A collection of enhancer trap insertional mutants for functional genomics in tomato.

With the completion of genome sequencing projects, the next challenge is to close the gap between gene annotation and gene functional assignment. Genomic tools to identify gene functions are based on the analysis of phenotypic variations between a wild type and its mutant; hence, mutant collections are a valuable resource. In this sense, T-DNA collections allow for an easy and straightforward identification of the tagged gene, serving as the basis of both forward and reverse genetic strategies. This study reports on the phenotypic and molecular characterization of an enhancer trap T-DNA collection in tomato (Solanum lycopersicum L.), which has been produced by Agrobacterium-mediated transformation using a binary vector bearing a minimal promoter fused to the uidA reporter gene. Two genes have been isolated from different T-DNA mutants, one of these genes codes for a UTP-glucose-1-phosphate uridylyltransferase involved in programmed cell death and leaf development, which means a novel gene function reported in tomato. Together, our results support that enhancer trapping is a powerful tool to identify novel genes and regulatory elements in tomato and that this T-DNA mutant collection represents a highly valuable resource for functional analyses in this fleshy-fruited model species.

Ethics competences in the undergraduate medical education curriculum: the Spanish experience.

Aim .To investigate if there are differences in medical ethics education between different schools of medicine in Spain, specifically between private and public schools and between recently founded schools and older ones. METHOD: The curricula of medical degrees from all Spanish faculties were reviewed for the 2014/2015 academic year, identifying subjects concerning bioethics, deontology, and ethics. We identified the type of teaching, format and method of the course, the number of credits and hours, and the school year of each subject. An analysis with descriptive parameters and the Cohen's coefficient (d) was performed. RESULTS: All medical schools in Spain (n=44) were included. A mean of 3.64 European Credit Transfer and Accumulation System (ECTS) credits was specifically devoted to ethical values teaching in Spain. Private medical schools offered more credits than public ones (6.51 ECTS vs 2.88 ECTS, relevant difference: d=2.06>>0.8), and the 10 most recently founded medical schools offered more credits than the 10 oldest (5.86 ECTS vs 2.63 ECTS, relevant difference: d=1.43>0.8). A mean of 36.75 hours was dedicated to ethics education. CONCLUSIONS: Although ethics education is incorporated into the training of future Spanish physicians, there is still notable heterogeneity between different medical schools in the time devoted to this topic.

[Communication skills in the curriculum of Medical students from Spain (1990-2014): From the Primary Health Care to the Bologna Plan. A descriptive study]. / Competencia en comunicación en el currículo del estudiante de Medicina de España (1990-2014): de la Atención Primaria al Plan Bolonia. Un estudio descriptivo.

OBJECTIVES: Analyse the currently situation of teaching in communication skills in the Spanish Medical Schools (MS) and the grade of implementation from the recommended by the National Agency of Evaluation of the Quality and Accreditation. DESIGN: Descriptive-comparative, transversal and quantitative-qualitative study. SITUATION: University teaching field. SAMPLE: The whole population of MS in Spain. INCLUSION CRITERIA: teaching during the academic year 2013-2014 the Degree in Medicine homologated by the National Agency of Evaluation of the Quality and Accreditation and offering information of their educational plans and teaching guides (online, telephonic or e-mail). VARIABLES: 1) Existence of courses about communication skills; 2) type of teaching; 3) format of the course; 4) credits; 5) length, and 6) grade. They were analysed with descriptive parameters and the Cohen's coefficient (d). RESULTS: Forty-two MS: 10 privates and 32 publics. Thirty MS (71.43%) presented at least one course of communication skills. The 78.12% of the public MS contained this kind of teaching as an entire course versus the 50.00% of the private MS. The national average by MS was 2.77 (σ=2.41), lower than the 5.0 credits recommended by the the National Agency of Evaluation of the Quality and Accreditation (relevant difference: d=1.63>>0,8). The 63.63% contained this teaching over the second cycle (mode: second degree). CONCLUSIONS: The course loads in terms of credits destined to this skills has increased considerably from 1990 to 2014 (d= 1,43>>0,8). However, it has not been completely established yet the recommendations about this aspect in most of the Spanish MS. The public universities seem to be more aware of this teaching. Furthermore, it would be interesting strengthen this skills in the sixth grade.

Production of engineered long-life and male sterile Pelargonium plants.

BACKGROUND: Pelargonium is one of the most popular garden plants in the world. Moreover, it has a considerable economic importance in the ornamental plant market. Conventional cross-breeding strategies have generated a range of cultivars with excellent traits. However, gene transfer via Agrobacterium tumefaciens could be a helpful tool to further improve Pelargonium by enabling the introduction of new genes/traits. We report a simple and reliable protocol for the genetic transformation of Pelargonium spp. and the production of engineered long-life and male sterile Pelargonium zonale plants, using the pSAG12::ipt and PsEND1::barnase chimaeric genes respectively. RESULTS: The pSAG12::ipt transgenic plants showed delayed leaf senescence, increased branching and reduced internodal length, as compared to control plants. Leaves and flowers of the pSAG12::ipt plants were reduced in size and displayed a more intense coloration. In the transgenic lines carrying the PsEND1::barnase construct no pollen grains were observed in the modified anther structures, which developed instead of normal anthers. The locules of sterile anthers collapsed 3-4 days prior to floral anthesis and, in most cases, the undeveloped anther tissues underwent necrosis. CONCLUSION: The chimaeric construct pSAG12::ipt can be useful in Pelargonium spp. to delay the senescence process and to modify plant architecture. In addition, the use of engineered male sterile plants would be especially useful to produce environmentally friendly transgenic plants carrying new traits by preventing gene flow between the genetically modified ornamentals and related plant species. These characteristics could be of interest, from a commercial point of view, both for pelargonium producers and consumers.

An insertional mutagenesis programme with an enhancer trap for the identification and tagging of genes involved in abiotic stress tolerance in the tomato wild-related species Solanum pennellii.

Salinity and drought have a huge impact on agriculture since there are few areas free of these abiotic stresses and the problem continues to increase. In tomato, the most important horticultural crop worldwide, there are accessions of wild-related species with a high degree of tolerance to salinity and drought. Thus, the finding of insertional mutants with other tolerance levels could lead to the identification and tagging of key genes responsible for abiotic stress tolerance. To this end, we are performing an insertional mutagenesis programme with an enhancer trap in the tomato wild-related species Solanum pennellii. First, we developed an efficient transformation method which has allowed us to generate more than 2,000 T-DNA lines. Next, the collection of S. pennelli T(0) lines has been screened in saline or drought conditions and several presumptive mutants have been selected for their salt and drought sensitivity. Moreover, T-DNA lines with expression of the reporter uidA gene in specific organs, such as vascular bundles, trichomes and stomata, which may play key roles in processes related to abiotic stress tolerance, have been identified. Finally, the growth of T-DNA lines in control conditions allowed us the identification of different development mutants. Taking into account that progenies from the lines are being obtained and that the collection of T-DNA lines is going to enlarge progressively due to the high transformation efficiency achieved, there are great possibilities for identifying key genes involved in different tolerance mechanisms to salinity and drought.

Efficient transformation of Kalanchoe blossfeldiana and production of male-sterile plants by engineered anther ablation.

Engineered male sterility in ornamental plants has many applications such as facilitate hybrid seed production, eliminate pollen allergens, reduce the need for deadheading to extend the flowering period, redirect resources from seeds to vegetative growth, increase flower longevity and prevent gene flow between genetically modified and related native plants. We have developed a reliable and efficient Agrobacterium-mediated protocol for the genetic transformation of different Kalanchoe blossfeldiana commercial cultivars. Transformation efficiency for cv. 'Hillary' was 55.3% whereas that of cv. 'Tenorio' reached 75.8%. Selection was carried out with the nptII gene and increasing the kanamycin concentration from 25 to 100 mg l(-1) allowed to reduced escapes from 50 to 60% to virtually 0%. This method was used to produce male-sterile plants through engineered anther ablation. In our approach, we tested a male sterility chimaeric gene construct (PsEND1::barnase) to evaluate its effectiveness and effect on phenotype. No significant differences were found in the growth patterns between the transgenic lines and the wild-type plants. No viable pollen grains were observed in the ablated anthers of any of the lines carrying the PsEND1::barnase construct, indicating that the male sterility was complete. In addition, seed set was completely abolished in all the transgenic plants obtained. Our engineered male-sterile approach could be used, alone or in combination with a female-sterility system, to reduce the invasive potential of new ornamentals, which has become an important environmental problem in many countries.

Communication skills training in undergraduate nursing programs in Spain.

OBJECTIVES: of the present cross sectional study were to analyze the offer of subjects with communication skills in the nursing degree in Spain and to describe the distribution of these subjects. Documentary, systematic and independent search of web pages from Spanish universities was carried out to collect the following variables: subjects with communication skills, course in which the subject is taught, type of teaching (exclusive or combined), type of subject (compulsory or optional) and number of credits on communication skills. Although an average of 3.6 subjects per center was offered, most of the subjects were combined and with little communication content load. In one third of the centers, the offer was below 2.3 credits. Only 1 in 6 centers had exclusive communication skills subjects, and a quarter of them were optional. The teaching load of communication contents was highest in optional subjects. The offer of communication skills contents in Spanish Nursing Schools was scarce and very heterogeneous between centers and between courses in a center, with excessive presence of combined and optional subjects. Our results may be useful when developing the teaching guides for subjects with communication skills, as well as when defining communication competencies in the different Nursing Schools.

[Risk factors of persistent disease at 5 years from diagnosis in differentiated thyroid cancer: study of 63 patients]. / Factores relacionados con la persistencia de la enfermedad a los 5 años del diagnóstico de cáncer diferenciado de tiroides: estudio de 63 pacientes.

OBJECTIVE: To identify possible risk factors associated with persistent disease 5 years after total or near-total thyroidectomy in patients with differentiated thyroid cancer (DTC). PATIENTS AND METHOD: Retrospective study evaluating data from 63 patients 5 years after they were first diagnosed of DTC. At this time of the study, 46 subjects were considered disease-free (F group) whereas 17 had evidence of persistent disease or had died from DTC (P group). We compared both groups of patients regarding the following variables: a) variables at diagnosis related to the patient (age, gender) and the tumor (histological type, size, extrathyroidal involvement, vascular invasion, multifocality, lymph node and distant metastases), and b) variables recorded during follow-up: percentage of subjects showing serum stimulated thyroglobulin > or = 10 ng/ml few weeks postoperatively (Tg0) and 6 to 12 months later (Tg1). RESULTS: Male gender, extrathyroidal involvement and lymph node metastases were more frequent in P group than in F group (41 vs. 11%, 60 vs. 18% and 50 vs. 5.5%; p < 0.05). During the follow-up the percentage of patients showing Tg > or = 10 ng/ml was higher in P group compared to F group, both at a few weeks postoperatively and 6 to 12 months later (Tg0, 75 vs. 13%; Tg1, 69% vs. 0; p < 0,05). CONCLUSIONS: In our patients, male gender, extrathyroidal involvement, and lymph node metastases at diagnosis were associated with persistent disease 5 years later. Serum stimulated thyroglobulin had a very high predictive value both just after surgery and in the next 6 to 12 months and could help identifying subjects who need a closer follow-up.

Overexpression of dehydrin tas14 gene improves the osmotic stress imposed by drought and salinity in tomato.

One strategy to increase the level of drought and salinity tolerance is the transfer of genes codifying different types of proteins functionally related to macromolecules protection, such as group 2 of late embryogenesis abundant (LEA) proteins or dehydrins. The TAS14 dehydrin was isolated and characterized in tomato and its expression was induced by osmotic stress (NaCl and mannitol) and abscisic acid (ABA) [Godoy et al., Plant Mol Biol 1994;26:1921-1934], yet its function in drought and salinity tolerance of tomato remains elusive. In this study, transgenic tomato plants overexpressing tas14 gene under the control of the 35SCaMV promoter were generated to assess the function of tas14 gene in drought and salinity tolerance. The plants overexpressing tas14 gene achieved improved long-term drought and salinity tolerance without affecting plant growth under non-stress conditions. A mechanism of osmotic stress tolerance via osmotic potential reduction and solutes accumulation, such as sugars and K(+) is operating in tas14 overexpressing plants in drought conditions. A similar mechanism of osmotic stress tolerance was observed under salinity. Moreover, the overexpression of tas14 gene increased Na(+) accumulation only in adult leaves, whereas in young leaves, the accumulated solutes were K(+) and sugars, suggesting that plants overexpressing tas14 gene are able to distribute the Na(+) accumulation between young and adult leaves over a prolonged period in stressful conditions. Measurement of ABA showed that the action mechanism of tas14 gene is associated with an earlier and greater accumulation of ABA in leaves during short-term periods. A good feature for the application of this gene in improving drought and salt stress tolerance is the fact that its constitutive expression does not affect plant growth under non-stress conditions, and tolerance induced by overexpression of tas14 gene was observed at the different stress degrees applied to the long term.

[About a case of multiple endocrine neoplasia type 1. Review of some clinical manifestations and treatment controversies]. / A propósito de un caso de neoplasia endocrina múltiple tipo 1. Revisión de algunas manifestaciones clínicas y controversias en el tratamiento.

The rare hereditary syndrome, multiple endocrine neoplasia type1 (MEN-1), is known to predispose affected individuals to endocrine neoplasms in a variety of tissues such as the parathyroid glands, the pituitary gland and the gastrointestinal tract. We describe the case of a man with traditionally-described manifestations (hyperparathyroidism and gastrinoma) and with other tumoral lesions arising from endocrine cells (insulinoma, gastric carcinoid, adrenal adenoma and pancreatic non-functioning neuroendocrine tumors) and non-endocrine cells (lipoma and collagenoma). Frequent recurrences in susceptible tissues that are not totally removed (as occurs in hyperparathyroidism and duodenal gastrinoma) and their unknown clinical significance have aroused current controversies in the therapeutic management of these entities, which is briefly reviewed.