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Biomedical subjects

E Ortega

Publications and source records attributed to E Ortega.

At least 37 records · Page 2Linked to original sources
PubMed2003

Alcohol intoxication increases allopregnanolone levels in male adolescent humans.

RATIONALE: Teenage drinking is a cause of growing concern in industrialized countries, where almost 35% of alcohol drinkers are under 16 years old. Increased anxiety, irritability and depression among adolescents may induce them to seek the anxiolytic and rewarding properties of alcohol. Allopregnanolone is rewarding in rodents, and therefore may contribute to the effects of alcohol. OBJECTIVE: In this paper, we studied the effects of acute alcohol intoxication on the plasma levels of allopregnanolone in male adolescents. METHODS: Blood samples were drawn from male adolescents who arrived at the Emergency Department of the Hospital. Two groups were studied: one study group was formed by adolescents who arrived with evident behavioral symptoms of acute alcohol intoxication (AAI) and the other by those arriving for mild trauma (contusions, sprains) after no consumption of alcohol (Controls). RESULTS: Our results demonstrate that AAI significantly increases serum allopregnanolone levels in male adolescents. CONCLUSIONS: Because alcohol and allopregnanolone positively modulate gamma-aminobutyric acid type A (GABAA) receptors, allopregnanolone may play a major role in the anxiolytic and rewarding effects of alcohol, either directly or by influencing the sensitivity of GABAA receptors to alcohol.

Adolescent↗
PubMed2003

Alcohol intoxication increases allopregnanolone levels in female adolescent humans.

Teenage drinking is a cause of growing concern in industrialized countries, where almost 35% of alcohol drinkers are under 16 years old. Increased anxiety, irritability, and depression among adolescents may induce them to seek the anxiolytic and rewarding properties of alcohol. We studied the effects of acute alcohol intoxication (AAI) on the plasma levels of progesterone and allopregnanolone in female adolescents. Blood samples were drawn from female adolescents who arrived at the emergency department. One study group was formed by those who arrived with evident behavioral symptoms of AAI and the other by those arriving for mild trauma (contusions, sprains) after no consumption of alcohol (controls). Our results demonstrate that AAI significantly increases serum progesterone and allopregnanolone levels in both follicular and luteal phases of the ovarian cycle. Since alcohol and allopregnanolone positively modulate gamma-aminobutyric acid type A (GABA(A)) receptors, allopregnanolone may play a major role in the anxiolytic and rewarding effects of alcohol, either directly or by influencing the sensitivity of GABA(A)-receptors to alcohol.

Adolescent↗
PubMed2003

Enantioseparation of basic pharmaceutical compounds by capillary electrophoresis using sulfated cyclodextrins. Application to E-6006, a novel antidepressant.

In this study, a chiral capillary electrophoresis method was optimized and validated for E-6006, a thienylpyrazolylethanamine derivative (pKa 8.9). Enantioselectivity of neutral and anionic cyclodextrins (CDs) was evaluated at acid pH (3), obtaining cathodic and anodic migration, respectively. Hydroxypropyl-beta-CD, carboxymethyl-beta-CD and sulfobutyl ether-beta-CD led to similar and partial selectivity, whereas sulfate (S)-beta-CD produced baseline separation of the enantiomers. Four types of sulfated CDs were compared considering: cavity size (alpha, beta, gamma) and random substitution versus unique derivative (S-beta-CD, 6-heptakis-S-beta-CD). Complete peak separation was obtained in all cases, but with different affinity and binding strength. Some factors that play a role in the complex formation include: position/region/degree of substitution, size of CD cavity and proportion of derivatives in mixtures. Enantioaffinity and enantioselectivity increased with the average of sulfate groups/mol. Beta cavity size complexed better, although alpha and gamma cavities did not compromise separation. 6-Heptakis-S-beta-CD had less affinity and separation efficiency, attributed to its lower degree and unique position of substitution. The method was optimized with S-beta-CD (Aldrich, randomly substituted, 7-11 groups/mol). With this selector, the effect of pH value (3-9) was evaluated. Around pH 7 the cross-over point with change in the direction and order of migration was observed, associated with great enantioselectivity and long migration times. Fine tuning was done by adjusting the CD concentration and the buffer counterion. Definitive conditions were: uncoated silica capillary, 10 mM S-beta-CD-25 mM sodium phosphate, pH 3. Validation parameters are included.

Antidepressive Agents↗
PubMed2003

Inheritance of self-compatibility in almond: breeding strategies to assure self-compatibility in the progeny.

To assure self-compatibility in the progenies, three different crosses were conducted for the first time in an almond breeding programme: self-pollination (266 descendants from 30 families), crosses between parents sharing an S-allele (108 descendants from five families) and crosses with homozygous self-compatible parents (62 descendants from five families). Depending on the cross, self-compatibility in the progenies was determined by observing pollen tube growth (by means of fluorescence microscopy), stylar S-RNases analysis or allele-specific PCR. The results obtained fit with the accepted hypothesis of inheritance of self-compatibility and the three crossing strategies used ensured 100% of self-compatible descendants. These strategies increase the efficiency of the breeding programme and avoid the laborious task of evaluating this characteristic. From the breeding point of view, self-fertilisation and crosses between relatives tend to produce inbreeding. Furthermore, these methods reduce the possibilities of choosing the parental combination. The use of homozygous self-compatible parents does not have any of these disadvantages. As far as we know, this is the first time that allele-specific PCR has been used for early selection of self-compatible seedlings. The advantages and disadvantages of the three methodologies used to determine self-compatibility are discussed.

Crosses, Genetic↗
PubMed2003

DHEA, PREG and their sulphate derivatives on plasma and brain after CRH and ACTH administration.

The term neurosteroids applies to steroids that are synthesized in the nervous system, either de novo from cholesterol or from steroid hormone precursors. RIA was used to determine plasma and brain levels of the neurosteroids pregnenolone (PREG), dehydroepiandrosterone (DHEA), and their sulfate derivatives (PREG-S and DHEA-S) in male and female rats after administration of two typical stress hormones: corticotropin-releasing hormone (CRH) and adrenocorticotropin hormone (ACTH). In all cases, the parameters measured were detectable in plasma and brain. PREG, PREG-S, and DHEA increased significantly in plasma and brain after CRH and ACTH administration in males and females. Because neurosteroids play an important role in mammalian physiology, including that of humans, stress situations may alter the physiological functions regulated by these neurosteroids.

Adrenocorticotropic Hormone↗
PubMed2003

Effect of some abiotic factors on the biological activity of Gluconacetobacter diazotrophicus.

AIMS: The effect of some abiotic factors, dryness, heat and salinity on the growth and biological activity of Gluconacetobacter diazotrophicus, and the influence of a salt stress on some enzymes involved in carbon metabolism of these bacteria is studied under laboratory conditions. METHODS AND RESULTS: Strain PAL-5 of G. diazotrophicus was incubated under different conditions of drying, heat and salinity. Cells showed tolerance to heat treatments and salt concentrations, and sensitivity to drying conditions. Higher NaCl dosage of 150 and 200 mmol l -1 limited its growth and drastically affected the nitrogenase activity and the enzymes glucose dehydrogenase, alcohol dehydrogenase, fumarase, isocitrate dehydrogenase and malate dehydrogenase. CONCLUSIONS: Gluconacetobacter diazotrophicus, despite its endophytic nature, tolerated heat treatments and salinity stress, but its nitrogenase activity and carbon metabolism enzymes were affected by high NaCl dosage. SIGNIFICANCE AND IMPACT OF THE STUDY: The investigation of the biological activity of G. diazotrophicus in response to different abiotic factors led to more knowledge of this endophyte and may help to clarify pathways involved in its transmission into the host plant.

Acetobacter↗
PubMed2003

Differential regulation of steroid 5alpha-reductase isozymes expression by androgens in the adult rat brain.

The enzyme 5alpha-reductase (5alpha-R) is present in many mammalian tissues, including the brain. The physiological importance of 5alpha-R in the brain derives from its capability to convert testosterone (T) to a more potent androgen, dihydrotestosterone (DHT), and to convert progesterone and deoxycorticosterone (DOC) to their respective 5alpha-reduced derivatives, precursors of allopregnanolone and tetrahydroDOC, potent allosteric modulators of the gamma-aminobutyric acid receptor (GABA(A)-R). 5alpha-R occurs as two isoforms, 5alpha-R type 1 (5alpha-R1) and 5alpha-R type 2 (5alpha-R2). We studied the effects of T and DHT on the mRNA levels of both 5alpha-R isozymes in the prefrontal cortex of the adult rat, using an accurate and precise method that combines the high specificity of one-step quantitative RT-PCR with the sensitivity of capillary electrophoresis. Our results demonstrate that both isozymes of 5alpha-R are expressed in the cerebral cortex of adult rats. The gene expression of 5alpha-R type 2 is under the positive control of T and DHT. The gene that codes for 5alpha-R type 1 is not constitutive, because its expression is negatively regulated by T and DHT. These results open up a new research line that may lead to a better understanding of the role of 5alpha-R isozymes in the physiology of the central nervous system.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed2003

Phagocytosis of Candida albicans and superoxide anion Levels in ring dove (Streptopelia risoria) heterophils: effect of melatonin.

We observed in previous studies on avian heterophils that incubation with either physiological or pharmacological concentrations of the neurohormone melatonin increased the phagocytosis of inert particles (latex beads), and also provoked a decline in superoxide anion levels of those phagocytes. In the present study, we wanted to corroborate whether melatonin acts on the oxidative metabolism that accompanies the respiratory burst during phagocytosis by inducing a more effective phagocytic activity at the same time as exerting an antioxidant effect to eliminate and/or scavenge the free radicals left over after the destruction of the foreign material. To this end, we evaluated the ingestion and destruction of Candida albicans (live particles) by ring dove (Streptopelia risoria) heterophils after different times of incubation (30 and 60 min) with physiological concentrations of melatonin (50 pg/ml diurnal and 300 pg/ml nocturnal), as well as with a pharmacological concentration 23 x 106 pg/ml (100 micro m) of the hormone. In parallel, using the same times of incubation, we evaluated the oxidative metabolism by determining the superoxide anion levels (O2-.). The results show that melatonin, at all the times and concentrations studied, increases both the phagocytosis index (number of C. albicans phagocytosed by 100 heterophils) and the candidicide power (percentage of C. albicans killed of those ingested by 100 heterophils). The effect was dose-dependent. With respect to the oxidative metabolism accompanying the digestion and destruction, there was a decline in superoxide anion levels after incubation with all of the concentrations of the hormone studied. The effect was dose-dependent and most pronounced at 60 min. These results thus corroborate the proposal that melatonin enhances the phagocytic function at the same time as neutralizing the oxidative stress derived from this immune function.

Animals↗
PubMed2003

Association of MICA-A5.1 allele with susceptibility to celiac disease in a family study.

OBJECTIVE: The aim of this study was to analyze the role of the major histocompatibility complex class I chain-related gene A (MICA) transmembrane polymorphism in celiac disease (CD) susceptibility. METHODS: Sixty-one celiac Spanish families were genotyped for MICA transmembrane polymorphism by a polymerase chain reaction method combined with fluorescent technology. A transmission disequilibrium test was performed to investigate the preferential transmission of MICA alleles to the affected offspring. RESULTS: The MICA A5.1 allele was shown to be significantly transmitted to the affected siblings. This association was independent of the CD-predisposing DQ2 haplotype. Additionally, we classified our celiac families into typical and atypical groups as we found a significant association with MICA A5.1 in typical celiac families. There was also an association tendency with atypical families. CONCLUSIONS: Our data suggest that the MICA A5.1 allele is associated with CD development independently of DQ2-extended haplotype and clinical forms of CD.

Alleles↗
PubMed2003

A comparative analysis of the modification of sexual desire of users of oral hormonal contraceptives and intrauterine contraceptive devices.

OBJECTIVE: To compare the influence of oral hormonal contraceptives (OCs) and the use of intrauterine contraceptive devices (IUDs) on the modification of sexual desire. MATERIALS AND METHODS: A prospective observational study of 1073 women using OCs or an IUD at the Family Planning Center 'Marina Alta' in Alicante, Spain. In order to evaluate the relative risk regarding the decrease in libido attributed to each contraceptive method, a logistic regression analysis was undertaken which considered the factors of age adjustment, level of studies, family planning information, relationship with partner, age when sexual relationships were initiated, parity, contraceptive method previously used and the duration of use of the contraceptive method. RESULTS: No differences in the decrease of sexual desire were observed between the use of the OC and IUD (odds ratio (OR) 1.32; 95% confidence interval (CI) 0.70-2.49), yet differences were noted, however, in relation to age (OR 1.05; 95% CI 1.01-1.10). Although these differences were not statistically significant, a high level of awareness regarding family planning was shown to increase sexual desire when compared to a lower level of information on this subject (OR 0.64; 95% CI 0.41-1.01). Sexual desire was seen to decrease if the quality of the relationship with the partner was average (OR 2.24; 95% CI 1.36-3.69) or poor (OR 4.69; 95% CI 1.93-11.4). Nulliparous women showed a greater decrease in sexual desire in relation to women who had already given birth (OR 1.57; 95% CI 1.00-2.47). Sexual desire was greater if the contraceptive method had already been in use for 6-12 months (OR 0.41; 95% CI 0.17-0.98). CONCLUSIONS: Sexual desire does not vary in relation to the use of OCs or IUDs, yet it does decrease with age, in nulliparous women and in those with an average or poor relationship with their partner. Furthermore, sexual desire shows an increase between the first 6 and 12 months of contraceptive treatment.

Adolescent↗
PubMed2002

Phagocytosis of apoptotic cells assessed by flow cytometry using 7-Aminoactinomycin D.

BACKGROUND: Apoptotic cells are recognized specifically by macrophages and are cleared rapidly by phagocytosis. However, the recognition mechanisms involved in the clearance of apoptotic cells by macrophages are still not fully understood. Therefore, new methods must be designed to better our understanding of the mechanisms of interaction between macrophages and apoptotic cells. 7-Aminoactinomycin D (7-AAD) is a fluorescent DNA-binding stain usually used as a single agent to detect apoptotic cells by flow cytometry. We propose the use of 7-AAD-stained apoptotic cells as targets for a new flow cytometry phagocytosis assay. METHODS: Murine T-cell lymphoma YAC-1 cells were treated with etoposide to induce apoptosis. Etoposide-treated YAC-1 target cells were stained subsequently with 7-AAD and then coincubated with resident peritoneal macrophages to allow phagocytosis. The samples were analyzed by flow cytometry. Macrophages that had phagocytosed 7-AAD-stained apoptotic cells were identified by their bright red fluorescence and the resulting values were expressed as the percentage of cells. RESULTS: The phagocytic cells appeared as a distinct population characterized by bright fluorescence, which could not be detected in the negative controls. The effects of a phagocytic enhancer (interferon-gamma [IFN-gamma]) or inhibitor (incubation at 4 degrees C) were assessed accurately with this flow cytometric method. CONCLUSIONS: We describe the use of 7-AAD in an assay that is easy and quick to perform. This flow cytometric-based assay allows the quantification of phagocytosis of apoptotic cells by macrophages.

Animals↗
PubMed2002

Suppression of splenic macrophage Candida albicans phagocytosis following in vivo depletion of natural killer cells in immunocompetent BALB/c mice and T-cell-deficient nude mice.

The resistance of mice to systemic infections caused by Candida albicans is associated with activated splenic macrophages. In addition, there is a correlation between natural killer (NK) cell activation and the resistance to systemic candidiasis. The present study was designed to clarify the role of NK cells in the control of splenic macrophage C. albicans phagocytosis by either depleting NK cells (anti-asialo GM(1) treatment) or maintaining them in an activated state (tilorone treatment) in both immunocompetent BALB/c mice and T-cell-deficient nude mice. The results of the in vitro phagocytosis assays were analyzed by flow cytometry and demonstrate the pivotal role of NK cells in controlling the capacity of splenic macrophages to phagocytose C. albicans. In summary, these data provide evidence that the NK cells are the main inducers of phagocytic activity of splenic macrophages and that they mediate the protection against C. albicans systemic infection.

Animals↗
PubMed2002

Use of SYTOX green dye in the flow cytometric analysis of bacterial phagocytosis.

BACKGROUND: Fluorescein isothiocyanate (FITC) is used widely to label the targets used in flow cytometric phagocytosis assays. Unfortunately, the fluorescence intensity of phagocytosed FITC-labeled targets is influenced by changes in intracellular pH level, making quantitative measurements with this fluorophore problematic. We describe the use of SYTOX green nucleic acid stain to measure phagocytosis by flow cytometry. METHODS: Suspensions of isopropyl alcohol-permeabilized Escherichia coli DH5alpha were stained with the SYTOX green dye and then incubated with resident peritoneal macrophages. The samples were analyzed by flow cytometry and phagocytosis was determined by gating the cells. RESULTS: Results are expressed as percentage of phagocyte-associated green fluorescent cells. The validity of the method was shown by the effects of a phagocytosis inhibitor (incubation at 4 degrees C) or enhancer (gamma interferon [IFN- gamma] treatment) being accurately assessed with this assay. CONCLUSIONS: The method described was reproducible and provides an advantageous alternative to the use of FITC to label bacteria for the flow cytometric measurement of target uptake by phagocytic cells.

Escherichia coli↗
PubMed2002

Simultaneous separation of the enantiomers of cizolirtine and its degradation products by capillary electrophoresis.

The simultaneous enantioselective separation of (+/-)-cizolirtine and its impurities: (+/-)-N-desmethylcizolirtine, (+/-)-cizolirtine-N-oxide and (+/- )-5-(alpha-hydroxybenzyl)-1-methylpyrazole was investigated by capillary electrophoresis. Electrokinetic chromatography with carboxymethyl-beta-CD (CM-beta-CD) and sulfobutyl-ether-beta-CD was tried, showing good enantioseparation but poor chemical selectivity. The four racemic pairs were baseline separated, in a single run, by cyclodextrin-modified micellar electrokinetic chromatography. The migration buffer composition was: (60 mM hydroxypropyl-beta-cyclodextrin-150 mM sodium dodecyl sulfate-50 mM disodium tetraborate, pH 9.2, in water)-butanol 95:5, v/v). Work was done to determine the effect of buffer components and their optimal concentration on selectivity. The method was validated with respect to enantioselectivity of cizolirtine as well as its degradation products and separation selectivity between the different components. Linearity, limit of detection, limit of quantitation and precision were also determined. This method is suitable for the enantiomeric purity determination and stability control of cizolirtine (racemic mixture or enantiomers) and its degradation products. Examples of electropherograms of (R)-cizolirtine degraded under stressed conditions are shown.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2002

Physiological concentrations of melatonin and corticosterone in stress and their relationship with phagocytic activity.

The aim of the present work was to investigate the possible connection between melatonin, corticosterone and the nonspecific immune response under both basal and stressed conditions. The concentrations used in the present study were as determined in an earlier work, and corresponded to the minimum and maximum concentrations of the hormones melatonin and corticosterone over one circadian cycle in mice subjected to stress and in a basal group of mice. The concentrations were 15 pg/ml and 90 pg/ml of melatonin and 50 ng/ml and 200 ng/ml of corticosterone for incubations of the basal group macrophages, and 15 pg/ml and 70 pg/ml and 200 ng/ml and 500 ng/ml of melatonin and corticosterone, respectively, for incubations of macrophages from the group of stressed mice. The results indicate that corticosterone stimulates antigen ingestion at all physiological doses tested in the macrophages from the unstressed mice. The greatest phagocytic stimulation was obtained in macrophages from mice in a basal situation, and which had been incubated with the diurnal concentrations of the two hormones (maximum corticosterone 200 ng/ml; minimum melatonin 15 pg/ml). When the macrophages were from stressed mice, neither corticosterone nor melatonin led to any change in their phagocytic activity.

Adjuvants, Immunologic↗
PubMed2002

In-vitro study of the effect of adrenaline on the functional capacity of human neutrophils: role during exercise.

We studied the effect of adrenaline on the capacity of human neutrophils to attach, ingest and destroy Candida albicans. The neutrophils were incubated in vitro in the presence of an adrenaline concentration taken as basal (10-10 m), and another that is referred to as being reached following physical exercise (10-9 m). Two higher concentrations (10-7 m and 10-5 m), which are seldom attained in blood but which can occur at specific locations in the organism, were tested as having a possible pharmacological application. At the two high concentrations (10-7 m and 10-5 m), the capacity to attach, ingest and destroy C. albicans was greater than at the physiological concentrations (10-10 m and 10-9 m). Indeed, the capacity to attach and ingest C. albicans was significantly less after incubation with these physiological concentrations than the control values (incubation in the absence of adrenaline). Hence, high concentrations of adrenaline seem to enhance neutrophils' phagocytic capacity compared to physiological plasma concentrations, whereas small variations such as those caused by physical exercise have no effect on this functional capacity.

Adolescent↗