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Biomedical subjects

A Pastor

Publications and source records attributed to A Pastor.

At least 19 recordsLinked to original sources

Development of a multianalyte method for the determination of anabolic hormones in bovine urine by isotope-dilution GC-MS/MS.

A sensitive, specific and selective multianalyte GC-MS/MS method has been developed for the determination of 11 anabolic hormones in bovine urine. After adjusting the urine pH to 4.8, the samples were spiked with deuterated internal standards and submitted to enzymatic hydrolysis with beta-glucuronidase/arylsulfatase. Hormones were eluted with methanol through a C18 solid phase cartridge and submitted to a liquid-liquid extraction. Analytes were derivatized by adding N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane and GC-MS data were obtained in the positive electron impact tandem mass mode. Under these conditions, no matrix effects were observed and limit of detection values were in the range of 0.005 ng/mL (diethylstilbestrol) to 0.38 ng/mL (17alpha-methyltestosterone and 17alpha-ethynylestradiol). Recoveries from 81% (alpha-zeranol) to 149% (17alpha-methyltestosterone) were found under the selected conditions. These results were better than those found using heptafluorobutyric anhydride (HFBA) as derivative reagent and those measured in full scan and selective ion monitoring modes.

Animals↗

Determination of acrylamide in foods by pressurized fluid extraction and liquid chromatography-tandem mass spectrometry used for a survey of Spanish cereal-based foods.

An automated and rapid method for the determination of acrylamide in different food products is presented. The method involves pressurized fluid extraction (PFE) of foods with acetonitrile and precipitation with Carrez reagents. The final extract is analysed by liquid chromatography coupled to electrospray ionization tandem mass spectrometry (ESI-MS-MS). The main parameters affecting the performance of ESI-MS-MS and PFE were optimized using a design of experiments approach. The limit of quantification of the method was 5 microg kg(-1), and recoveries from incurred samples ranged between 93 and 101%. The accuracy was evaluated using the reference test materials FAPAS T3002, T3005 and T3011. Using the optimized method, 62 food samples of potato chips, snacks, biscuits, breakfast cereals and crisp bread sampled from Valencia, Spain, supermarkets were surveyed for acrylamide levels. The levels were similar to those reported in the European Union and USA.

Acrylamide↗

Comparative toxicokinetics of cadmium in Artemia.

The toxicokinetics of cadmium was determined for five populations belonging to four species of Artemia (A. salina, A. parthenogenetica, A. franciscana, and A. persimilis) using a bicompartmental model. The effects of sublethal cadmium concentration on the kinetic parameters in A. parthenogenetica were determined. The BCF values are inversely related to the concentration, as is the uptake rate (ku), whilst the elimination rate (ke) constant seems to be directly related to the exposure concentration. Values corresponding to the rate of metal influx (I) remain relatively constant (0.2477 to 0.4455 microg/g.h) in the concentration range from 0.1 to 1 mg Cd/L, and are higher (1.098 microg/g.h) at an exposure to 10 mg Cd/L. The cadmium accumulation pattern seems well conserved in the genus and is characterized by a fast elimination of the metal with Ke ranging from 0.0050 to 0.0231 h(-1). A. persimilis displays a different model to that corresponding to the other studied species exposed to the same cadmium concentration, presenting a low uptake rate constant (1.0564 mL/g. h) and a low BCF (211.3 mL/g).

Animals↗

Organochlorinated pesticides in sediments from the Lake Albufera of Valencia (Spain).

Bottom sediment samples from 121 sites of the Lake Albufera of Valencia were analyzed. Dieldrin, endrin, heptachlor and op'-DDT were not detected (<0.01 ng g(-1)) in 88-93% of the sites. Aldrin and HCB concentration ranges were between <0.01 and 0.1 ng g(-1) in 86% and 94% of the sites, respectively. Heptachlor-epoxide and lindane 95% confidence intervals were 0.2-0.5 and 0.06-0.12, respectively. The greatest average concentration corresponds to pp'-DDE, pp'-DDD and pp'-DDT. The sum of six isomers and derivatives of the DDT average concentration reaches 2.1 ng g(-1), as opposed to 2.7 ng g(-1) for the sum of 13 pesticides considered. In the site with a major contamination, 27.0 ng g(-1) of pp'-DDD and 12.8 ng g(-1) of pp'-DDT were accumulated. The DDE:DDT proportion average was 0.37, indicating an aged DDT contamination. Concentrations of pesticides in sediments were compared to three sediment quality guidelines, and indicated that a low biological effects level can be expected in either sediments or aquatic organisms.

DDT↗

Lymphoproliferative disease after lung and heart-lung transplantation: first description in Spain.

Lymphoproliferative syndromes are the most common tumors in transplant recipients. More than 90% of posttransplantation lymphoproliferative syndromes (PTLS) are considered to be associated with Epstein-Barr virus, and 86% are of the B-cell line. Histopathology ranges from polymorphic-reactive to monomorphic forms. Clonality should be studied using molecular biology techniques. Clinically, a differentiation is usually made between early PTLS (occurring within 1 year after transplantation) and late PTLS, which occur as localized or disseminated nodal lymphomas. In localized forms, immunosuppression should be discontinued or decreased, and the involved area should be subsequently resected or irradiated. In disseminated cases, immunosuppression should be decreased and administration of acyclovir/ganciclovir should be considered. If this is not effective, treatment should be started with anti-CD20 monoclonal antibodies (rituximab). If no response occurs, use of chemotherapy, possibly with interferon, should be considered. Our aim was to report the incidence, clinical signs, and treatment in a series of patients undergoing lung transplantation (LTx).

Adolescent↗

Pulmonary tuberculosis in lung and heart-lung transplantation: fifteen years of experience in a single center in Spain.

The increase in the number of solid organ transplants has resulted in an increased incidence of opportunistic infections, including infection by typical and atypical mycobacteria, with risk of developing tuberculosis. Pretransplant chemoprophylaxis with isoniazid has become increasingly common in an attempt to prevent the disease. The source of infection in tuberculosis (TB) may be difficult to identify. Infection may be caused by reactivation of a primary infection in the recipient, reactivation of a lesion from the donor lung, or primary infection. There are few reports on TB in lung transplantation. Incidence in the reported series ranges from 6.5% to 10%. Our series of 7 patients out of a total 271 patients (2.58%) represents a rate higher than reported for the general Spanish population, 26.7/10(5) inhabitants and for lung transplant candidates (0.18%). Our aim was to evaluate the incidence, clinical signs, and outcome of TB in our series of patients undergoing lung transplantation in the 15 years since inception of the program (February 1990 to December 2004). Morbidity and mortality was high (42.8%), but limited to patients in whom treatment was not administered or could not be successfully completed. However, early detection and treatment are essential.

Adult↗

Steroid withdrawal in lung transplant recipients.

OBJECTIVE: Many of the long-term complications in lung transplantations are secondary effects of immunosuppression. Corticosteroids are partially responsible for the development of osteoporosis, raised blood pressure, diabetes, muscular disorders, gastric ulcers, and other conditions. We analyzed the long-term result of steroid withdrawal in our lung transplant recipients. MATERIALS AND METHODS: When respiratory function stabilized, to avoid secondary effects, steroid treatment was withdrawn in 34 of the 375 lung transplant patients in our centers We evaluated the characteristics of the donors and recipients, their compatibility, the pre, and post-steroid withdrawal complications, and type of immunosuppressant. RESULTS: The mean age of patients was 42 +/- 7 years and of donors, 25 +/- 9 years. The primary diseases were: 15 emphysema, six pulmonary fibrosis, 10 cystic fibrosis, and three primary pulmonary hypertension. Twenty seven patients had double lung transplants and seven single lung. The mean steroid withdrawal period was 881 +/- 237 days posttransplantation. The most frequent treatment regimen at the time of steroid withdrawal was cyclosporine, azathioprine, and minimal steroid doses. Six recipients had to be restarted on steroids one patient who required a kidney transplant, three cases due to an infectious process with a differential diagnosis of rejection, and two cases due to loss of FEV1 (forced expiratory volume in 1 s), suggestive of chronic rejection. There was an improvement in blood pressure in five patients, in plasma cholesterol and triglyceride levels in eight patients, and insulin withdrawal in two diabetic patients. CONCLUSIONS: Steroid treatment may be suspended 2 to 3 years, posttransplant in selected lung transplant recipients. The usual patient profile shows few rejection episodes with cyclosporine and azathioprine immunosuppression. What is notable is the low mean age of donors. Close clinical monitoring and lung function testing are of major importance in the weeks following steroid withdrawal.

Adolescent↗

Application of accelerated solvent extraction followed by gel performance chromatography and high-performance liquid chromatography for the determination of polycyclic aromatic hydrocarbons in mussel tissue.

Accelerated solvent extraction (ASE) has been evaluated as a fast alternative to methanolic saponification for the extraction of 12 polycyclic aromatic hydrocarbons (PAHs) from mussel tissue. Several solvent systems and different operating conditions were investigated. The mixture dichloromethane-acetone (1:1, v/v) gave the best recoveries at 125 degrees C and 1500 psi, in a total time of 10 min. No yield difference was found between freeze-drying (Fd) or drying the wet mussel with diatomaceous earth (Ded) prior to extraction. The ASE method was validated using the standard reference material SRM 2977, a freeze-dried mussel tissue with naturally present organic contaminants. The performance characteristics of the ASE method (trueness: 70-110%; precision: 4-14% and limit of quantification (LOQ): 0.1-0.25 microg/kg) meet the criteria established by the European Union for quantitative methods of analysis for official control of organic residues and contaminants. ASE provides a 24 times faster extraction than MSE and reduces 12 times the volume of solvent required.

Animals↗

[Evaluation of microsurgical treatment in a series of 121 intracranial aneurysms].

OBJECTIVE: The results obtained with therapy of intracranial aneurysms, in terms of morbidity and mortality, are very important when the patient has to choose between microsurgical techniques or endovascular management. The aim of this paper is to review the information regarding current microsurgical treatment of intracranial aneurysms, and presenting our experience over the last five years. MATERIAL AND METHODS: We studied 101 consecutive patients with 121 intracranial aneurysms admitted between 1996 and 2000 with the initial diagnosis of subarachnoid hemorrhage. We paid special attention to the day of admission from the onset of the symptomatic hemorrhage to the grade of Hunt&Hess scale and the possibility of early or delayed microsurgical treatment. The diagnosis was based on four vessels cerebral angiography and in a few cases with CT-angiography. All patients were treated by microsurgical technique and such treatment was completed by nimodipine, intensive care unit management and in some cases of postoperative suspected vasospasm, induced arterial hypertension was applied. Post surgical angiography was carried out in all patients to confirm the clipping of the cerebral aneurysm. The 12 months assessment was based on the Glasgow Outcome Scale (GOS). RESULTS: The 92.1% of the patients were admitted with a grade equal or below III in the Hunt&Hess scale. A 80% were operated within the 72 hours of admission and in the remaining cases, the surgical treatment was delayed due to a grade IV or V or to a medical contraindication. Four patients died (3.9%). At 12 months follow up, 88.9% presented a score I or II in the GOS. CONCLUSION: According to our results, there are a substantial improvements in the microsurgical treatment of cerebral aneurysms, specially in patients admitted early after the onset of the symptoms of their hemorrhage, who have a grade I to III in the Hunt&Hess scale and showed a good level of consciousness. We think that the improvement of our results are due to: l. the high percentage of patients admitted with grades I to III. 2. the high percentage of patients operated within the first 72 hours from the onset of their symptomatic hemorrhage. 3. surgery was always carried out by the same two experienced vascular neurosurgeons. 4. intraoperative measures taken to prevent the rupture of the aneurysm. 5. early administration of nimodipine, ICU management, doppler studies and in seldom cases, induced hypertension therapy to treat the vasospasm and postoperative hypotension.

Adolescent↗

Localization of functional calcitonin gene-related peptide binding sites in a subpopulation of cultured dorsal root ganglion neurons.

In this study we investigated whether cultured dorsal root ganglion (DRG) neurons from the adult rat express binding sites for calcitonin gene-related peptide (CGRP). These were identified on fixed cells by using CGRP labeled at the N-terminal site with 1.4-nm gold particles. After 1 day in culture, about 20% of small to medium-sized DRG neurons showed CGRP-gold binding. Binding of CGRP-gold was dose-dependently reduced by coadministration of CGRP. The calcium imaging technique in living cells revealed that the bath administration of CGRP evoked an increase of the intracellular calcium in up to 30% of the DRG neurons tested. Both depletion of intracellular calcium stores by thapsigargin or using a calcium-free medium blocked the CGRP-mediated increase of cytosolic calcium in most neurons. Thus intracellular and extracellular sources of calcium are relevant for the CGRP response. Using the whole-cell patch-clamp technique, about 30% of the neurons were found to exhibit an inward current and a depolarization upon administration of CGRP close to the neurons. Immunocytochemical double-labeling techniques showed that most of the CGRP-gold binding sites were expressed in unmyelinated (neurofilament 200-negative) DRG neurons. Most of the neurons with CGRP-gold binding sites also expressed the tyrosine kinase A receptor, and all of them showed CGRP-like immunoreactivity. This study shows, therefore, that a subpopulation of unmyelinated, peptidergic primary afferent neurons express CGRP binding sites that can be activated by CGRP in an excitatory direction. The binding sites may serve as autoreceptors because all of these neurons also synthesize CGRP. The activation of CGRP binding sites may sensitize primary afferent neurons and influence the release of mediators.

Animals↗

[Lumbar root compression secondary to juxtafacet cysts: review of 10 cases].

OBJECT: We report a series of 10 patients with 11 juxtafacet cysts of the lumbar spine treated in our center from 1994 to 2000. METHODS: The clinical histories, radiographic images, surgical protocols and pathological records of the 10 patients diagnosed of lumbar juxtafacet cyst have been analyzed. RESULTS: Six patients were women and four were men. The average age of presentation was 54 years. The most frequent clinical presentation was radicular pain, and motor or sensitive deficits were not very common. Computed tomography and magnetic resonance are essential in establishing the diagnosis. The cysts were located at L4-L5 in eight patients, at L2-L3 in one patient, and at L3-L4 in other patient. Eight patients were treated by means of decompressive laminectomy and excision of the cyst, and the remaining two underwent conservative treatment. Degenerative phenomena adjacent to the juxtafacet cysts are easy to evidence. All the patients were improved of their radicular pain. CONCLUSIONS: Analysing our series, we conclude that the juxtafacet cysts appear more frequently at L4-L5 level in people of advanced age, and produce commonly low back pain and radiculopathy. Although they can be treated initially in a conservative way, in the presence of a progressive sciatic syndrome, surgical management is preferable.

Adult↗

Asymptomatic brugada syndrome case unmasked during dimenhydrinate infusion.

Typical ECG of that described for Brugada syndrome was elicited in a patient diagnosed with labyrinthopathy during infusion of dimenhydrinate, a first-generation antihistamine usually used to treat motion sickness. Although the patient had no history of syncope or palpitations, and there was no family history of cardiac disease or sudden death, the ECG abnormality was reproduced later with intravenous flecainide, and an asymptomatic Brugada syndrome was diagnosed.

Adult↗

Factors influencing the inhibition of biliary glutathione efflux induced by biliary obstruction.

The aim of this study was to investigate mechanisms responsible for the inhibition of biliary glutathione efflux in rats with secondary biliary cirrhosis. Rats were studied after bile duct obstruction for 28 days. The biliary secretion of reduced glutathione (GSH), oxidised glutathione (GSSG) and cysteine were completely inhibited in biliary obstructed rats. Hepatic gamma glutamyltranspeptidase (gamma-GT) activity increased significantly, but following its inhibition by acivicin administration GSH, GSSG and cysteine were still absent in bile. Biliary obstruction resulted in a significant increase of the permeability of the paracellular pathway, as shown by the higher bile/plasma ratio and hepatic clearance of [14C]sucrose. GSH and GSSG were, however, significantly lower in the carotid artery and hepatic vein of obstructed animals and the arteriovenous difference across the liver was reduced. The concentration of GSH was significantly reduced and that of GSSG increased in the liver of obstructed rats. Biliary obstruction induced an increase in the hepatic concentration of cysteine and an inhibition of both gamma glutamylcysteine synthetase and methionine adenosyl transferase activities. Dichlorofluorescein (DCF) and the GSSG/GSH ratio and thiobarbituric acid reactive substances (TBARS) concentration, markers of reactive oxygen species production and lipid peroxidation, respectively, were significantly increased. Our data indicate that increased degradation or blood reflux of glutathione do not participate in the disruption of its secretion into bile and support the view that impairment of glutathione synthesis and oxidative stress could contribute to the decline in biliary glutathione output.

Animals↗

Electrothermal atomic absorption spectrometric diagnosis of familial hypercholesterolemia.

We have developed a new nonradioactive assay to identify human low-density lipoprotein receptor defects. It is based on the incubation of cultured cells with colloidal gold-LDL conjugates and quantitation of the gold associated with the cells by electrothermal atomic absorption spectrometry. After an oxidative treatment with nitric and hydrochloric acids, the biological matrix interferes neither with the gold recovery nor with the gold measurements, which are linear, at least from 0.15 to 3 ng of gold. When cells expressing a functional LDL receptor are incubated with increasing amounts of colloidal-gold LDL conjugates, the obtained saturation curve parallels that described when [125I]LDL is used as ligand. Moreover, this new assay allows us to clearly distinguish among fibroblasts from normal subjects or from heterozygous or homozygous patients of familial hypercholesterolemia, a very common autosomal disease. The assay is easy to perform, is sensitive, and avoids the use of radioactive compounds. Therefore, it could be successfully employed in the clinical diagnosis of this disease. Furthermore, since the methodology developed here can be applied to quantify the association of other gold-conjugated ligands to cells, it could have a widespread use in a variety of clinical and basic research studies.

Animals↗

Direct amination of olefins through sequential triazolinedione ene reaction and carbanion-assisted cleavage of the N-N urazole bond

[formula: see text] Allylic amines 5 are obtained in 30-55% overall yields by the base-catalyzed hydrolysis of trialkylated allylic urazoles 3; the latter are prepared by the TAD ene reaction of the appropriate olefin and further N-alkylation with alpha-bromoacetophenone. The proposed mechanism for this novel urazole rupture is based on the generation of a carbanion adjacent to the hydrazide functionality, which induces urazole ring-opening by cleavage of the N-N bond.

Journal Article↗