53 and inhibition of cell proliferation are observed after NCS treatment. These results suggest that HPV-positive cervical cancer cells are capable of responding efficiently to DNA damage provoked by NCS treatment through a p53-dependent pathway in spite of the presence of E6 protein.

Modification of actin, myosin and tubulin distribution during cytoplasmic granule movements associated with platelet adhesion.

BACKGROUND AND OBJECTIVES: Cytoskeletal elements determine the changes in platelet cell shape which occur during adhesion, aggregation and release of granular contents as part of the activation process. The aim of this study was to characterize the changes in the distribution of actin filaments, myosin and tubulin molecules during several stages of platelet adhesion to glass and their association with granule displacement, as assessed by confocal microscopy. DESIGN AND METHODS: Platelets obtained from healthy donors were adhered to glass and cytoskeleton distribution was characterized and correlated to changes of cell shape and intracellular granule displacement by immunofluorescence assays and phase contrast microscopy. Treatment with specific cytoskeleton inhibitors such as cytochalasin D, butanedione monoxime and colchicine were used before and after the adhesion process. The spatial distribution of the cytoskeleton in association with cytoplasmic granules was analyzed in both confocal microscopy projections and three-dimensional images obtained by merging the respective projections. RESULTS: Our experiments revealed that as platelets contact the substrate, a sequential and simultaneous rearrangement of actin filaments, myosin and tubulin molecules occurred and this was related to cell shape, as well as to movements of cytoplasmic granules. Treatment of platelets with cytoskeleton inhibitors, modified not only the target molecule but also other cytoskeletal components with consequent alterations in the studied platelet functions. INTERPRETATION AND CONCLUSIONS: During platelet adhesion to glass and granule displacement, a close spatial and functional relation between actin filaments, myosin molecules and microtubules was observed suggesting that these different cytoskeleton components interact in supporting the platelet functions here studied.

Frecuencia de la dismenorrea en adolescentes.Politécnico América Libre, Contramaestre ,Santiago de Cuba / Frequency of it dismenorrhea in adolescents. Politécnic Amrica Libre, Boatswain, Santiago of Cuba

Este estudio se realiza por la frecuencia de la dismenorrea en los adolescentes, encontramos en la muestra estudiada que se acompaña de otros signos y síntomas, que pueden ser la hipermenorrea, molestias posmenstruales e inestabilidad psíquica, evidentemente constituye un problema de salud.El grupo etareo se encuentra entre los 12 y 15 años, con un patrón menstrual dentro de los límites normales.Los resultados mustran que no se le presta debida atención a la dismennorrea y que se interrelacionados factores psicogénos, ginecológicos y endocrino

Frecuencia de la dismenorrea en adolescentes: Politécnico América Libre, Contramaestre, Santiago de Cuba / Frequency of it dismenorrhea in adolescents: Politécnic Amrica Libre, Boatswain, Santiago of Cuba

Este estudio se realiza por la frecuencia de la dismenorrea en los adolescentes, encontramos en la muestra estudiada que se acompaña de otros signos y síntomas, que pueden ser la hipermenorrea, molestias posmenstruales e inestabilidad psíquica, evidentemente constituye un problema de salud.El grupo etareo se encuentra entre los 12 y 15 años, con un patrón menstrual dentro de los límites normales.Los resultados mustran que no se le presta debida atención a la dismennorrea y que se interrelacionados factores psicogénos, ginecológicos y endocrino