A short in-frame deletion in NTRK1 tyrosine kinase domain caused by a novel splice site mutation in a patient with congenital insensitivity to pain with anhidrosis

Background: Congenital insensitivity to pain with anhidrosis (CIPA) is a rare autosomal recessive genetic disease characterized by the lack of reaction to noxious stimuli and anhidrosis. It is caused by mutations in the NTRK1 gene, which encodes the high affinity tyrosine kinase receptor I for Neurotrophic Growth Factor (NGF). -- Case Presentation: We present the case of a female patient diagnosed with CIPA at the age of 8 months. The patient is currently 6 years old and her psychomotor development conforms to her age (RMN, SPECT and psychological study are in the range of normality). PCR amplification of DNA, followed by direct sequencing, was used to investigate the presence of NTRK1 gene mutations. Reverse transcriptase (RT)-PCR amplification of RNA, followed by cloning and sequencing of isolated RT-PCR products was used to characterize the effect of the mutations on NTRK1 mRNA splicing. The clinical diagnosis of CIPA was confirmed by the detection of two splice-site mutations in NTRK1, revealing that the patient was a compound heterozygote at this gene. One of these alterations, c.574+1G > A, is located at the splice donor site of intron 5. We also found a second mutation, c.2206-2 A > G, not previously reported in the literature, which is located at the splice acceptor site of intron 16. Each parent was confirmed to be a carrier for one of the mutations by DNA sequencing analysis. It has been proposed that the c.574+1G > A mutation would cause exon 5 skipping during NTRK1 mRNA splicing. We could confirm this prediction and, more importantly, we provide evidence that the novel c.2206-2A > G mutation also disrupts normal NTRK1 splicing, leading to the use of an alternative splice acceptor site within exon 17. As a consequence, this mutation would result in the production of a mutant NTRK1 protein with a seven aminoacid in-frame deletion in its tyrosine kinase domain. --Conclusions: We present the first description of a CIPA-associated NTRK1 mutation causing a short interstitial deletion in the tyrosine kinase domain of the receptor. The possible phenotypical implications of this mutation are discussed.

A Multiscale-Domain Decomposition Method for High Reynolds Number Flows

The high Reynolds number flow contains a wide range of length and time scales, and the flow domain can be divided into several sub-domains with different characteristic scales. In some sub-domains, the viscosity dissipation scale can only be considered in a certain direction; in some sub-domains, the viscosity dissipation scales need to be considered in all directions; in some sub-domains, the viscosity dissipation scales are unnecessary to be considered at all. For laminar boundary layer region, the characteristic length scales in the streamwise and normal directions are L and L Re-1/ 2 , respectively. The characteristic length scale and the velocity scale in the outer region of the boundary layer are L and U, respectively. In the neighborhood region of the separated point, the length scale l<domain decomposition method is developed for the high Reynolds number flow. First, the whole domain is decomposed to different sub-domains with the different characteristic scales. Then the different dominant equation of all sub-domains is defined according to the diffusion parabolized (DP) theory of viscous flow. Finally these different equations are solved simultaneously in whole computational region. For numerical tests of high Reynolds numerical flows, two-dimensional supersonic flows over rearward and frontward steps as well as an interaction flow between shock wave and boundary layer were solved numerically. The pressure distributions and local coefficients of skin friction on the wall are given. The numerical results obtained by the multiscale-domain decomposition algorithm are well agreement with those by NS equations. Comparing with the usual method of solving the Navier-Stokes equations in the whole flow, under the same numerical accuracy, the present multiscale domain decomposition method decreases CPU consuming about 20% and reflects the physical mechanism of practical flow more accurately.

Towards multilingual domain module acquisition

Máster y Doctorado en Sistemas Informáticos Avanzados, Informatika Fakultatea - Facultad de Informática

Semi-automatic generation of learning domain modules for technology supported learning systems

In a time when Technology Supported Learning Systems are being widely used, there is a lack of tools that allows their development in an automatic or semi-automatic way. Technology Supported Learning Systems require an appropriate Domain Module, ie. the pedagogical representation of the domain to be mastered, in order to be effective. However, content authoring is a time and effort consuming task, therefore, efforts in automatising the Domain Module acquisition are necessary.Traditionally, textbooks have been used as the main mechanism to maintain and transmit the knowledge of a certain subject or domain. Textbooks have been authored by domain experts who have organised the contents in a means that facilitate understanding and learning, considering pedagogical issues.Given that textbooks are appropriate sources of information, they can be used to facilitate the development of the Domain Module allowing the identification of the topics to be mastered and the pedagogical relationships among them, as well as the extraction of Learning Objects, ie. meaningful fragments of the textbook with educational purpose.Consequently, in this work DOM-Sortze, a framework for the semi-automatic construction of Domain Modules from electronic textbooks, has been developed. DOM-Sortze uses NLP techniques, heuristic reasoning and ontologies to fulfill its work. DOM-Sortze has been designed and developed with the aim of automatising the development of the Domain Module, regardless of the subject, promoting the knowledge reuse and facilitating the collaboration of the users during the process.

Calpain-Mediated Processing of Adenylate Cyclase Toxin Generates a Cytosolic Soluble Catalytically Active N-Terminal Domain

Bordetella pertussis, the whooping cough pathogen, secretes several virulence factors among which adenylate cyclase toxin (ACT) is essential for establishment of the disease in the respiratory tract. ACT weakens host defenses by suppressing important bactericidal activities of the phagocytic cells. Up to now, it was believed that cell intoxication by ACT was a consequence of the accumulation of abnormally high levels of cAMP, generated exclusively beneath the host plasma membrane by the toxin N-terminal catalytic adenylate cyclase (AC) domain, upon its direct translocation across the lipid bilayer. Here we show that host calpain, a calcium-dependent Cys-protease, is activated into the phagocytes by a toxin-triggered calcium rise, resulting in the proteolytic cleavage of the toxin N-terminal domain that releases a catalytically active "soluble AC''. The calpain-mediated ACT processing allows trafficking of the "soluble AC'' domain into subcellular organella. At least two strategic advantages arise from this singular toxin cleavage, enhancing the specificity of action, and simultaneously preventing an indiscriminate activation of cAMP effectors throughout the cell. The present study provides novel insights into the toxin mechanism of action, as the calpain-mediated toxin processing would confer ACT the capacity for a space- and time-coordinated production of different cAMP "pools'', which would play different roles in the cell pathophysiology.