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Biomedical subjects

R Montes

Publications and source records attributed to R Montes.

At least 19 recordsLinked to original sources

Rapid screening of polychlorinated biphenyls in sediments using non-equilibrium solid-phase microextraction and fast gas chromatography with electron-capture detection.

A rapid method for the screening of polychlorinated biphenyls (PCBs) in sediment samples is described. Analytes are concentrated on a poly(dimethylsiloxane) (PDMS) solid-phase microextraction (SPME) fibre, separated using a narrow-bore gas chromatography column (10 m x 0.1mm I.D.) and determined with a micro-electron-capture detector (micro-ECD). The narrow-bore column showed a similar separation efficiency to a conventional capillary one (30 m x 0.32 mm I.D.) and allowed a 3 times reduction in the analysis time. Moreover, thermal desorption of SPME fibres in the narrow-bore column did not produce any degradation in the quality of separations when compared to the split injection mode. The use of an oxidative sample treatment was mandatory to remove sulphur interferences in the chromatograms and increase the yield of PCBs during SPME for short sampling times. Under optimised conditions, a fraction of sediment (0.1-0.5 g) was mixed with 5 ml of a 0.2M potassium permanganate solution and 0.5 ml of sulphuric acid 6M in a 22 ml closed vessel. Sample treatment and headspace SPME of PCBs were performed simultaneously for 10 min at 100 degrees C. Globally, the whole analytical procedure takes about 20 min and presents quantification limits between 0.5 and 1.3 ng/g. Accuracy was evaluated by analysing a reference sediment sample with certified concentrations of several PCBs.

Chromatography, Gas↗

Association of increased fibrinogen concentration with impaired activation of anticoagulant protein C.

BACKGROUND: Low levels of activated protein C (APC) are a risk factor for venous thrombosis. The mechanisms leading to interindividual differences in APC are not totally elucidated. Protein C is activated by the thrombin-thrombomodulin complex. As thrombin binds to fibrinogen and thrombomodulin through a common region, it is conceivable that fibrinogen influences the activation of protein C. This would help to explain the association between high levels of fibrinogen and an increased thrombotic risk. METHODS: We analyzed the association between circulating APC levels and fibrinogen concentration in 382 healthy subjects. Subsequently, we studied the effect of increasing fibrinogen concentrations on the APC generation on cultured endothelial cells. RESULTS: An independent inverse association between circulating APC levels and fibrinogen was found [betacoefficient, -0.16; 95% confidence interval (95% CI) -0.26, -0.06; P = 0.001]. For each 100 mg dL(-1) increase in fibrinogen, the independent risk of having low APC levels (<0.7 ng mL(-1)) was almost three times higher (OR 2.8; 95% CI 1.1, 7.2; P = 0.04). Accordingly, a notable association between increasing fibrinogen concentrations and the reduction in the thrombin-thrombomodulin dependent activation of protein C on endothelial cells was found (r = -0.57; P = 0.002). CONCLUSIONS: We present evidence of an inverse association between circulating APC and fibrinogen levels. According to this finding together with the results of our in vitro experiments, we propose that the impairment in the generation of APC on endothelial cells constitutes a new prothrombotic mechanism of fibrinogen.

Adult↗

Development of a matrix solid-phase dispersion method for the screening of polybrominated diphenyl ethers and polychlorinated biphenyls in biota samples using gas chromatography with electron-capture detection.

A low cost method for the screening of six polybrominated diphenyl ethers (PBDEs) and seven polychlorinated biphenyls (PCBs) in biological samples containing up to 100% of fat is presented. Compounds are extracted from the sample and isolated from lipids using a matrix solid-phase dispersion (MSPD) cartridge and 20 ml of n-hexane as elution solvent. PBDEs and PCBs are fractionated on a second cartridge containing 2 g of a normal phase sorbent. The potential of neutral silica, Florisil and basic alumina to separate PBDEs and PCBs in two independent fractions has been evaluated. The best results were obtained using silica. PCBs are eluted, in a first fraction, using n-hexane. PBDEs are further recovered with n-hexane-dichloromethane. The applicability of the method for the screening of PBDEs and PCBs, in samples containing both groups of compounds, has been demonstrated using spiked, certified and real polluted samples from different biota materials. Globally, recoveries higher than 75% and quantification limits around 0.4 ng/g have been achieved using gas chromatography with electron-capture detection (GC-ECD).

Animals↗

Autoantibodies against the endothelial receptor of protein C are associated with acute myocardial infarction in young women.

BACKGROUND: Acute myocardial infarction (AMI) is rare among young women. The search for unknown risk factors is warranted. Endothelial protein C receptor (EPCR) is largely present at the endothelial surface of large arteries. No studies about association of anti-EPCR autoantibodies (anti-EPCR) with AMI are available. METHODS: Plasma IgA, IgM and IgG anti-EPCR levels were measured by enzyme-linked immunosorbent assay in 165 women younger than 45 years who survived a first AMI and 165 healthy women, matched by age and geographical origin. RESULTS: Using the 90th percentile of IgA anti-EPCR in the control group, IgA anti-EPCR were independently associated with AMI after adjustment for cardiovascular risk factors (OR 5.1; 95% CI 1.7-15.6; P = 0.004). The risk apparently conferred by IgA anti-EPCR increased dose-dependently (P for trend =0.0002). IgM anti-EPCR were less consistently associated with AMI: a significant increase in the risk was found when women above the 90th percentile were compared with those in the lowest quartile (OR 3.6; 95% CI 1.2-11.5; P = 0.03). IgG anti-EPCR were similar in patients and controls. A total of 145 patients underwent coronary arteriography. IgA or IgM anti-EPCR were not different among patients with different degrees of atherosclerotic lesion (anova, P = 0.77 and 0.24, respectively). CONCLUSIONS: High levels of IgA and, to a lesser extent, IgM anti-EPCR, are associated with AMI in young women.

Adult↗

[Octreotide in the treatment of angiodysplasia in patients with advanced chronic renal failure].

Angiodysplasia is an important cause of gastrointestinal bleeding in patients with chronic renal failure. Octreotide, a long-acting synthetic somatostatin analogue that reduces splachnic blood flow have been used to treat esophageal varicose hemorrhage, but its efficacy for bleeding vascular ecstasies is awaiting support. We present three patients with chronic renal failure (two with diabetic nephropaty and the third with mesangiocapilar glomerulonephritis and hepatic cirrosis), seric creatinine 3-4,5 mg/dl, and recurrent gastrointestinal bleeding due to diffuse angiodysplasia and vascular ecstasies, diagnosed by oral endoscopy, colonoscopy and video capsule. They all were treated with octreotide, administered subcutanesly 0.1 mg twice a day for six months, with significantly decreased blood requirements in all of them, as well as the occurrence of bleeding episodes. It was well tolerated and none side-effects occurred in any subject. In our experience, octreotide is an effective and safe drug in bleeding angiodysplasia and ecstasies vascular of the gastrointestinal tract in patients with chronic renal failure, and it may be a good option especially in patients who are not candidates for surgery or endoscopic treatment due to inaccessible sites, spread of the lesion, old age and/or concomitant disorders.

Adult↗

[Treatment of hyperphosphatemia with sevelamer in patients with chronic renal failure].

UNLABELLED: Sevelamer is a recent phosphate binder that is mineral-free, and represents a great advance in the treatment of hyperphosphatemia in patients with hypercalcemia and/or gastric intolerance to calcium-based phosphate binders. The communications about the experience with the use of sevelamer in patients non-yet in dialysis is scanty. The aim of our study is to investigate retrospectively the gastrointestinal tolerance of sevelamer, their efficacy as phosphate binder and other parameters in a group of 89 patients with chronic renal failure in predialysis. We have analysed the effects of sevelamer at baseline and after 1, 3 and 6 months on the following data and parameters: calcium, phosphate, intact PTH, venous bicarbonate, urea, creatinine, creatinine clearance, side-effects, number of patients that were discontinued, and co-treatment during the study period with phosphate-based binders, calcitriol, lipid-lowering drugs and sodium bicarbonate. RESULTS: 19 patients (21.3%) refused to continue with sevelamer at the first month (16 patients had digestive intolerance and 3 several symptoms). Serum phosphate fell at 3 months (5 +/- 0.8 mg/dl basal vs 4.8 +/- 0.7 mg/dl, p = 0.02) and 6 months (5 +/- 0.8 mg/dl basal vs 4.7 +/- 0.9 mg/dl, p = 0.07). Serum calcium fell at 6 months (9.8 +/- 0.7 mg/dl basal vs 9.4 +/- 0.6 mg/dl, p = 0.03). Venous bicarbonate and iPTH were unchanged, but the quantity of sodium bicarbonate administered increased significantly. Blood cholesterol fell at 1 months (193 +/- 49 mg/dl basal vs 173 +/- 52 mg/dl, p = 0.001) and 3 months (205 +/- 49 mg/dl basal vs 170 +/- 49 mg/dl, p = 0.004), in spite of a significant reduction of the dose of statins. CONCLUSIONS: Sevelamer is an effective phosphate binder in predialysis patients and also reduces significantly the serum cholesterol, improving the blood lipid profile. The levels of venous bicarbonate remained unchanged, at expenses of an increment in the dose of sodium bicarbonate supplementation.

Acid-Base Equilibrium↗

[Peritoneal equilibrium test using icodextrin and glucose at different concentrations].

UNLABELLED: The aim of this study was to compare, under standard conditions (Peritoneal Equilibrium Test, "PET"), the peritoneal permeability to water and several solutes using icodextrin and glucose (3.86% and 1.36%) dialysates. The study includes 14 patients (3 women and 11 men), mean age 64 +/- 13 years, average time on peritoneal dialysis 23.5 +/- 17 months. PETs with icodextrin were performed in all of them (n = 14); PETs with 3.86% glucose were carried out in 7, and PETs with all the three solutions were performed in 5 patients. Samples were taken at 0, 30, 60, 120, 240 minutes, and after the rinsing procedure using 1.36% glucose in order to calculate the residual volume. RESULTS: Sodium concentration in the effluent and D/P sodium did not change significantly from minute 0 to 240 with icodextrin and 1.36% glucose; but with 3.86% glucose both sodium and D/P sodium decreased at thirty minutes, remained at the same levels till the 120 minutes and then had a tendency to increase. Glucose concentration and osmolarity in the effluent did not vary throughout the time with icodextrin, but progressively decreased during the 4-hour period with 3.86% and 1.36% glucose solutions. The drainage after the 4-hour period was higher for the 3.86% glucose (2,608 +/- 388 ml, p = 0.03) than for the 1.36% glucose (2,070 +/- 120 ml) or the icodextrin (2,212 +/- 213 ml). Low molecular weight permeability: D/P creatinine after the 4-hour dwell was significantly lower for the icodextrin (0.66 +/- 0.1, p = 0.05) than for the 3.86% glucose (0.71 +/- 0.1) or the 1.36% glucose (0.72 +/- 0.1). The creatinine clearance for 3.86% glucose (7.4 +/- 0.4 ml/min p = 0.007) was higher than for icodextrin (5.6 +/- 0.5 ml/min) or for 1.36% glucose (5.8 +/- 0.6 ml/min). The clearances for total protein, albumin and beta 2-microglobulin did not show significant differences between the solutions. CONCLUSIONS: Our study confirms that the icodextrin solution remains iso-osmolar with plasma during the 4-hour dwell. The sodium profile suggests that the ultrafiltration induced by icodextrin and 1.36% glucose depends on small pore-mediated sodium and water transport; on the other hand, 3.86% glucose also induces transport of water without solutes throughout the ultra-small aquaporin-mediated, pores, producing sodium dilution in the effluent. Ultrafiltration and solute clearances for icodextrin are lower than for 3.86 glucose during a 4-hour dwell.

Adult↗

[Clinical experience with icodextrin. Multicenter study].

UNLABELLED: The aim of this study was to analyse our experience with icodextrin in Andalusia, Spain. The study includes 51 patients (30 women and 21 men) on peritoneal dialysis (21 on CAPD and 30 on Automated Peritoneal Dialysis) treated with icodextrin for 10.3 +/- 7 months (0-41 months). Their mean age was 57 +/- 18 years (18-86 years). We have recorded the appearance of side effects, and the evolution of several biochemical parameters at baseline and after 6, 12 ans 18 months from initiation of icodextrin. We also studied drainage fluid from 12 patients after an icodextrin exchange. RESULTS: There were side effects (all cutaneous) in 4 out of 51 patients (7.8%). Two of the affected suffered from cutaneous hypersensitivity reactions, and icodextrin had to be suspended; the other two had exfoliative dermatitis affecting hands and feet that disappeared without have to withdraws icodextrin. Biochemical parameters: Serum sodium levels decreased from baseline to six months (138 +/- 6 mEq/l vs 136 +/- 3 mEq/l; p = 0.006), and then persisted at the same levels throughout the rest of the study period. There was a slight but significant decreased of serum HDL-cholesterol at six months vs baseline (55 +/- 26 mg/dl vs 51 +/- 20 mg/dl, p = 0.04), and a further decrease at twelve months vs six months (42 +/- 15 mg/dl vs 51 +/- 13 mg/dl, p = 0.054). There were no significant variations of glucose, osmolality, cholesterol, LDL-cholesterol (tendency to increase), triglycerides, beta 2 m and weight (tendency to increase; p = 0.08). In relation with the icodextrin exchange: average ultrafiltration 296 +/- 119 ml (ranging from 104 to 480 ml), creatinine clearance 1.9 +/- 0.5 litres (20.5% of daily creatinine clearance), urea clearance 2.08 +/- 0.5 litres (18.7% of daily urea clearance), total protein losses 3.2 +/- 0.9 g, albumin losses 1.4 +/- 0.5 g; urea and creatinine clearances were negatively correlated with ratios D/P4 of urea and creatinine of PET and positively correlated with ratio G4/G0. In conclusion, side effects are scarce with the use of icodextrin. As described in other studies, there is a trend to a slight decrease in serum sodium. The long-term use of icodextrin does not-prevent weight gain or deterioration of patients on peritoneal dialysis, despite the diminution of glucose load.

Adolescent↗

Hemostatic disturbances in patients with systemic inflammatory response syndrome (SIRS) and associated acute renal failure (ARF).

Endothelial damage plays a central role in the development of an SIRS-related Multiple Organ Dysfunction Syndrome (MODS) as a consequence of the establishment of a hemostatic imbalance between coagulation and fibrinolysis systems. Until now, sepsis is the SIRS model that has been most studied. The aim of this study was to assess the endothelial damage and the hemostatic imbalance in early stages of an SIRS of different origins, and to study if there are any differences in these disturbances between infectious and noninfectious SIRS. The endothelial damage and hemostatic changes were studied in 40 patients with SIRS (with less than 12 h of evolution) and an acute renal failure. Infectious SIRS was diagnosed in 19 cases and noninfectious SIRS in the remaining 21 patients. Patients with SIRS presented significantly higher values (p<0.001) for factors related to endothelial damage [von Willebrand factor (vWF), thrombomodulin, tissue plasminogen activator (t-PA), and plasminogen activator inhibitor type 1 (PAI-1) antigen], hypercoagulability [prothrombin fragment 1+2 (F1+2) and thrombin-antithrombin complexes (TAT)], and fibrinolysis (D-dimer and PAI activity) with respect to the control group. However, although the group with infectious SIRS presented higher values for all the factors except for the t-PA and D-dimer with respect to SIRS of other origins, none of these differences reached statistical significance (p>0.05). Our data show that patients with SIRS and associated acute renal failure, irrespective of the origin (infectious or noninfectious), show signs of intense endothelial damage and hypercoagulability throughout the process.

Acute Disease↗

Prevention of renal fibrin deposition in endotoxin-induced DIC through inhibition of PAI-1.

Plasminogen activator inhibitor-1 (PAI-1) increases in endotoxemia thus possibly cooperating in altering the hemostatic balance in a prothrombotic direction. The effect of the inhibition of PAI-1 with the monoclonal antibody MA-33B8 was studied systemically and in kidneys in a lapine model of endotoxin-induced disseminated intravascular coagulation (DIC). The increase in plasmatic PAI activity in the control group (n = 9) was inhibited in the MA-33B8 treated rabbits (n = 5). Control rabbits showed renal fibrin deposits, whereas only one of the MA-33B8 rabbits did so. These results were confirmed immunohistochemically in kidneys as PAI-1 immunostaining was seen inside the glomeruli and larger vessels in the control group, whereas MA-33B8 rabbits showed a remarkable decrease, demonstrating that MA-33B8 successfully inhibited PAI-1 in the kidneys as well. Therefore evidence for the important role of PAI-1 in fibrin generation in endotoxin-induced DIC is presented, suggesting that strategies aiming at its reduction can be useful in this pathology.

Animals↗

Do the low molecular weight heparins improve efficacy and safety of the treatment of deep venous thrombosis? A meta-analysis.

BACKGROUND AND OBJECTIVES: We compared the efficacy and safety of low molecular weight heparins (LMWH) and unfractionated heparin (UFH) in the treatement of deep venous thrombosis (DVT). A comparison between two daily subcutaneous injections of LMWH against a single injection was also performed. DESIGN AND METHODS: The study was performed by a meta-analysis. Clot improvement in venography, recurrency, total mortality and major hemorrhages were assessed in 4,472 patients with DVT from 21 studies treated with subcutaneous LMWH or UFH. RESULTS: An improvement in clot reduction (odds ratio 0.73, 95% confidence interval 0.59 to 0.90, p = 0.004), a decrease in total mortality (0. 68, 0.50 to 0.91, p = 0.012) and a lower incidence of hemorrhage (0. 65, 0.43 to 0.98, p = 0.047) were observed in LMWH treated patients. There were no differences in recurrences (0.78, 0.59 to 1.04, p = 0. 10). A single dose of LMWH was better than two in reducing major bleeding (c2 = 4.99, p = 0.025); however, the two dose regimen was more effective in clot reduction (c2 = 8.56, p = 0.004). INTERPRETATION AND CONCLUSIONS: LMWH is superior to UFH in terms of safety and efficacy. A single daily dose of LMWH dose is a suitable therapeutic regimen and could facilitate the outpatient treatment of venous thromboembolism.

Hemorrhage↗

[Evidence for glutamatergic neurotransmission in the geniculo-cortical pathway in the rat].

PURPOSE: To investigate the possible participation of glutamate in the neurotransmission of the rat geniculo-cortical pathway. METHODS: In a first set of experiments, we studied the effects of microiontophoresis of kynurenate, a broad spectrum antagonist of glutamate receptors, on the excitation evoked in visual cortex cells by electrical stimulation of the dorsal lateral geniculate nucleus and the optic tract. Cell activity was determined by extracellular unit recordings. In a second set of experiments, we studied the effects of electrical stimulation of the dorsal lateral geniculate nucleus and the optic tract on the release of glutamate and aspartate in the visual cortex in vivo. Push pull perfusions were made in the visual cortex and the amino acids contents of the perfusates were analysed by high performance liquid chromatography. RESULTS: Microiontophoresis of kynureate in area 17 cells selectively supressed the excitatory response to dorsal geniculate nucleus and optic tract stimulation. On the other hand, stimulation of these structures significantly increased glutamate, but not aspartate, release in area 17. CONCLUSIONS: These results demonstrate that glutamate is the neurotransmitter of the geniculo-cortical pathway in the rat.

Animals↗

Evidence that heparin but not hirudin reduces PAI-1 expression in cultured human endothelial cells.

Heparin and other antithrombotic drugs besides their anticoagulant action could have a profibrinolytic effect. We have analyzed the effect of unfractionated heparin (UFH) and hirudin on PAI-1 gene expression in human umbilical vein endothelial cells (HUVEC). Cells were stimulated with UFH (1 and 10 IU/ml) and hirudin (20 and 100 TIU/ml). Samples were obtained before and 2, 6, and 24 hours after stimulation. mRNA analysis was conducted by reverse transcription followed by polymerase chain reaction, and PAI-1 antigen was determined by ELISA. Addition of UFH (10 IU/ml) to HUVEC resulted in a decrease of PAI-1 mRNA at 6 hours (40% reduction) and 24 hours (60% reduction) and PAI-1 antigen. Hirudin, however, did not modify significantly the PAI-1 mRNA nor the inhibitor secretion. The addition of UFH (10 or 100 IU/ml) to endotoxin-stimulated HUVEC also reduced the increased PAI-1 mRNA and antigen secretion (45%), whereas no effect could be observed with hirudin. Our results suggest that UFH, but not hirudin, by reducing the endothelial expression of PAI-1 might have a profibrinolytic effect.

Cells, Cultured↗

Effects of low molecular weight heparin, alone or combined with antithrombin III, on mortality, fibrin deposits and hemostatic parameters in endotoxin-induced disseminated intravascular coagulation in rabbits.

The effect of low molecular weight heparin (LMWH) with or without antithrombin III (AT III) has been studied in a rabbit model of disseminated intravascular coagulation (DIC) induced by continuous infusion of 100 microg/kg/hr of Escherichia coli endotoxin for 6 hr. LMWH (5 and 10 IU/kg/hr/6 hr), alone or in combination with AT III (20 U/kg/hr/6 hr), or saline were administered simultaneously with endotoxin. Hemostatic markers at 0, 2, and 6 hr as well as kidney fibrin deposits and the mortality rate at 24 hr were determined. Rabbits receiving only endotoxin showed an impairment in hemostasis, as well as high kidney fibrin deposits and a high mortality rate. LMWH alone did not exert any effect. The simultaneous infusion of LMWH and AT III exerted a beneficial effect on the hemostatic markers and reduced the kidney fibrin deposits as well as the mortality rate in a LMWH dose-dependent manner. Fibrinogen and protein C consumption were significantly higher and renal fibrin deposits more intense in the rabbits that had died in the first 24 hr. There was also a significant positive correlation between kidney fibrin deposits and platelets, fibrinogen, and protein C consumption, taking the whole rabbit population. It is concluded that the simultaneous infusion of LMWH and AT III is useful in this DIC model and would make it possible to reduce significantly the AT III doses used when AT III is given alone.

Animals↗

Vaccination with the extracellular domain of p185neu prevents mammary tumor development in neu transgenic mice.

The HER2/neu oncogene product, p185(HER2/neu), is overexpressed on the surface of many human breast cancers. Strains of transgenic mice have been developed that express the rat neu oncogene in mammary epithelial cells and develop spontaneous mammary tumors that overexpress p185neu. This model provides an ideal system for testing interventions to prevent tumor development. In this study, we immunized neu-transgenic mice with a vaccine consisting of the extracellular domain of p185neu (NeuECD). Immunized mice developed Neu-specific humoral immune responses, as measured by circulating anti-Neu antibodies in their sera, and cellular immune responses, as measured by lymphocyte proliferation to NeuECD in vitro. In addition, the subsequent development of mammary tumors was significantly lower in immunized mice than in controls and vaccine treatment was associated with a significant increase in median survival.

3T3 Cells↗

Changes in the fibrinolytic components of cultured human umbilical vein endothelial cells induced by endotoxin, tumor necrosis factor-alpha and interleukin-1alpha.

BACKGROUND AND OBJECTIVE: Vascular fibrinolysis, a major natural defense mechanism against thrombosis, is a highly regulated process. The aim of this study was to evaluate the effect of endotoxin, tumor necrosis factor-alpha (TNFalpha) and interleukin-1alpha (IL-1alpha), on the fibrinolytic potential of cultured human umbilical vein endothelial cells (HUVEC). DESIGN AND METHODS: Samples of stimulated conditioned media were collected over a period of 24 hours to determine: plasminogen activator (PA) and plasminogen activator inhibitor (PAI) activity, PAI-1 mRNA, tissue-type plasminogen activator (t-PA) antigen and urokinase-type plasminogen activator (u-PA) antigen. RESULTS: Similar changes were observed after endotoxin and cytokine stimulation: there was a significant increase of PAI activity (p<0.01), starting at 6 hours, which remained 24 hours after stimulation. PAI-1 mRNA also showed an important rise with these agents, although cytokines induced an earlier and more intense inhibitor response (up to 6-fold increase). PA activity increased significantly at 6 hours (p<0.01) to drop at 24 hours and was mainly related to the presence of u-PA. INTERPRETATION AND CONCLUSIONS: We conclude that endotoxin,+TNFalpha and IL-1alpha induce profound alterations in the fibrinolytic potential of HUVEC, characterized by an initial rise of activators (u-PA) followed by a strong increase of PAI-1. These changes may be of pathophysiologic significance for thrombosis and inflammatory reactions.

Cells, Cultured↗