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Biomedical subjects

J A Pascual

Publications and source records attributed to J A Pascual.

At least 19 recordsLinked to original sources

Effectiveness of municipal waste compost and its humic fraction in suppressing Pythium ultimum.

The effect of addition of a municipal solid waste (MSW) compost and its water-soluble and humic fraction to suppress the effect of Pythium ultimum on pea plants was studied and compared with that of a chemical pesticide (metalaxyl). The biotic and abiotic characteristics of compost involved in the biocontrol effects of these materials were also evaluated. The addition into soil of whole composts and their humic fractions reduced the effect of the pathogen on pea plants, significantly reducing the number of root lesions and Pythium populations and avoiding reductions of plant growth. The greatest pathogen suppression was achieved with the chemical pesticide. However, it also caused a significant decrease in the number of nontarget bacteria and fungi and on beneficial soil microorganisms such as Trichoderma and Pseudomonas. Addition of organic amendments increased population size of nontarget and specific biocontrol microorganisms. The humic fraction showed similar results to compost. All this suggests that metalaxyl has a nonspecific effect, producing adverse effects on aspects of soil quality. This was avoided if the chemical pesticide was reduced and replaced by organic amendments such as an MSW compost or its humic fraction.

Agriculture↗

Fast screening of anabolic steroids and other banned doping substances in human urine by gas chromatography/tandem mass spectrometry.

A fast and sensitive method for the comprehensive screening of anabolic agents and other banned doping substances using gas chromatography/tandem mass spectrometry (GC/MS/MS) with an external ionization ion trap mass spectrometer is presented. The method takes advantage of the resolving power of MS/MS to eliminate background interferences, thus speeding up the chromatographic analysis. For each compound, different fragmentation reactions were studied and their collision energies optimized to obtain the best sensitivity in terms of their signal-to-noise ratio (S/N). A dramatic reduction in overall analysis time was achieved compared with other common approaches. More than 50 substances could finally be monitored in less than 7.4 min with detection limits (S/N >3) lower than 0.5 ng ml(-1) for most of the compounds with special sensitivity requirements according to the International Olympic Committee (IOC). A validation procedure for qualitative analysis was performed. The selectivity of the method showed that no interfering peaks were observed at the retention time of the analytes. Good intermediate precision, below 25% for most of the compounds, and robustness were observed. The optimized method was successfully applied to analyse more than 100 real human urine samples with optimum sensitivity and specificity rates.

Anabolic Agents↗

13C/12C isotope ratio MS analysis of testosterone, in chemicals and pharmaceutical preparations.

The 13C/12C ratio can be used to detect testosterone misuse in sport because (semi)-synthetic testosterone is supposed to have a 13C abundance different from that of endogenous natural human testosterone. In this study, gas chromatography/combustion isotope ratio mass spectrometry (GC/C/IRMS) analysis for the measurement of the delta 13C/1000 value of testosterone from esterified forms of 13 pharmaceutical preparations, six reagent grade chemicals and three bulk materials (raw materials used in pharmaceutical proarations) obtained world-wide was investigated after applying a strong acidic solvolytic procedure. Mean delta 13C/1000 values of non esterified (free) testosterone from chemicals and bulk materials of several testosterone esters were in the range: -25.91/-32.82/1000 while the value obtained for a (semi)-synthetic, reagent grade, free testosterone was -27.36/1000. The delta 13C/1000 results obtained for testosterone from the pharmaceuticals investigated containing testosterone esters were quite homogeneous (mean and S.D. of delta 13C/1000 values of free testosterone: 27.43 +/- 0.76/1000), being the range between -26.18 and -30.04/1000. Values described above were clearly different from those reported by several authors for endogenous natural human testosterone and its main metabolites excreted into the urine in non-consumers of testosterone (delta 13C/1000 range: from -21.3 to -24.4/1000), while they were similar to those of urinary testosterone and metabolites from individuals treated with testosterone esters and testosterone precursors. This finding justifies the fact that administration of these pharmaceutical formulations led to a statistical decrease of carbon isotope ratio of urinary testosterone and its main metabolites in treated subjects.

Carbon↗

Effect of biocontrol strains of Trichoderma on plant growth, Pythium ultimum polulations, soil microbial communities and soil enzyme activities.

Five strains of Trichoderma with known biocontrol activities were assessed for their effect upon pea growth and their antagonistic activity against large Pythium ultimum inocula. The effect of Trichoderma inocula upon the indigenous soil microflora and soil enzyme activities in the presence and absence of Pythium is assessed. In the absence of Pythium, Trichoderma strain N47 significantly increased the wet shoot weight by 15% but did not significantly affect the dry weight, whilst strains T4 and N47 significantly increased the root weights by 22% and 80%) respectively. Strains TH1 and N47 resulted in significantly greater root lengths. Pythium inoculation significantly reduced the root length and the number of lateral roots and nodules, and significantly increased the root and rhizosphere soil fungal populations. Pythium inoculation significantly reduced the plant wet and dry shoot weights and significantly increased the wet and the dry shoot/root ratio. All the Trichoderma strains reduced the number of lesions caused by Pythium and increased the number of lateral roots. The effect of the Pythium on emergence and shoot growth was significantly reduced by all the Trichoderma strains except strain To10. Inoculation with Trichoderma strains TH1 and T4 resulted in significantly greater wet root weights (62% and 57%, respectively) in the presence of Pythium compared to the Pythium control. Strain N47 significantly increased the shoot/root ratio compared to the Pythium control. Inoculation with Trichoderma strains T4, T12 and N47 significantly reduced Pythium populations. Pythium increased the activity of C, N and P cycle enzymes, whilst four Trichoderma strains reduced this effect, indicating reduced plant damage and C leakage. Overall, strains T4 and N47 had the greatest beneficial characteristics, as both these strains improved plant growth in the absence of Pythium and reduced plant damage in the presence of Pythium. The dual properties of these strains improve the commercial application, giving them an advantage over single action inocula, especially in the absence of plant pathogens.

Acetylglucosaminidase↗

Fully automated analytical method for codeine quantification in human plasma using on-line solid-phase extraction and high-performance liquid chromatography with ultraviolet detection.

A simple, sensitive and fully automated analytical method for the analysis of codeine in human plasma is presented. Samples are added with oxycodone, used as internal standard (I.S.), and directly loaded in the autosampler tray. An on-line sample clean-up system based on solid-phase extraction (SPE) cartridges (Bond-Elut C2, 20 mg) and value switching (Prospekt) is used. Isocratic elution improved reproducibility and allowed the recirculation of the mobile phase. A Hypersil BDS C18, 3 microns, 10 x 0.46 cm column was used and detection was done by UV monitoring at 212 nm. Retention times of norcodeine (codeine metabolite), codeine and oxycodone (I.S.) were 5.5, 6.4 and 9.1 min, respectively. Morphine was left to elute in the chromatographic front. Detection limit for codeine was 0.5 microgram l-1 and inter-assay precision (expressed as relative standard deviation) and accuracy (expressed as relative error) measured at 2 micrograms l-1 were 5.03% and 1.82%. Calibration range was 2-140 micrograms l-1.

Automation↗

Quantitative determination of tricyclic antidepressants and their metabolites in plasma by solid-phase extraction (Bond-Elut TCA) and separation by capillary gas chromatography with nitrogen-phosphorous detection.

An analytical method for the simultaneous determination of amitriptyline, nortriptyline, imipramine, desipramine, clomipramine, and desmethylclomipramine in human plasma using promazine as internal standard is described. The method is based on a solid-phase extraction procedure using the Bond-Elut TCA columns followed by separation and detection using capillary gas chromatography with a specific nitrogen phosphorous detector (GC/NPD). Using the new extraction procedure, the problem of adsorption losses was overcome, and good recoveries were achieved for all compounds tested (>87%). Furthermore, clean extracts free of chromatographic interferences were obtained. Complete separation of underivatized, tricyclic antidepressant compounds was achieved in <11 minutes with reliable chromatographic performance. The limits of detection ranged from 1.2 to 5.8 microg/l. Calibration curves, showing good linearity, were prepared covering the therapeutic concentrations range expected (20 to 500 microg/l). The interassay precision values (RSD) ranged from 4.5% to 9.8%. It is concluded that extraction of amitriptyline, nortriptyline, imipramine, desipramine, clomipramine, and desmethylclomipramine, with Bond Elut TCA solid-phase columns followed by their detection using GC/NPD provides a sensitive and reproducible method that can be easily automated for immediate and routine analysis of clinical samples.

Amitriptyline↗

Rapid and sensitive determination of zidovudine and zidovudine glucuronide in human plasma by ion-pair high-performance liquid chromatography.

A rapid, simple and sensitive method is described for the simultaneous determination in human plasma of the well known antiviral agent zidovudine (AZT) and its main metabolite, zidovudine-5'-O-glucuronide (G-AZT), using a solid-phase extraction method for sample preparation and a rapid ion-pair HPLC separation method with diode-array ultraviolet detection. The method overcomes the problems experienced in other procedures because of the large difference in polarity of the two compounds and the pH-sensitive retention of G-AZT by using n-octylamine to increase the retention of the glucuronide and improve the overall chromatographic behaviour. AZT and G-AZT are eluted at 3.6 and 5 min, respectively, with 7-ethyltheophylline used as internal standard eluting at 4.2 min. Caffeine, a good marker for the elution of related compounds present in plasma, appears well before, at 2.6 min. Quantification limits were established at 0.025 and 0.050 micrograms/ml for AZT and G-AZT, respectively. The improvement in method reproducibility due to the late elution of G-AZT could be observed even at the quantification limit at which an inter-assay relative standard deviation of only 6.4% was found after 3 months of routine use of the method.

Antiviral Agents↗

Direct determination of anabolic steroids in human urine by on-line solid-phase extraction/liquid chromatography/mass spectrometry.

A fully automated method is presented for the determination of anabolic steroids in human urine. On line solid-phase extraction with automatic cartridge exchange is used for sample clean-up followed by an high-performance liquid chromatographic isocratic separation. A narrow-bore 5 microns reversed-phase column (250 x 2.1 mm i.d.) at a flow rate of 0.2 ml min-1 is connected to a particle beam interface linked to a single-quadrupole mass spectrometer. No derivatization is needed and normal electron impact mass spectra are obtained for a series of up to 28 steroids and metabolites. Urine samples collected following the ingestion of therapeutic doses of some of the steroids were analysed and the detection and confirmation of excreted compounds accomplished. A dramatic reduction in human intervention and overall analysis time was achieved compared with other common approaches.

Anabolic Agents↗

International cooperation in analytical chemistry: experience of antidoping control at the XI Pan American Games.

We describe the experience of the international cooperation carried out for antidoping control at the XI Pan American Games. A temporary accreditation was granted by the International Olympic Committee to the accredited laboratory of Barcelona (Spain) to set up a Doping Control Laboratory in Havana. Two other laboratories from Mexico and Cuba contributed personnel, materials, and instrumentation. The main issues associated with the preparation and organization of the project are described. During 16 days, 741 urine samples were tested for stimulants, narcotics, anabolic steroids, beta-blockers, diuretics, cannabinoids, local anesthetics, and human chorionic gonadotropin by gas-liquid chromatography, gas chromatography/mass spectrometry, high-performance liquid chromatography, and immunoassay techniques. Analytical methodologies, quality-control strategies, and the main results are reported.

Acebutolol↗

Variability of venous anatomy of rat testis: application to experimental testicular surgery.

The venous drainage of the testis of the laboratory rat was observed in 31 animals. The right testicular (internal spermatic) vein drained directly into the right common iliac vein in 77.4%, and into the inferior vena cava in 22.6% of the animals. The left testicular vein drained into the left common iliac vein in all animals, but in 90.3% there was also an accessory branch of the testicular vein draining into the left renal vein. These observations suggest that in the rat the exact anatomy of the venous drainage of each testis should be identified prior to undertaking any surgical procedure on the testis where the venous vasculature plays a major role such as testicular transplantation or the creation of an experimental varicocele.

Animals↗

Long-term effects of prepubertal testicular vessel ligation on testicular function in the rat.

To determine the effects of unilateral testicular vein and artery ligation in the immature rat on the function and final location of the testis at adulthood, 10-day-old male rats underwent either a sham operation or unilateral ligation of these vessels of the still undescended testis. Testicular location, blood flow, size and histology as well as ventral prostate weights were measured 50 days later at adulthood. At age 60 days, it was determined that all testes were descended into the scrotum, and there were no differences in testis and ventral prostate weights, intratesticular sperm counts and mean seminiferous tubular area between the control and sham operated animals. However, there was an 18% reduction in testicular blood flow (ml. per 100 gm. per minute +/- standard error of mean) in the operated animals when compared to the sham (20.43 +/- 1.10 versus 16.69 +/- 0.74, p less than 0.02). These data indicate that although there is a slight but significant reduction in testicular blood flow at adulthood when the testicular artery and vein are ligated early in life, this diminution is not sufficient to alter the ultimate location, testicular weight and spermatogenic function of the testis. This would suggest that after ligation of the main testicular vessels to the immature testis, the collateral blood supply is able to compensate with time to allow normal growth and development of the testis. These experimental observations provide additional support for the 2-staged approach to the high undescended testis whereby the testicular vessels are initially ligated and a subsequent procedure is performed to place the undescended testis into the scrotum.

Animals↗

Recovery of testicular blood flow following ligation of testicular vessels.

To determine whether initial ligation of the testicular vessels of the high undescended testis followed by a delayed secondary orchiopexy is a viable alternative to the classical Fowler-Stephens procedure, a series of preliminary experiments were conducted in the rat in which testicular blood flow was measured by the 133xenon washout technique before, and 1 hour and 30 days after ligation of the vessels. In addition, testicular histology, and testis and sex-accessory tissue weights were measured in 6 control, 6 sham operated and 6 testicular vessel ligated rats 54 days after vessel ligation. The data demonstrate that ligation and division of the testicular blood vessels produce an 80 per cent decrease in testicular blood flow 1 hour after ligation of the vessels. However, 30 days later testis blood flow returns to the control and pre-treatment value. There were no significant changes in testis or sex-accessory tissue weights 54 days after vessel ligation. Histologically, 4 of the surgically operated testes demonstrated necrosis of less than 25 per cent of the seminiferous tubules while 1 testis demonstrated more than 75 per cent necrosis. The rest of the tubules in all 6 testes demonstrated normal spermatogenesis. From this study we conclude that initial testicular vessel ligation produces an immediate decrease in testicular blood flow but with time the collateral vessels are able to compensate and return the testis blood flow to its normal pre-treatment value. These preliminary observations lend support for the concept that initial ligation of the testicular vessels followed by a delayed secondary orchiopexy in patients with a high undescended testis may be a possible alternative to the classical Fowler-Stephens approach.

Animals↗

Corticosteroid-binding globulin in depression.

We undertook this study to evaluate the role of plasma free and total cortisol, and corticosteroid-binding globulin (CBG) in depression. Plasma was collected from 15 drug-free depressed patients and 15 age and sex-matched control subjects at 0900 and 2300 and at 1600 following an oral dose of dexamethasone. Depressed patients had elevated concentrations of both free and total cortisol at all time points studied. The binding capacity and affinity of plasma CBG for cortisol was similar in both groups at all times studied. No differences in CBG measures were noted if the depressed subjects were divided into elderly or younger subjects and dexamethasone suppression test (DST) suppressors or non-suppressors.

Aged↗

[Mucinous cystadenoma of ileocecal appendix].

We present two patients with mucinous cystadenoma and mucinous cystadenocarcinoma of the appendix, respectively. These are very uncommon lesions in that localization. We point out the lack of specificity of the term "mucocele" and also the need of strict criteria to define this type of lesions in order to establish the adequate treatment. Usually the diagnosis is not possible before surgery. The treatment consists of simple appendectomy for the benign lesions and right hemicolectomy for mucinous carcinoma.

Adult↗

[The development of the torsion angle of the tibia-fibula unit in various age groups].

We studied the torsion angle of the tibiofibular unit in both lower legs of 1588 schoolchildren of both sexes (998 girls and 590 boys), aged between 3 and 18 years divided into four age groups. The torsion angle of the tibio-fibular unit progressively increases with age. Stable values were measured only in the 11-15 years age group. Statistically significant differences of the torsion angle between one lower limb and the other were detected in the girls of the 6-10 and 11-15 age groups, and in the boys of the 6-10 age groups.

Adolescent↗