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Biomedical subjects

N Fernández

Publications and source records attributed to N Fernández.

At least 19 recordsLinked to original sources
PubMed2006

Liquid versus solid phase bioassays for dredged material toxicity assessment.

Since 1994 the results of the analyses of key chemical compounds (trace metals, polychlorinated biphenyls and polycyclic aromatic hydrocarbons) and the comparison with the corresponding sediment quality guidelines (SQGs) are used in decision-making for dredged material management in Spain. Nonetheless in the last decades a tiered testing approach is promoted for assessing the physical and chemical characteristics of dredged sediments and their potential biological effects in the environment. Bioassays have been used for sediment toxicity assessment in Spain but few or no experiences are reported on harbour sediments. We studied the incidence of toxicity in the 7 d bioassay using rotifers (Brachionus plicatilis) and the 48 h bioassay using sea urchin (Paracentrotus lividus) embryos over a series of experiments employing 22 different elutriates. The relative performance of this exposure phase was not comparable to data on the 10-d acute toxicity test using the burrowing amphipod Corophium volutator and the polychaete Arenicola marina, carried out on the whole sediments. These results evidence the importance of the exposure route and the test selected in decision-making, as the toxicity registered for the undiluted elutriates was largely due to the different solubility of sediment-bound contaminants. This work and other studies indicate that for many sediments, a complete battery of test is recommended together with physico-chemical analyses to decide whether dredged sediments are suitable for open water disposal or not.

Animals↗
PubMed2006

Coronary reactive hyperaemia and arterial pressure in anaesthetized goats.

To study the effects of arterial pressure on coronary reactive hyperaemia, left circumflex coronary artery flow was measured, and reactive hyperaemia was determined after 5, 10 or 20 s of occlusion of this artery in anaesthetized goats during normotension, hypertension and hypotension. During hypertension induced by aortic constriction (mean arterial pressure, MAP = 140 +/- 6 mmHg) coronary vascular resistance (CVR), reactive hyperaemia (ratio of peak in hyperaemic flow to control flow and ratio of repayment to debt) and the decrease in CVR during the peak in hyperaemic flow were comparable to those during normotension. During hypertension induced by noradrenaline (MAP = 144 +/- 6 mmHg) CVR was 16% lower (P < 0.05), reactive hyperaemia was reduced by 14-25% (P < 0.05) and the decrease in CVR during the peak in hyperaemic flow was lower than the values of these parameters during normotension. During hypotension induced by constriction of the caudal vena cava (MAP = 40 +/- 4 mmHg) CVR was 22% lower (P < 0.05), reactive hyperaemia was reduced by 25-65% (P < 0.05) and the decrease in CVR during the peak in hyperaemic flow was less compared to the values of these parameters during normotension. During hypotension induced by isoprenaline (MAP = 45 +/- 4 mmHg) CVR was 59% lower, reactive hyperaemia was reduced by 55-100% (P < 0.01) and the decrease in CVR during the peak in hyperaemic flow was less compared to the values of these parameters during normotension. Arterial pressure is a main determinant of coronary reactive hyperaemia after brief periods of ischaemia, and the relationship between arterial pressure and reactive hyperaemia may depend in part on changes in CVR after variations in arterial pressure. These changes in CVR may be related to the action on coronary vessels of myocardial factors and vascular myogenic mechanisms.

Adrenergic beta-Agonists↗
PubMed2006

Plasmid-mediated VEGF gene transfer induces cardiomyogenesis and reduces myocardial infarct size in sheep.

We have recently reported that in pigs with chronic myocardial ischemia heart transfection with a plasmid encoding the 165 isoform of human vascular endothelial growth factor (pVEGF165) induces an increase in the mitotic index of adult cardiomyocytes and cardiomyocyte hyperplasia. On these bases we hypothesized that VEGF gene transfer could also modify the evolution of experimental myocardial infarct. In adult sheep pVEGF165 (3.8 mg, n=7) or empty plasmid (n=7) was injected intramyocardially 1 h after coronary artery ligation. After 15 days infarct area was 11.3+/-1.3% of the left ventricle in the VEGF group and 18.2+/-2.1% in the empty plasmid group (P<0.02). The mechanisms involved in infarct size reduction (assessed in additional sheep at 7 and 10 days after infarction) included an increase in early angiogenesis and arteriogenesis, a decrease in peri-infarct fibrosis, a decrease in myofibroblast proliferation, enhanced cardiomyoblast proliferation and mitosis of adult cardiomyocytes with occasional cytokinesis. Resting myocardial perfusion (99mTc-sestamibi SPECT) was higher in VEGF-treated group than in empty plasmid group 15 days after myocardial infarction. We conclude that plasmid-mediated VEGF gene transfer reduces myocardial infarct size by a combination of effects including neovascular proliferation, modification of fibrosis and cardiomyocyte regeneration.

Animals↗
PubMed2006

Microbiological and structural aspects of granular sludge from autotrophic denitrifying reactors.

Denitrification is applied in the tertiary treatment of wastewater to reduce N-pollutants. Fluorescence in situ hybridisation (FISH), CARD (catalyzed reporter deposition)-FISH, cloning, and scanning electron microscopy (SEM) were applied to follow the evolution of the microbial composition and structure of granular sludge in autotrophic denitrifying bioreactors fed with nitrate and thiosulfate. With this goal, FISH oligonucleotide probes for the autotrophic denitrifiers, Thiobacillus denitrificans and Thiomicrospira denitrificans, were designed and their utility tested. CARD-FISH and cloning data showed that bacterial diversity changed with bioreactor operation time. After 110 days of operation, the abundance of Thiobacillus denitrificans cells increased considerably: from 1 to 35% of total DAPI-stained cells and from no isolated clones to 30% of the total positives clones. This fact strongly suggests that this microorganism played a dominant role in the autotrophic denitrification. The Archaeal diversity remained almost unchanged and it was mainly represented by Methanosaeta soehngenii. Scanning electron microscopy results indicated a considerable loss in the integrity of the sludge granules during the operation, with risk of sludge buoyancy.

Base Sequence↗
PubMed2004

Effect of milking pipeline height on machine milking efficiency and milk quality in sheep.

This experiment studied the effect of milking pipeline height (mid- vs. low-level milking system) on milking efficiency and milk composition. The experiment was of 8 wk duration: 2 wk preexperimental period and 6 wk experimental, in crossover design (2 x 2). Ewes were milked in a 2 x 12 milking parlor with 2 milking pipelines set at a milking vacuum of 36 kPa with a pulsation rate of 180 cycle/min and ratio of 50%. Height of the milkline had no effect on yield of milk at the time of milking, yield after stripping, milk composition, SCC, and number of teatcup fall-offs. Nor did milkline height have any effect on milk lipolysis or on the distribution of fatty acids. The level of free fatty acids was higher in evening than in morning milk (60.5 vs. 25.6 mg/L). Likewise, the increase in the degree of lipolysis between the receiver (40.4 mg/L) and the refrigeration tank (45.8 mg/L) underlines the importance of the milk delivery line design. The parameters (time and flow rate) that define the first peak in the milk emission kinetics were statistically different between lines, so care must be taken when comparing milk emission curves from both types of pipeline.

Animals↗
PubMed2003

Integrative assessment of marine pollution in Galician estuaries using sediment chemistry, mussel bioaccumulation, and embryo-larval toxicity bioassays.

An integrative assessment of environmental quality was carried out in selected sites along the Galician coast (NW Iberian Peninsula) combining analytical chemistry of seawater and sediments, bioaccumulation in the marine mussel, and embryo-larval sediment toxicity bioassays, in order to link biological and chemical criteria for the assessment of coastal pollution. Maximum values of Hg and Cu in seawater, sediment and mussels, were found in the inner part of Ria of Pontevedra, while maximum levels of organics (polychlorinated biphenyls, hexachlorobenzene and aldrin) were found in mussels from A Coruña. Outstanding values of Cu, Pb and Zn have been found in seawater and sediment from a single site, P3, which also was the most toxic in the embryo-larval bioassays performed with four different phyla of marine organisms: mollusks, echinoderms, arthropods and chordates. Sediment quality effects range-median values provided a valuable reference to predict biological effects from sediment chemistry data, while effects range-low values were too conservative. Sediment toxicity could also be predicted by using a toxic-unit model based on published EC50 values for trace metals and mobilization factors independently obtained from measurements of metal contents in sediments and their elutriates. When chemical and toxicological data are independently used to arrange sampling sites by using non-metric multidimensional scaling, a remarkable degree of concordance between both types of configurations could be observed.

Animals↗
PubMed2003

Assessment of coastal marine pollution in Galicia (NW Iberian Peninsula); metal concentrations in seawater, sediments and mussels (Mytilus galloprovincialis) versus embryo-larval bioassays using Paracentrotus lividus and Ciona intestinalis.

Sediments from three Galician Rias were tested for toxicity using sea-urchin and ascidian sediment elutriate embryo-larval bioassays. Trace metal contents in seawater, sediments and mussels were also determined and subjected to multidimensional scaling methods which grouped stations according to chemical contamination. High metal contents were found in seawater, sediments and mussels from the Ria of Pontevedra, and moderate levels were detected in the Ria of Vigo and Ria of Arousa. The results revealed that samples assessed as toxic, according to the sea-urchin and ascidian embryo-larval bioassays, were among the most polluted by trace metals. A good agreement was reported between ordination plots resulting from applying multidimensional scaling to the chemical data, and the results of the biological endpoints tested.

Animals↗
PubMed2003

Measurements of associations of cell-surface receptors by single-particle fluorescence imaging.

SPFI (single-particle fluorescence imaging) uses the high sensitivity of fluorescence to visualize individual molecules that have been selectively labelled with small fluorescent particles. The images of particles are diffraction-limited spots that are analysed by fitting with a two-dimensional Gaussian function. The spot intensities depend on whether they arise from one or more particles; this provides the basis for determining self-association of cell-surface receptors. We have used this approach to determine dimerization of MHC class II molecules and its disruption by interface peptides. We have also exploited the positional information obtained from SPFI to detect co-localization of cell-surface molecules. This involves labelling two different molecules with different coloured fluorophores and determining their positions separately by dual wavelength imaging. The images are analysed to quantify the overlap of the particle images and hence determine the extent of co-localization of the labelled molecules. The technique provides quantification of the extent of co-localization and can detect whether co-localized molecules occur singly or in clusters. We have obtained preliminary data for co-localization of lipopolysaccharide and CD14 on intact cells. We also show that HLA-DR (human leukocyte antigen-DR) and CD74 are partially co-localized and that interaction between these molecules involves the peptide-binding groove of HLA-DR.

Animals↗
PubMed2003

Anaerobic biodegradability of water separated from extracted crude oil.

A study of the anaerobic biodegradability of the three categories of water separated from extracted crude oil (Venezuelan oilfields)--light, medium or heavy crude--was carried out at laboratory scale using UASB reactors working at mesophilic conditions. Chemical oxygen demand (COD) removal in a low loaded UASB reactor fed with water separated from extracted light crude was high, with an average 87% purification efficiency. The remaining COD was made up of the nonbiodegradable and the very slowly biodegradable fractions of the organic matter in the water. During a second period, the hydraulic retention time was reduced in stages, thus increasing the loading rate. In the experimental conditions used, COD concentration at the outlet remained below the Venezuelan standard limit for discharge into the environment (350 mg COD l(-1)) when the hydraulic retention time (HRT) was above 10 hours and the OLR under 3 g COD l(-1) d(-1). For HRT less than 10 hours, or organic loading rate (OLR) greater than 3 g COD l(-1) d(-1), COD at the outlet of the reactor rose as a consequence both of increased volatile fatty acids (VFA) concentrations (indicating an overloading of the methanogenic population) and, also, of the increase in the non-VFA COD (indicating a decrease in the acidification efficiency). On the other hand, results with UASB reactors operated at a low loading rate and fed with water separated from extracted medium and heavy crude oil showed that purification efficiency was low, with only 20% and 37% COD removal respectively. Continuing the operation of the UASB reactor fed with water separated from medium oil over a prolonged period did not bring any improvement, indicating that no adaptation of the sludge occurred.

Bacteria, Anaerobic↗
PubMed2003

Influence of pulsation rate on udder health and teat thickness changes in dairy ewes.

In this work on machine milking of ewes, pulsation rates of 120 and 180 cycles per min were compared, both with a pulsation ratio of 50:50 and a vacuum level of 36 kPa, comparing intramammary infection (IMI), somatic cell count (SCC) and teat end thickness changes. To this end, two groups of 20 Manchega ewes were used in a crossover experimental design with two experimental periods of 24 d for each. Bacterial exposure of all teats was increased by dipping them in a suspension of Staphylococcus simulans at four consecutive milkings of each period. Pulsation rate of 180 cycles per min, compared with 120 cycles per min, had no negative effect upon new IMI (11 and 16% of ewes infected, respectively) and SCC. No teat end lesions were observed in those animals milked with the two pulsations assayed. Also, teat thickness changes (-0.38 and -0.36 mm at 120 and 180 cycles per min, respectively) were not affected significantly. Finally, in absence of IMI, the two pulsation rates assayed did not affect the SCC.

Animals↗
PubMed2003

Influence of vacuum level and overmilking on udder health and teat thickness changes in dairy ewes.

The effects of vacuum level and overmilking on udder health were studied in ewes. Vacuum levels of 36 and 42 kPa were assigned to two groups of 23 Manchega ewes in a crossover study design with two experimental periods of 5 wk for each. Moreover, for each ewe, one teat was overmilked 1.5 to 2 min at all milkings during these 10 wk. The milking machine used had a midlevel milkline and pulsation was fixed at 180 cycles per min and a pulsation ratio of 50:50. Bacterial exposure of all teats was increased by dipping them in a suspension of Staphylococcus simulans at eight milkings of each period. New intramammary infections (IMI) were not significantly affected by the vacuum level used (18 and 23% of ewes infected, at 36 and 42 kPa, respectively) or application of overmilking (9 and 11% of half udders infected without and with overmilking, respectively). Likewise, neither factor significantly affected the somatic cell count (SCC) of the milk. Teat thickness changes after milking varied significantly due to the presence of overmilking (-13.6 and -7.4%, in teats not overmilked and overmilked, respectively) but were not affected by vacuum level. At no time were any lesions or variations visibly noted in the teat walls or orifice. So, in this work we were unable to demonstrate that the vacuum and overmilking levels assayed, both used with a pulsation rate of 180 cycles/min, have an important effect on the state of udder health in the short term. Furthermore, it was also observed that, in absence of IMI, the two factors studied did not cause irritation of any kind in the gland that might influence the SCC of the milk.

Animals↗
PubMed2002

Therapeutic effects of psyllium in type 2 diabetic patients.

OBJECTIVE: The aim of this study was to evaluate the effects of psyllium in type 2 diabetic patients. DESIGN: The study included three phases: phase 1 (1 week), phase 2 (treatment, 14 g fibre/day, 6 weeks) and phase 3 (4 weeks). At the end of each phase a clinical evaluation was performed after the ingestion of a test breakfast of 1824.2 kJ (436 kcal). Measurements included concentrations of blood glucose, insulin, fructosamine, GHbA(1c), C-peptide and 24 h urinary glucose excretion. In addition, uric acid, cholesterol and several mineral and vitamin concentrations were also evaluated. SETTING: The study was performed at the Department of Pharmacology, Toxicology and Nursing at the University of León (Spain). SUBJECTS: Twenty type 2 diabetic patients (12 men and 8 women) participated in the study with a mean age of 67.4 y for men and 66 y for women. The mean body mass index of men was 28.2 kg/m(2) and that of women 25.9 kg/m(2). RESULTS: Glucose absorption decreased significantly in the presence of psyllium (12.2%); this reduction is not associated with an important change in insulin levels (5%). GHbA(1c), C-peptide and 24 h urinary glucose excretion decreased (3.8, 14.9 and 22.5%, respectively) during the treatment with fibre (no significant differences) as well as fructosamine (10.9%, significant differences). Psyllium also reduced total and LDL cholesterol (7.7 and 9.2%, respectively, significant differences), and uric acid (10%, significant difference). Minerals and vitamins did not show important changes, except sodium that increased significantly after psyllium administration. CONCLUSIONS: The results obtained indicate a beneficial therapeutic effect of psyllium (Plantaben) in the metabolic control of type 2 diabetics as well as in lowering the risk of coronary heart disease. We also conclude that consumption of this fibre does not adversely affect either mineral or vitamin A and E concentrations. Finally, for a greater effectiveness, psyllium treatment should be individually evaluated.

Aged↗
PubMed2002

The need for continuous immunosuppression with cyclosporin A to maintain an experimental model of uveal melanoma.

We investigated the need for continuous immunosuppression to maintain experimental tumours derived from human uveal melanoma cells implanted in the choroid of pigmented rabbits. Two groups of pigmented rabbits immunosuppressed with cyclosporin A (CsA) were implanted with human uveal melanoma cells in the suprachoroidal space. After 5 weeks, CsA was discontinued in group 2. Animals were treated with prophylactic antibiotics and examined weekly for tumour growth, weight and secondary effects; blood urea nitrogen levels were measured every two weeks. Autopsies and histopathological studies were performed after death or euthanasia at the end of week 12. The difference between the groups in the development of ophthalmoscopic tumours was not statistically significant 5 weeks after implantation. Tumours in group 1 grew progressively throughout the experiment, whereas group 2 tumours showed marked regression 3-4 weeks after discontinuing CsA. Tumours in group 1 were significantly larger and had greater mitotic activity and showed more ciliary body, optic nerve and extrascleral invasion than tumours in group 2, which showed massive fibrosis, minimal mitotic activity and marked inflammatory cell infiltration. Continuous immunosuppression with CsA seems to be necessary to maintain tumour growth in this experimental model of uveal melanoma.

Animals↗
PubMed2001

Diabetes abolishes the gender difference in vasopressin-mediated potentiation of sympathetic vasoconstriction.

Electrical field stimulation (4 Hz, 0.2 ms pulse duration, at a supramaximal voltage of 70 V, for 1 s) of isolated rat tail artery segments produced contraction which was lower in female than in male rats, and was reduced by streptozotocin-induced diabetes in both genders. This contraction was potentiated by vasopressin (10(-12)-10(-10) M) more in normoglycemic male than in normoglycemic female rats, and this effect of vasopressin was increased by the cyclooxigenase inhibitor meclofenamate (10(-5) M) in female control rats, but not in diabetic female, or control and diabetic male rats, and it was not modified by the nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 10(-4) M). Endothelin-1 (10(-10)-3 x 10(-9) M) also potentiated the contraction to electrical stimulation. This potentiation was similar in all experimental groups, and it was not modified by meclofenamate or L-NAME. These results suggest that the potentiating effect of vasopressin, but not that of endothelin-1, on the sympathetic vasoconstriction, is lower in females than in males, probably by an increased release of vasodilating prostanoids, and this release may be reduced by diabetes in females.

Animals↗
PubMed2001

Adrenergic vasoconstrictor activity in the cerebral circulation after inhibition of nitric oxide synthesis in conscious goats.

The interaction between nitric oxide (NO) and adrenergic activity in the cerebral circulation was studied using conscious goats, where blood flow to one brain hemisphere (cerebral blood flow) was electromagnetically measured, and the effects of phentolamine and hexamethonium on cerebrovascular resistance were evaluated before (control) and after inhibition of NO synthesis with NW-nitro-L-arginine methyl ester (L-NAME). L-NAME (12 goats, 40 mg kg(-1) administered i.v.) reduced cerebral blood flow from 62 +/- 3 to 44 +/- 2 ml min(-1), increased mean systemic arterial pressure from 100 +/- 3 to 126 +/- 4 mm Hg, decreased heart rate from 79 +/- 5 to 50 +/- 4 beats min(-1) and increased cerebrovascular resistance from 1.63 +/- 0.08 to 2.91 +/- 0.016 mm Hg ml(-1)min(-1) (all P < 0.01). These hemodynamic variables normalized 48-72 h after L-NAME administration. Phentolamine (six goats, 1 mg), injected into the cerebral circulation. increased cerebral blood flow without changing systenic arterial pressure, but its cerebrovascular effects were augmented for about 24 h after L-NAME. The decrements in cerebrovascular resistance induced by phentolamine, in mm Hg ml(-1) min(-1), were: under control, 0.42 +/- 0.05; immediately after L-NAME, 1.38 +/- 0.09 (P < 0.01 compared with control); by about 24 h after L-NAME, 0.71 +/- 0.09 (P < 0.05 compared with control); and by about 48 h after L-NAME, 0.40 +/- 0.07 (P > 0.05 compared with control). Hexamethonium (six goats, 0.5-1 mg kg(-1) min(-1) i.v.) decreased mean systemic arterial pressure to about 75 mm Hg and caused tachycardia similarly before and after L-NAME, but the decrements in cerebrovascular resistance were augmented for about 24 h after L-NAME. The decrements in cerebrovascular resistance induced by hexamethonium, in mm Hg ml(-1).min(-1), were: under control. 0.61 +/- 0.09, immediately after L-NAME, 1.33 +/- 0.16 (P < 0.01 compared with control); by about 24 h after L-NAME, 1.18 +/- 0.10 (P < 0.01 compared with control): and by about 48 h after L-NAME, 0.99 +/- 0.10 (P > 0.05 compared with control). Therefore, these results suggest that adrenergic vasoconstrictor tone in cerebral vasculature may be augmented after inhibition of NO synthesis, and that this increment may contribute to the reduction of cerebral blood flow after inhibition of NO formation.

Adrenergic alpha-Antagonists↗
PubMed2001

FcgammaRIIA exogenously expressed in HeLa cells activates the mitogen-activated protein kinase cascade by a mechanism dependent on the endogenous expression of the protein tyrosine kinase Syk.

HeLa cells transfected to express the human Fc receptor FcgammaRIIA were stimulated with aggregates of IgG, IgG-ovalbumin equivalence immune complexes and monoclonal antibody reacting with FcgammaRIIA. All of these stimuli activated the cells as judged from the band-shift characteristic of the activation of the p42-MAP/ERK kinase. Since this response is currently associated with the activation of the protein tyrosine kinase Syk, the expression of which is currently thought to be restricted to hemopoietic cells, the results were considered as an indirect evidence of the expression in HeLa cells of either Syk or another protein tyrosine kinase accounting for the same function. Transfection with a dominant negative Syk mutant abrogated the response to FcgammaRIIA cross-linking, whereas overexpression of Syk did not increase the extent of the response. Further evidence of the expression of syk was obtained by the reverse transcription PCR approach and sequencing of the DNA bands. Moreover, immunoprecipitation with anti-Syk antibody of the cell lysates obtained after cross-linking of FcgammaRIIA followed by immunoblotting with anti-phosphotyrosine antibody showed the phosphorylation of a protein band migrating as Syk. These data indicate that expression of FcgammaRIIA on epithelial HeLa cells conveys signals to the p42-MAP/ERK kinase by a mechanism involving the recruitment of Syk. In contrast, cross-linking of this receptor does not yield productive signals coupled to other responses associated to the FcgammaR system such as triggering of the arachidonic acid cascade, activation of the NF-kappaB system and production of chemotactic cytokines.

Animals↗