The role of the mammalian DNA end-processing enzyme polynucleotide kinase 3'-phosphatase in spinocerebellar ataxia Type 3 pathogenesis

DNA strand-breaks (SBs) with non-ligatable ends are generated by ionizing radiation, oxidative stress, various chemotherapeutic agents, and also as base excision repair (BER) intermediates. Several neurological diseases have already been identified as being due to a deficiency in DNA end-processing activities. Two common dirty ends, 3'-P and 5'-OH, are processed by mammalian polynucleotide kinase 3'-phosphatase (PNKP), a bifunctional enzyme with 3'-phosphatase and 5'-kinase activities. We have made the unexpected observation that PNKP stably associates with Ataxin-3 (ATXN3), a polyglutamine repeat-containing protein mutated in spinocerebellar ataxia type 3 (SCA3), also known as Machado-Joseph Disease (MJD). This disease is one of the most common dominantly inherited ataxias worldwide; the defect in SCA3 is due to CAG repeat expansion (from the normal 14-41 to 55-82 repeats) in the ATXN3 coding region. However, how the expanded form gains its toxic function is still not clearly understood. Here we report that purified wild-type (WT) ATXN3 stimulates, and by contrast the mutant form specifically inhibits, PNKP's 3' phosphatase activity in vitro. ATXN3-deficient cells also show decreased PNKP activity. Furthermore, transgenic mice conditionally expressing the pathological form of human ATXN3 also showed decreased 3'-phosphatase activity of PNKP, mostly in the deep cerebellar nuclei, one of the most affected regions in MJD patients' brain. Finally, long amplicon quantitative PCR analysis of human MJD patients' brain samples showed a significant accumulation of DNA strand breaks. Our results thus indicate that the accumulation of DNA strand breaks due to functional deficiency of PNKP is etiologically linked to the pathogenesis of SCA3/MJD.

Lightweight adjustable partition system: evaluation of resistance to horizontal loads and functional failure from impacts

This paper presents and discusses the results of the serviciability and use condition tests carried on an innovative solution for partitions, designated AdjustMembrane developed within a research project. The proposed system is a modular non-loadbearing wall, tensioned between the pavements and ceiling slabs, which are used as anchoring elements. It allows several advantages, related with the weight reduction to achieve a good sustainable performance, such as the reduction of construction costs, energy, and materials, and it is easy to recycle and to reuse, allowing self-construction. Apart from a general presentation of the partition technology, this paper presents and discusses the results of experimental tests carried out. From the results obtained, it is possible to conclude that the solution fulfils the requirements for this typology of wall in terms of resistance to horizontal loads induced by soft and hard body impacts.

Mitigating the impact of occupational noise exposure for elderly workers: Setting the functional requirements for an ANC system

Noise affects people in very different aspects and in almost every aspect of our daily life. The most prominent impact of noise exposure is hearing loss. However, it can also impair people at their work settings due to other effects rather than hearing loss. Older works tend to be more susceptible to noise exposure effects at work, firstly because most of them already have some ‘natural’ hearing loss, as a results of the ageing process, and secondly because they also tend to be more susceptible at an psychological level. The current study is an attempt to describe the potential problem and to make a survey to identify the available active noise cancelation systems, as well as to specific the main requirements of this type of systems to be applied in such contexts. Several aspects of characteristics of the ANC systems were identified and are presented in this study. From the obtained results it was possible to have a clearer idea about the potential of this technology, and to confirm that this type of solution can be extremely important as a component of an active ageing program, as the preservation of hearing will also impact on the social life of the exposed workers.

Development of computational tools for the integrated analysis of DNA microarray data with applications in cancer research

The MAP-i Doctoral Program of the Universities of Minho, Aveiro and Porto

Integrating data from heterogeneous DNA microarray platforms

DNA microarrays are one of the most used technologies for gene expression measurement. However, there are several distinct microarray platforms, from different manufacturers, each with its own measurement protocol, resulting in data that can hardly be compared or directly integrated. Data integration from multiple sources aims to improve the assertiveness of statistical tests, reducing the data dimensionality problem. The integration of heterogeneous DNA microarray platforms comprehends a set of tasks that range from the re-annotation of the features used on gene expression, to data normalization and batch effect elimination. In this work, a complete methodology for gene expression data integration and application is proposed, which comprehends a transcript-based re-annotation process and several methods for batch effect attenuation. The integrated data will be used to select the best feature set and learning algorithm for a brain tumor classification case study. The integration will consider data from heterogeneous Agilent and Affymetrix platforms, collected from public gene expression databases, such as The Cancer Genome Atlas and Gene Expression Omnibus.

Systematics, functional morphology and distribution of a bivalve (Apachecorbula muriatica gen. et sp. nov.) from the rim of the ‘Valdivia Deep’ brine pool in the Red Sea

The deep brine pools of the Red Sea comprise extreme, inhospitable habitats yet house microbial communities that potentially may fuel adjacent fauna. We here describe a novel bivalve from a deep-sea (1525 m) brine pool in the Red Sea, where conditions of high salinity, lowered pH, partial anoxia and high temperatures are prevalent. Remotely operated vehicle (ROV) footage showed that the bivalves were present in a narrow (20 cm) band along the rim of the brine pool, suggesting that it is not only tolerant of such extreme conditions but is also limited to them. The bivalve is a member of the Corbulidae and named Apachecorbula muriatica gen. et sp. nov. The shell is atypical of the family in being modioliform and thin. The semi-infaunal habit is seen in ROV images and reflected in the anatomy by the lack of siphons. The ctenidia are large and typical of a suspension feeding bivalve, but the absence of guard cilia and the greatly reduced labial palps suggest that it is non-selective as a response to low food availability. It is proposed that the low body mass observed is a consequence of the extreme habitat and low food availability. It is postulated that the observed morphology of Apachecorbula is a result of paedomorphosis driven by the effects of the extreme environment on growth but is in part mitigated by the absence of high predation pressures.

Towards high performance and multi-functional structural membranes using advanced fibrous and textile materials

Scientific and technological advancements in the area of fibrous and textile materials have greatly enhanced their application potential in several high-end technical and industrial sectors including construction, transportation, medical, sports, aerospace engineering, electronics and so on. Excellent performance accompanied by light-weight, mechanical flexibility, tailor-ability, design flexibility, easy fabrication and relatively lower cost are the driving forces towards wide applications of these materials. Cost-effective fabrication of various advanced and functional materials for structural parts, medical devices, sensors, energy harvesting devices, capacitors, batteries, and many others has been possible using fibrous and textile materials. Structural membranes are one of the innovative applications of textile structures and these novel building skins are becoming very popular due to flexible design aesthetics, durability, lightweight and cost benefits. Current demand on high performance and multi-functional materials in structural applications has motivated to go beyond the basic textile structures used for structural membranes and to use innovative textile materials. Structural membranes with self-cleaning, thermoregulation and energy harvesting capability (using solar cells) are examples of such recently developed multi-functional membranes. Besides these, there exist enormous opportunities to develop wide varieties of multi-functional membranes using functional textile materials. Additionally, it is also possible to further enhance the performance and functionalities of structural membranes using advanced fibrous architectures such as 2D, 3D, hybrid, multi-layer and so on. In this context, the present paper gives an overview of various advanced and functional fibrous and textile materials which have enormous application potential in structural membranes.

Politically driven cycles in fiscal policy: in depth analysis of the functional components of government expenditures

NIPE - WP 02/2016

Clinical characteristics of women diagnosed with carcinoma who tested positive for cervical and anal high-risk human papillomavirus DNA and E6 RNA

High-risk human papillomavirus (hrHPV) is an essential cause of cervical carcinoma and is also strongly related to anal cancer development. The hrHPV E6 oncoprotein plays a major role in carcinogenesis. We aimed to evaluate the frequency of hrHPV DNA and E6 oncoprotein in the anuses of women with cervical carcinoma. We analyzed 117 women with cervical cancer and 103 controls for hrHPV and the E6 oncogene. Positive test results for a cervical carcinoma included 66.7 % with hrHPV-16 and 7.7 % with hrHPV-18. One case tested positive for both HPV variants (0.9 %). The samples from the anal canal were positive for HPV-16 in 59.8 % of the cases. Simultaneous presence of HPV in the cervix and anal canal was found in 53.8 % of the cases. Regarding expression of E6 RNA, positivity for HPV-16 in the anal canal was found in 21.2 % of the cases, positivity for HPV-16 in the cervix was found in 75.0 %, and positivity for HPV-18 in the cervix was found in 1.9 %. E6 expression in both the cervix and anal canal was found in 19.2 % of the cases. In the controls, 1 % tested positive for HPV-16 and 0 % for HPV-18. Anal samples from the controls showed a hrHPV frequency of 4.9 % (only HPV16). The presence of hrHPV in the anal canal of women with cervical cancer was detected at a high frequency. We also detected E6 RNA expression in the anal canal of women with cervical cancer, suggesting that these women are at risk for anal hrHPV infection.

Microscopy- or DNA-based analyses: Which methodology gives a truer picture of stream-dwelling decomposer fungal diversity?

We assessed aquatic hyphomycete diversity in autumn and spring on oak leaves decomposing in five streams along a gradient of eutrophication in the Northwest of Portugal. Diversity was assessed through microscopy-based (identification by spore morphology) and DNA-based techniques (Denaturing Gradient Gel Electrophoresis and 454 pyrosequencing). Pyrosequencing revealed five times greater diversity than DGGE. About 21% of all aquatic hyphomycete species were exclusively detected by pyrosequencing and 26% exclusively by spore identification. In some streams, more than half of the recorded species would have remained undetected if we had relied only on spore identification. Nevertheless, in spring aquatic hyphomycete diversity was higher based on spore identification, probably because many species occurring in this season are not yet connected to ITS barcodes in genetic databases. Pyrosequencing was a powerful tool for revealing aquatic hyphomycete diversity on decomposing plant litter in streams and we strongly encourage researchers to continue the effort in barcoding fungal species.

Polymer electrolyte based on DNA and N,N,N-trimethyl-N-(2-hydroxyethyl)ammonium bis(trifluoromethylsulfonyl)imide

Combining ionic liquids (ILs) with polymers offers the prospect of new applications, where they surpass the performance of conventional media, such as organic solvents, giving advantages in terms of improved safety and a higher operating temperature range. In this work we have investigated the morphology, thermal and electrochemical properties of polymer electrolytes prepared through the addition of con- trolled quantities of the cholinium based IL N,N,N-trimethyl-N-(2-hydroxyethyl)ammonium bis(trifluo- romethylsulfonyl)imide ([N1 1 1 2(OH)] [NTf2]) to a deoxyribonucleic acid (DNA) host network. These novel IL-based electrolytes have been analyzed aiming at applications in electrochemical devices. An optimized sample showed good thermal stability up to 155 °C and a wide electrochemical window of ~3.5 V. The highest conductivity was registered for the DNA[N1 1 1 2(OH)][NTf2] (1:1) (2.82 × 10-5 and 1.09 × 10-3 S cm-1 at 30 and 100 °C, respectively).

Saprotrophic fungi Hypholoma fasciculare effect on the fungal community associated to Castanea sativa

Dissertação de mestrado em Biologia Molecular, Biotecnologia e Bioempreendedorismo em Plantas

Fluorescence-based approaches towards the assessment of structural and functional alterations in lysosomes and vacuoles induced by lactoferrin and acetic acid

Dissertação de mestrado em Biofísica e Bionanossistemas

Unnatural amino acids as functional building blocks for fluorophore-labelled peptides

Comunicação oral convidada - IL4

Highly functional and biodegradable nanofibrous scaffolds combined with stem cells for bone and cartilage tissue engineering

Among the various possible embodiements of Advanced Therapies and in particular of Tissue Engineering the use of temporary scaffolds to regenerate tissue defects is one of the key issues. The scaffolds should be specifically designed to create environments that promote tissue development and not merely to support the maintenance of communities of cells. To achieve that goal, highly functional scaffolds may combine specific morphologies and surface chemistry with the local release of bioactive agents. Many biomaterials have been proposed to produce scaffolds aiming the regeneration of a wealth of human tissues. We have a particular interest in developing systems based in nanofibrous biodegradable polymers1,2. Those demanding applications require a combination of mechanical properties, processability, cell-friendly surfaces and tunable biodegradability that need to be tailored for the specific application envisioned. Those biomaterials are usually processed by different routes into devices with wide range of morphologies such as biodegradable fibers and meshes, films or particles and adaptable to different biomedical applications. In our approach, we combine the temporary scaffolds populated with therapeutically relevant communities of cells to generate a hybrid implant. For that we have explored different sources of adult and also embryonic stem cells. We are exploring the use of adult MSCs3, namely obtained from the bone marrow for the development autologous-based therapies. We also develop strategies based in extra-embryonic tissues, such as amniotic fluid (AF) and the perivascular region of the umbilical cord4 (Whartonâ s Jelly, WJ). Those tissues offer many advantages over both embryonic and other adult stem cell sourcess. These tissues are frequently discarded at parturition and its extracorporeal nature facilitates tissue donation by the patients. The comparatively large volume of tissue and ease of physical manipulation facilitates the isolation of larger numbers of stem cells. The fetal stem cells appear to have more pronounced immunomodulatory properties than adult MSCs. This allogeneic escape mechanism may be of therapeutic value, because the transplantation of readily available allogeneic human MSCs would be preferable as opposed to the required expansion stage (involving both time and logistic effort) of autologous cells. Topics to be covered: This talk will review our latest developments of nanostructured-based biomaterials and scaffolds in combination with stem cells for bone and cartilage tissue engineering.