Biomedical subjects
Publications and source records attributed to E Bastida.
A 37-year-old man who had suffered a thoracic trauma presented night release of whitish urine 2 years later. Thoracic computed tomography and aortography demonstrated an aneurysm of the thoracic aorta. Lymphography confirmed the compression of the thoracic duct by the aneurysm. After surgical repair the patient has remained asymptomatic.
A total of 15 chronic renal failure patients on hemodialysis therapy underwent some kind of cardiovascular surgery between August 1984 and March 1993. Ten had a valve abnormality, and the remaining five had coronary artery disease. All of them were hemodialyzed the day before surgery and 24-48 h after the operation. Eleven recovered well after surgery, four died of septic shock: two of these were in septic shock prior to surgery; one was in acute congestive heart failure, and one was operated during an acute myocardial infarction. All operative deaths occurred in the patients who underwent non-elective surgery or were preoperatively in New Heart Association (NYHA) class IV. The factors having an impact on morbidity and mortality seem to be more related to the previous clinical situation and to the urgency of the operation than to the status of chronic renal failure. An early and adequate assessment of the candidates, when possible avoiding emergency surgery and acute left ventricular dysfunction, as well as careful management during cardiopulmonary bypass procedures (CPB) and the immediate post-surgical period will certainly improve the result of cardiac surgery in these patients, making it similar to those who are not in chronic renal failure.
PURPOSE: To assess the value of routine intraoperative transesophageal echocardiography (TEE) in unselected patients (P) undergoing cardiac surgery. DESIGN: Routine intraoperative TEE in unselected patients undergoing cardiac surgery and evaluation of its usefulness and contribute to a successful surgery. PATIENTS: In-hospital patients from cardiology, cardiac surgery and intensive care departments of a general hospital classified as a reference medical centre in cardiovascular pathology. MATERIAL AND METHODS: From February 1994 to May 1994 an intraoperative TEE was routinely performed in patients undergoing cardiac surgery. A total of 130 TEE studies were made corresponding to 128 patients, 69 males and 61 females, with a mean age of 49.4 +/- 12.1 years. Indications for surgery were as following: a) Coronary artery bypass grafting (C.A.B.G.)-39; b) Valvular surgery-76; c) C.A.B.G. and valvular surgery-5; d) Thoracic aorta pathology associated or not to aortic valve surgery and/or coronary arteries re-implantation-6; e) Other-4. TEE performed using either a monoplane, biplane or multiplane probe and the images acquisition was made before thoracotomy and cardiopulmonary bypass (CPB) and after CPB with hemodynamic stabilization. RESULTS: Pre CPB imaging yielded unsuspected findings in 11 P (8.5%) that changed the planned surgery in 7 cases (5.4%). Post CPB echo study, which was performed after hemodynamic stabilization, modified the surgical strategy in 13 cases (10.9%) avoiding 1 predetermined surgical procedure (tricuspid ring annuloplasty) and a new surgery in another case, leading to a 3 non-predetermined surgical procedures (by showing significative tricuspid regurgitation), to a further surgery in the same valve in 6 patients (mitral paravalvular leak with significative regurgitation in 1P, another mitral prosthesis dysfunction in 1P, residual mitral valve insufficiency after mitral valve repair in 3 P and aortic prosthesis dysfunction in 1 P), to a intraaortic counterpulsation balloon device in 1 P and to new CPB due to an insufficient extraction of calcified pericardium in one case of constrictive pericarditis. In 23 cases (17.7%) TEE has modified the anesthetic procedure by demonstrating signs of hemodynamic changes before Swan-Ganz catheter, thus prompting changes in the administrations of fluids and either in inotropic or vasodilator agents. In total, intraoperative TEE has changed the surgical and/or anesthetic plan in 43 cases (33.0%). There was no difficulties or complications related to the procedure. CONCLUSION: These data indicate that intraoperative TEE is useful in formulating the surgical plan and assessing immediate operative results as well as a guide to anesthetic procedures. Its high rentabillity in modifying the surgical and/or anesthetic plans lead us to believe that it must be used as a routine procedure in patients undergoing cardiac surgery.
Evidence indicates that complex interactions occur between blood platelets and polymorphonuclear leukocytes (PMN) referring to both activation and inhibition phenomena. We studied the influence of co-incubating activated PMN and platelets at varying cell counts in the production of two active metabolites, thromboxane A2 (TxA2) measured as thromboxane B2 (TxB2) and platelet activating factor (PAF). The decrease observed in the quantity of TxA2 synthesized by 2 x 10(8) platelets/ml in the presence of physiological PMN counts clearly indicates an inhibitory effect of PMN in AA-derived metabolites. This suggests there is an influence of PMN-released products on activated platelets, decreasing their capacity to synthesize TxA2. Moreover, we found that the addition of platelets, which do not make measurable quantities of PAF, to PMN suspensions significantly decreased the capacity of PMN to synthesize this mediator. Together with this inhibiting action we also observed an activating effect, in such a way that the more PMN were added to platelet suspensions the more TxA2 was produced. Conversely, the maximal amounts of PAF were synthesized when the highest platelet count suspensions were added to PMN. In summary, the results we present here clearly show cellular interactions between platelets and PMN that directly influence the amounts of metabolites synthesized by both cells.
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A parallel-plate perfusion chamber has been used to evaluate the contribution of the adhesive membrane glycoprotein CD36 (GPIV) to platelet adhesion on type I collagen in flowing whole blood at a shear rate of 800 s-1. In one series of experiments, reconstituted normal blood (hematocrit 0.4; platelet count 1.5 x 10(5)/microL) was prepared from washed red blood cells, plasma, and washed platelets that had been incubated with Fab fragments of a monospecific polyclonal anti-CD36 antibody (50 micrograms/mL, 30 minutes, 37 degrees C). Percent surface coverage of collagen-coated coverslips using reconstituted blood with antibody-blocked platelets, as compared with paired reconstituted controls (100%), was 50% at 2 minutes, 87% at 5 minutes, and 90% at 10 minutes. Further studies were performed by perfusion of whole blood from a healthy donor of the Naka-negative phenotype, whose platelets constitutively lack CD36, over collagen-coated coverslips. In this case, percent surface coverage was 55% of normal controls at 2 minutes, 76% of controls at 5 minutes, and 72% of controls at 10 minutes. In both preparations, platelets lacking functional CD36 had a statistically significant decrease (P < .005) in adhesion after 2 minutes and 10 minutes perfusion but not at 5 minutes. These results show that functional CD36 facilitates the rapid adhesion of platelets to collagen and that this effect is seen at the earliest time points of their interaction.
Previously, we have demonstrated that stimulation of endothelial cells (ECs) with interleukin-1 alpha (IL-1 alpha) enhances the synthesis and expression of the vitronectin receptor (VnR), promotes VnR-dependent adhesion of human A549 adenocarcinoma cells to ECs, and is associated with decreased EC 13-hydroxyoctadecadienoic acid (13-HODE) synthesis in vitro. To determine whether these observations are relevant in vivo, we examined the acute retention and subsequent metastasis of intravenously-injected B16F10 melanoma cells in murine lungs, in relation to vessel wall 13-HODE. In C57BL/6 mice pretreated with IL-1 alpha, vessel wall 13-HODE was decreased and B16F10 lung entrapment and metastasis were increased. The latter two events were blocked by pretreating the animals with the GRGDS peptide. These data suggest a relationship between vessel wall 13-HODE synthesis, adhesion molecule expression, and adhesion of B16F10 cells to the endothelium.
PURPOSE: To assess, by means of a morphometric method, the interaction of platelets with subendothelium, as induced by perfusion with the Baumgartner system, and to compare the antiplatelet effect of acetylsalicylic acid (ASA) and dipyridamole (DIP) and the combination of both drugs. MATERIAL AND METHODS: Blood samples were drawn from healthy volunteers and platelet-rich plasma specimens were prepared at a concentration of 4 x 10(8)/mL. Abdominal aorta segments were obtained from New Zealand rabbits, they being deprived of endothelium by alpha-chymotrypsin. Red cells and platelets were separately or conjointly treated on reconstituted whole blood using ASA and DIP at therapeutic doses, separately or in association. Perfusion experiences were carried out in Baumgartner's annular chamber. Upon finishing the studies, the aorta segments were morphometrically evaluated by an optic analysis system attached to a computer with an automatic image recognition programme. The statistical analysis was performed with Student's t test, differences between groups of p < 0.05 being assigned significance. RESULTS: Treatment with the association ASA-DIP was the only one to significantly reduce interaction, as both the size of aggregates and the extent of the covered surface decreased when whole blood was treated, and only the covered surface did when red cells were separately treated. When treating only platelets, the reduction of aggregates had no significance. CONCLUSION: The synergism of ASA and DIP in antiplatelet therapy was confirmed, as well as the important enhancing effect exerted by red cells.
PURPOSE: To evaluate the direct action of PAF on the pro-coagulant activity of cultured vascular endothelial cells; to analyse by photometric methods the thrombogenic effect of PAF on platelets, and to assess platelet deposition on vascular endothelium. MATERIAL AND METHODS: Human endothelial cells were isolated from umbilical cord vein and incubated for three minutes at 22 degrees C with different PAF concentrations (10(9) M to 10(-7) M) in order to assess the influence of this lipidic mediator on the procoagulant activity of the cells. The effect of PAF on platelet aggregation was assessed by aggregation studies using arachidonic acid (AA) and different PAF and Lyso-PAF concentrations (10(-8) M to 10(-4) M). Serotonin (5-HT) release was tested in platelet rich plasma (PRP) samples highly sensitive to PAF. PRP samples were incubated for 30 minutes at 22 degrees C with 100 microCi 3H-5-HT. Platelets were activated with 10(-7) M to 10(-9) M PAF concentrations, the percentage of 3H-5-HT released into the extra-platelet medium being calculated. Baumgartner's continuous perfusion model was used to study platelet deposition on the vascular endothelium. The morphometric evaluation was carried out by a planimeter assembled to a data processor with a programme devised for analysing the platelet-subendothelium interaction. RESULTS: Evaluation of the procoagulant activity on the cell surface: The expression of the procoagulant activity showed no variations with respect to the controls under different concentrations of PAF. Platelet aggregation and release studies: Normal values of platelet aggregation (80% to 100%) were found when using AA, however, there were great case to case variations under different PAF concentrations. Serotonin release was less marked than aggregation itself. The use of Lyso-PAF failed to elicit platelet aggregation and serotonin release in any case. Morphometric evaluation: The results attained showed that perfusion carried out with 10(-8) M PAF concentration showed contact, adhesiveness and thrombus formation figures similar to those of control perfusion. CONCLUSIONS: Human platelets are not too sensitive to PAF activity, only high PAF concentrations being capable of inducing platelet aggregation and 5-HT release with ample variability. This suggests the existence of a heterogeneous platelet population with PAF receptors. Low PAF concentrations do not modify the haemostatic function, and only those PAF concentrations inducing maximal release and aggregations could reduce the interaction of platelets with vascular subendothelium and the formation of thrombi.
Laminin (LM) is a basement membrane glyco-protein which exhibits a number of biological activities, including the promotion of cell attachment and migration. In a static system, platelets attach to LM without spreading. In order to elucidate the mechanisms mediating platelet-LM interactions under flow conditions, we studied the effects of blocking subendothelial LM and its platelet receptor. We measured the extent of platelet adhesion to the endothelial cell extracellular matrix (ECM) using a parallel plate perfusion chamber. To do this, we performed the following experiments: i) blockade of subendothelial LM by incubation of ECM with an anti-LM antibody (Ab); ii) blockade of the platelet receptor for LM with: a) an anti-67 KDa receptor Ab; and b) three different LM-derived peptides (CFALRGDNP, IKVAV and CDPGYIGSR). In i) perfusates consisted of whole blood, whereas in ii) perfusates were prepared with washed platelets pre-incubated with Ab or peptides and reconstituted with plasma and washed red blood cells. Perfusions were carried out at 800 s-1 shear rate. Platelet deposition was morphometrically evaluated by a computerized system. Our results indicated that when ECM was pre-incubated with an anti-LM Ab (100 micrograms/ml, 30 min, 37 degrees C) a reduction of the surface coverage (SC) of 23.3 +/- 2.3% (p less than 0.01) was observed. When washed platelets were pre-incubated with an anti-67 KDa receptor Ab (40 micrograms/ml, 30 min, 37 degrees C) their interaction with ECM was decreased by 22.3 +/- 2.1% of SC (p less than 0.005). In other experiments platelets were pre-incubated with CFALRGDNP, IKVAV and CDPGYIGSR (200 micrograms/ml, 30 min, 37 degrees C).(ABSTRACT TRUNCATED AT 250 WORDS)
In order to elucidate whether a single dose of 150 mg acetylsalicylic acid three times a day, provokes significant changes into platelet aggregation, we analyzed a group of 50 patients who underwent coronary revascularization. During 48 hours before surgery, all patients received 100 mg dipyridamole orally every 6 hours and 1 hour after the end of the operation 100 mg dipyridamole via nasogastric tube. Patients were then randomized into three groups to receive 3 times a day a capsule containing: 50 mg acetylsalicylic acid, 50 mg acetylsalicylic acid plus 75 mg dipyridamole or placebo. Before the antiplatelet therapy was started, the day of the operation and 5 days after the onset of the treatment, blood samples were taken to determine platelet aggregation (turbidimetric technique) and the thromboxane A2 liberation. The three groups were comparable for all the studied clinical and angiographic variables. Preoperative dipyridamole did not provoke significant changes in platelet aggregation (acetylsalicylic acid group 86.5 +/- 10% vs 71 +/- 28.8%; acetylsalicylic acid + dipyridamole group 81.5 +/- 7.8% vs 75.3 +/- 22.7%; placebo group 83.7 +/- 8.9% vs 70 +/- 21.5%). After 5 days of treatment with acetylsalicylic acid, both groups under active treatment showed a significant (p less than 0.001) decrease of aggregability (acetylsalicylic acid 12.8 +/- 4.4%; acetylsalicylic acid + dipyridamole 20.5 +/- 17%; placebo 72 +/- 21.5%).(ABSTRACT TRUNCATED AT 250 WORDS)
Dipyridamole (DIP) is a drug widely used as an antiplatelet agent, which also has effects on endothelial cells. In this study, the effects of treating confluent endothelial cell monolayers (EC) with DIP on EC viability (trypan blue exclusion test) and metabolic activity (3H-thymidine incorporation) were examined. Platelet reactivity of the extracellular matrix (ECM) produced by untreated and DIP-treated ECs was determined morphometrically by a perfusion technique. Levels of ECM-associated von Willebrand factor (vWF) and fibronectin (FN) were also quantified (ELISA). The present results indicate that treatment of EC with 10 microM DIP did not reduce EC viability but that the incorporation of labelled nucleotides was significantly decreased (p less than 0.01). Platelet deposition onto the ECM generated by DIP-treated cells, perfused at a shear rate of 1300 sec-1, differed significantly with respect to controls (p less than 0.05), and platelet adhesion was also reduced (25% less, p less than 0.05). This effect was shear rate dependent, as no differences were noted when the ECMs were perfused at 300 sec-1 shear rate. Levels of VWF and FN associated with ECM remained unchanged with respect to controls. These results suggest that treatment with DIP alters EC metabolic activity, which in turn, influences the reactivity of the ECM generated by treated cells.
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.
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.
We present here the clinical results with a second-generation porcine bioprosthesis, the Carpentier-Edwards supra-annular valve (CESA). Two-hundred and twenty-two CESA bioprostheses were implanted in 189 patients during a four-year period (from 1984 to 1987), either as an isolated procedure or associated to mitral or tricuspid repair. The mid-term clinical results have been evaluated after a mean follow-up of 3.4 years, being 96% complete. There were 16 in-hospital deaths (8.4%) and 6 late, potentially valve-related, cardiac deaths (1.1% patients/year). Overall, 86.7 +/- 2% of the patients were free from cardiac death at 6 years (95.1 +/- 2% of the patients surviving the operative period). Linearized rates of valve related complications were the following: 1.4% patients/year for thromboembolism (including valve thrombosis), 0.5% patients/year for treatment-related hemorrhage and 0.7% patients/year for endocarditis. We did not found any case of either intrinsic or extrinsic valve failure, unrelated to infection of thrombosis. Two patients were reoperated, one because of valve thrombosis and the other due to prosthetic valve endocarditis (reoperation rate of 0.3% patients/year). When lethal and nonlethal valve-related complications (including in-hospital deaths) were considered all together, 75.8 +/- 8.4% of the patients remained alive and free of morbid events at 6 years. When patients were grouped according to the valve replaced (aortic, mitral and multiple), best results were found with patients submitted to isolated aortic valve replacement. We conclude that the CESA bioprosthesis has an excellent mid-term clinical performance. However, longer follow-up is necessary to know if improvement in valve design and manufacturing results in increased valve durability.
Modulator is an endogenous low-mol wt regulator of the glucocorticoid-receptor complex. Structural analysis of purified modulator suggested that it was a novel ether aminophosphoglyceride (Bodine and Litwack 1988b). Analogs of the putative modulator structure have now been synthesized. The synthetic compounds are 1-O-(6-carboxylhexyl)-glycero-3-phosphoserine and the sn-2-methoxy and sn-1-ethylester derivatives. Like modulator, these novel synthetic compounds are water soluble. However, thin-layer chromatography and spectroscopic analysis of these phosphoglycerides indicated significant structural differences between modulator and the synthetic analogs. In particular, the chromatographic behavior of the compounds suggests that modulator is more highly charged than the synthetic derivatives. The synthetic compounds, as well as lysophosphatidylserine, were also tested for in vitro modulator activity using the glucocorticoid-receptor complex activation inhibition and steroid-binding stabilization assays. None of the analogs exhibited modulator activity in these assays. However, the synthetic compounds were generally less detrimental to receptor steroid-binding than lysophosphatidylserine. From this work, we conclude that although modulator is not mimicked by one of these synthetic phosphoglycerides, a starting point for future structure-function studies has nonetheless been established.
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.
The 15-omega-lipoxygenase enzyme in endothelial cells metabolizes endogenous linoleic acid (18:2) into 13-hydroxyoctadecadienoic acid (13-HODE) under basal conditions, i.e., in unstimulated endothelial cells. 13-HODE is thought to regulate the non-adhesivity of the endothelium, contributing to vessel wall/blood cell biocompatibility. We performed experiments, therefore, to determine the relationship between basal levels of cAMP, 13-HODE synthesis, and platelet/endothelial cell adhesion. We found that 13-HODE synthesis increased with elevated cAMP levels and that the elevated 13-HODE levels correlated with increased 18:2 turnover in the triacylglycerol pool. In contrast, neither 18:2 nor arachidonic acid (20:4) turnover in the phospholipid nor prostacyclin (PGI2) production were changed with elevated cAMP levels. Platelet/endothelial cell adhesion was inversely proportional to 13-HODE synthesis. We conclude that intracellular 13-HODE influences platelet/vessel wall interactions, is synthesized from 18:2 released from the endogenous triacylglycerol pool, and that this pathway is modulated by intracellular cAMP levels.