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Biomedical subjects

I Rodríguez

Publications and source records attributed to I Rodríguez.

At least 19 recordsLinked to original sources

Determination of acidic drugs in sewage water by gas chromatography-mass spectrometry as tert.-butyldimethylsilyl derivatives.

A procedure is described for the determination of five acidic non-steroidal anti-inflammatory pharmaceuticals (ibuprofen, naproxen, ketoprofen, tolfenamic acid and diclofenac) in sewage water. The analytical method involves the concentration of water samples using a solid-phase extraction polymeric sorbent, functionalized with N-vinylpyrrolidone. Analytes were eluted with ethyl acetate. derivatized using N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) and analyzed by GC-MS. Influence of time, temperature and volume of MTBSTFA in the yield of the derivatization step were studied in detail using a factorial central composite design. Quantification limits of the analytical procedure for 500 ml of sewage water ranged from 20 to 50 ng/l. Recoveries from 90 to 115% were found for sewage water samples spiked with the studied compounds at the low ng/ml level. Results obtained for real samples show the presence of ibuprofen and naproxen in both influent and effluent of a sewage water treatment plant.

Anti-Inflammatory Agents, Non-Steroidal↗

Helium-3 MRI diffusion coefficient: correlation to morphometry in a model of mild emphysema.

Hyperpolarised gases have been most recently used in magnetic resonance imaging to demonstrate new image-derived pulmonary function parameters. One of these parameters is the apparent diffusion coefficient, which reflects the sizes of the structures that compartmentalise gas within the lung (i.e. alveolar space). In the present study, noninvasive parameters were compared to microscopic measurements (mean linear intercept and mean alveolar internal area). Nonselective helium-3 gas density coronal ex vivo images and apparent diffusion maps were acquired in control and elastase-induced panacinar emphysema rats. Total lung capacity was considered the reference for both imaging experiments and lung fixation. A mild degree of emphysema was found based on mean linear intercept (134 +/- 25 microm) versus control (85 +/- 14 microm). The apparent diffusion coefficients were significantly different between the two groups (0.18 +/- 0.02 and 0.15 +/- 0.01 cm2 x s(-1) for elastase and control, respectively). A significant correlation between the apparent diffusion coefficient and corresponding morphometric parameters in mild emphysema was demonstrated for the first time. This study opens the possibility of estimating absolute airspace size using noninvasive techniques.

Animals↗

Effect of vigabatrin on the pharmacokinetics of carbamazepine.

OBJECTIVE: To evaluate a possible interaction between vigabatrin and carbamazepine in epileptic patients. METHODS: Steady-state serum concentrations of carbamazepine with and without vigabatrin were compared. The study group consisted of 15 patients (eight females, seven males, and mean age 31 +/- 12 years), with refractory partial epilepsy. They received vigabatrin as add-on therapy. Patients received carbamazepine monotherapy for at least 6 months and the carbamazepine-vigabatrin combination for at least 3 months. Blood samples were obtained in the morning, before the first daily dose and the carbamazepine plasma concentrations were analysed by fluorescence polarization immunoassay (TDx System). RESULTS: No statistically significant differences were found in mean carbamazepine daily dose. Mean trough concentrations were 7.9 +/- 1.4 microg/mL with carbamazepine alone, and 6.5 +/- 2.0 microg/mL with carbamazepine-vigabatrin association (P < 0.03). The mean values of pharmacokinetic parameters were: level/dose ratio (L/D) = 0.59 +/- 0.20 vs. 0.45 +/- 0.15 (P < 0.05) and plasma clearance (Cl) = 78.5 +/- 25.8 vs. 105.8 +/- 38.9 mL/h/kg (P < 0.05), with carbamazepine alone and carbamazepine-vigabatrin combination, respectively. CONCLUSION: Vigabatrin produced a statistically significant increase in the plasma clearance of carbamazepine when the two drugs were given simultaneously.

Administration, Oral↗

Subclinical pain and thermal sensory dysfunction in children and adolescents with Type 1 diabetes mellitus.

AIMS: To assess cutaneous thermal and pain thresholds in upper and lower limbs in neurologically asymptomatic children and adolescents, and to study the relationships of clinical parameters and these sensory thresholds in subclinical diabetic neuropathy. METHODS: Thirty-five neurologically asymptomatic patients, aged 8-16 years, diagnosed with Type 1 diabetes mellitus (DM), and a control group of 35 healthy age-matched subjects participated in the study. Warmth, cold, and heat-induced pain thresholds were measured in the dorsum of the right arm and foot, using quantitative sensory testing (QST). Relevant clinical parameters, retrieved from medical records or measured at the QST session, were obtained for each patient. RESULTS: Compared with controls, diabetic patients had increased thresholds for warmth in the hand (P = 0.002), and cold and warmth in the foot (P = 0.015 and P = 0.004, respectively). Of the diabetic patients, 43% showed abnormality of at least one sensory threshold. A significant correlation was observed for duration of diabetes and heat-induced pain threshold in the hand (P = 0.045), but no correlation was found for age, height, weight, pre-test blood glucose, age of onset, current insulin dose, and mean of glycosylated haemoglobin determinations of the previous 18 months. No significant correlation was found for other sensory thresholds. CONCLUSIONS: Using QST, abnormal cutaneous thermal perception is a common finding, in both upper and lower limbs, in neurologically asymptomatic young diabetic patients. Heat-induced pain threshold in the hand was correlated with the duration of the diabetes.

Adolescent↗

Chimeric synthetic peptides from the nucleocapsid p24 protein of human immunodeficiency virus type-1.

Two chimeric synthetic peptides incorporating antigenic sequences from N-terminal (peptide C14) (134-163) and C-terminal (peptide C15) (335-364) of the p24 protein of human immunodefiency virus (HIV- 1), were synthesized. Peptides C14-GG-C15 and C15-GG-C14 represented sequences from the p24 protein in both possible orders, separated by two glycine residues as arm spacers. These peptides were evaluated as antigen in an Ultramicroenzyme-linked immunosorbent assay (UMELISA) using sera of HIV-1-infected individuals (n = 16) with different titers of antibodies and the specificity was evaluated with healthy blood donors (n = 20). The results were compared to plates coated with monomeric peptides C14 and C15. The chimeric peptide C14-GG-C15 was the most antigenic. Those results may be related to the peptide structure, the sequence order in the chimeric peptide, and epitope accessibility to the antibodies. This chimeric peptide would be very useful for HIV-1 diagnostics.

Amino Acid Sequence↗

Simultaneous determination of butyltin and phenyltin species in sediments using ultrasound-assisted leaching.

A fast and simple procedure is presented for the simultaneous leaching of butyl (mono, di and tributyl) and phenyl organotin species from sediment samples. Leached compounds are further ethylated with sodium tetraethylborate in aqueous medium, and analyzed by gas chromatography. After testing the stability of triphenyltin under different extraction conditions, ultrasound-assisted leaching at room temperature in the presence of acetic acid was been proposed as an extraction procedure compatible with the simultaneous determination of phenyl- and butyltin compounds in sediments. Recoveries between 70 and 90% were obtained for phenyl species in spiked samples prepared in the laboratory. Results for butyltin species were validated by use of the reference material PACS-2. Quantification limits, using GC-MIP-AES as measurement technique, were approximately 5-10 ng g(-1). Precision in the consecutive analysis of three sediment samples varied between 3 and 10%.

Chromatography, Gas↗

Regulation of serine-type exoproteinases by endogenous inhibitors present in exoantigens of the mycelial form of Paracoccidioides brasiliensis.

We have partially characterized some biochemical properties of exoproteinases secreted into culture medium by the mycelial form of Paracoccidioides brasiliensis, a dimorphic fungus that causes human disease in Latin America. Proteinase activity was analyzed in solid- and liquid-phase systems using zymography and Azocoll, respectively. Minimal or no gelatinase activity was observed by zymography in the crude filtrates among proteins with a relative mobility greater than 200 kDa. When the crude filtrate was fractionated by isoelectric focusing or ion exchange chromatography, we observed striking activation of gelatinases, both those of high apparent molecular mass and alkaline isoelectric points (pI), as well as those of lower molecular mass and acidic pI. The apparent high molecular mass gelatinases, pI 10, showed optimal activity at pH 7.0. They were totally inhibited by phenylmethylsulfonylfluoride and partially inhibited by incubation with previously neutralized fractions of pI 5.4 and 6.1. The latter inhibition could be reversed by exposure to 10% isopropanol. These results provide evidence of regulatory mechanisms controlling proteinase activity in secreted proteins. The principal mechanism appears to be the formation of reversible complexes with endogenous inhibitors.

Antigens, Fungal↗

Pneumococcal pyomyositis.

Pyomyositis is most commonly caused by Staphylococcus aureus. A 25-month-old child developed infection of the biceps brachialis muscle caused by Streptococcus pneumoniae. The child had no underlying immune or anatomic defect.

Child, Preschool↗

Solid-phase extraction of phenols.

Sample preparation for phenol analysis using solid-phase extraction (SPE) is reviewed. The scope of the review has been restricted to the literature dealing with the analysis of phenols as the main objective. The use, advantages and disadvantages of silica sorbents, polymeric, functionalized, carbon-based and mixed available sorbents, when applied to the separation and preconcentration of phenols, as well as the available experimental devices, are discussed. Other aspects such as phenol derivatisation prior to SPE, solid-phase microextraction, matrix effects and the storage of phenols in SPE cartridges, have been also discussed.

Chromatography, Liquid↗

Very high expression of an anti-carcinoembryonic antigen single chain Fv antibody fragment in the yeast Pichia pastoris.

In this paper we report the development of a recombinant strain of the yeast Pichia pastoris, which secretes an anti-carcinoembryonic antigen single chain Fv (scFv) antibody fragment to the culture supernatant as a biologically active protein, at levels of 1.2 g l(-1). The yeast scFv was purified by IMAC, with a final yield of approximately 0.440 g of 93% pure scFv per liter of culture supernatant. The specific activity in ELISA of the yeast scFv was almost three times higher than that of a bacterial periplasmic counterpart. These results reaffirm that the yeast P. pastoris is a suitable host for high level production of scFv antibody fragments with potential in vivo diagnostic and therapeutic applications.

Adenosine Triphosphatases↗

High-speed chiral separations on microchip electrophoresis devices.

High-speed electrophoretic chiral separations have been successfully performed in a microfabricated device by employing cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC). Utilizing short separation channels and relatively high field strengths in combination with small volume-defined injection plugs, and operating in counter-electroosmotic flow conditions, fast and efficient separations of fluorescein insothiocyanate (FITC)-labeled amino acid enantiomers were obtained. Analysis time ranged from 75 s for the most basic amino acids to 160 s for the most acidic ones with associated efficiencies from 7000 up to 28 000 effective plates (100 000 to 395 000 plates/m). Buffer parameters were varied in order to study the effect on chiral resolution. A buffer system consisting of 100 mM borate (pH 9.4), 30 mM of SDS, and 10 mM gamma-CD as chiral selector provided adequate resolution of the majority of FITC-amino acid enantiomers tested.

Animals↗

Microchannel electrophoretic separation of biogenic amines by micellar electrokinetic chromatography.

Fast, efficient separation of most common biogenic amines was successfully performed on a glass microchip capillary electrophoresis device. The amines putrescine, histamine, tyramine, cadaverine, phenethylamine, tryptamine, spermidine and spermine were derivatized prior to fluorescence detection with fluorescein isothiocyanate. Separation was carried out using a channel length of 28 mm, a cross section of 50 x 8 microm, and a field strength of 600 V/cm. After optimization of buffer electrolyte conditions (120 mM boric acid, pH 9.4, modified with 40 mM SDS), fluorescein thiocarbamyl amine derivatives were successfully resolved. Analysis time was as short as 75 s. Determination of the biogenic amines was achieved in soy sauce samples.

Biogenic Amines↗

Ion-pair solid-phase extraction of biogenic amines before micellar electrokinetic chromatography with laser-induced fluorescence detection of their fluorescein thiocarbamyl derivatives.

Extraction conditions by ion-pair solid-phase extraction of biogenic amines from soy sauce samples were studied. The type of surfactant used as ion-pair reagent, the mode of extraction and the pH were factors studied to enhance the extraction efficiency. High recoveries were obtained using dodecylbenzenesulfonic acid as ion-pair reagent. Octanesulfonate and decanesulfonate did not provide satisfactory recoveries. A dynamic ion-pair solid-phase extraction method was also tested for comparison. The extracted amines were derivatized with fluorescein isothiocyanate, separated by micellar electrokinetic chromatography and detected by laser-induced fluorescence (MEKC-LIF). Because of sample clean-up after extraction and the very high number of theoretical plates obtained with the MEKC-LIF method, identification and quantification of biogenic amines in soy sauce samples could be accomplished. Biogenic amines in soy sauce samples could be separated and quantified with detection limits of the order of 0.1 microg/mL, depending on the sample source.

Biogenic Amines↗

Coexistence of primary hyperparathyroidism and thyroid disease.

Primary hyperparathyroidism (HPT) and thyroid disease are both relatively common diseases, which can coexist in the same patient. However, mere coincidence does not seem to be a satisfactory explanation for this association. Some factors like prior exposure to radiation may play a role in the association. The aim of this study was to determine the frequency of thyroid disease associated with HPT in 54 consecutive patients who underwent parathyroidectomy in our center from January 1990 to December 1997. Twenty-eight (52%) patients had thyroid disease associated to HPT. The patients had a mean age of 61+/-13 yr; they were predominantly postmenopausal women. Thyroid disease was detected preoperatively in 20 (71%) patients and during the surgical procedure in 8 (29%). Two patients had previous radiation exposure; one of them with papillary carcinoma of the thyroid. Two patients had hypothyroidism. UItrasonography was performed in the majority of patients in their preoperative evaluation. A multinodular goiter was seen to be the most frequent finding (76%). In addition to parathyroidectomy, 23 (82%) patients were also thyroidectomized. In conclusion, patients with HPT showed a high prevalence of thyroid disease, especially in postmenopausal women. Unsuspected thyroid lesions were found with sufficient frequency to warrant careful preoperative and intraoperative evaluation of both glands, in order to obviate reoperation. In experienced hands, combined surgery can be safely performed. Cervical ultrasonography is useful in the preoperative detection of nodular thyroid disease in these patients.

Adolescent↗

Effects of physical exercise in pre-and postmenopausal women on lipid peroxidation and antioxidant systems.

The effects of moderate physical activity were studied in 19 postmenopausal and 25 premenopausal women on the enzymatic (superoxide dismutase and catalase) and non-enzymatic (vitamins A and E) antioxidant systems and on the processes of lipid peroxidation (malondialdehyde), and to see whether there is any variation in oxidative stress indicators between the postmenopausal and non-menopausal women. The subjects were also studied anthropometrically. At the end of a training period, there was a decrease in body fat percentage and in various skin folds. Neither group presented a rise in lipid peroxidation levels, although there was a significant (p<0.05) rise in superoxide dismutase activity in the postmenopausal women.

Adult↗

Use of the C3H/He Lyme disease mouse model for the recovery of a Spanish isolate of Borrelia garinii from erythema migrans lesions.

A skin biopsy from a patient with erythema migrans was inoculated into C3H/He mice and into culture medium. A Borrelia garinii strain named Rio1 was isolated from both a direct BSK medium culture and a mouse ear-punch biopsy culture. Inoculating human tissue into mice produced a disease resulting in severe inflammation of the left tibio-tarsal joint, development of perivascular infiltrates as seen in ear-punch biopsies and the spread of spirochaetes along the skin, far from the inoculation site. The isolation of this strain confirms the circulation of this Borrelia species in Spain as a human pathogen, as well as its arthrogenicity in an animal model. The method used to recover strain Rio1 from human tissue is described as rapid and sensitive compared to direct inoculation of tissue into BSK medium.

Animals↗