Dissecting the role of CLASP1 in the mammalian development

A mitose é o evento celular, através do qual uma células transmite uma cópias do seu DNA às células filhas. Este processo é mediado pelo fuso mitótico, o qual consiste numa rede bipolar microtubulos. A dinâmica dos microtubulos é regulada por proteínas associadas a estes (MAPs – Microtubule-Associated Proteins), tais como as proteínas associadas às extremidades positivas dos microtubulos (+TIPs – Plus-ends Tracking proteins). As proteínas associadas às CLIPs (CLASPs – CLIP-associated proteins) pertencem a esta família e estão altamente conservadas nos eucariotas. Estas interagem com os microtubulos regulando o fuso mitótico, a segregação dos cromossomas e o comportamento dos microtubulos ao nível do cinetocoro. Assim, as CLASPs têm sido descritas como essenciais à manutenção da integridade genética durante a divisão celular. Um modelo animal knockout para o gene Clasp1 é uma ferramenta indispensável à descoberta do papel da CLASP1 a nível fisiológico. Nos animais knockout foi observado um fenótipo letal, no qual 100% dos recém-nascidos morreram poucos minutos após o nascimento, no decurso de falência respiratória. Após análise histopatológica, observamos que os pulmões dos animais knockout apresentam um atraso no desenvolvimento. Porém, a análise da expressão de marcadores de diferenciação celular, mostrou que os pneumócitos tipo I e II estão presente e diferenciados nos animais knockout aquando do seu nascimento. No entanto, um defeito primário a nível pulmonar ainda não pode ser excluído. Níveis elevados de glicogénio no parênquima alveolar dos animais knockout sugerem imaturidade pulmonar ou deficiente produção do líquido surfactante. Adicionalmente, ainda não está esclarecido de que forma pode este atraso explicar a letalidade observada nos recémnascidos knockout. Verificamos também que expressão de CLASP1 é transiente ao longo do desenvolvimento, sendo particularmente elevada no cérebro, o que pode explicar o seu papel já descrito na biologia dos neurónios. A CLASP1 é ubiquamente expressa em mamíferos adultos, o que sugere que esta proteína é também importante em tecidos diferenciados. Nesta fase, o significado biológico da CLASP1 em mamíferos ainda não foi descortinado. No entanto, nenhum animal knockout para Clasp1 foi capaz de sobreviver ex uterus, o que sugere um papel fundamental desta proteína na fase final do desenvolvimento dos mamíferos.

Supporting leadership development by exposure to role models: two case studies

The idiomatic expression “In Rome be a Roman” can be applied to leadership training and development as well. Leaders who can act as role models inspire other future leaders in their behaviour, attitudes and ways of thinking. Based on two examples of current leaders in the fields of Politics and Public Administration, I support the idea that exposure to role models during their training was decisive for their career paths and current activities as prominent characters in their profession. Issues such as how students should be prepared for community or national leadership as well as cross-cultural engagement are raised here. The hypothesis of transculturalism and cross-cultural commitment as a factor of leadership is presented. Based on current literature on Leadership as well as the presented case studies, I expect to raise a debate focusing on strategies for improving leaders’ training in their cross-cultural awareness.

Language and trade: the (new) route of portuguese in the 21st century

Globalisation has eliminated frontiers and in the case of Europe helped the crossing of borders and changed forever the concept of social (and I would also say individual) geography: the rest of the world is out there; we can pretend not to see it, but we cannot avoid it. Moreover, Europe is undergoing a serious crisis, also economic, and new markets and business partners are welcome. In this context, cultural bonds like a common language can open new investment paths and give rise to successful stories. In this paper we intend to present an example of how low linguistic distance can lead to good business, even if a) in the internationalization process of the companies’ language is often forgotten as a management element and b) consumers of language products (like User Guides) are also not stimulating investment in language by the companies. Through the results of 2 studies carried out in 2010 and 2011 we will show how a pluricentric language like Portuguese is managed in multinational companies (MC) and Small and medium Enterprises (SMEs). The second study is based on an online survey questioning the effectiveness, efficiency and general quality of User Guides and the reaction of consumers to language. Results show that although playing a role in the internationalization process of companies in the same linguistic space, language is opportunistically used. On the other hand, Portuguese and Brazilian consumers show a very functional perception of the Portuguese language...

Development of a biosensor for urea assay based on amidase inhibition, using an ion-selective electrode

A biosensor for urea has been developed based on the observation that urea is a powerful active-site inhibitor of amidase, which catalyzes the hydrolysis of amides such as acetamide to produce ammonia and the corresponding organic acid. Cell-free extract from Pseudomonas aeruginosa was the source of amidase (acylamide hydrolase, EC 3.5.1.4) which was immobilized on a polyethersulfone membrane in the presence of glutaraldehyde; anion-selective electrode for ammonium ions was used for biosensor development. Analysis of variance was used for optimization of the biosensorresponse and showed that 30 mu L of cell-free extract containing 7.47 mg protein mL(-1), 2 mu L of glutaraldehyde (5%, v/v) and 10 mu L of gelatin (15%, w/v) exhibited the highest response. Optimization of other parameters showed that pH 7.2 and 30 min incubation time were optimum for incubation ofmembranes in urea. The biosensor exhibited a linear response in the range of 4.0-10.0 mu M urea, a detection limit of 2.0 mu M for urea, a response timeof 20 s, a sensitivity of 58.245 % per mu M urea and a storage stability of over 4 months. It was successfully used for quantification of urea in samples such as wine and milk; recovery experiments were carried out which revealed an average substrate recovery of 94.9%. The urea analogs hydroxyurea, methylurea and thiourea inhibited amidase activity by about 90%, 10% and 0%, respectively, compared with urea inhibition.

Chitosan-alginate microparticulate delivery system for an alternative route of administration of BCG vaccine

Immunisation against M. tuberculosis with current available BCG vaccine lacks efficacy in preventing adult pulmonary tuberculosis. Targeting nasal mucosa is an attractive option for a more effective immunization. The delivery of BCG via the intranasal route involves overcoming barriers such as crossing the physical barrier imposed by the mucus layer and ciliar remotion, cellular uptake and intracellular trafficking by antigen presenting cells. Due to its biodegradable, immunogenic and mucoadhesive properties, chitosan particulate delivery systems can act both as vaccine carrier and adjuvant, improving the elicited immune response. In this study, different combinations of Chitosan/Alginate/TPP microparticles with BCG were produced as vaccine systems. The developed microparticle system successfully modulates BCG surface physicochemical properties and promotes effective intracellular uptake by human macrophage cell lines Preliminary immune responses were evaluated after s.c. and intranasal immunisation of BALB/c mice. BCG vaccination successfully stimulated the segregation of IgG2a and IgG1, where intranasal immunisation with chitosan/alginate particulate system efficiently elicited a more equilibrated cellular/humoral immune response.

Development of novel ionic liquids based on ampicillin

Novel ionic liquids containing ampicillin as an active pharmaceutical ingredient anion were prepared with good yields by using a new, efficient synthetic procedure based on the neutralization of a moderately basic ammonia solution of ampicillin with different organic cation hydroxides. The relevant physical and thermal properties of these novel ionic liquids based on ampicillin were also evaluated.

International sustainable development cooperation of physiotherapy course from Escola Superior de Tecnologia da Saúde de Lisboa

Objectives - To share with delegates the international experience in several Program of Physiotherapy course from Escola Superior de Tecnologia da Saúde de Lisboa (ESTeSL).

On the development of textual competence in primary education

The aim of this article is to present the results of an action research project, which has been put into practice in Primary Education. This project was intended to develop students’ textual competence, considering both comprehension and textual production. Our starting hypothesis was that teaching the schematisation of text types, focusing on linguistic devices that underlie text production, would promote the development of textual competence, leading to the production of more coherent and cohesive texts. In order to test this hypothesis we implemented the project in three phases. First, before the intervention, we collected texts produced by the students. Secondly, we implemented a didactic program designed to develop students’ textual competence. Lastly, after the intervention, we collected students’ texts once again. Data was analyzed according to categories that confer cohesion and coherence to different types of texts. Narrative, descriptive, and explanatory texts were assessed in terms of 1) building an autonomous text; 2) hierarchisation of information, and 3) textual organisation. Overall, results indicate that students developed their text conceptualisations, their understanding of the different structures of texts, and produced better writing. Indeed, their written work shows a marked progression from the beginning of the intervention program to the end of the program.

Development of novel ionic liquids based on valproate anion

Valproic acid (2-propyl pentanoic acid) is a pharmaceutical drug used for treatment of epileptic seizures absence, tonic-clonic (grand mal), complex partial seizures, and mania in bipolar disorder [1]. Valproic acid is a slightly soluble in water and therefore as active pharmaceutical ingredient it is most commonly applied in form of sodium or magnesium valproate salt [1].However the list of adverse effects of these compounds is large and includes among others: tiredness, tremor, sedation and gastrointestinal disturbances [2]. Ionic liquids (ILs) are promising compounds as Active Pharmaceutical Ingredients (APIs)[3]. In this context, the combinations of the valproate anion with appropriate cation when ILs and salts are formed can significantly alter valproate physical, chemical and thermal properties.[4] This methodology can be used for drug modification (alteration of drug solubility in water, lipids, bioavailability, etc)[2] and therefore can eliminate some adverse effect of the drugs related to drug toxicity due for example to its solubility in water and lipids (interaction with intestines). Herein, we will discuss the development of ILs based on valproate anion (Figure 1) prepared according a recent optimized and sustainable acid-base neutralization method [4]. The organic cations such as cetylpyridinium, choline and imidazolium structures were selected based on their biocompatibility and recent applications in pharmacy [3]. All novel API-ILs based on valproate have been studied in terms of their physical, chemical (viscosity, density, solubility) and thermal (calorimetric studies) properties as well as their biological activity.

Development and biological evaluation of API-ILs based on anti-bacterial and anti-fungal drugs

In recent years Ionic Liquids (ILs) are being applied in life sciences. ILs are being produce with active pharmaceutical drugs (API) as they can reduce polymorphism and drug solubility problems [1] Also ILs are being applied as a drug delivery device in innovative therapies What is appealing in ILs is the ILs building up platform, the counter-ion can be carefully chosen in order to avoid undesirable side effects or to give innovative therapies in which two active ions are paired. This work shows ILs based on ampicillin (an anti-bacterial agent) and ILs based on Amphotericin B. Also we show studies that indicate that ILs based on Ampicillin could reverse resistance in some bacteria. The ILs produced in this work were synthetized by the neutralization method described in Ferraz et. al. [2] Ampicillin anion was combined with the following organic cations 1-ethyl-3-methylimidazolium, [EMIM]; 1-hydroxy-ethyl-3-methylimidazolium, [C2OHMIM]; choline, [cholin]; tetraethylammonium, [TEA]; cetylpyridinium, [C16pyr] and trihexyltetradecylphosphonium, [P6,6,6,14]. Amphotericin B was combined with [C16pyr], [cholin] and 1-metohyethyl-3-methylimidazolium, [C3OMIM]. The ILs-APIs based on ampicillin[2] were tested against sensitive Gram-negative bacteria Escherichia coli ATCC 25922 and Klebsiella pneumonia (clinical isolated), as well as on Gram positive Staphylococcus Aureus ATCC 25923, Staphylococcus epidermidis and Enterococcus faecalis. The arising resistance developed by bacteria to antibiotics is a serious public health threat and needs new and urgent measures. We study the bacterial activity of these compounds against a panel of resistant bacteria (clinical isolated strains): E. coli CTX M9, E. coli TEM CTX M9, E. coli TEM1, E. coli CTX M2, E. coli AmpC Mox2. In this work we demonstrate that is possible to produce ILs from anti-bacterial and anti-fungal compounds. We show here that the new ILs can reverse the bacteria resistance. With the careful choice of the organic cation, it is possible to create important biological and physic-chemical properties. This work also shows that the ion-pair is fundamental in ampicillin mechanism of action.

Development of an injectable grout for concrete repair and strengthening

This paper deals with the coupled effect of temperature and silica fume addition on rheological, mechanical behaviour and porosity of grouts based on CEMI 42.5R, proportioned with a polycarboxylate-based high range water reducer. Preliminary tests were conducted to focus on the grout best able to fill a fibrous network since the goal of this study was to develop an optimized grout able to be injected in a mat of steel fibers for concrete strengthening. The grout composition was developed based on criteria for fresh state and hardened state properties. For a CEMI 42.5R based grout different high range water reducer dosages (0%, 0.2%, 0.4%, 0.5%, 0.7%) and silica fume (SF) dosages (0%, 2%, 4%) were tested (as replacement of cement by mass). Rheological measurements were used to investigate the effect of polycarboxylates (PCEs) and SF dosage on grout properties, particularly its workability loss, as the mix was to be injected in a matrix of steel fibers for concrete jacketing. The workability behaviour was characterized by the rheological parameters yield stress and plastic viscosity (for different grout temperatures and resting times), as well as the procedures of mini slump cone and funnel flow time. Then, further development focused only on the best grout compositions. The cement substitution by 2% of SF exhibited the best overall behaviour and was considered as the most promising compared to the others compositions tested. Concerning the fresh state analysis, a significant workability loss was detected if grout temperature increased above 35 degrees C. Below this temperature the grout presented a self-levelling behaviour and a life time equal to 45 min. In the hardened state, silica fumes increased not only the grout's porosity but also the grout's compressive strength at later ages, since the pozzolanic contribution to the compressive strength does not occur until 28 d and beyond. (C) 2012 Elsevier Ltd. All rights reserved.

Development and validation of the Portuguese version of the WHOQOL-OLD module

OBJECTIVE: The increasing proportion of older adults in the general population and the specific characteristics of this age group show the need for the development of specific instruments to measure quality of life in older adults. The study aimed at describing the development and validation of the Portuguese version of the World Health Organization Quality of Life for Older Persons (WHOQOL-OLD) module. METHODS: The WHOQOL-OLD instrument was administered in a sample of 424 older adults in the city of Porto Alegre, Southern Brazil, in 2005. The questionnaire comprises 24 items divided into six facets: sensory abilities; autonomy; past, present and future activities; social participation; death and dying; and intimacy. Besides the WHOQOL-OLD module, the WHOQOL-BREF, BDI and BHS instruments were also applied. The instrument's internal consistency was assessed using Cronbach's alpha coefficient. RESULTS: The instrument showed adequate internal consistency (Cronbach's coefficients ranging from 0.71 to 0.88), discriminant validity (p<0.01), concurrent validity (correlation coefficients ranging from -0.61 to -0.50) and test-retest reliability (correlation coefficients ranging from 0.58 to 0.82). Findings concerning criterion validity need further studies. CONCLUSIONS: The WHOQOL-Old module is a useful alternative with good psychometric performance in the investigation of quality of life in older adults.

Development of conceptacles in Amphiroa (Corallinales, Rhodophyta)

Here, we describe the development of sporangial and gametangial conceptacles for Amphiroa beauvoisii and A. vanbosseae; sporangial conceptacles only for A. misakiensis; and gametangial conceptacles only for A. cryptarthrodia and A. rigida. The descriptions are based on the observation of histological preparations obtained from 112 specimens collected from the Gulf of California, in Mexico, and the Azores archipelago of Portugal. Information on the development of the sporangial conceptacle pore and conceptacle senescence is here described and illustrated for the first time. Four development patterns were observed: two for sporangial conceptacles; one for spermatangial conceptacles; and one for carposporangial conceptacles. The phases of development of the sporangial conceptacle were found to be useful in delimiting species within the genus. Based on the sporangium location on the cavity floor and the pore canal anatomy, the species A. beauvoisii, A. misakiensis and A. vanbosseae can be distinguished from each other.