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A Ordinas

Publications and source records attributed to A Ordinas.

At least 145 records · Page 8Linked to original sources
PubMed1991

Comparison between human umbilical artery and rabbit abdominal aorta as substrata for platelet adhesion and platelet thrombus formation under flow conditions.

Rabbit abdominal aortas and human umbilical arteries are currently used as substrata for the study of platelet adhesion and aggregate formation under flow conditions. Using immunohistochemical and ultrastructural methods, we have analyzed both vessel surfaces. The reactivity towards platelets of the subendothelium (SE) exposed on these vessels after mechanical or enzymatic digestion (alpha-chymotrypsin) was morphometrically quantified and the nature of the interaction studied in the electron microscope. After mechanical damage, the ultrastructural study of rabbit aortas showed a clearly defined internal elastic lamina (IEL). In contrast, umbilical vessels lacked a consistent IEL and masses of amorphous material often located deeper in the media were the main constitutents of the SE. Immunohistochemical labeling of the von Willebrand factor bound to both types of vessel differed considerably. Quantification of platelet interactions after perfusion of citrated blood showed qualitative differences between mechanically damaged rabbit or human vessels. Enzymatic digestion produced a more thrombogenic surface on rabbit aortas (p less than 0.01 vs. nondigested), but decreased their reactivity towards platelets on umbilical arteries (p less than 0.01 vs. nondigested). The ultrastructural study of the interacting platelets revealed that aggregates, when present, were found on the extracellular matrix underlying endothelial cells of rabbit aortas, but interacting with fibrillar structures probably derived from cell elements of the media in the case of umbilical arteries. These findings indicate that rabbit aortas and umbilical arteries possess structural characteristics that result in different thrombogenic properties with respect to circulating platelets.

Aorta, Abdominal↗
PubMed1991

Effects of endothelial cell treatment on 13-HODE and prostacyclin synthesis and its correlation with tumor cell-vascular endothelial cell adhesion.

Adhesion of tumor cells to vascular endothelial surfaces is one of the key steps in metastatic dissemination. Several factors are believed to be implicated in the regulation of the adhesive properties of tumor cells. We show that the adhesion of five different tumor cell lines, all of them of human origin, to human umbilical vascular endothelial cells (ECs) significantly increases following pretreatment of ECs with the cytokines interleukin-1 and tumor necrosis factor, whereas tumor cell/EC interactions remained unchanged after incubation with interferon-gamma. Significant augmentation in tumor cell adhesion was also observed when ECs were treated with the lipoxygenase inhibitors salicylate and the compound BW755C. In all cases, increased tumor cell adhesion was concomitant with significant decreases in the EC levels of linoleic acid, lipoxygenase-derived metabolite 13-hydroxy-octadecadienoic acid (13-HODE). On the contrary, pretreatment of the EC monolayers with aspirin did not result in any changes towards tumor cell adhesion. These results suggest that tumor cell/EC interaction is modulated, at least in part, by intracellular levels of 13-HODE and is independent of prostacyclin (PGI2) production by the ECs.

6-Ketoprostaglandin F1 alpha↗
PubMed1990

[Effects of the treatment with human recombinant erythropoietin on primary hemostasis in uremic patients].

The effect of human recombinant erythropoietin (r-EPO) on primary hemostasia was studied in 9 patients undergoing hemodialysis. The analysis was performed before and after the hematocrit reached a value of 30%. The most important complications observed during the study period were a death for acute myocardial infarction in a patient with previous severe ischemic cardiopathy and a thrombosis of the venous line. The bleeding time shortened in four patients although the mean value did not change significantly. Platelet count showed a non significant increase. There was a significant improvement in in vitro platelet aggregability with ADP (p less than 0.05), arachidonic acid (p less than 0.05), adrenaline (p less than 0.05), and ristocetin (p less than 0.05) as well as in the parameters that quantify the interaction between platelets and subendothelium in in vitro experiments using perfused models (p less than 0.05). There were no significant changes in coagulation and fibrinolysis tests. The treatment with r-EPO improved primary hemostasia in uremic patients. This beneficial effect was due to the increased hematocrit and to the improvement of platelet function induced by r-EPO.

Adult↗
PubMed1990

Uremic platelets have a functional defect affecting the interaction of von Willebrand factor with glycoprotein IIb-IIIa.

Uremic patients have an impaired platelet function that has been related to membrane glycoprotein (GP) abnormalities. Using a perfusion system, we have studied the interaction of normal and uremic platelets with vessel subendothelium (SE) under flow conditions. Reconstituted blood containing washed platelets, purified von Willebrand factor (vWF) (1 U/mL), and normal washed red blood cells was exposed to de-endothelialized rabbit segments for 10 minutes at two different shear rates (800 and 1,600 seconds-1). In some experiments a monoclonal antibody to the GPIIb-IIIa complex (EDU3) was added to the perfusates. With normal platelets, the percentage of the vessel covered by platelets (%CS) was 23.1% +/- 3.7% at 800 seconds-1 and 30% +/- 4.3% at 1,600 seconds-1. Platelets were observed in contact or forming monolayers on vessel SE. EDU3 inhibited the spreading of normal platelets. The %CS (11.1% +/- 3.3%) was statistically decreased (P less than .01) and most of the platelets were observed in contact with the vessel surface. These data indicate that, under flow conditions, the interaction of vWF with GPIIb-IIIa can support the spreading of normal platelets in the absence of exogenous fibrinogen. Under the same experimental conditions, the interaction of uremic platelets with SE was markedly impaired at both shear rates studied (P less than .01 v normal platelets). The presence of EDU3 did not modify the interaction of uremic platelets. These results confirm the impairment of the platelet adhesion observed in uremic patients. Furthermore, they indicate the presence of a functional defect in the interaction of vWF with GPIIb-IIIa. The fact that perfusions with normal and uremic platelets in the presence of an antibody to the GPIIb-IIIa complex did not show any differences gives indirect evidence on a functionally normal interaction vWF/GPIb in uremic patients.

Blood Platelets↗
PubMed1990

Investigation of plasma von Willebrand factor and circulating platelet aggregating activity in mitomycin C-related hemolytic-uremic syndrome.

We have studied the ability of the plasma to induce aggregation of both homologous and heterologous platelets in four patients with hemolytic-uremic syndrome (HUS) associated with chemotherapy with mitomycin C (MMC). Neither platelet aggregation was elicited by patients' plasmas nor the in vitro addition of purified von Willebrand factor (vWF) had any effect on the aggregation pattern. In addition, ristocetin-induced binding of patients' vWF to formaldehyde-fixed platelets was normal, and multimeric vWF analysis revealed a normal structure of patients' plasmatic vWF whatever the clinical stage in which it was studied. These findings suggest that, in spite of the existence of common clinical and biological features in the various forms of HUS, the pathogenesis of MMC-related HUS may be, at least in part, different from that of the other forms of HUS in which both platelet-aggregating activity and alterations in the vWF are found.

Blood Platelets↗
PubMed1990

Differential localization of von Willebrand factor, fibronectin and 13-HODE in human endothelial cell cultures.

Von Willebrand factor (vWF), fibronectin (FN) and 13-hydroxy-octadecadienoic acid (13-HODE) are known to influence the regulation of the adhesive properties of vascular surfaces. In the present study vWF, FN and 13-HODE were comparatively localized in endothelial cells (EC) and in the extracellular matrix (ECM) produced by EC. An indirect immunofluorescent technique was applied to coverslips containing human EC cultures previously fixed and permeabilized following different procedures: A. Alcohol/acetone; B. Paraformaldehyde alone and C. Paraformaldehyde followed by Triton X-100. vWF was observed inside EC (A), on the ECM produced by EC (B) or in EC and ECM (C) depending on the fixation procedures used. FN was mainly localized in the ECM despite the fixation procedures employed. FN was only seen in relation to cell bodies after strong permeabilization (A). Under our experimental conditions 13-HODE was never found in ECM. This latter antigen was observed randomly dispersed in those preparations fixed with alcohol/acetone, indicating that it is probably extracted by this fixative. 13-HODE was detected in granular shaped structures in EC after permeabilization with detergent (C). These results suggest that the cellular localization of vWF and FN is compatible with an adhesive role related to the abluminal side of ECs. 13-HODE was readily observed after mild permeabilization. This finding would be morphologically consistent with its contribution to the regulation of the vessel wall thromboresistance.

Cells, Cultured↗
PubMed1990

[Update on the role of vascular, plasmatic and cellular components in the regulation of the interaction of platelets with the subendothelium].

The interaction of platelets (Plts) with subendothelium (SE) or with extracellular matrices (ECM) generated by endothelial cells was studied under flow conditions. The role of: a) platelet membrane glycoproteins (GPS) or subunits; b) plasma adhesive proteins such as von Willebrand factor (VWF) and fibronectin (Fn); and c) structural proteins of the SE such as laminin (Lm). Specific antibodies or purified proteins were added to the perfusates. GPS. Monoclonal antibodies (MoAb) directed against distinct epitopes of the GPIIb-IIIa affected in different manner the deposition of platelets. MoAb EDU3 decreased (p less than 0.01) the surface of the vessel covered by platelets (% CS) and increased the presence of platelets in contact (% C) with the SE. MoAb C17 did not modify the % CS but altered the morphological characteristics of the aggregates. A monoclonal antibody against GPIIb, decreased the % CS (p less than 0.01) without influencing % C; VWF. A linear progression of the % CS was observed for VWF levels ranging from 0.1 to 0.6 U/mL. A plateau was reached for concentrations of vWF above 0.6 U/mL; Fn. Perfusions in which blood was reconstituted with Fn-depleted plasma showed the cooperation of this protein facilitating Plt-Plt interaction. No statistical differences were observed in the % CS when ECM were incubated with a MoAb against Fn (3E3); Lm. The % CS decreased statistically (p less than 0.01) when SE or ECM were incubated with an antibody to Lm. These results show the critical role of vascular, plasmatic and cellular components in the maintenance of and adequate platelet function.

Animals↗
PubMed1990

The role of platelets in cancer metastasis.

Clinical, experimental and ultrastructural studies strongly suggest a role for platelets in metastatic dissemination. Several mechanisms have been proposed to explain the potential contribution of blood platelets to the metastatic cascade. Experimentally, many tumour cells of either animal or human origin have the capacity to activate platelets, although the mechanisms by which malignant cells exert this effect is not yet fully understood. Possible mechanisms include: (1) generation of thrombin; (2) activation by ADP; (3) release of cathepsin B; (4) eicosanoid metabolism. A number of observations also indicated that tumour-cell-induced platelet aggregation required specific receptor sites. We have shown that platelet glycoprotein GPIb and the complex GPIIb/IIIa are necessary for tumour-cell-induced platelet aggregation. We and others reported the isolation of a microparticulate aggregating material from different types of tumour cell lines. This material has been identified as a sialolipoprotein complex which possesses tissue-factor-like activity. The role of sialic acid in the metastatic potential of cells is also believed to be important and may partly modulate their interactions with platelets. In vivo, rheological factors may also regulate the interactions of tumour cells with blood and vascular structures and an alternative approach to the evaluation of platelet-tumour-cell interaction under dynamic conditions has been the use of perfusion systems. Thus, we have established the crucial role of Ca2+ in supporting tumour-cell-platelet activation and subsequent thrombus formation. More recently we investigated the patterns of adhesion of a highly metastatic human adenocarcinoma of the lung to exposed extracellular matrix generated by human vascular endothelial cells in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Diphosphate↗
PubMed1990

A rapid embedding procedure for the study of platelet interactions with extracellular matrices in a flowing system. Effect of aspirin on platelet activity.

The cultured endothelial cell (EC) is currently used as a model for the study of the interaction of platelets with the vascular wall. Described is a method for rapid quantitative and qualitative evaluation of platelet interactions with extracellular matrices (ECM) produced by human cultured ECs growing on plastic coverslips. Morphometric calculations can be performed on the same perfused coverslips. A very good correlation (r = 0.96) was found between results of a morphometric method en face and those obtained from analysis of cross sections of the perfused coverslips. A shear rate-dependent increase on platelet deposition onto ECMs was observed with both morphometric procedures. The method is sensitive enough to detect drug-related changes of platelet function. An impairment of the interaction of platelets with the ECM was observed when blood obtained from healthy volunteers who took 500 mg aspirin/day for five days was perfused. Aspirin showed a marked effect, decreasing platelet spreading onto the subendothelium (p less than 0.05). The embedding method described benefits from the use of plastic coverslips that are easily detected from the glycol methacrylate compound used for the embedding procedure. Quantitative analysis en face (covered surface) and qualitative evaluation of platelet interactions (contact, adhesive and aggregated platelets) in cross sections are performed on the same coverslip. This embedding procedure provides a useful tool for the study not only of platelet interactions with ECMs but also for the investigation of interactions of blood elements with other cultured cells.

Aspirin↗
PubMed1990

[Variations in the thrombogenic properties of the extracellular matrix produced by endothelial cells in consecutive passages].

Cultured endothelial cells are currently used as a model for the study of the interaction of platelets with the vascular wall under flow conditions. Little is known about the influence of cell-culture conditions on the preservation of the original characteristics of primary tissues. In our study, the reactivity of platelets towards extracellular matrices produced by endothelial cells from different passage numbers was morphometrically evaluated. The presence and distribution of the extracellular von Willebrand factor was analyzed by immunofluorescent techniques. Morphology of endothelial cells in culture varied dramatically along the culture and polymorphism increased clearly at fourth or fifth passages. Extracellular matrix thrombogenicity decreased significantly from the fifth passage number. Simultaneously, von Willebrand factor distribution in ECM changed its typical pattern. Our results confirm that the extracellular matrix produced by human umbilical vein derived-endothelial cells constitutes a good model for the study of platelet adhesion onto vascular subendothelium. The limited viability of ECs and the variation in the thrombogenic properties of ECM formed after multiple passages must be taken into account when these cells or their products are used in experimental research.

Cell Division↗
PubMed1989

Platelet and shear rate promote tumor cell adhesion to human endothelial extracellular matrix--absence of a role for platelet cyclooxygenase.

Blood platelets are thought to be involved in certain aspects of malignant dissemination. To study the role of platelets in tumor cell adherence to vascular endothelium we performed studies under static and flow conditions, measuring tumor cell adhesion in the absence or presence of platelets. We used highly metastatic human adenocarcinoma cells of the lung, cultured human umbilical vein endothelial cells (ECs) and extracellular matrices (ECM) prepared from confluent EC monolayers. Our results indicated that under static conditions platelets do not significantly increase tumor cell adhesion to either intact ECs or to exposed ECM. Conversely, the studies performed under flow conditions using the flat chamber perfusion system indicated that the presence of 2 x 10(5) pl/microliters in the perfusate significantly increased the number of tumor cells adhered to ECM, and that this effect was shear rate dependent. The maximal values of tumor cell adhesion were obtained, in presence of platelets, at a shear rate of 1,300 sec-1. Furthermore, our results with ASA-treated platelets suggest that the role of platelets in enhancing tumor cell adhesion to ECM is independent of the activation of the platelet cyclooxygenase pathway.

Adenocarcinoma↗
PubMed1989

Influence of shear stress on tumor-cell adhesion to endothelial-cell extracellular matrix and its modulation by fibronectin.

The interaction of malignant cells with blood-vessel endothelial cells and their underlying basement membrane is an important step in the development of secondary metastases. We investigated the interactions of highly metastatic human tumor cells, the A-549 adenocarcinoma of the lung, with cultured endothelial cells (EC) and their extracellular matrix (ECM). We studied the adhesion patterns of the A-549 tumor cells to EC and ECM under static and flow conditions. Our results provide evidence that tumor-cell adhesion depends not only on the characteristics of the tumor cells themselves, but also on the properties of the EC and ECM. Our results also indicate that tumor-cell adhesion to ECM is shear-rate-dependent, and that it is partially modulated by fibronectin. Moreover, our results suggest that the arg-gly-asp (RGD) common adhesion receptor site is also involved in the adhesion of the A-549 cells to EC and ECM.

Adenocarcinoma↗
PubMed1989

Role for platelet von Willebrand factor in supporting platelet-vessel wall interactions in von Willebrand disease.

Twelve infusions of plasma concentrates of von Willebrand factor (vWF) were given to four patients with severe (type III) von Willebrand disease (vWD). Their prolonged bleeding times were either completely or partially corrected after five infusions and had not changed after the remaining seven. In contrast, the low platelet coverage of the subendothelial surface of rabbit aorta perfused with normal washed platelets and red cells resuspended in preinfusion patient plasma was completely or partially corrected in ten instances by replacing preinfusion plasma with postinfusion plasma and remained unchanged in two. Postinfusion improvement in surface coverage was greater than that in bleeding time, suggesting that vWF from normal platelets is needed to support optimal platelet-vessel wall interactions in vWD. This possibility was further explored through other perfusion experiments. The subendothelial surface covered by platelets from an untreated patient with type III vWD (containing no measurable vWF) or from a type IIA vWD patient (containing dysfunctional vWF) resuspended in normal plasma was much smaller than that covered by normal platelets resuspended in normal plasma. These results establish that platelet vWF is important in supporting platelet-vessel wall interactions in vWD and also provide experimental support in favour of the therapeutic transfusion of normal platelets in addition to vWF concentrates to correct the bleeding time in vWD patients.

Blood Platelets↗
PubMed1989

Uremic plasma after infusion of desmopressin (DDAVP) improves the interaction of normal platelets with vessel subendothelium.

Effects of 1-deamino-8-D-arginine vasopressin (DDAVP; 0.4 microgram/kg iv) were studied in 11 patients with uremia. Bleeding time, platelet retention on glass beads, factor VIII activities, plasma catecholamine levels, and studies on platelet interaction with the subendothelium were performed before, 1 hour after, and 6 hours after DDAVP infusion. Perfusates consisting of normal washed platelets, uremic platelet-poor plasma (u-PPP) and washed red blood cells were perfused through the Baumgartner perfusion system at a shear rate of 800 sec-1. One hour after DDAVP infusion, a shortening in the bleeding time and an increase in platelet retention on glass beads were noticed in these patients (p less than 0.01). Simultaneously, plasma levels of noradrenaline, factor VIII coagulant (FVIII:C), and von Willebrand factor (vWF) activities were statistically increased. Platelet deposition and platelet aggregate formation on subendothelium were consistently increased (p less than 0.05) in perfusions carried out with blood reconstituted with u-PPP obtained 1 hour after DDAVP. The "in vitro" addition of 1 U/ml vWF or 1 U/ml vWF plus 1 U/ml factor VIII to the pretreatment u-PPP had no significant influence on the parameters that quantify platelet-subendothelium interaction. However, after the addition of 10 ng/ml noradrenaline to a similar system containing basal u-PPP, a clear improvement (p less than 0.05) in platelet deposition was noticed. Our results confirm the hemostatic effectiveness of DDAVP in patients with uremia and reveal an increased platelet interaction with subendothelium mediated by a factor present in uremic plasma after DDAVP administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1988

Asialo von Willebrand factor enhances platelet adhesion to vessel subendothelium.

Native von Willebrand factor (N-vWF) binds to platelets activated by thrombin, ADP or ristocetin. Asialo vWF (As-vWF) induces platelet aggregation in absence of platelet activators. N-vWF mediates platelet adhesion to vessel subendothelium at high shear rates. We have investigated the role of As-vWF in supporting platelet deposition to rabbit vessel subendothelium at a shear rate of 2,000 sec-1, using the Baumgartner perfusion system. We have studied the effects of the addition of As-vWF (from 2 to 12 micrograms/ml) to perfusates consisting of washed red blood cells, 4% human albumin and washed platelets. Our results show a significant increase in platelet deposition on subendothelium (p less than 0.01) in perfusions to which As-vWF had been added. Blockage of the platelet glycoproteins Ib and IIb/IIIa (GPIb and GPIIb/IIIa) by specific monoclonal antibodies (LJIb1 and LJCP8, respectively) resulted in a decrease of platelet deposition in both types of perfusates prepared with N-vWF and As-vWF. Our results indicate that As-vWF enhances platelet deposition to vessel subendothelium under flow conditions. Furthermore, they suggest that this effect is mediated by the binding of As-vWF to platelet membrane receptors, which in turn, promote platelet spreading and adhesion to the subendothelium.

Antibodies, Monoclonal↗
PubMed1988

Ristocetin induces platelet aggregation: a morphological demonstration.

Changes in the morphology of human platelets induced by ristocetin in platelet-rich plasma (PRP) have been analysed at the ultrastructural level by means of a tannic acid procedure. Studies were also undertaken to measure the release of serotonin. Modifications of the aggregation tests induced by apyrase, a monoclonal antibody (Mab) to GPIIb/IIIa and by EDTA were also investigated. Transmission electron microscopy revealed that ristocetin precipitated adhesive proteins on the platelet membrane. An electron-dense deposit was seen within 20 s after ristocetin was added. When experiments were carried out in the aggregometer cuvette during stirring, groups of platelets became activated, changed their shape, and finally aggregated releasing part of their contents. The morphology of aggregates did not differ from those formed in the presence of ADP. Aggregation studies demonstrated that a Mab to GPIIb/IIIa modified the extent and the rate of the aggregation curve when RIPA was performed in citrated platelet-rich plasma (c-PRP), while apyrase modifies the extent, but not the slope, of the curve. Neither the antibody nor apyrase modified RIPA when it was performed in PRP obtained in the presence of EDTA. All this evidence suggests that RIPA in c-PRP, besides reflecting the interaction of GPIb with vWF, may also test other mechanisms of the platelet function including: assembly of GPIIb/IIIa complex, interaction of fibrinogen with this glycoprotein complex, and possibly the release reaction.

Antibodies, Monoclonal↗
PubMed1988

Effects of a fish oil enriched diet on aspirin intolerant asthmatic patients: a pilot study.

The effect of a fish oil enriched diet containing about 3 g of eicosapentaenoic acid was studied in 10 patients with aspirin intolerant asthma. Subjects were studied during six weeks on a control diet followed by six weeks on the fish oil diet in a single blind study design. They were asked to record their peak expiratory flow (PEF) twice daily, bronchodilator and steroid doses, and subjective ratings of pulmonary symptoms on diary cards. There were no significant changes in symptom scores over the six weeks of either the control diet or the fish oil diet. PEF values, however, were significantly lower during the fifth and sixth week of the fish oil diet than during the control diet (308 v 262 l/min week 5 and 306 v 256 l/min week 6). Bronchodilator usage was also greater during the fifth and sixth week of the fish oil diet than during the control period (12.0 v 7.4 and 13.0 v 7.4 puffs a day in weeks 5 and 6). This pilot study suggests that fish diets may have a deleterious effect on patients with aspirin intolerant asthma.

Adult↗