Activin B promotes the initiation and progression of liver fibrosis.

The role of activin B, a transforming growth factor ß (TGFß) superfamily cytokine, in liver health and disease is largely unknown. We aimed to investigate whether activin B modulates liver fibrogenesis. Liver and serum activin B, along with its analog activin A, were analyzed in patients with liver fibrosis from different etiologies and in mouse acute and chronic liver injury models. Activin B, activin A, or both was immunologically neutralized in mice with progressive or established carbon tetrachloride (CCl4 )-induced liver fibrosis. Hepatic and circulating activin B was increased in human patients with liver fibrosis caused by several liver diseases. In mice, hepatic and circulating activin B exhibited persistent elevation following the onset of several types of liver injury, whereas activin A displayed transient increases. The results revealed a close correlation of activin B with liver injury regardless of etiology and species. Injured hepatocytes produced excessive activin B. Neutralizing activin B largely prevented, as well as improved, CCl4 -induced liver fibrosis, which was augmented by co-neutralizing activin A. Mechanistically, activin B mediated the activation of c-Jun-N-terminal kinase (JNK), the induction of inducible nitric oxide synthase (iNOS) expression, and the maintenance of poly (ADP-ribose) polymerase 1 (PARP1) expression in injured livers. Moreover, activin B directly induced a profibrotic expression profile in hepatic stellate cells (HSCs) and stimulated these cells to form a septa structure. Conclusions: We demonstrate that activin B, cooperating with activin A, mediates the activation or expression of JNK, iNOS, and PARP1 and the activation of HSCs, driving the initiation and progression of liver fibrosis.

Correlation between platelet thrombus formation on collagen-coated beads and platelet aggregation induced by ADP.

BACKGROUND: The thrombus-forming ability is a critical in vitro parameter to assess platelets (PLTs), but flow-based methods using collagen-coated materials generally require multistep, proficiency, and advanced analysis. STUDY DESIGN AND METHODS: Commercially available collagen-coated bead columns were examined to assess thrombus-forming ability of PLTs. The retention rate as an index of thrombus formation was calculated using the PLT count before and after column passage. Thrombi were imaged by anti-CD41 using a fluorescent microscope. PLT aggregation was measured by light-transmitting aggregometry. RESULTS: The retention rate was low when apheresis-collected PLT concentrates (PCs) were suspended in plasma either with or without Ca2+. Reconstitution of PCs with red blood cells (RBCs) increased the retention rate with good reproducibility on repeated-measurements, and therefore, PLT samples were reconstructed with RBCs in subsequent experiments. The retention rate of PCs varied widely in a product-dependent manner, and was correlated with the aggregation rate induced by ADP, but not that by collagen. Using platelet-rich-plasma, antagonists of P2Y1 or P2Y12 receptors for ADP reduced both the retention and aggregation of PLTs. Acetylsalicylic acid reduced retention, although it had no effect on ADP-induced aggregation. Prostaglandin E1 significantly inhibited both retention and aggregation. These anti-PLT reagents resulted in reduced or no thrombus formation on the beads. CONCLUSION: The collagen-coated bead column was useful to readily examine the thrombus-forming ability of PLTs. Variance of the PLT retention rate was correlated with responsiveness to ADP. Results from anti-PLT reagents revealed that thrombus formation on collagen-coated beads was similar to in vivo thrombus development.

Synthesis of a Novel Series of Amino Acid Prodrugs Based on Thienopyridine Scaffolds and Evaluation of Their Antiplatelet Activity.

The thienopyridines class of drugs used as P2Y12 receptor antagonists plays a vital role in antiplatelet therapy. To further optimized this compound class, we designed and synthesized a series of amino acid prodrugs of 2-hydroxytetrahydrothienopyridine. All compounds were then evaluated for their inhibitory effect on ADP-induced platelet aggregation in rats and then ED50 and bleeding time of the most potent compounds were compared with commercial drugs. The results showed compound 5c could be a potent and safe candidate for further research.

The novel compound MP407 inhibits platelet aggregation through cyclic AMP-dependent processes.

Platelet hyperactivity plays a critical role for initiating several vascular diseases such as atherothrombosis. Therefore, development of effective antiplatelet agents is necessary for ameliorating platelet-related diseases. In this study, we investigated the effects of the new synthesized compound, MP407 on platelet aggregation and further elucidated the underlying mechanisms. Our results demonstrated that MP407 dose-dependently inhibited collagen-induced platelet aggregation, thromboxane B2 (TXB2) production, intracellular Ca2+ mobilization, platelet membrane GPIIb/IIIa expression, and the phosphorylation of Akt, GSK3ß, p38MAPK, and phospho (Ser) PKC substrate (p47). Moreover, MP407 is able to increase the cyclic AMP formation both in resting and activated platelets. However, blocking cyclic AMP formation with 2'5'-ddAdo, an inhibitor of adenylate cyclase, greatly reversed the antiplatelet activity of MP407 and related platelet-activating pathways. MP407 also enhanced VASP phosphorylation at Ser157 in collagen-stimulated platelets, which was attenuated by addition of 2'5'-ddAdo. Therefore, the antiplatelet activity of MP407 may be modulated by cyclic AMP-dependent regulation of Akt, GSK3ß, p38MAPK and VASP phosphorylation. Notably, treatment with MP407 markedly reduced the pulmonary thrombosis and the numbers of paralysis and death in mice induced by ADP injection, but did not affect the bleeding time. Taken together, MP407 may be a potential candidate or lead compound for developing novel antiplatelet or antithrombotic agents for platelet hyperactivity-triggered disease therapy.

Quantification of the Blood Platelet Reactivity in the ADP-Induced Model of Non-Lethal Pulmonary Thromboembolism in Mice with the Use of Laser Doppler Flowmetry.

INTRODUCTION: The paper describes an alternative method for quantification of in vivo ADP-induced thromboembolism. The aim of the studies was to develop a method of quantification which would not require either extravasation or labelling of platelets. Our proposed approach is based on the monitoring of changes of blood flow with the use of laser Doppler flowmetry. MATERIALS AND METHODS: Mice of C57Bl strain were used in the study. ADP was injected to the vena cava and blood flow was monitored with the use of a laser Doppler flowmeter in the mesentery. Measurements in platelet-depleted mice, mice pretreated with cangrelor, an ADP receptor antagonist, and eptifibatide, a blocker of fibrinogen binding to GPIIbIIIa, were conducted as the proof-of-concept in the performed experiments. Intravital microscopy and ex vivo imaging of organs was performed to identify the sites of aggregate formation resulting from ADP injection. RESULTS: The injection of ADP resulted in a dose-dependent reduction of the blood flow in the mesentery. These responses were fully attributable to blood platelet aggregation, as shown by the lack of the effect in platelet-depleted mice, and significantly reduced responses in mice pretreated with cangrelor and eptifibatide. No platelet aggregate formation in mesenteric vessels was revealed by intravital microscopy, while ex vivo imaging showed accumulation of fluorescent labelled platelets in the lung. CONCLUSIONS: Injection of ADP to the venous system results in the formation of platelet aggregates predominantly in the lung. This results in reversible blood flow cessation in peripheral blood vessels. The measurement of this blood flow cessation in the mesentery allows indirect measurement of ADP-induced pulmonary thromboembolism. We suggest that this approach can be useful for in vivo screening for antiplatelet drug candidates.

[Effect of different fractions of Taohong Siwu decoction on ADP-induced platelet aggregation and thrombin activity].

To evaluate the effect of different fractions of Taohong Siwu decoction on ADP-induced platelet aggregation and thrombin activity, and to exploit the bioactive constituents, ADP-induced platelet aggregation rate in rabbits was determined by using the method of turbidity method. A bioassay called thrombin time was developed for determining anti-thrombin activities. UHPLC-Q-TOF-MS method was used to qualitatively analyze the chemical constituents of different parts. Alcohol precipitation deposition fraction, alcohol precipitation supernatant fraction and 20% to 30% alcohol elution fraction could significantly inhibit ADP-induced platelet aggregation. Alcohol precipitation supernatant fraction, water insoluble fraction and 40% to 70% alcohol elution fraction could significantly inhibit thrombin activity. The main components of alcohol precipitation deposition fraction, alcohol precipitation supernatant fraction and 20% to 40% alcohol elution fraction were analyzed and identified as aromatic acids, glycosides and phthalides. The bioactive constituents of Taohong Siwu decoction for inhibiting ADP-induced platelet aggregation and thrombin activity include aromatic acids, glycosides and phthalides. This experiment provides scientific basis to further explore the bioactive constituents and mechanism of Taohong Siwu decoction in treating blood stasis syndrome.

A novel role of Eruca sativa Mill. (rocket) extract: antiplatelet (NF-κB inhibition) and antithrombotic activities.

BACKGROUND: Epidemiological studies have shown the prevention of cardiovascular diseases through the regular consumption of vegetables. Eruca sativa Mill., commonly known as rocket, is a leafy vegetable that has anti-inflammatory activity. However, its antiplatelet and antithrombotic activities have not been described. METHODS: Eruca sativa Mill. aqueous extract (0.1 to 1 mg/mL), was evaluated on human platelets: (i) P-selectin expression by flow cytometry; (ii) platelet aggregation induced by ADP, collagen and arachidonic acid; (iii) IL-1ß, TGF-ß1, CCL5 and thromboxane B2 release; and (iv) activation of NF-κB and PKA by western blot. Furthermore, (v) antithrombotic activity (200 mg/kg) and (vi) bleeding time in murine models were evaluated. RESULTS: Eruca sativa Mill. aqueous extract (0.1 to 1 mg/mL) inhibited P-selectin expression and platelet aggregation induced by ADP. The release of platelet inflammatory mediators (IL-1ß, TGF-ß1, CCL5 and thromboxane B2) induced by ADP was inhibited by Eruca sativa Mill. aqueous extract. Furthermore, Eruca sativa Mill. aqueous extract inhibited NF-κB activation. Finally, in murine models, Eruca sativa Mill. aqueous extract showed significant antithrombotic activity and a slight effect on bleeding time. CONCLUSION: Eruca sativa Mill. presents antiplatelet and antithrombotic activity.

Reduction of death rate due to acute myocardial infarction in subjects with cancers through systemic restoration of impaired nitric oxide.

INTRODUCTION: Excessive aggregation of platelets at the site of plaque rupture on the coronary artery led to the formation of thrombus which is reported to precipitate acute myocardial infarction (AMI). Nitric oxide (NO) has been reported to inhibit platelet aggregation and induce thrombolysis through the in situ formation of plasmin. As the plasma NO level in AMI patients from two different ethnic groups was reduced to 0 µM (median) compared to 4.0 µM (median) in normal controls, the effect of restoration of the NO level to normal ranges on the rate of death due to AMI was determined. METHODS AND RESULTS: The restoration of plasma NO level was achieved by a sticking small cotton pad (10×25 mm) containing 0.28 mmol sodium nitroprusside (SNP) in 0.9% NaCl to the abdominal skin of the participants using non-toxic adhesive tape which was reported to normalize the plasma NO level. The participants (8,283) were volunteers in an independent study who had different kinds of cancers and did not wish to use any conventional therapy for their condition but opted to receive SNP "pad" for their condition for 3 years. The use of SNP "pad" which normalized (≈4.0 µM) the plasma NO level that in consequence reduced the death rate due to AMI, among the participants, was found to be significantly reduced compared to the death due to AMI in normal population. CONCLUSION: Our data suggested that the use of SNP "pad" significantly reduced the death due to AMI. TRIAL REGISTRATION: www.ctri.nic.in CTRI/2013/12/004236.

Flavan-3-ol-enriched dark chocolate and white chocolate improve acute measures of platelet function in a gender-specific way--a randomized-controlled human intervention trial.

SCOPE: We examined whether flavan-3-ol-enriched dark chocolate, compared with standard dark and white chocolate, beneficially affects platelet function in healthy subjects, and whether this relates to flavan-3-ol bioavailability. METHODS AND RESULTS: A total of 42 healthy subjects received an acute dose of flavan-3-ol-enriched dark, standard dark or white chocolate, in random order. Blood and urine samples were obtained just before and 2 and 6 h after consumption for measurements of platelet function, and bioavailability and excretion of flavan-3-ols. Flavan-3-ol-enriched dark chocolate significantly decreased adenosine diphosphate-induced platelet aggregation and P-selectin expression in men (all p ≤ 0.020), decreased thrombin receptor-activating peptide-induced platelet aggregation and increased thrombin receptor-activating peptide-induced fibrinogen binding in women (both p ≤ 0.041), and increased collagen/epinephrine-induced ex vivo bleeding time in men and women (p ≤ 0.042). White chocolate significantly decreased adenosine diphosphate-induced platelet P-selectin expression (p = 0.002) and increased collagen/epinephrine-induced ex vivo bleeding time (p = 0.042) in men only. Differences in efficacy by which flavan-3-ols affect platelet function were only partially explained by concentrations of flavan-3-ols and their metabolites in plasma or urine. CONCLUSION: Flavan-3-ols in dark chocolate, but also compounds in white chocolate, can improve platelet function, dependent on gender, and may thus beneficially affect atherogenesis.

[Dose-effect relationship of traditional Chinese medicine formula for promoting blood circulation to remove stasis on ADP-induced platelet aggregation and rabbit plasma thrombin time].

OBJECTIVE: To investigate the effect of five traditional Chinese medicine formula for promoting blood circulation to remove stasis (Xuefuzhuyu Tang, Shaofuzhuyu Tang, Gexiazhuyu Tang, ShentongZhuyu Tang, Tongqiaohuoxue Tang) on platelet aggregation and clotting time in a dose-response relationship. METHOD: The platelet aggregation was tested with Born's method and thrombin time (TT) method was established to determine the clotting time. RESULT: The formula for promoting blood circulation to remove stasis showed significant inhibitory effects on the platelet aggregation and prolonged clotting time, the supernate of 80% alcohol solution showed the most potent inhibitory activity among the three kinds of samples from one formula (P < 0.01), especially the supernate of Shaofuzhuyu Tang and Gexiazhuyu Tang were better than other formula on platelet aggregation. About the anticoagulant, the whole formula showed the evidently prolonging the clotting time than other two fractions (P < 0.01) and the precipitation were the worst. CONCLUSION: The formula for promoting blood circulation to remove stasis showed significant effect on platelet aggregation and clotting time, and different samples of the formula had different efficacy. The study established a foundation for further bio-activity evaluation of these formulae and provided evidence for the treatment of cardiovascular and cerebrovascular diseases.

The effects of imidazoline agents on the aggregation of human platelets.

Clonidine (2-[(2,6-dichlorophenyl)amino]-2-imidazoline), an imidazoline alpha(2)-adrenoceptor agonist, is known to exert complex effects on human platelet aggregation distinct from those of the catecholamines, which are non-imidazoline alpha-adrenoceptor agonists. This study has investigated the aggregatory/anti-aggregatory effects of various imidazolines on human platelets. Blood samples were taken from normal volunteers and platelet aggregation was assessed by a turbidimetric method using a Chronolog aggregometer. Noradrenaline (2 microM) and adenosine diphosphate (1 microM) were used as aggregating agents. The results showed that, with the exception of moxonidine, all of the imidazoline agents used (with or without alpha(2)-adrenoceptor activity) were able to inhibit noradrenaline-induced platelet aggregation. Compared with the non-imidazoline alpha(2)-adrenergic antagonist, yohimbine, the rank order of potency was: efaroxan (IC50 = 3.07 x 10(-8) M) > idazoxan (IC50 = 1.74 x 10(-7) M) > tolazoline (IC50 = 3.90 x 10(-7) M) > clonidine (IC50 = 1.49 x 10(-6) M) congruent with antazoline (IC50 = 1.77 x 10(-6) M) > yohimbine (IC50 = 3.19 x 10(-6) M) > rilmenidine (IC50 = 1.27 x 10(-5) M) > moxonidine (IC50 > 10(-4) M). Clonidine-displacing substance (CDS), a putative endogenous ligand at imidazoline receptors, was found to inhibit noradrenaline-induced platelet aggregation. Harmane, norharmane and agmatine, putative candidates for the active principle of CDS, had no effect on noradrenaline-induced platelet aggregation. In contrast to noradrenaline-induced aggregation, ADP-induced platelet aggregation was neither potentiated nor inhibited by the imidazoline agents, with the exceptions of clonidine and moxonidine. In conclusion, most imidazoline agents effectively inhibit noradrenaline-induced human platelet aggregation. The lack of effect of moxonidine and the proposed endogenous ligands suggested this effect was mediated by an 'atypical' non-adrenoceptor imidazoline-binding site. The results indicated an anti-aggregatory role of imidazoline compounds on noradrenaline-induced human platelet aggregation. In addition, CDS might be an endogenous modulator that prevented platelet hyper-reactivity to catecholamine stimulation.

[Phenomenon of pseudomutagenesis: experiments with modifiers of cell metabolism in human lymphocytes]. / Iavlenie psevdomutageneza: opyty s modifikatorami kletochnogo metabolizma na limfotsitakh cheloveka.

The term "pseudomutagens" is suggested for factors that increase the spontaneous mutation level not by induction of DNA molecule changes but as a result of inhibition of the repair of spontaneous pre-mutation lesions. The expected regularities of pseudomutagen effects, which are suitable as criteria for their identification, are considered. The experimental results on the effects of six modifiers of cell metabolism in human lymphocytes are analyzed. Four of the modifiers (5-fluorodeoxyuridine, sodium cyanide, sodium fluoride and monoiodoacetic acid) behaved as pseudomutagens according to all the criteria applied. 2,4-Dinitrophenol proved to be a true mutagen. Adenosine diphosphate revealed unusual properties: it inhibited the repair of spontaneous lesions but decreased the frequency of structural mutations induced by radiation. The significance of the phenomenon of the pseudomutagenesis for the problem of environmental mutagens/carcinogens is discussed.

Formation of occlusive platelet aggregates in whole blood caused by low concentrations of ADP.

Minute concentrations of ADP are released when platelets are exposed to shear stress during extracorporeal flow. However, based on current methods, these low concentrations have not been shown to have a significant impact on platelet function. We report here the formation of rigid microaggregates (MA) in response to low concentrations of ADP. A newly developed light scattering whole blood aggregometer (LSWBA) was used to detect an aggregation dose response to ADP (0-2 microM) in heparinized (1.5 U/ml) human blood. Although the LSWBA showed that ADP induced MA were reversible, evidence provided by constant pressure filtration (50 mm Hg) suggested that aggregates existed as rigid particles in the blood for up to 6 minutes. The possible implications of these findings to extracorporeal circulation are discussed.

Influência da cafeína no comportamento da pressäo arterial e da agregaçäo plaquetária / Influence of caffeine on blood pressure and platelet aggregation

OBJECTIVE: Studies have demonstrated that methylxanthines, such as caffeine, are A1 and A2 adenosine receptor antagonists found in the brain, heart, lungs, peripheral vessels, and platelets. Considering the high consumption of products with caffeine in their composition, in Brazil and throughout the rest of the world, the authors proposed to observe the effects of this substance on blood pressure and platelet aggregation. METHODS: Thirteen young adults, ranging from 21 to 27 years of age, participated in this study. Each individual took 750mg/day of caffeine (250mg tid), over a period of seven days. The effects on blood pressure were analyzed through the pressor test with handgrip, and platelet aggregation was analyzed using adenosine diphosphate, collagen, and adrenaline. RESULTS: Diastolic pressure showed a significant increase 24 hours after the first intake (p<0.05). This effect, however, disappeared in the subsequent days. The platelet aggregation tests did not reveal statistically significant alterations, at any time during the study. CONCLUSION: The data suggest that caffeine increases diastolic blood pressure at the beginning of caffeine intake. This hypertensive effect disappears with chronic use. The absence of alterations in platelet aggregation indicates the need for larger randomized studies.

Agregaçäo de plaquetas em crianças com asma crônica grave / Platelet aggregation in children with grave chronic asthma

As plaquetas podem estar envolvidas na fisiopatologia da asma liberando mediadores inflamatórios e/ou espasmogênicos, bem como interagindo com outras células inflamatórias. O objetivo deste trabalho foi investigar a agregaçao plaquetária de crianças com asma crônica grave. Foram selecionadas 16 crianças entre 7 e 15 anos de idade, tratadas com brocodilatadores inalatórios e beclometasona (no máximo 750 mug/dia). Doadores de sangue foram selecionados como controles-sadios. Plasma em plaquetas (PRP) e plasma pobre em plaquetas (PPP) foram preparados de acordo com procedimento padrao. A agregaçao de plaquetas foi estudada em agregômetro Payton de dois canais, calibrados com PRP (0 por cento) e PPP (100 por cento) segundo a transmitância de luz. Os estímulos utilizados para induzir a agregaçao plaquetária foram adrenalina (1-30 muM) ou adenosina bifosfato (ADP,1-30 muM). Os resultados foram expressos como percentagem da transmitância máxima. As medianas da agregaçao plaquetária dos pacientes foram respectivamente (adrenalina/ADP 1,3,10 e 30 muM): 0/0, 30/48/41/52, 45/58. As medianas da agregaçao plaquetária dos controles-sadios foram respectivamente (adrenalina/ADP 1,3,10 e 30 muM): 0/0, 12/44, 29/65, 54/74. A análise estatística demontra que a agregaçao de plaquetas dos pacientes foi significativamente menor que a de controles-sadios quando o estímulo foi ADP nas concentraçoes 10 e 30 muM. Em contrapartida, frente aos estímulos adrenalina (1-30 muM) ou mesmo ADP em concentraçoes menores (1-3 muM), a agregaçao de plaquetas dos pacientes foi similar a de controles-sadios. Concluímos que plaquetas de crianças com asma crônica grave sao hiporresponsivas quando o estímulo é ADP em altas concentraçoes. Os mecanismos bioquímicos e molecular envolvidos nesse fenômeno encontram-se sob investigaçao.

Impaired responsiveness of platelets from patients with stable angina pectoris to antiaggregating and cyclicAMP-elevating effects of prostaglandin E1.

The antiplatelet effects of prostacyclin (PGI2) and prostaglandin E1 (PGE1) are mediated by the same receptor and are secondary to intraplatelet cyclicAMP formation. Therefore, any dysfunction in PGI2/PGE1-stimulated cyclicAMP generation might lead to pathologically increased platelet aggregation. This possible consequence has not yet been studied. We examined antiaggregating effects of PGE1 in comparison with its cyclicAMP-elevating potency in platelets obtained from normal subjects and patients with stable angina pectoris; platelet hyperaggregability in such patients has been documented by us previously. ADP-induced aggregation was measured in platelet-rich plasma (PRP); PGE1 was added to platelets 0.5 min after ADP for assessment of reversal of incipient aggregation. Concentrations of PGE1 associated with 50% reversal of aggregation (C50) were 2.4 +/- 0.3 x 10(-8) M in normal subjects and 6.3 +/- 1.6 x 10(-7) M in patients (p < 0.01). PGE1 produced a concentration-dependent increase in intraplatelet cyclicAMP, and there was a strong correlation between cyclicAMP-stimulating and antiaggregating effects of PGE1. Maximal increases in cyclicAMP with PGE1 10(-4) M were 330 +/- 10% for normal subjects and 220 +/- 20% for patients (p < 0.01). Thus, the observed decrease in PGE1-induced reversal of platelet aggregation in patients can be attributed to a suppressed cyclicAMP response to PGE1. These results are likely also to imply reduced platelet sensitivity in vivo to endogenous PGE1 and PGI2, which in turn might contribute to platelet hyperaggregability observed in cardiovascular diseases.

Alguns aspectos da funçäo endotelial em cirurgia cardíaca / Some aspects of the endothelial function in cardiac surgery

Este estudo mostra alguns aspectos da funçäo endotelial relacionados, diretamente, com a cirurgia cardíaca: 1) Após isquemia miocárdica global seguida de reperfusäo, o endotélio coronariano perde a habilidade de expressar vasodilataçäo endotélio-dependente mediada por receptores, ao passo que o relaxamento endotélio-dependente mediado pelo cálcio ionóforo A23187 e a fosfolipasec C, que näo dependem de estimulaçäo de receptores, encontra-se inalterada. O relaxamento produzido pelo fluoreto de sódio, o qual atua através de G-proteína(s), encontra-se comprometido. Estes experimentos indicam que o comprometimento da produçäo de EDRF/NO mediada por receptores após a lesäo de reperfusäo possa ser devido a uma disfunçäo de G-proteínas que liga os receptores da célula endotelial à via da sínese de EDRF/NO; 2) Quarenta e cinco minutos de parada cardioplégica de coraçöes de cäes, pela soluçäo St. Thomas näo comprometem a produçäo de EDRF/NO em artérias epicárdicas coronárias. Estudos farmacológicos in vitro semelhantes, testando-se os efeitos da soluçäo UW, suportaram o conceito de que ela näo lesa o endotélio coronariano, sendo segura para a preservaçäo cardíaca durantes transplantes cardíacos; 3) Em segmentos de artérias coronárias, renais, femorais, e em segmentos de artéria pulmonar, a protamina induziu vasodilataçäo endotélio-dependente, mediada pela estimulaçäo da liberaçäo de EDRF/NO. Nas circulaçöes coronariana e sistêmica, ao contrário do que se verificou nos experimentos envolvendo a circulaçäo pulmonar, este efeito foi independente da presença de heparina; 4) Em 83 por cento dos ensaios biológicos, o efluente da AMI esquerda induziu um relaxamento maior do anel coronariano bioensaiado do que o efluente da AMI direita, por liberaçäo basal de EDRF/NO. Este inibe a adesividade e a agregaçäo plaquetárias e a aterogêne, contribuindo para os resultados superiores obtidos quando se utiliza esta artéria para a revascularizaçäo do miocárdi. Quando expostos à hipoxia, as atividades vasodilatadoras da AMI e da veia safena foram maiores. Esta acentuaçäo da vasodilataçäo causada pela hipóxia foi inibida pelo tratamento com a indometacina, e, rapidamente, revertida, quando se restabeleceu a normóxia.

Vagal and sympathetic nerve activities influenced by posterior cerebral circulation in rabbits.

Vagal and sympathetic nervous activities in the rabbit were recorded while vertebral blood flow was partially blocked by the injection of adenosine 5'-diphosphate (ADP; platelet aggregator). When a small dose of ADP (0.2 mg/kg) was administered into a unilateral vertebral artery, sympathetic nerve (SN) activity increased, and its magnitude was inversely correlated with the extent of the decrease in blood pressure (BP). A larger dose (2 mg/kg) of ADP suppressed SN activity on the injected side, whereas the change was small on the non-injected side. Vagal nerve (VN) activity showed a monophasic excitatory response on both sides, although the change was larger on the injected than on the non-injected side. As a result, asymmetry in autonomic nerve activity was more distinct in SN than in VN. The present study demonstrated that asymmetry of autonomic nervous function can result from changes in blood flow in the cerebellum and brainstem.

The effect of calcitonin gene-related peptide on arrhythmia caused by adenosine diphosphate and desacetyldigilanide-C in rats.

We investigated the effect of calcitonin gene-related peptide on the arrhythmia induced by two drugs in rats. The results of our experiments have proved that calcitonin gene-related peptide could reduce the degree of atrioventricular block, protect against the attacks of sinus standstill and ventricular fibrillation produced by adenosine diphosphate, and improve restoration of sinus rhythm. Calcitonin gene-related peptide was able to eliminate sinus standstill and ventricular fibrillation resulting from administration of desacetyldigilanide-C. These results demonstrate that calcitonin gene-related peptide has strong antiarrhythmic effects in experimental animals.

Effect of long-term bisphosphonate treatment on morbidity due to bone metastases in breast cancer patients.

The effect of long-term bisphosphonate (APD) treatment on the morbidity from bone metastases in breast cancer patients was studied in a controlled clinical trial. 131 patients were randomized between treatment with APD (300 mg/day orally) or control. Systemic treatment for breast cancer was left to the discretion of the physician. The distribution of cases according to age, receptor status and previous treatment was similar in both groups. Patients were examined at 3-month intervals, while bone scans and radiography of relevant lesions in the skeleton were performed every 6 months. After a median follow-up of 13 months, the morbidity in the treated group was significantly less than in the controls. This concerned the occurrence of hypercalcemia, bone pain and fractures, and the need for radiotherapy of osteolytic lesions. In this interim analysis, APD treatment more than halved the requirement for specific treatment of bone lesions. The treatment is simple and well tolerated at a relatively low dosage. A higher oral dose was precluded due to gastrointestinal toxicity. Because the effect of APD on skeletal morbidity was not complete, efforts should be made to develop more effective and less toxic bisphosphonates.