Defective regulatory volume decrease in human cystic fibrosis tracheal cells because of altered regulation of intermediate conductance Ca2+-dependent potassium channels

The cystic fibrosis transmembrane conductance regulator (CFTR) protein has the ability to function as both a chloride channel and a channel regulator. The loss of these functions explains many of the manifestations of the cystic fibrosis disease (CF), including lung and pancreatic failure, meconium ileus, and male infertility. CFTR has previously been implicated in the cell regulatory volume decrease (RVD) response after hypotonic shocks in murine small intestine crypts, an effect associated to the dysfunction of an unknown swelling-activated potassium conductance. In the present study, we investigated the RVD response in human tracheal CF epithelium and the nature of the volume-sensitive potassium channel affected. Neither the human tracheal cell line CFT1, expressing the mutant CFTR-ΔF508 gene, nor the isogenic vector control line CFT1-LC3, engineered to express the βgal gene, showed RVD. On the other hand, the cell line CFT1-LCFSN, engineered to express the wild-type CFTR gene, presented a full RVD. Patch-clamp studies of swelling-activated potassium currents in the three cell lines revealed that all of them possess a potassium current with the biophysical and pharmacological fingerprints of the intermediate conductance Ca2+-dependent potassium channel (IK, also known as KCNN4). However, only CFT1-LCFSN cells showed an increase in IK currents in response to hypotonic challenges. Although the identification of the molecular mechanism relating CFTR to the hIK channel remains to be solved, these data offer new evidence on the complex integration of CFTR in the cells where it is expressed.

Comparative Analysis of the Regulation of Expression and Structures of Two Evolutionarily Divergent Genes for Δ1-Pyrroline-5-Carboxylate Synthetase from Tomato1

We isolated two tomato (Lycopersicon esculentum) cDNA clones, tomPRO1 and tomPRO2, specifying Δ1-pyrroline-5-carboxylate synthetase (P5CS), the first enzyme of proline (Pro) biosynthesis. tomPRO1 is unusual because it resembles prokaryotic polycistronic operons (M.G. García-Ríos, T. Fujita, P.C. LaRosa, R.D. Locy, J.M. Clithero, R.A. Bressan, L.N. Csonka [1997] Proc Natl Acad Sci USA 94: 8249–8254), whereas tomPRO2 encodes a full-length P5CS. We analyzed the accumulation of Pro and the tomPRO1 and tomPRO2 messages in response to NaCl stress and developmental signals. Treatment with 200 mm NaCl resulted in a >60-fold increase in Pro levels in roots and leaves. However, there was a <3-fold increase in the accumulation of the tomPRO2 message and no detectable induction in the level of the tomPRO1 message in response to NaCl stress. Although pollen contained approximately 100-fold higher levels of Pro than other plant tissues, there was no detectable increase in the level of either message in pollen. We conclude that transcriptional regulation of these genes for P5CS is probably not important for the osmotic or pollen-specific regulation of Pro synthesis in tomato. Using restriction fragment-length polymorphism mapping, we determined the locations of tomPRO1 and tomPRO2 loci in the tomato nuclear genome. Sequence comparison suggested that tomPRO1 is similar to prokaryotic P5CS loci, whereas tomPRO2 is closely related to other eukaryotic P5CS genes.

Modulation of promoter occupancy by cooperative DNA binding and activation-domain function is a major determinant of transcriptional regulation by activators in vivo.

Binding of transcriptional activators to a promoter is a prerequisite process in transcriptional activation. It is well established that the efficiency of activator binding to a promoter is determined by the affinity of direct interactions between the DNA-binding domain of an activator and its specific target sequences. However, I describe here that activator binding to a promoter is augmented in vivo by the effects of two other determinants that have not been generally appreciated: (i) the number of activator binding sites present in a promoter and (ii) the potency of activation domains of activators. Multiple sites within a promoter can cooperatively recruit cognate factors regardless of whether they contain an effective activation domain. This cooperativity can result in the synergistic activation of transcription. The second effect is the enhancement of activator binding to a promoter by the presence of activation domains. In this case, activation domains are not simply tethered to the promoter by the DNA-binding domain but instead assist the DNA-binding domain being tethered onto the promoter. This effect of activation domains on DNA binding is instrumental in determining how potent activators can induce steep transcriptional increases at low concentrations.

Serve analysis of professional players in beach volleyball

This study has been developed for the European Beach Volleyball Championship in 2005. The video-recorded analysis was held using Sportcode Pro v.8.5.2 software. The aim of study was to determine the types of serve used, depending on the time of the set in which they occur. Quantitative analysis with a sample of 10 players that make up 5 teams with a total of four meetings with a total of 327 serves analyzed. The serves were classified depending on the period which they occurred, the period being 1 (items 1 to 7), period 2 (from point 8 to 14) and period 3 (point 15 to 21). he statistical analysis was conducted using the statistical software SPSS 19, Chi-square test established significant differences between the different types of serve for period 1 and 2 (p<0.05), but no significant differences were established in the period 3 to floating serve and power jump (p>0.05). The results showed a decrease of using a serve with jump power at period 1 (89.7%) compared to the period 3 (27.3%), while the floating and floating serve jump respectively increase at period 1 (6.3% -4%) in the period 3 (23.4% -49.4%).

Development of multidisciplinary workshops between different subjects as an innovative teaching methodology for the acquisition of professional skills of the future architect

The implantation of the new Architecture Degree and the important normative changes in the building sector imply the need to use new teaching methodologies that enhance skills and competences in order to response to the increasing requirements demanded by society to the future architect. The aim of this paper is to present, analyze and discuss the development of multidisciplinary workshops as a new teaching methodology used in several Construction subjects of the Architecture Degree in the University of Alicante. Workshops conceived with the aim to synthesize and complement the technical knowledge acquired by the students during the Degree and to enhance the skills and competencies necessary for the professional practice. With that purpose, we decided to experiment on current subjects of the degree during this academic year, by applying the requirements defined in the future Architecture Degree in a practical way, through workshops between different subjects, superposing the technical knowledge with the resolution of constructive problems in the development of an architectural project. Developing these workshops between subjects we can dissolve the traditional boundaries between different areas of the Degree. This multidisciplinary workshop methodology allows the use of all the global knowledge acquired by students during their studies and at the same time, it enhances students’ ability to communicate and discuss their ideas and solutions in public. It also increases their capacity of self-criticism, and it foments their ability to undertake learning strategies and research in an autonomous way. The used methodology is based on the development of a practical work common to several subjects of different knowledge areas within the "Technology Block" of the future Architecture Degree. Thus, students work approaching the problem in a global way discussing simultaneously with teachers from different areas. By using these new workshops we stimulate an interactive class versus a traditional lecture. Work is evaluated continuously, valuing the participative pupil´s attitude, working in groups in class time, reaching weekly objectives and stimulating the individual responsibility and positive interdependence of the pupil inside the working group. The exercises are designed to improve students’ ability to transmit their ideas and solutions in public, knowing how to discuss and defend their technical resolutions to peers and teachers (Peer Reviewing), their capacity for self-criticism and their capacity to undertake strategies and autonomous learning processes at the same time they develop a personal research into new technologies, systems and materials. Students have shown their majority preference for this teaching methodology by the multidisciplinary workshops offered in the last years, with very satisfactory academic results. In conclusion, it can be verified nowadays the viability of the introduction of new contents and new teaching methodologies necessary for the acquisition of the skills in the future Architecture Degree, through workshops between several subjects that have had a great acceptance in students and positive contrasted academic results.

Common trees / by J. Francis Macbride, Asst. Curator, Taxonomy, Dept. of Botany.

no.11(1936)

A classification of the Conularida / G. Winston Sinclair, Dept. of Geology, University of Michigan.

v.10:no.13(1952)