Application of genomic tools in plant breeding.

Plant breeding has been very successful in developing improved varieties using conventional tools and methodologies. Nowadays, the availability of genomic tools and resources is leading to a new revolution of plant breeding, as they facilitate the study of the genotype and its relationship with the phenotype, in particular for complex traits. Next Generation Sequencing (NGS) technologies are allowing the mass sequencing of genomes and transcriptomes, which is producing a vast array of genomic information. The analysis of NGS data by means of bioinformatics developments allows discovering new genes and regulatory sequences and their positions, and makes available large collections of molecular markers. Genome-wide expression studies provide breeders with an understanding of the molecular basis of complex traits. Genomic approaches include TILLING and EcoTILLING, which make possible to screen mutant and germplasm collections for allelic variants in target genes. Re-sequencing of genomes is very useful for the genome-wide discovery of markers amenable for high-throughput genotyping platforms, like SSRs and SNPs, or the construction of high density genetic maps. All these tools and resources facilitate studying the genetic diversity, which is important for germplasm management, enhancement and use. Also, they allow the identification of markers linked to genes and QTLs, using a diversity of techniques like bulked segregant analysis (BSA), fine genetic mapping, or association mapping. These new markers are used for marker assisted selection, including marker assisted backcross selection, 'breeding by design', or new strategies, like genomic selection. In conclusion, advances in genomics are providing breeders with new tools and methodologies that allow a great leap forward in plant breeding, including the 'superdomestication' of crops and the genetic dissection and breeding for complex traits.

[Contribution of (99m)Tc-mebrofenin scintigraphy to the diagnosis of ectopic liver. A case report]. / Contribución de la gammagrafía hepatobiliar con (99m)Tc-mebrofenina al diagnóstico del hígado ectópico. A propósito de un caso.

We present the case of a woman with a parasplenic mass who underwent abdominal ultrasounds, computed tomography and magnetic resonance imaging. The liver scintigraphy with (99m)Tc-Mebrofenin confirmed the diagnosis of ectopic liver.

Evolution of the bioavailability and other pharmacokinetic parameters of levodopa (with carbidopa) in rabbits.

Levodopa pharmacokinetics show important inter- and intraindividual differences when it is administered by the oral route. As a result of fluctuating drug plasma concentrations, patients may develop motor fluctuations and dyskinesias. Therefore, it is important to perform studies on levodopa pharmacokinetics in the same individual. The aim of this study was to contribute to a better knowledge of the evolution of the pharmacokinetics of levodopa administered with carbidopa. The study involved the oral administration of 20/5 mg/kg levodopa/carbidopa to rabbits for two different time periods (7 or 14 days), due to the fact that inhibition of aromatic L-amino-acid decarboxylase by carbidopa is not immediate. After 7 days of treatment, the levodopa AUC increased by 12.6% from day 1 (range: 114.2-150.7 microg.min/ml) to day 7 (range: 131.1-166.0 microg.min/ml) and C(max) increased by 9.6% (range: 1.90-2.86 microg/ml on day 1 and 2.12-3.13 microg/ml on day 7). After 14 days of treatment, the increase in AUC was 17.0% (range: 119.6-160.1 microg.min/ml on day 1 and 142.9-172.7 microg.min/ml on day 14) and C(max) increased by 6.5% (range: 2.29-2.96 microg/ml on day 1 and 2.41-3.07 microg/ml on day 14). The values obtained for C(min) (sample obtained immediately before levodopa/carbidopa administration) in both groups increased progressively with the duration of the treatment. C(max) and AUC values were very similar after 7 or 14 days of treatment. The time needed for C(min) stabilization was slightly higher, because we found significant differences until day 11 of treatment.

Therapeutic effects of psyllium in type 2 diabetic patients.

OBJECTIVE: The aim of this study was to evaluate the effects of psyllium in type 2 diabetic patients. DESIGN: The study included three phases: phase 1 (1 week), phase 2 (treatment, 14 g fibre/day, 6 weeks) and phase 3 (4 weeks). At the end of each phase a clinical evaluation was performed after the ingestion of a test breakfast of 1824.2 kJ (436 kcal). Measurements included concentrations of blood glucose, insulin, fructosamine, GHbA(1c), C-peptide and 24 h urinary glucose excretion. In addition, uric acid, cholesterol and several mineral and vitamin concentrations were also evaluated. SETTING: The study was performed at the Department of Pharmacology, Toxicology and Nursing at the University of León (Spain). SUBJECTS: Twenty type 2 diabetic patients (12 men and 8 women) participated in the study with a mean age of 67.4 y for men and 66 y for women. The mean body mass index of men was 28.2 kg/m(2) and that of women 25.9 kg/m(2). RESULTS: Glucose absorption decreased significantly in the presence of psyllium (12.2%); this reduction is not associated with an important change in insulin levels (5%). GHbA(1c), C-peptide and 24 h urinary glucose excretion decreased (3.8, 14.9 and 22.5%, respectively) during the treatment with fibre (no significant differences) as well as fructosamine (10.9%, significant differences). Psyllium also reduced total and LDL cholesterol (7.7 and 9.2%, respectively, significant differences), and uric acid (10%, significant difference). Minerals and vitamins did not show important changes, except sodium that increased significantly after psyllium administration. CONCLUSIONS: The results obtained indicate a beneficial therapeutic effect of psyllium (Plantaben) in the metabolic control of type 2 diabetics as well as in lowering the risk of coronary heart disease. We also conclude that consumption of this fibre does not adversely affect either mineral or vitamin A and E concentrations. Finally, for a greater effectiveness, psyllium treatment should be individually evaluated.

[Usefulness of 201T1 gammagraphy in the diagnosis of carcinoma of the larynx]. / Utilidad de la gammagrafía con 201Tl en el diagnóstico del carcinoma de laringe.

For the evaluation of a patient with a laryngeal tumor we need the clinical exam and other exams as the CT scan or MRI. Those studies have a sensitivity of less than 80%. For that reason in the last years there has been a development of new techniques trying to increase the accuracy. The 201Tl SPECT is one of them although it was developed for cardiological purposes. We present our experience in 46 patients with laryngeal tumor in whom we did a 201Tl SPECT as part of the extension study. The sensitivity of the study was 81.6% in the diagnosis of the primary tumor and 46.1% for the neck adenopathies. The 201Tl SPECT can be a good method for the evaluation and detection of recurrences in patients with pharyngo-laryngeal tumor.

Effects of ispaghula husk and guar gum on postprandial glucose and insulin concentrations in healthy subjects.

OBJECTIVE: The aim of this study was to evaluate, under the same experimental conditions and in the same subjects, the effects of ispaghula husk and guar gum on postprandial glucose and insulin concentrations in healthy female subjects. DESIGN: An oral glucose load with and without fiber was administered in the morning after an overnight fast. The study of the fiber effect was planned according to a randomized and cross-over design. SETTING: The study was performed at the Department of Pharmacology, Toxicology and Nursing at the University of León (Spain). SUBJECTS: Ten healthy female volunteers aged 30-48 y with normal body mass indices participated in this study. RESULTS: A significant decrease in mean serum insulin concentrations was observed from 30 to 90 min in the presence of both fibers. The area under the insulin curve was significantly reduced by 36.1% for ispaghula husk and 39.4% for guar gum. The area under the glucose curve was reduced by 11.1% (significant difference) for ispaghula husk and 2.6% for guar gum (no significant difference). CONCLUSIONS: According to the results obtained in this study, the administration of ispaghula husk may be beneficial due to its ability to reduce glucose postprandial concentration and especially insulin requirements. Individualization of the treatment would be advisable due to large individual variations observed in glycemic and insulinemic postprandial responses.

Influence of two dietary fibers in the oral bioavailability and other pharmacokinetic parameters of ethinyloestradiol.

Dietary fibers are widely used in hypoglycaemic, hypolipidemic, slimming diets. It is probable that their ingestion coincides with the oral administration of drugs and a modification of their pharmacokinetics can appear. In the present study, the influence of two soluble fibers (guar gum and psyllium) was evaluated on the pharmacokinetics of ethinyloestradiol (EE) when they were administered together to female rabbits via the oral route. Three groups of rabbits were used. All animals received 1 mg/kg of EE; this compound was administered alone in the control group and with 3.5 g of guar gum or psyllium in the other two groups. When guar gum was administered, there was a decrease in the extent of EE absorbed, but no change was observed in the rate of absorption. When psyllium was administered, the extent of EE absorbed increased slightly and the rate of absorption was slower.

Organochlorine pesticide residues in muscle tissue of rainbow trout, Oncorhynchus mykiss, taken from four fish farms in León, Spain.

Levels of nine organochlorine pesticides (lindane, heptachlor epoxide, aldrin, dieldrin, endrin, o,p'-TDE, p,p'-TDE, p,p'-DDE and p,p'-DDT) were determined in muscle samples of rainbow trout, Onchorhynchus mykiss, collected from four fish farms in the province of León, in the north-west of Spain (Europe). The highest incidence percentage was for lindane (67.5%) and heptachlor epoxide (55.0%). Organochlorine residue levels detected contributed slightly to acceptable daily intakes (ADIs) established by the Food and Agriculture Organization of the United Nations and the World Health Organization (lindane 0.22-2.3%; sigma DDT 0.05-0.46%; heptachlor epoxide 10.0-71.4% and sigma dieldrin 16.7-33.3% assuming 300 g of trout muscle as a mean daily intake). The highest concentration found was for heptachlor epoxide (0.043 microgram/g).

Influence of two commercial fibers in the pharmacokinetics of ethinylestradiol in rabbits.

Fiber formulations are used in human nutrition owing to their beneficial properties for health. It is probable that ingestion of fiber coincides with the oral administration of drugs, and a modification of its oral absorption, and therefore of its pharmacokinetics, can appear. In the present study, the compartmental and noncompartmental pharmacokinetic parameters of ethinylestradiol (EE) in rabbits after oral administration were determined. It was also studied whether the presence of two different fiber formulations [A, wheat bran (76.5%), fruit fiber (12%) and guar gum (2%) and B, Plantago ovata seeds (65%) and P. ovata seed cuticles (2.2%)] in the gastrointestinal tract modified the pharmacokinetics of EE when administered at the same time. Three groups of rabbits were used: control, fiber A and fiber B. The animals in all three groups received 1 mg/kg b. wt. EE. The estrogen was administered alone in the control group and in the presence of 4 g of fiber A and fiber B, respectively, in the other two groups. After compartmental (two-compartment open model) and noncompartmental analyses of plasma concentrations, statistical analysis revealed that the presence of fiber (both A and B) decreased between 29% and 35% the extent of EE absorbed (represented by the pharmacokinetic parameters area under the curve and the maximum plasma concentration) without affecting the rate of the absorption process (represented by the time to reach maximum concentration and the absorption rate constant).

Bioavailability of levamisole after intramuscular and oral administration in sheep.

AIMS: To determine the bioavailability of levamisole in sheep. METHODS: Levamisole was administered to three groups of six Merino sheep orally and intramuscularly at three dose levels of 5, 7.5 and 10 mg/kg. There was a washout period of 1 week between treatments. Blood samples were collected by jugular venepuncture and plasma was separated immediately by centrifugation and stored at 20 degrees C until analysed. The levamisole concentration in plasma was determined by high performance liquid chromatography with a U.V. detection method. Individual plasma levamisole concentration-time data were analysed using the compartmental method. RESULTS: The values obtained for k(a), C(max), t(max) and F show a moderate rate and extent of absorption after oral administration of levamisole while, after intramuscular administration, these values demonstrate a high rate and extent of absorption of levamisole. The intramuscular bioavailability was higher than the oral bioavailability (rate of absorption three-fold faster, extent of absorption 25-33% higher and C(max) two-fold higher). The Friedman test involving dose and route of administration showed that the route of administration affects k(a), C(max), t(max) and F; significant differences were found in these parameters. CLINICAL RELEVANCE: On the basis of these data, the recommended routes for the administration of levamisole in sheep are oral for gastro-intestinal nematodiasis and intramuscular for extragastric nematodiasis.

[Study of the demand for home care by the Pamplona Extrahospital Emergency Services in 1995]. / Estudio de la demanda de atención domiciliaria en 1995 del Servicio de Urgencias Extrahospitalario de Pamplona.

The present paper is a retrospective descriptive study, carried out in the Special Casualty Service (Servicio Especial de Urgencias) of Pamplona. It evaluates 8,411 cases of calls from homes, extracted from an aleatory sample stratified by day of the week and season of the year, with a N=647. The diagnoses were codified according to the ICHPPC-2 of the WONCA and the statistical study with SPSS/PC+V 4.0. The monthly average was 701 calls from homes, with a daily average of 19 (from Monday to Friday), with 38 on Sundays and holidays. With respect to distribution by time, on working days 66.5% were distributed between 15.00 and 22.00 hours, while on Sundays and holidays 51% fell between 8.00 and 13.00 hours. The overall rate of use was situated at 32.71/1,000 inhabitants and year (with extreme rates in Mendillorri=7.42 and II Ensanche=57.35). In 74% of the cases a doctor or nurse visited the home. The most frequent cause for home care are illnesses of the respiratory apparatus with 23.5%. Some 40.9% of the patients attended were over 75 years of age. The conclusions are: 1. There is a low rate of home care. 2. Those requiring such visits are of an advanced age. 3. Illnesses of the respiratory apparatus are the most frequent cause of demand. 4. The zones of most demand are those where a Health Centre has not been set up as such.

Plasma protein binding of levamisole in several species.

The binding of levamisole to total plasma proteins of 6 animal species was determined in vitro by equilibrium dialysis. The percentage of bound drug protein was independent of levamisole concentration within the range studied, 5-50 micrograms/ml (ANOVA). Levamisole was bound to a low extent to plasma proteins of each animal species (19.40-25.91%). There were significant differences in the extent of levamisole binding among species (ANOVA). Owing to the low degree of protein binding and the high volume of distribution of levamisole, the variations in protein binding due to different factors would not be of major clinical importance in its therapeutic application.

Pharmacokinetics of levamisole in sheep after intravenous administration.

The pharmacokinetics of levamisole at doses of 5, 7.5 and 10 mg/kg were determined after its intravenous administration to eighteen healthy Merino sheep. Using compartmental analysis, the disposition of the drug best fitted a two-compartmental open model. The mean values for the compartmental volume of distribution at steady state (Vss) were 2.034 +/- 0.23 I, 2.347 +/- 0.720 and 2.001 +/- 0.367 I/kg for each dose, respectively, and values obtained using the statistical moment theory were 2.141 +/- 0.269,2.390 +/- 0.536 and 2.140 +/- 0.345 l/kg for each dose, respectively. There were no dose-related differences (one-way ANOVA) in the constants describing distribution and elimination phases (alpha and beta) or Vss, but significant differences were detected in the total body clearance (Cl) and the area under the plasma concentration-time curve (AUC). After non-compartmental analysis, no significant differences were found when the parameters lambda (the linear terminal slope) and Vss were compared, but significant differences were detected in Cl and AUC. There were no significant differences between the values obtained using the compartmental and non-compartmental analysis when lambda -beta, Cl, Vss, and AUC were compared.

Study of the pharmacokinetic interaction between ethinylestradiol and amoxicillin in rabbits.

Several antibiotics have been implicated in oral contraception failure when they are administered at the same time as the oral contraceptive (OC) pill. In the present paper, a study about amoxicillin-ethinylestradiol (EE2) pharmacokinetic potential interaction was studied. Two rabbit groups were utilized, the first group received amoxicillin (10 mg/kg) and EE2 (30, 50 and 100 micrograms/kg, respectively), both by intravenous (i.v.) route. The second group received amoxicillin (oral route, 10 mg/kg/day) and EE2 (i.v. route, 100 mu/kg) on day 1, 4 and 8 of antibiotic treatment, respectively. After compartmental (two-compartment open model) and non-compartmental analysis of plasma concentrations, the statistical study (ANOVA p < or = 0.05) revealed that the presence of amoxicillin did not modify the EE2 distribution and elimination pharmacokinetic parameters (by comparison with those obtained in a previous study where EE2 was administered alone). There also were no significant differences with the time of amoxicillin oral treatment.

Pharmacokinetics of ethinyloestradiol in rabbits after intravenous administration.

The pharmacokinetics of ethinyloestradiol (EE2) after intravenous administration of 30, 50 and 100 micrograms.kg-1 was investigated in rabbits. A high-performance liquid chromatographic (HPLC) method with electrochemical (EC) detection was used to measure EE2 in plasma samples in order to avoid the interferences of natural oestrogens. After compartmental analysis, the disposition of EE2 was well described by a two-compartmental open model with mean values of: alpha = 0.3448 +/- 0.2922, 0.1965 +/- 0.1755, 0.3058 +/- 0.1225 min-1, and beta = 0.0137 +/- 0.0018, 0.0140 +/- 0.0065, 0.0198 +/- 0.0066 min-1, for the three doses studied, respectively. There were no dose-related differences (ANOVA, P < 0.05) in a, b or Vss, but significant differences were detected in clearance (90.9 +/- 18.7; 80.6 +/- 17.6; 116.3 +/- 21.5 ml.min-1.kg-1) between the 100 micrograms.kg-1 group and lower dose groups. The AUC increased significantly with the doses (341.7 +/- 67.1; 645.8 +/- 143.9; 892.2 +/- 211.9 ng.min.ml-1). After non-compartmental analysis there were no significant differences in lambda, MRT or Vss as a function of dose, but these differences were significant when Cl or AUC were compared. There were no significant differences in AUC or Cl values obtained by compartmental and non-compartmental analysis.

Organochlorine pesticide residues in bovine milk from Leon (Spain).

Residue levels of the organochlorine pesticides (alpha-HCH, lindane, heptachlor-epoxide, aldrin, endrin, dieldrin, o,p'-TDE, p,p'-TDE, p,p'-DDE and p,p'-DDT) were determined in raw bovine milk and compared with the maximum levels allowed by the European Union (EU) in these foods. The highest incidence percentage of the ten insecticides measured was for lindane, followed by alpha-HCH and aldrin. Moreover, the highest mean residue level was for alpha-HCH. None of the samples analyzed exceeded the maximum levels allowed by the EU.

Bioavailability of levamisole administered by subcutaneous and oral routes in rabbits.

The bioavailability of levamisole in rabbits was determined after subcutaneous and oral administration at three dose levels of 12.5, 16.0 and 20.0 mg/kg. After non-compartmental analysis the mean values obtained were: Cmax = 3.54, 4.51 and 5.39 micrograms/ml; tmax = 12.0, 22.0 and 20.0 min; F = 134.8, 105.4 and 124.1% after subcutaneous administration for each dose, respectively, and Cmax = 0.71, 1.32 and 1.77 micrograms/ml; tmax = 46.0, 96.0 and 84.0 min; F = 53.0, 62.0 and 80.7% after oral administration. The extent and rate of absorption from the two routes differed significantly, except for tmax at the 12.5 mg/kg dose. After compartmental analysis the pharmacokinetics of levamisole was characteristic of a two-compartment open model in 13 rabbits and of a one-compartment open model in two rabbits after subcutaneous administration, while it was two compartmental in nine and one compartmental in six rabbits after oral administration. The ka values were 0.321, 0.145 and 0.145 min-1 after subcutaneous administration and 0.054, 0.023 and 0.027 min-1 after oral administration. There were no significant differences between the values of Cmax, tmax and AUC calculated by compartmental and non-compartmental analysis.