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Biomedical subjects

F J López

Publications and source records attributed to F J López.

At least 19 recordsLinked to original sources
PubMed2006

Comparison of two quantitative GC-MS methods for analysis of tomato aroma based on purge-and-trap and on solid-phase microextraction.

Two analytical procedures, one based on purge-and-trap and the other on solid phase microextraction, both followed by GC-MS measurement using an ion-trap mass spectrometer in the electron impact mode, have been developed for determination and quantitation of up to 39 aroma compounds in fresh tomatoes. The method based on purge-and-trap for isolation of the volatile compounds uses Tenax as adsorbent and a hexane-diethyl ether mixture as solvent for elution. The method was validated for linearity, precision (better than 20% for most compounds), and limit of detection, which was approximately 1 ng g(-1). This method enabled identification of up to 30 compounds in real samples. Use of SPME was considered as an alternative, to simplify sample treatment while maintaining the information level for the samples (e.g. the number of compounds detected) and quality of quantitation. A procedure based on SPME using a Carboxen/polydimethylsiloxane fibre was developed and validated for determination of 29 aroma compounds; precision was better than 20% and limits of detection ranged from 4 to 30 ng g(-1).

Gas Chromatography-Mass Spectrometry↗
PubMed2006

Effects of polyphenolic antioxidants on exercise-induced oxidative stress.

Polyphenols are of increasing interest to consumers and food manufacturers for several reasons. Commonly referred to as antioxidants (they are the most abundant antioxidants in our diets), they may prevent various oxidative stress-related diseases, such as cancer, cardiovascular disease, inflammation and others. Physical activity is known to induce oxidative stress in individuals after intensive exercise. In this study, the effect of the flavonoid contents (which are the most abundant polyphenols) was investigated, as the only antioxidant in a replacement drink designed for sportsmen on various oxidative stress biomarkers after two identical trials of sub-maximal aerobic exercise, in a group of 30 sportsmen. In one of the trials, the cyclists consumed the antioxidant supplement (with 2.3g polyphenols/trial), and in another they consumed a placebo. Blood samples were collected both at rest and after exercise immediately and 45 minutes (min) later, for measurements of plasmatic indices of oxidative stress: lipid oxidation (TBARS), total antioxidant status (TAS); protein oxidation (carbonyl groups, CO) and the lactate dehydrogenase (LDH) and creatine kinase (CK) enzymes for each trial. All values were adjusted for changes in plasma volume. No changes were detected in plasma TAS and LDH after exercise or after the polyphenolic supplement. CK and TBARS increased after exercise in both tests. However, in response to strenuous exercise, the polyphenol-supplemented test showed a smaller increase in plasma TBARS and CK than the placebo test. CO increased by 12% in response to the placebo test, whereas it decreased by 23% in the polyphenol-supplement test. This may indicate that the antioxidant supplement offered protection against exercise-induced oxidative stress.

Adult↗
PubMed2006

Residue levels of captan and trichlorfon in field-treated kaki fruits, individual versus composite samples, and after household processing.

The dissipation of residue levels of captan and trichlorfon in field-treated kaki crops was studied according to good laboratory practices to propose maximum residue limits (MRLs). Residue levels of captan and trichlorfon were analysed by GC/MS and LC-MS/MS, respectively. Residue levels of captan and trichlorfon permitted one to propose MRLs in kaki of 3 and 5 mg kg(-1), respectively. The behaviour of these residues was also studied after peeling and cooking, and in individual fruits versus composite samples. Residue levels of these compounds for individual fruits suggested that a variability factor up to three could be set for the acute risk assessment. Levels of captan decreased by more than 90% after peeling and completely after cooking. Trichlorfon penetrates into the flesh in a proportion of 70% of the residue at the pre-harvest interval. Cooking resulted in a decrease of 27% of residue levels of trichlorfon.

Captan↗
PubMed2005

The effects of an antioxidant-supplemented beverage on exercise-induced oxidative stress: results from a placebo-controlled double-blind study in cyclists.

The objective of this study was to test the effects of an antioxidant-supplemented beverage on exercise-induced oxidative stress in moderately trained cyclists. A double-blind study was conducted in moderately trained cyclists. They were randomly allocated to receive either an antioxidant (13 subjects) or a placebo (13 subjects) 15 min pre-exercise (30 ml.kg(-1)) and during a 90 min constant-load test (30 ml.kg(-1).15 min(-1)) on a bicycle ergometer at 70% VO2max The supplemented beverage contained black grape (81 g/l), raspberry (93 g/l) and red currant (39 g/l) concentrates, and its total antioxidant capacity, measured by the ABTS(.+) technique, was 0.41 mM Trolox. No significant difference from basal to post-exercise period was detected for plasma TBARS in either the placebo group or the group receiving the antioxidant supplemented beverage. Post-exercise carbonyls decreased by 29% in the group receiving antioxidants, and the pattern of change was significantly different between antioxidant and placebo conditions. The urinary excretion of 8-OHdG increased significantly by 21% in the placebo group. Again differences in the pre- to post-exercise change were significant between both conditions These results suggest that in moderately trained cyclists, antioxidant supplementation counters oxidative stress induced by a 90 min exercise at 70% VO2max.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed2004

Determination of tridemorph and other fungicide residues in fruit samples by liquid chromatography-electrospray tandem mass spectrometry.

A rapid and sensitive liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) method for the determination of tridemorph and other pre- and post-harvest fungicides (carbendazim, thiabendazole, imazalil, propiconazole and bitertanol) in banana and orange samples has been developed and validated. The sample preparation was a simple extraction step with acetone using a high-speed blender prior to the injection of the five-fold diluted extract into the LC system with no other previous sample pre-treatment. Quantification was carried out using a matrix matched calibration curve which was linear in the range of 1-100 ng ml(-1) for all the compounds. The limit of quantification was 0.05 mg kg(-1) for all studied compounds, whereas limits of detection ranged between 0.005 and 0.025 mg kg(-1) (0.01 mg kg(-1) for tridemorph). Recoveries for tridemorph from spiked banana and orange samples at 0.05 and 1 mg kg(-1) were satisfactory, with values between 83 and 99% and relative standard deviations (R.S.D.s) lower than 13% (n = 5). For the other fungicides, recoveries between 75 and 95% with R.S.D.s lower than 12% were obtained. The developed method has been applied to the determination of selected fungicides in real samples of bananas and oranges from different origin. Thiabendazole and imazalil have been detected in almost all orange samples analyzed, and in around of 30% of banana samples. Bitertanol residues exceeded the maximum residue level (0.05 mg kg(-1)) in three banana samples while tridemorph was only detected in one sample.

Calibration↗
PubMed2003

Application of solid-phase microextraction for the determination of pyrethroid residues in vegetable samples by GC-MS.

A solid-phase microextraction (SPME) method has been developed for the determination of 7 pyrethroid insecticides (bifenthrin, lambda-cyhalothrin, permethrin, cyfluthrin, cypermethrin, fenvalerate, and tau-fluvalinate) in water, vegetable (tomato), and fruit (strawberry) samples, based on direct immersion mode and subsequent desorption into the injection port of a GC/MS. The SPME procedure showed linear behavior in the range tested (0.5-50 microg L(-1) in water and 0.01-0.1 mg kg(-1) in tomato) with r(2) values ranging between 0.97 and 0.99. For water samples limits of detection ranged between 0.1 and 2 microg L(-1 )with relative standard deviations lower than 20%. Detection limits for tomato samples were between 0.003 and 0.025 mg kg(-1) with relative standard deviations around 25%. Finally, the SPME procedure has been applied to vegetable (tomato) and fruit (strawberry) samples obtained from an experimental plot treated with lambda-cyhalothrin, and in both cases the analyte was detected and quantified using a calibration curve prepared using blank matrix. SPME has been shown to be a simple extraction technique which has a number of advantages such as solvent-free extraction, simplicity, and compatibility with chromatographic analytical systems. Difficulties with the correct quantification in a complex matrix are also discussed.

Food Analysis↗
PubMed2003

Rapid multiresidue determination of organochlorine and organophosphorus compounds in human serum by solid-phase extraction and gas chromatography coupled to tandem mass spectrometry.

A rapid analytical method for the multiresidue determination of several organochlorine and organophosphorus pesticides and polychlorinated biphenyls in human serum samples has been developed. Analytes were isolated by solid-phase extraction using C18 cartridges with subsequent analysis by GC-MS/MS using a glass liner packed with CarboFrit in the GC injection port. Labelled surrogate internal standards (fenitrothion D6, HCB (13)C6, p, p'-DDE D8 and PCB 138 (13)C12) were added to the samples before the extraction and were used for quantitation and for quality control in the analysis of real-world samples. Accuracy and precision were evaluated by using serum samples fortified at two concentration levels for the three families of compounds, with satisfactory results in the majority of cases. The high selectivity and sensitivity of GC-MS/MS allowed low detection limits of 0.05-0.5 ng mL(-1) for most of the analytes investigated. The developed procedure improves other current methodologies for the analysis of pesticides and PCBs in biological fluids, especially as regards to analysis time and simplicity of sample treatment. The method was applied to several serum samples obtained from farmers devoted to citrus crop production. Chlorpyrifos, HCB, p, p'-DDE and the higher chlorinated PCBs (153, 138 and 180) were the most frequently detected compounds.

Gas Chromatography-Mass Spectrometry↗
PubMed2003

Estradiol prevents amyloid-beta peptide-induced cell death in a cholinergic cell line via modulation of a classical estrogen receptor.

The pathology of Alzheimer's disease includes amyloid-beta peptide aggregation that contributes to degeneration of cholinergic neurons. Even though the underlying molecular mechanisms remain unclear, recent in vitro evidence supports a protective role for estrogens against several neurotoxic agents. Here we report that, in a murine cholinergic cell line (SN56), the massive cell death induced by 1-40 fragment of amyloid-beta peptide was prevented by 17beta-estradiol through a mechanism that may involve estrogen receptor activation. The protective effect of estradiol was observed in a dose-dependent manner, and was completely blocked by the pure antiestrogen ICI 182,780. In contrast, the inactive isomer 17alpha-estradiol consistently showed weaker neuroprotection than the native hormone that was unaffected by ICI 182,780 treatment. In addition, equivalent concentrations of 17beta-estradiol enhanced luciferase activity in cells transfected with a luciferase reporter gene driven by tandem estrogen response elements. Estrogen-induced luciferase activity was blocked by ICI 182,780, indicating estrogen receptor-dependent transcriptional activity. We also observed by reverse transcription-polymerase chain reaction, Western blot and immunocytochemistry that increasing concentrations of 17beta-estradiol enhanced the expression of estrogen receptor alpha mRNA and protein during amyloid-beta-induced toxicity. Under these conditions, it was found by confocal microscopy that the localization of estrogen receptor alpha in the absence of hormone was mainly extranuclear. However, the receptor was consistently observed also at the nuclear region after estrogen exposure. Overall, these data suggest that estrogen may exert neuroprotective effects against amyloid-beta-induced toxicity by activation of estrogen receptor-mediated pathways. In addition, intracellular estrogen receptors are up-regulated by their cognate hormone even during exposure to neurotoxic agents.

Acetylcholine↗
PubMed2003

Effect of treatment with the selective oestrogen receptor modulator LY117018-HCl on pituitary sensitivity to GnRH and subsequent ovulation.

The effects of LY117018-HCl (LY; a benzothiophene similar to raloxifene) were examined on various reproductive parameters in female rats. Four-day cyclic rats were treated (10:00 h on dioestrus) with LY (0.01, 0.1, 0.5, 1, 2, 4 or 16 mg kg(-1) p.o.) and assessed for ovulation at oestrus. LY inhibited ovulation at doses as low as 0.5 mg kg(-1), and ovulation did not occur at doses of 4 and 16 mg kg(-1). LY (16 mg kg(-1)) reduced wet uterine mass and LH concentrations at the time of the expected ovulatory surge. Ovulation induced by hCG in pentobarbital-treated rats was not altered by LY treatment, indicating normal ovarian sensitivity to gonadotrophins. LY, however, completely blocked the effects of oestradiol (under either negative or positive feedback modes) on LH secretion in ovariectomized rats. GnRH secretion into hypophyseal portal blood during pro-oestrus was not affected by treatment with LY, whereas the concentrations of serum LH remained reduced. Finally, treatment with LY markedly reduced pituitary sensitivity to GnRH during pro-oestrus, as it completely blocked GnRH-induced LH secretion. These results demonstrate that LY inhibits oestradiol action in the uterus and prevents ovulation in normal cyclic rats. LY-induced inhibition of ovulation is not caused by an alteration of the ovarian response to gonadotrophins or an impairment of GnRH secretion at the hypothalamus, but by a reduction in the sensitivity of gonadotrophs to the stimulatory effects of GnRH during pro-oestrus.

Animals↗
PubMed2002

Headspace solid-phase microextraction in combination with gas chromatography and tandem mass spectrometry for the determination of organochlorine and organophosphorus pesticides in whole numan blood.

A method for the determination of several organochlorine and organophosphorus pesticides in human whole blood samples was developed. The combination of solid-phase microextraction in headspace mode with gas chromatography with tandem mass spectrometry allowed the determination of 11 selected pesticides at ppb levels, minimizing the sample treatment. Quantitation was carried out by means of calibration curves prepared in blood using labelled surrogate/internal standards. The method showed good linearity between 1 and 50 ng ml(-1) (0.5-25 ng ml(-1) for HCB) using second-order calibration curves. Precision was found to be better than 20% at the three concentration levels assayed in the range of ng ml(-1). The detection limits obtained were in the range 0.02-0.7 ng ml(-1), except for p,p'-DDT (3 ng ml(-1)). The developed procedure was applied to blood and serum samples obtained from agricultural workers. HCB. beta-HCH and p,p'-DDE were most frequently detected in the samples analyzed.

Gas Chromatography-Mass Spectrometry↗
PubMed2002

Bioaccumulation of chlorpyrifos through an experimental food chain: study of protein HSP70 as biomarker of sublethal stress in fish.

The accumulation and transfer of the organophosphorus pesticide chlorpyrifos has been studied in an experimental aquatic two-level food chain using two species of the crustacean Artemia (A. franciscana and A. parthenogenetica) and the small fish Aphanius iberus. Artemia adults contaminated by exposure to the pesticide in water were used as live prey for Aphanius, the next trophic level. During the experimental bioaccumulation phase, fish were fed chlorpyrifos-contaminated Artemia pools with concentrations between 6.5 and 14.5 ng/g fresh weight for 32 days. Both concentrations accumulated in fish, and biomagnification factor (BMF) values showed a continuous decrease during the bioaccumulation phase, probably due to the physicochemical characteristics of the organophosphorus pesticide, to the biotransformation ability of fish and to the progressive adaptation of fish metabolism to toxic exposure. The first day that fish were fed uncontaminated preys, the pesticide accumulated via food was rapidly eliminated and was not detected. The effect of chlorpyrifos exposure through the food chain on stress protein (HSP70) synthesis was measured as a general biochemical response of stress in the fish (A. iberus). The levels of HSP70 were significantly higher in fish fed on contaminated Artemia than in the control fish fed on uncontaminated Artemia. Results showed that the HSP70 induction in fish could be associated to exposure of chlorpyrifos via food.

Animals↗
PubMed2001

Multielemental determination of arsenic, selenium and chromium(VI) species in water by high-performance liquid chromatography-inductively coupled plasma mass spectrometry.

A new method for the simultaneous chromatographic separation and determination of arsenite, arsenate, mono-methylarsonic acid, dimethylarsinic acid, selenite, selenate and hexavalent chromium in water is presented. Speciation was achieved by on-line coupling of anion-exchange LC and inductively coupled plasma mass spectrometry (ICP-MS). Optimisation of the chromatographic conditions led to baseline separation of the seven species in 14 min using gradient elution with NH4NO3 20 mM, pH 8.7-NH4NO3 60 mM, pH 8.7 as mobile phase. Detection limits are in the range 40-60 ng l(-1) for arsenic species, around 130 ng l(-1) for Cr(VI), and higher for Se(IV) and Se(VI) (1.2 and 1.4 microg l(-1) respectively). The method showed good accuracy and repeatability, and no interference of chloride on 75As, 77Se or 53Cr was observed. The developed method was applied to the analysis of several environmental surface water samples.

Arsenic↗
PubMed2001

Simultaneous determination of arsenic species and chromium(VI) by high-performance liquid chromatography-inductively coupled plasma-mass spectrometry.

The simultaneous determination of As(III), As(V), monomethylarsenic acid (MMA), dimethylarsinic acid (DMA) and Cr(VI) in fresh water has been carried out by coupling an anion-exchange column to an inductively coupled plasma-mass spectrometer. Optimisation of chromatographic conditions led to baseline separation of signals from the five species in approximately 9 min using gradient elution. Detection limits were 0.02-0.05 microg As l(-1) and 5.5 microg Cr l(-1). Repeatability was 2-3% for arsenic species and higher, i.e., 8%, for Cr(VI) due to the higher background for this species. Arsenic species and hexavalent chromium stability in surface water samples was evaluated, and storage conditions were set to 1 day at 4 degrees C in polyethylene flasks (without acidification) in order to avoid As(III)-As(V) conversions. The method was applied to the analysis of surface water.

Arsenic↗
PubMed2001

Multiresidue determination of organophosphorus and organochlorine pesticides in human biological fluids by capillary gas chromatography.

Two multiresidue analytical methods for the simultaneous determination of organophosphorus and organochlorine pesticides in human urine and serum samples are described. The first approach is based on liquid-liquid microextraction with dichloromethane, and the second uses solid-phase extraction with C18. In both methods, the extracts are analyzed by capillary gas chromatography using nitrogen-phosphorus detection (NPD) and electron-capture detection (ECD). Limits of detection of the overall procedure of analysis are at the low ng mL(-1) level. Stability experiments have been performed with spiked urine and serum samples stored at 4 degrees C for 1 month. Finally, the solid-phase extraction procedure was applied to real-world samples. Quantification was performed by NPD or ECD, and peak identity was confirmed by use of mass-selective detection (MSD).

Chromatography, Gas↗
PubMed2001

Androgen responsiveness of the pituitary gonadotrope cell line LbetaT2.

Androgens have a profound effect on the hypothalamic-pituitary axis by reducing the synthesis and release of the pituitary gonadotropin LH. The effect on LH is partly a consequence of a direct, steroid-dependent action on pituitary function. Although androgen action has been well studied in vivo, in vitro cell models of androgen action on pituitary gonadotropes have been scarce. Recently, an LH-expressing cell line, LbetaT2, was generated by tumorigenesis targeted to the LH-producing cells of the mouse pituitary. The purpose of these studies was to determine the presence of androgen receptor (AR) and establish its function in this cell line. RT-PCR analysis indicated that the LbetaT2 cell line expresses AR mRNA. Transient transfection assays, using the mouse mammary tumor virus (MMTV) promoter, showed that a functional AR is also present. Testosterone (TEST), dihydrotestosterone (DHT), 7alpha-methyl-19-nortestosterone (MENT), and fluoxymesterone (FLUOXY) increased reporter gene activity in the rank order of potencies MENT>DHT> TEST>FLUOXY. Additionally, activation of MMTV promoter activity by DHT in LbetaT2 cells was diminished by the AR antagonists casodex and 2-hydroxy-flutamide, indicating that the effects of DHT are mediated through AR. In summary, these studies showed that the LbetaT2 cell line is a useful model for the evaluation and molecular characterization of androgen action in pituitary gonadotropes.

Androgen Receptor Antagonists↗
PubMed2000

Solid-phase microextraction in pesticide residue analysis.

The applications of solid-phase microextraction (SPME) for sample preparation in pesticide residue analysis are reviewed in this paper taking into account the different approaches of this technique coupled mainly to gas chromatography but also to high-performance liquid chromatography. A complete revision of the existing literature has been made considering the different applications divided according to the pesticide families (organochlorine, organophosphorus, triazines, thiocarbamates, substituted uracils, urea derivatives and dinitroanilines among others) and the sample matrices analysed which included environmental samples (water and soil), food samples and biological fluids. Details on the analytical characteristics of the procedures described in the reviewed papers are given, and new trends in the applications of SPME in this field are discussed.

Chromatography, Gas↗
PubMed2000

Effects of added caffeine on results following artificial insemination with fresh and refrigerated rabbit semen.

Lactating rabbits (n=1335) were artificially inseminated to study the effect of the addition of caffeine on rabbit semen stored for up to 96 h. Concentration of 0-5 mM/l were tested. Whereas a concentration of 0.2 mM/l increased spermatozoa motility, higher concentration values adversely affected reproductive parameters. Spermatozoa stored at 18 degrees C for 72-96 h did not have the capacity to react with caffeine when it was added before insemination. Caffeine did not enhance fertility or prolificacy, regardless of its ability to increase sperm motility.

Animals↗