PubMed Health⌕ Search

Biomedical subjects

E Moyano

Publications and source records attributed to E Moyano.

At least 37 records · Page 2Linked to original sources
PubMed2000

On-line ion-pair solid-phase extraction-liquid chromatography-mass spectrometry for the analysis of quaternary ammonium herbicides.

An ion-pair on-line solid-phase extraction procedure using C8 extraction disks, suitable for liquid chromatography-mass spectrometry analysis is developed to determine quaternary ammonium herbicides (quats) in water samples. The separation of these compounds was performed using ion-pair chromatography with heptafluorobutyric acid (15 mM, pH 3.3) and acetonitrile gradient elution. Detection was carried out using a quadrupole mass spectrometer. Water sample volumes up to 50 ml can be preconcentrated with recoveries higher than 70%. Good precision and accuracy (day-to-day and run-to-run) were obtained and the detection limits ranged from 6 to 85 ng l(-1). The proposed on-line ion-pair solid-phase method enables compliance with European Community directives for drinking waters (100 ng l(-1)).

Chromatography, Liquid↗
PubMed2000

Determination of heterocyclic aromatic amines in meat extracts by liquid chromatography-ion-trap atmospheric pressure chemical ionization mass spectrometry.

When protein-rich foods are processed under normal cooking conditions, heterocyclic aromatic amines (HAAs) can be generated at a few parts per billion level. In this work, we have analyzed the HAAs present in a lyophilized meat extract by means of a simplified solid-phase extraction procedure. All the analytes were collected in a single extract with recoveries in the range of 45.6-75.2%, so the analysis time has been greatly reduced. Problems derived from the less exhaustive purification of the extract have been solved by using MS(ion trap) detection. The RSD for quantification ranged from 2.1% to 5.1% for run-to-run precision and from 5.2% to 11% for day-to-day precision. The limits of detection for standard solutions ranged from 20 to 150 pg injected. For the meat extract analyzed limits of detection from 0.9 to 11.2 ng g(-1) were obtained. Results of the quantification are in agreement with those obtained using different clean-up procedures.

Amines↗
PubMed2000

The potential utility of the Semi-Nested Multiplex PCR technique for the diagnosis and investigation of congenital malaria.

We report three cases of congenital malaria involving two malarial immune mothers living in Spain. Diagnostic PCR and Genotyping PCR for merozoite surface proteins 1 and 2 were essential to show that mothers and new-borns had different Plasmodium population parasites at the moment of the delivery, and that the infection was acquired earlier in gestation by transplacental transmission. In the first case the Plasmodium species founded in both, mother and child were different. Malaria in the twins showed a mixed infection (P. falciparum plus P. malariae) while the mother presented a P. falciparum infection. These facts were confirmed studying the polymorphisms for MSP1 and MSP2. Blood samples of the newborns were analyzed an half hour after delivery excluding the possibility of re-infection by mosquito bite and indicating a vertical transmission during pregnancy.

Adult↗
PubMed2000

[Self-training in needle-knife sphincterotomy].

AIM: To analyze our experience in self-training in needle-knife sphincterotomy and to estimate the number of procedures necessary to reach competence in this technique. METHOD: Retrospective analysis of the first 60 needle-knife sphincterotomies performed, paying attention to success rates, the need for one or two attempts and the incidence and severity of early complications. The results were analyzed and divided into 2-year periods. The training period was subsequently determined using a qualitative method which combines the three above-mentioned criteria and sets competence level at 90%. RESULTS: Of the 60 attempted needle-knife sphincterotomies (five in Billroth-II patients), 54 (90%) were completed. The procedure was required in 20% of all the sphincterotomies completed. Needle-knife sphincterotomy was completed at the first attempt in 39 (65%) patients and complications appeared in 13 (22%), three (5%) of which were serious but not fatal. Analysis of 2 year periods showed an almost 100% increase in the total number of needle-knife sphincterotomies completed and in those completed at the first attempt, as well as a reduction in the complication rate, which subsequently stabilized at below 15%. Forty needle-knife sphincterotomies were needed to reach competence level. CONCLUSION: Needle-knife sphincterotomy can be learned without training from an endoscopist with experience in this technique. Competence can be reached after 40 procedures. Nevertheless, we propose an alternative method to our own to achieve competence in less time and with fewer complications.

Clinical Competence↗
PubMed1999

Ion-pair liquid chromatography--atmospheric pressure ionization mass spectrometry for the determination of quaternary ammonium herbicides.

High-performance liquid chromatography coupled with atmospheric pressure ionization mass spectrometry (electrospray and atmospheric pressure chemical ionization) has been used to characterize some quaternary ammonium herbicides (quats). The separation of these compounds was carried out using ion-pair chromatography with heptafluorobutyric acid (15 mM, pH 3.3) and acetonitrile gradient elution for successful coupling to mass spectrometry. Detection limits down to 0.1-4 micrograms l-1 were obtained for spiked tap water following a preconcentration step. Good reproducibilities (day-to-day and run-to-run) were also obtained.

Atmospheric Pressure↗
PubMed1998

Two bZIP proteins from Antirrhinum flowers preferentially bind a hybrid C-box/G-box motif and help to define a new sub-family of bZIP transcription factors.

Two genes encoding bZIP proteins are expressed in flowers of Antirrhinum majus, predominantly in vascular tissues, carpels and anthers. The sequences of cDNA clones encoding these proteins show them to belong to a distinct sub-family of bZIP proteins which also includes LIP19 from rice and MLIPI5 and OBF1 from maize. The sub-family is characterized by the inclusion of very small proteins consisting of essentially a basic domain and a long leucine zipper. Members also have a conserved upstream open reading frame (uORF) in their 5' leader sequences, implying a common mode of post-transcriptional control. In vitro, the Antirrinum bZIP proteins preferentially bind to a novel hybrid C-box/G-box motif which is found in the promoters of some plant histone genes and of some nuclear-encoded genes with plastidial protein products. Expression of the bZIP proteins in transgenic tobacco under control of the CaMV 35S promoter supports the view that they can regulate expression of genes which contain the preferred target motif within their regulatory sequences, although both enhancement and repression of transcript levels of target genes were observed, indicating that the bZIP proteins probably interact with other factors to modulate transcription in different ways, as has been observed for the small MAF family of bZIP proteins in vertebrates.

Amino Acid Sequence↗
PubMed1998

A fruit-specific putative dihydroflavonol 4-reductase gene is differentially expressed in strawberry during the ripening process.

A cDNA clone encoding a putative dihydroflavonol 4-reductase gene has been isolated from a strawberry (Fragaria x ananassa cv Chandler) DNA subtractive library. Northern analysis showed that the corresponding gene is predominantly expressed in fruit, where it is first detected during elongation (green stages) and then declines and sharply increases when the initial fruit ripening events occur, at the time of initiation of anthocyanin accumulation. The transcript can be induced in unripe green fruit by removing the achenes, and this induction can be partially inhibited by treatment of de-achened fruit with naphthylacetic acid, indicating that the expression of this gene is under hormonal control. We propose that the putative dihydroflavonol 4-reductase gene in strawberry plays a main role in the biosynthesis of anthocyanin during color development at the late stages of fruit ripening; during the first stages the expression of this gene could be related to the accumulation of condensed tannins.

Alcohol Oxidoreductases↗
PubMed1997

Liquid chromatography-atmospheric pressure ionization mass spectrometry for the determination of chloro- and nitrophenolic compounds in tap water and sea water.

Liquid chromatography coupled to atmospheric-pressure ionization mass spectrometry (LC-API-MS) with negative ion detection was studied for the determination of a variety of phenolic compounds in environmental waters. An isocratic mobile phase of 0.05% acetic acid-acetonitrile (50:50, v/v) was used. The influence of post-column addition of different bases on the sensitivity of the detection in electrospray (ES) was studied. The [M-H]-ion was the base peak for all the compounds using both ES and atmospheric-pressure chemical-ionization (APCI) ion sources. Moreover, abundant structural information was obtained by increasing the extraction voltage. Detection limits for standard solutions ranging from 2 to 13 ng injected for LC-ES-MS and from 0.02 to 20 ng for LC-APCI-MS were obtained. Good reproducibilities (day-to-day and run-to-run) were observed. The optimum LC-ES-MS and LC-APCI-MS conditions thus determined were used for a quantitative analysis of some phenolic compounds in spiked tap water and sea water samples.

Chlorophenols↗
PubMed1997

Liquid chromatography-atmospheric-pressure chemical ionization mass spectrometry as a routine method for the analysis of mutagenic amines in beef extracts.

A liquid chromatography-mass spectrometry (LC-MS) method using atmospheric-pressure chemical ionisation as interface was developed for the simultaneous determination of 14 heterocyclic aromatic amines and related compounds in beef extracts. The separation was performed on a conventional C18 column using a binary mobile phase composed of acetonitrile and 50 mM ammonium acetate at pH 5.7, and elution was carried out in gradient mode. Several parameters influencing the mass spectra were optimized, and the effect of the variation of cone voltage on the mass spectra was studied. The [M+H]+ ions and some fragments produced in the source were observed in the mass spectra when several extraction voltages were applied. Quality parameters (run-to-run and day-to-day reproducibility, intervals of linearity, and limits of detection) were studied in the optimum working conditions. The method was used to analyze the heterocyclic amines present in a commercial beef extract. Therefore, a solid-phase extraction clean-up procedure was performed prior the LC-MS analysis due to the complexity of the sample and the compounds Glu-P-1, Harman, Norharman and A alpha C were identified in the samples at ppb levels and successfully confirmed using in-source fragmentation.

Amines↗
PubMed1997

Liquid chromatography-electrospray mass spectrometry with in-source fragmentation for the identification and quantification of fourteen mutagenic amines in beef extracts.

Conditions for the liquid chromatography-electrospray mass spectrometric (LC-ES-MS) determination of the fourteen most abundant heterocyclic amines and related compounds that can be produced in thermally processed foods were established. The simultaneous separation of all HAs on an octadecylsilane microbore column using a gradient elution with 5 mM ammonium acetate, pH 4.0, and acetonitrile was optimized. The mass spectra obtained at different extraction voltages were studied. The highest extraction voltages provided structural information for the characterization of these amines. Good detection limits comparable with those of LC and electrochemical detection were obtained. The method was applied to the analysis of these mutagens in beef extracts after a solid-phase extraction clean-up and in-source fragmentation had been applied for confirmation purposes. The levels of several heterocyclic amines measured were estimated to be in the range of 7.8-135.2 ng g-1.

Amines↗
PubMed1997

Cloning, molecular characterization and expression pattern of a strawberry ripening-specific cDNA with sequence homology to pectate lyase from higher plants.

A strawberry fruit cDNA showing sequence similarity to higher-plant pectate lyase genes has been isolated by differential screening of a strawberry fruit cDNA subtractive library. The cDNA contains a 396 amino acids open reading frame corresponding to a 44.8 kDa protein. The transcript is predominantly expressed in ripe fruits and was not detected at high levels in any other plant tissues. The removal of the achenes from unripe green fruits induced the expression of this putative pectate lyase gene. In common with other ripening related genes in strawberry, this induction was partially inhibited by treatment of de-achened fruit with the auxin NAA. Southern blot analysis of genomic DNA indicates that in strawberry there is more than one putative pectate lyase gene. We propose that the ripe fruit expression of this strawberry gene with similarity to pectate lyases could be related to cell wall pectin degradation contributing to strawberry fruit softening.

Amino Acid Sequence↗
PubMed1996

Determination of heterocyclic amines by pneumatically assisted electrospray liquid chromatography-mass spectrometry.

Electrospray ionization mass spectrometry was applied to the study of the amines IQ, Trp-P-1, Trp-P-2, PhIP and A alpha C and the co-mutagens harman and norharman. The results obtained on a triple quadrupole mass spectrometer equipped with a pneumatically assisted electrospray source are reported. The chromatographic conditions were optimized with a reversed-phase column (1 mm I.D.) using acetonitrile-5 mM ammonium acetate (pH 6.7) (50:50) as the mobile phase at a flow-rate of 50 microliters min-1. Different parameters influencing the mass spectra were investigated. For these compounds [M + H]+ in the positive-ion mode and also some fragments produced through collisionally activated decomposition in the interface were observed. Detection limits of 5.4-44 pg were obtained for standard solutions of these amines. Analysis of a meat extract was performed by HPLC-MS using single-ion monitoring after a solid-phase extraction clean-up.

Amines↗
PubMed1996

Apparent redundancy in myb gene function provides gearing for the control of flavonoid biosynthesis in antirrhinum flowers.

Two Myb-related transcription factors, Myb305 and Myb340, are expressed specifically in flowers of Antirrhinum. The proteins are structurally very similar throughout their DNA binding domains, implying that they bind to common target motifs. This binding has been demonstrated experimentally. Myb305 has been shown to activate the gene encoding the first enzyme of phenylpropanoid metabolism, phenylalanine ammonia-lyase. We show that Myb340 can also activate transcription from the phenylalanine ammonia-lyase gene promoter and that both transcription factors can activate two other genes involved in flavonoid metabolism, thereby linking early and later steps in plant secondary metabolism. Myb340 is a stronger activator than Myb305, but relatively more Myb305 than Myb340 protein is able to bind to target promoters when both proteins are synthesized in yeast or Escherichia coli, probably as a result of inhibition of Myb340 binding by phosphorylation. This means that Myb305 can compete with Myb340 to reduce its effective transcriptional activation when both transcription factors are expressed in the same cell. This competitive interaction has been demonstrated in plant cells. Expression patterns determined by in situ hybridization showed that the two transcription factors are expressed within the same cells of the flower and imply that the detailed specializations in function of these two apparently redundant transcription factors may be used to provide gears that adjust the rate of induction of secondary metabolism to floral development.

Binding Sites↗
PubMed1994

A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes.

Synthesis of flavonoid pigments in flowers requires the co-ordinated expression of genes encoding enzymes in th phenylpropanoid biosynthetic pathway. Some cis-elements involved in the transcriptional control of these genes have been defined. We report binding of petal-specific activities from tobacco and Antirrhinum majus (snapdragon) to an element conserved in promoters of phenylpropanoid biosynthetic genes and implicated in expression in flowers. These binding activities were inhibited by antibodies raised against Myb305, a flower-specific Myb protein previously cloned from Antirrhinum by sequence homology. Myb305 bound to the same element and formed a DNA-protein complex with the same mobility as the Antirrhinum petal protein in electrophoretic mobility shift experiments. Myb305 activated expression from its binding site in yeast and in tobacco protoplasts. In protoplasts, activation also required a G-box-like element, suggesting co-operation with other elements and factors. The results strongly suggest a role for Myb305-related proteins in the activation of phenylpropanoid biosynthetic genes in flowers. This is consistent with the genetically demonstrated role of plant Myb proteins in the regulation of genes involved in flavonoid synthesis.

Base Sequence↗
PubMed1992

Purification and properties of three NAD(P)+ isozymes of L-glutamate dehydrogenase of Chlamydomonas reinhardtii.

Three isozymes of glutamate dehydrogenase (GDH) of Chlamydomonas reinhardtii, induced under different trophic and stress conditions, have been purified about 800-1000-fold to electrophoretic homogeneity. They are hexamers of Mr 266,000-269,000 as deduced from gel filtration and sedimentation coefficient data. GDH1 consisted of six identical subunits of 44 kDa each, whereas both GDH2 and GDH3 consisted of six similar-sized monomers (4 of 44 kDa and 2 of 46 kDa). Optimum pH for the three activities with each pyridine nucleotide was identical (8.5 with NADH; 7.7 with NADPH; and 9.0 with NAD+). The isozymes exhibited similar high optimum temperature values (60-62 degrees C) and isoelectric points (7.9-8.1). Activity was enhanced in vitro by Ca2+ ions and strongly inhibited by pyridoxal 5'-phosphate, KCN, o-phenanthroline and EDTA, and to a lesser extent by pHMB and methylacetimidate. In the aminating reaction the three isozymes were inhibited in a concentration-dependent process by both NADH and NADPH, with apparent Km values for NH4+ ranging from 13-53 mM; 0.36-1.85 mM for 2-oxoglutarate and 0.07-0.78 mM for NADH and NADPH. In the deaminating reaction apparent Km values ranged from 0.64-3.52 mM for L-glutamate and 0.20-0.32 for NAD+. In addition, the three isozymes exhibited a non-hyperbolic kinetics for NAD+ with negative cooperativity (n = 0.8).

Animals↗
PubMed1992

Intracellular Localization of Three l-Glutamate Dehydrogenase Isozymes from Chlamydomonas reinhardtii.

The intracellular localization of the activity and synthesis of three isozymes of NAD(P)(+)-glutamate dehydrogenase from the unicellular green alga Chlamydomonas reinhardtii cw-92 has been established. Isozyme activities have been located within mitochondria by using differential centrifugation techniques and discontinuous Percoll gradient separations. Experiments with protein synthesis inhibitors cycloheximide, rifampicin, chloramphenicol, and actinomycin D, under dark and carbon starvation conditions, revealed that synthesis of the three isozymes was likely to occur in cytosol as precursor proteins that are then transported and processed inside the mitochondria.

Journal Article↗