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Biomedical subjects

J A Fernández

Publications and source records attributed to J A Fernández.

At least 19 recordsLinked to original sources

Testing the use of juvenile Salmo trutta L. as biomonitors of heavy metal pollution in freshwater.

Individual specimens of Salmo trutta were captured, from four sampling sites in Galician rivers (NW Spain) affected by different types of contamination: diffuse urban waste, run-off from an unrestored dump at a copper mine and waste from a fish farm. The ages of the captured trouts were established and only those belonging to the 1+ age class were selected for study. The liver and kidney were removed from each fish and analysed to determine the tissue concentrations of Cu, Fe and Zn. The results obtained showed that: (i) the use of 1+ individuals allowed differentiation of contamination scenarios on the basis of the tissue concentrations of metal; (ii) the use of 1+ individuals allowed standardization of the time of exposure, which was sufficiently long for differential uptake to have taken place; (iii) liver tissue provided the best results as, less effort was required than for processing kidney tissue, and significant differences between sampling sites were detected because the intrapopulational variability in metal levels was lower than for kidney, and (iv) the levels of elements detected were not affected by basal tissue concentrations or residual concentrations due to past contamination, which older trouts may have been exposed to. In addition, the use of 1+ trout may provide better results in annual environmental sampling surveys.

Aging↗

Plasma contains protein S monomers and multimers with similar direct anticoagulant activity.

BACKGROUND: Protein S (PS) has activated protein C-independent, direct anticoagulant activity (PS-direct). We reported that both multimers and monomers of affinity-purified PS have PS-direct similar to that in plasma, in contrast to another report. OBJECTIVE: We extended our studies to establish the molecular forms and activity of plasma PS. METHODS: Novel ELISAs were developed that could detect only multimeric, not monomeric, PS because they employed the same monoclonal antibody for capture and detection. PS forms were also examined on native PAGE immunoblots. A new activity assay for PS-direct was applied to plasma and gel-filtered plasma fractions. RESULTS: Plasma PS multimers were clearly demonstrated using the ELISAs; 30-60% of free plasma PS appeared to be multimeric, a proportion similar to that of affinity-purified PS. On immunoblots, plasma PS multimers were more easily detected after gel filtration; plasma PS monomers and several apparent multimers comigrated with respective forms of affinity-purified PS. Antigen elution profiles after gel filtration of plasma revealed at least one major peak of apparent PS multimers (40-55% of free PS appeared multimeric). Biotin-factor Xa could bind to both plasma PS monomers and multimers. Strong plasma PS-direct was demonstrated, and plasma PS monomers, multimers, and PS-C4b-binding protein complexes each reconstituted PS-depleted plasma to similar levels of PS-direct. CONCLUSION: Our data are in disagreement with a report that monomeric purified PS has little PS-direct and that only monomeric PS exists in plasma. We find that both affinity-purified and plasma PS exist as monomers and multimers with similar PS-direct.

Antibodies, Monoclonal↗

Considerations on the sample size of wood mice used to biomonitor metals.

The concentrations of various metals (Zn, Cu, Mn and Cr) in liver, kidneys and brain from specimens of the wood mouse Apodemus sylvaticus captured from 5 sites were measured. Two of the sites were in a restored mine dump, another in an area characterized by serpentized soils and the remaining two were control sites. The sample size required for statistical differentiation of the sampling sites was calculated from the mean values and the variability in bioaccumulation corresponding to each of the sites. The relationship between the sample size and the results of the statistical test used to reveal significant differences between the mean concentrations of metals in organs from A. sylvaticus is demonstrated. The homeostatic control exerted by the wood mice on the tissue levels of metals reduced the interpopulational variability, thereby homogenizing the mean bioaccumulation corresponding to the different sampling stations. The sample size, tissue regulation of the levels of heavy metals and the patterns of variability are of vital importance in evaluating the usefulness of A. sylvaticus as a biomonitor of heavy metals.

Animals↗

[Hereditary pancreatitis caused by a new mutation in the trypsinogen gene. Report of a family].

Hereditary pancreatitis is an uncommon autosomal dominant disease secondary to a mutation normally located in the trypsinogen gene, preventing trypsin deactivation. This mutation translates clinically into recurrent attacks of acute pancreatitis and an increased risk of pancreatic cancer. We report a case of acute hereditary pancreatitis due to a trypsinogen mutation that has previously been described in only one family.

Child↗

Limited generation of activated protein C during infusion of the protein C activator thrombin analog W215A/E217A in primates.

Anticoagulation with activated protein C (APC) reduces the mortality of severe sepsis. We investigated whether the circulating protein C (PC) pool could be utilized for sustained anticoagulation by endogenous APC. To generate APC without procoagulant effects, we administered the anticoagulant thrombin mutant W215A;E217A (WE) to baboons. In preliminary studies, administration of high dose WE (110 microg kg(-1) i.v. bolus every 120 min; n = 2) depleted PC levels by 50% and elicited transient APC bursts and anticoagulation. The response to WE became smaller with each successive injection. Low dose WE infusion (5 microg kg(-1) loading + 5 microg kg(-1) h(-1) infusion; n = 5) decreased plasma PC activity by 15%, from 105% to 90%, to a new equilibrium within 60 min. APC levels increased from 7.5 ng mL(-1) to 86 ng mL(-1) by 40 min, then declined, but remained elevated at 34 ng mL(-1) at 240 min. A 22-fold higher dose WE (n = 5) decreased PC levels to 60% by 60 min without significant further depletion in 5 h. The APC level was 201 ng mL(-1) at 40 min and decreased to 20 ng mL(-1) within 120 min despite continued activator infusion. Co-infusion of WE and equimolar soluble thrombomodulin (n = 5) rapidly consumed about 80% of the PC pool with significant temporal increase in APC generation. In conclusion, low-grade PC activation by WE produced sustained, clinically relevant levels of circulating APC. Limited PC consumption in WE excess was consistent with the rapid depletion of cofactor activity before depletion of the PC zymogen. Reduced utilization of circulating PC might have similar mechanism in some patients.

Amino Acid Substitution↗

Definition and number of subsamples for using mosses as biomonitors of airborne trace elements.

We collected 50 subsamples of the moss Scleropodium purum from each of three sampling sites and determined the concentrations of As, Cd, Cu, K, Hg, Ni, Pb, Se, and Zn in each subsample. We then calculated the number of subsamples required to determine significant differences in the mean concentrations of two sampling sites. We found that to differentiate between an uncontaminated sampling site and another, slightly contaminated site, 30 subsamples are required from each. On the basis of these results and because, to date, there are no studies that justify the application of the previously proposed recommendations, studies of local variability in other areas, and under different conditions, with other contaminants and moss species must be undertaken. For posterior comparison of data on variability as well as application of the recommendations associated with such results, the general use of a single definition of a subsample is required. We propose the following definition of a subsample: "1 g dry weight (approx.), collected within a 25-cm radius of a node, selected at random from a 1 x 1-m sampling grid placed in a sampling site."

Air Pollution↗

Modelling the extra and intracellular uptake and discharge of heavy metals in Fontinalis antipyretica transplanted along a heavy metal and pH contamination gradient.

Samples of the aquatic bryophyte Fontinalis antipyretica Hedw. were transplanted to different sites with the aim of characterizing the kinetics of the uptake and discharge of heavy metals in the extra and intracellular compartments. The accumulation of metals in extracellular compartments, characterized by an initial rapid accumulation, then a gradual slowing down over time, fitted perfectly to a Michaelis-Menten model. The discharge of metals from the same compartment followed an inverse linear model or an inverse Michaelis-Menten model, depending on the metal. In intracellular sites both uptake and discharge occurred more slowly and progressively, following a linear model. We also observed that the acidity of the environment greatly affected metal accumulation in extracellular sites, even when the metals were present at relatively high concentrations, whereas the uptake of metals within cells was much less affected by pH.

Aluminum↗

Mapping the results of extensive surveys: the case of atmospheric biomonitoring and terrestrial mosses.

In this paper we discuss some difficulties associated with the process of constructing maps of pollution from data obtained in surveys covering extensive areas. As we show here, these problems may be wide-ranging but are seldom recognized by investigators. The origin of the problems is the existence of multiple sources of pollution in the study area, each of different intensity and affecting areas of different extent. The particular spatial structure of the pollution sources interacts with the spatial layout of the samples, resulting in data sets with distributions that are very different from the usually assumed normal distribution, and characterized by heavy tails and gross outliers. These distributions arise because of incomplete sampling of small-scale pollution processes (i.e. those occurring on a spatial scale smaller than the spatial scale of the sampling grid). After discussion of the potential problems and appropriate techniques for analyzing this kind of data, we applied the proposed techniques to a real data set of heavy metal contents in terrestrial mosses. From the exercise we concluded that a) the first step in analysis of this kind of data must be to check for the presence of spatial structure on scales larger than the sampling grid, to avoid mapping noise, and b) the map generated must not contain information about pollution sources with a spatial scale smaller than the spatial scale of the sampling grid. We present and discuss the performance of robust statistical methods of testing for spatial structure (based on robust variograms and randomization testing) and of filtering the small-scale spatial processes (using median-polishing) prior to mapping.

Atmosphere↗

The effect of sampling design on extensive bryomonitoring surveys of air pollution.

In this study, we investigated if the different sampling designs normally used in extensive bryomonitoring surveys of air pollution allow compliance with the objectives proposed in the 'Heavy Metals in European Moss' project. For this, we have used data on concentrations of As, Cu, Se and Zn from five moss sampling surveys carried out in Galicia (NW Spain), using three types of sampling design: regular grid, sampling sites inside squares and the strategy recommended in the guidelines for European surveys. To study the effect of the interaction between the sampling design and the spatial pattern of the presence of contaminants, we have used a simulated and a real pattern onto which we superimposed the three sampling designs considered. We found that only random sampling and grid sampling allow compliance with the proposed objectives, and we discuss the advantages of the latter over the former. The data corresponding to studies using different sampling designs are not comparable, and in future large-scale surveys, the same design must be used if the results are to be compared and if valid conclusions are to be reached. The scale of the grid determines the scale to which the objectives are complied with, so that for correct standardization of the technique, and taking into account the most commonly used grid sizes, we recommend a grid size of 30 x 30 or 32 x 32 km. When additional samplings are required, the size of the grid size should be 30/2(n) or 32/2(n) km.

Air Pollution↗

Physiological evidence for a sodium-dependent high-affinity phosphate and nitrate transport at the plasma membrane of leaf and root cells of Zostera marina L.

Zostera marina L. is an angiosperm that grows in a medium in which inorganic phosphate (P(i)) and nitrate (NO(3)(-)) are present in micromolar concentrations and must be absorbed against a steep electrochemical potential gradient. The operation of a Na(+)-dependent NO(3)(-) transport was previously demonstrated in leaf cells of this plant, suggesting that other Na(+)-coupled systems could mediate the uptake of anions. To address this question, P(i) transport was studied in leaves and roots of Z. marina, as well as NO(3)(-) uptake in roots. Electrophysiological studies demonstrated that micromolar concentrations of P(i) induced depolarizations of the plasma membrane of root cells. However, this effect was not observed in leaf cells. P(i)-induced depolarizations showed Michaelis-Menten kinetics (K(m)=1.5+/-0.6 microM P(i); D(max)=7.8+/-0.8 mV), and were not observed in the absence of Na(+). However, depolarizations were restored when Na(+) was resupplied. NO(3)(-) additions also evoked depolarizations of the plasma membrane of root cells only in the presence of Na(+). Both NO(3)(-)- and P(i)-induced depolarizations were accompanied by an increase in cytoplasmic Na(+) activity, detected by Na(+)-sensitive microelectrodes. P(i) net uptake (measured in depletion experiments) was stimulated by Na(+). These results strongly suggest that P(i) uptake in roots of Z. marina is mediated by a high-affinity Na(+)-dependent transport system. Both NO(3)(-) and P(i) transport systems exploit the steep inwardly directed electrochemical potential gradient for Na(+), considering the low cytoplasmic Na(+) activity (10.7+/-3.3 mM Na(+)) and the high external Na(+) concentration (500 mM Na(+)).

Biological Transport, Active↗

Active biomonitoring of element uptake with terrestrial mosses: a comparison of bulk and dry deposition.

Moss (Scleropodium purum) transplants were used to study bioconcentration originating from dry and bulk deposition, by measuring the tissue contents of Al, As, Ca, Cu, Fe, Hg, Mn, Ni, Pb, Se and Zn. Furthermore, a laboratory experiment was carried out to determine the sequence of maximum concentration and affinity of Al, Cu, Fe, Hg, Ni, Pb and Zn, in S. purum. We found that in many of the transplants, for the same period of exposure, higher levels of metals were accumulated via dry deposition than via bulk deposition. This result may be explained by the simple washing action of the rain on the surface of the moss, and by the existence of processes that provoke the loss of some of the accumulated elements: intercationic displacement and leaching caused by acid precipitation. Modelling of the final bioconcentration observed, as a balance of inputs and outputs of elements, revealed that this terrestrial moss does not integrate, but rather concentrates atmospheric deposition, and there exists a state of unstable equilibrium between inputs and outputs of elements, a state that is determined by the characteristics of the surrounding environment. On the basis of the results obtained in this study, we can conclude that at present, it is not possible to extrapolate the calibrations between the concentrations of elements accumulated in a certain species of moss to values of atmospheric deposition (bulk deposition) from one place to another with different environmental conditions.

Acid Rain↗

A new method for testing the sensitivity of active biomonitoring: an example of its application to a terrestrial moss.

For the first time, the concepts of limit of detection and limit of quantification, commonly used in analytical chemistry, are applied to the field of active biomonitoring with terrestrial mosses, using the controls as blanks so that the limits indicate the error associated with the transplant technique. The application of these concepts to data corresponding to Hg concentrations in the surroundings of a chlor-alkali plant and a power plant, makes interpretation of the results easier by providing better spatial and temporal coherence. This procedure may allow improvement in the standardization of active biomonitoring techniques as it is applicable to all kinds of biomonitors. One disadvantage of the application of the limit of detection and limit of quantification is that they only take into account the alpha error, or risk of false positives, and do not take into account the beta error, or risk of false negatives.

Adaptation, Physiological↗

Identification of pollution sources by means of moss bags.

Moss bags containing Scleropodium purum were exposed for 30 days in an industrial area in which a chlor-alkali plant (CAP) and a concrete factory (CF) are located close to one another (121 m). Following the exposure, the transplanted mosses were analyzed to determine levels of Hg, Zn, Ni, and Cr. The enrichment factors (EFs) of the metals (final concentration/initial concentration) were calculated; the mean values obtained were 26.4 for Hg, 6.6 for Zn, 1.07 for Ni, and 1.0 for Cr. Dispersion of Cr and Ni was closely related, probably because they were dispersed together on the same particle. The spatial structure of the data was examined, and significant fits to the semivariograms were found for Hg, Zn, and Cr, using kriging to interpolate the data. Using this technique, it was possible to estimate where the maximum concentrations of elements occurred, allowing us to distinguish from which focal point they were emitted: Hg from the CAP, and Zn and Cr from the CF. The combined use of moss bags and geostatistical techniques allowed characterization of the emissions and identification of focal points of contamination in the study area.

Air Pollutants↗

Natural gamma radiation map (MARNA) and indoor radon levels in Spain.

During the last decade, the Department of Applied and Medical Physics has been involved in the development of a radiation protection programme. In the framework of this programme, measurements of indoor radon, principally, have been carried out nationwide. Geometric mean radon concentrations of 45 Bq m(-3) in the whole country and 130 Bq m(-3) in the high natural radiation area have been estimated. On the other hand, the so-called MARNA Project is developed into the framework of an agreement subscribed between the Spanish Nuclear Safety Council (CSN) and the National Uranium (ENUSA), the first phase of which has been the elaboration of the Natural Gamma Radiation Map of Spain on the scale of 1:1,000,000 using radiometric data generated in the 30 years of the lifetime of the ancient National Uranium Exploration and Investigation Plan mainly through airborne, carborne, and by foot surveys, within the MARNA Project itself. The lowest averaged dose rate from external gamma radiation (19.3 nGyh(-1)) was found in carbonate bedrock and the highest (87.7 nGyh(-1)) was found in granite and clay bedrock. This paper summarizes the main results obtained from the measurements performed in both projects, with special interest in those concerning the correlation between the data reported in order to conclude about the potential benefit of the MARNA maps in the definition of affected areas in the country.

Air Pollution, Indoor↗

Cardiovascular changes during drainage of pericardial effusion by thoracoscopy.

BACKGROUND: Cardiovascular changes during drainage of pericardial effusion are not well understood, and most studies are of systemic effects and not of right ventricular performance. Thoracoscopy is not widely used to drain pericardial effusions because of haemodynamic changes in relation to the use of single lung ventilation. PATIENTS AND METHODS: We studied 16 patients undergoing partial pericardiectomy for pericardial effusion, using videothoracoscopy with a low-pressure pneumothorax (6 mm Hg). Cardiac output was measured by thermodilution with the patient anaesthetized in the supine position before the procedure; in the right lateral position after a low-pressure pneumothorax had been established; and after drainage of the pericardial effusion. RESULTS: Before the procedure, cardiac output was low and central venous pressure and pulmonary artery occlusion pressure were increased. Systemic vascular resistance and arterial blood pressure were within normal limits. Cardiac filling pressure and pulmonary arterial pressure increased during the pneumothorax. After the drainage cardiac index increased and systemic and pulmonary vascular resistances were reduced. CONCLUSIONS: Pericardial effusion reduces right ventricular distensibility, right and left systolic ventricular function, and cardiac output. Anaesthesia with mechanical ventilation and a low-pressure pneumothorax do not affect the circulation greatly. Drainage of the pericardial effusion allows cardiac distensibility to increase and cardiac performance changes to allow increased ejection.

Adolescent↗

Direct anticoagulant activity of protein S-C4b binding protein complex in Heerlen heterozygotes and normals.

BACKGROUND: Plasma protein S normally circulates free (40%) or complexed with C4b-binding protein (PS-C4BP); only free protein S is a cofactor for activated protein C during factor (F) Va inactivation. Protein S-Heerlen lacks a carbohydrate group, leading to low plasma free protein S levels, but normal levels of PS-C4BP. OBJECTIVES: Because protein S-Heerlen is not associated with thrombosis, we investigated whether PS-C4BP is directly anticoagulant in plasma and whether PS-Heerlen-C4BP has enhanced direct anticoagulant activity. METHODS: An assay for protein S direct activity was applied to Heerlen-heterozygous plasmas. Free and complexed protein S were repeatedly isolated from normal and Heerlen-heterozygous plasmas and tested for direct anticoagulant activity in prothrombinase assays and in plasma. RESULTS: Heerlen-heterozygous plasmas were deficient in free and total protein S antigen but had normal to high protein S direct anticoagulant activity. Purified Heerlen-heterozygous PS-C4BP was 7-fold more potent than normal PS-C4BP in inhibiting full prothrombinase activity, and 22-fold more potent in inhibiting prothrombin activation in the absence of FVa; it also specifically prolonged plasma clotting times 14-fold more than normal PS-C4BP. Heerlen-heterozygous PS-C4BP did not compete for limiting phospholipids any better than normal PS-C4BP. However, ligand blots and surface plasmon resonance studies showed that Heerlen-heterozygous PS-C4BP bound more avidly to FXa than did normal PS-C4BP (apparent Kd = 4.3 nm vs. 82 nm). CONCLUSIONS: Plasma-derived PS-C4BP has direct anticoagulant activity in plasma and in purified systems. Enhanced direct activity of PS-Heerlen-C4BP may compensate for low free protein S levels and low cofactor activity in individuals with protein S-Heerlen.

Adult↗