Identifica����o de compostos que modulam a patog��nese da doen��a de Machado-Joseph em C.elegans: autofagia como alvo terap��utico: identification of small compounds that modulate pathogenesis of Machado-Joseph disease in a C.elegans model: targeting autophagy

A doen��a de Machado-Joseph (DMJ) ou ataxia espinocerebelosa do tipo 3 (SCA3), conhecida por ser a mais comum das ataxias heredit��rias dominantes em todo o mundo, �� uma doen��a neurodegenerativa autoss��mica dominante que leva a uma grande incapacidade motora, embora sem alterar o intelecto, culminando com a morte do doente. Atualmente n��o existe nenhum tratamento eficaz para esta doen��a. A DMJ �� resultado de uma altera����o gen��tica causada pela expans��o de uma sequ��ncia poliglutam��nica (poliQ), na regi��o C-terminal do gene que codifica a prote��na ataxina-3 (ATXN3). Os mecanismos celulares das doen��as de poliglutaminas que provocam toxicidade, bem como a fun����o da ATXN3, n��o s��o ainda totalmente conhecidos. Neste trabalho, usamos, pela sua simplicidade e potencial gen��tico, um pequeno animal invertebrado, o nem��tode C. elegans, com o objetivo de identificar f��rmacos eficazes para o combate contra a patog��nese da DMJ, analisando simultaneamente o seu efeito na agrega����o da ATXN3 mutante nas c��lulas neuronais in vivo e o seu impacto no comportamento motor dos animais. Este pequeno invertebrado proporciona grandes vantagens no estudo dos efeitos t��xicos de prote��nas poliQ nos neur��nios, uma vez que a transpar��ncia das suas 959 c��lulas (das quais 302 s��o neur��nios) facilita a dete����o de prote��nas fluorescentes in vivo. Para al��m disso, esta esp��cie tem um ciclo de vida curto, �� econ��mica e de f��cil manuten����o. Neste trabalho test��mos no nosso modelo transg��nico da DMJ com 130Qs em C.elegans dois compostos potencialmente moduladores da agrega����o da ATXN3 mutante e da resultante disfun����o neurol��gica, atuando pela via da autofagia. De modo a validar a poss��vel import��ncia terap��utica da ativa����o da autofagia os compostos candidatos escolhidos foram o Lit��o e o an��logo da Rapamicina CCI-779, testados independentemente e em combina����o. A neuroprote����o conferida pelo Lit��o e pelo CCI-779 independentemente sugere que o uso destes f��rmacos possa ser considerado uma boa estrat��gia como terapia para a DMJ, a testar em organismos evolutivamente mais pr��ximos do humano. A manipula����o da autofagia, segundo v��rios autores, parece ser ben��fica e pode ser a chave para o desenvolvimento de novos tratamentos para v��rias doen��as relacionadas com a agrega����o proteica e o envelhecimento.

Identification of neuronal alterations induced by Shank3 mutations using iPS cells from Phelan-McDermid Patients' Fibroblasts

A fam��lia de prote��nas Shank �� o principal conjunto de proteinas de suporte e est�� localizada na densidade p��s-sin��ptica das sinapses excitat��rias. Existem 3 genes na fam��lia Shank, Shank1, Shank2 e Shank3 e s��o caracterizados por m��ltiplos dom��nios repetidos de anquirina pr��ximo ao N-terminal seguido pelos dom��nios Src homologo 3 e PDZ, uma regi��o longa rica em prolina e um dom��nio de motivo �� est��ril pr��ximo ao C-terminal. Shank prote��nas conectam duas subunidades de receptors glutamat��rgicos, recetores NMDA e recetores metabotr��picos de glutamato do tipo-I (mGluRs). O dom��nio PDZ da Shank conecta-se ao C-terminal do GKAP e este, liga-se, ao complexo recetor PSD-95-NMDA. Por outro lado, a prote��na Homer interage com o dom��nio rico em prolina para confirmar a associa����o entre a prote��na Shank com o mGluR tipo-I. A prote��na espec��fica em estudo, Shank3, �� haploinsuficiente em pacientes com sindrome Phelan-McDermid devido �� dele����es no bra��o comprido do cromossoma 22 levando �� danos intelectuais, aus��ncia ou atraso no discurso, comportamentos semelhantes ao autismo, hipotonia e caracter��sticas dism��rficas. Neste trabalho, investigamos o papel da Shank3 na fun����o sin��ptica para compreender a rela����o entre altera����es nesta prote��na e as caracter��sticas neurol��gicas presente em Pacientes com s��ndrome Phelan-McDermid. Foram utilizados dois modelos diferentes, ratinhos knockout Shank3 e hiPSC de pacientes com PMS. Ratinhos geneticamente modificados s��o ferramentas uteis no estudo de genes e na compreens��o dos mecanismos que experi��ncias in vitro n��o s��o capazes de reproduzir, mas de maneira a compreender melhor as patologias humanas, decidimos trabalhar tamb��m com c��lulas humanas. Os fibroblastos dos pacientes com s��ndrome Phelan-McDermid fora reprogramados em hiPS cells, diferenciados em neur��nios e comparados com os neur��nios obtidos a partir de doadores saudav��is e da mesma idade. A reprograma����o em iPSC foi realizada por infec����o de lentivirus com quatro genes de reprograma����o OCT4, c-MYC, SOX2 e KFL4 para posteriormente serem diferenciados em neur��nios, com cada passo sendo positivamente confirmado atrav��s de marcadores neuronais. Atrav��s dos neur��nios diferenciados, analisamos a express��o de prote��nas sin��pticas. Pacientes com haploinsuficiencia na prote��na Shank3 apresentam n��veis elevados de prote��na mGluR5 e decrescidos de prote��na Homer sugerindo que a haploinsuficiencia leva a desregula����o do complexo mGluR5-Homer-Shank3 conduzindo tamb��m, a defeitos na matura����o sin��ptica. Assim, a express��o da prote��na mGluR5 est�� alterada nos pacientes com PMS podendo estar relacionada com defeitos encontrados na diferencia����o neuronal e matura����o sin��ptica observados nos neur��nios de pacientes. Conclusivamente, iPS cells representam um modelo fundamental no estudo da prote��na Shank3 e a sua influ��ncia no sindrome de Phelan-McDermid.

Contribution of hospital effluents to the load of pharmaceuticals in urban wastewaters: Identification of ecologically relevant pharmaceuticals

The impact of effluent wastewaters from four different hospitals: a university (1456 beds), a general (350 beds), a pediatric (110 beds) and a maternity hospital (96 beds), which are conveyed to the same wastewater treatment plant (WWTP), was evaluated in the receiving urban wastewaters. The occurrence of 78 pharmaceuticals belonging to several therapeutic classes was assessed in hospital effluents and WWTP wastewaters (influent and effluent) as well as the contribution of each hospital in WWTP influent in terms of pharmaceutical load. Results indicate that pharmaceuticals are widespread pollutants in both hospital and urban wastewaters. The contribution of hospitals to the input of pharmaceuticals in urban wastewaters widely varies, according to their dimension. The estimated total mass loadings were 306 g d��� 1 for the university hospital, 155 g d��� 1 for the general one, 14 g d��� 1 for the pediatric hospital and 1.5 g d��� 1 for the maternity hospital, showing that the biggest hospitals have a greater contribution to the total mass load of pharmaceuticals. Furthermore, analysis of individual contributions of each therapeutic group showed that NSAIDs, analgesics and antibiotics are among the groups with the highest inputs. Removal efficiency can go from over 90% for pharmaceuticals like acetaminophen and ibuprofen to not removal for ��-blockers and salbutamol. Total mass load of pharmaceuticals into receiving surface waters was estimated between 5 and 14 g/d/1000 inhabitants. Finally, the environmental risk posed by pharmaceuticals detected in hospital and WWTP effluents was assessed by means of hazard quotients toward different trophic levels (algae, daphnids and fish). Several pharmaceuticals present in the different matrices were identified as potentially hazardous to aquatic organisms, showing that especial attention should be paid to antibiotics such as ciprofloxacin, ofloxacin, sulfamethoxazole, azithromycin and clarithromycin, since their hazard quotients in WWTP effluent revealed that they could pose an ecotoxicological risk to algae.

Brewer's spent grain from different types of malt: Evaluation of the antioxidant activity and identification of the major phenolic compounds

The antioxidant activity and phenolic composition of brewer's spent grain (BSG) extracts obtained by microwave-assisted extraction from twomalt types (light and darkmalts) were investigated. The total phenolic content (TPC) and antioxidant activity among the light BSG extracts (pilsen, melano, melano 80 and carared)were significantly different (p b 0.05) compared to dark extracts (chocolate and black types), with the pilsen BSG showing higher TPC (20 �� 1 mgGAE/g dry BSG). In addition, the antioxidant activity assessed by 2,2-diphenyl- 1-picrylhydrazyl, 2,2���-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and deoxyribose assays decreased as a result of increasing kilning temperatures in the following order: pilsen N melano N melano 80 N carared N chocolate N black. HPLC-DAD/ESI-MS/MS analysis indicated the presence of phenolic acids, such as ferulic, p-coumaric and syringic acids, as well as several isomeric ferulate dehydrodimers and one dehydrotrimer. Chocolate and black extracts, obtained frommalts submitted to the highest kilning temperatures, showed the lowest levels of ferulic and p-coumaric acids. These results suggested that BSG extracts from pilsen malt might be used as an inexpensive and good natural source of antioxidants with potential interest for the food, pharmaceutical and/or cosmetic industries after purification.

PAH air pollution at a Portuguese urban area: carcinogenic risks and sources identification

This study aimed to characterize air pollution and the associated carcinogenic risks of polycyclic aromatic hydrocarbon (PAHs) at an urban site, to identify possible emission sources of PAHs using several statistical methodologies, and to analyze the influence of other air pollutants and meteorological variables on PAH concentrations.The air quality and meteorological data were collected in Oporto, the second largest city of Portugal. Eighteen PAHs (the 16 PAHs considered by United States Environment Protection Agency (USEPA) as priority pollutants, dibenzo[a,l]pyrene, and benzo[j]fluoranthene) were collected daily for 24 h in air (gas phase and in particles) during 40 consecutive days in November and December 2008 by constant low-flow samplers and using polytetrafluoroethylene (PTFE) membrane filters for particulate (PM10 and PM2.5 bound) PAHs and pre-cleaned polyurethane foam plugs for gaseous compounds. The other monitored air pollutants were SO2, PM10, NO2, CO, and O3; the meteorological variables were temperature, relative humidity, wind speed, total precipitation, and solar radiation. Benzo[a]pyrene reached a mean concentration of 2.02 ngm���3, surpassing the EU annual limit value. The target carcinogenic risks were equal than the health-based guideline level set by USEPA (10���6) at the studied site, with the cancer risks of eight PAHs reaching senior levels of 9.98��10���7 in PM10 and 1.06��10���6 in air. The applied statistical methods, correlation matrix, cluster analysis, and principal component analysis, were in agreement in the grouping of the PAHs. The groups were formed according to their chemical structure (number of rings), phase distribution, and emission sources. PAH diagnostic ratios were also calculated to evaluate the main emission sources. Diesel vehicular emissions were the major source of PAHs at the studied site. Besides that source, emissions from residential heating and oil refinery were identified to contribute to PAH levels at the respective area. Additionally, principal component regression indicated that SO2, NO2, PM10, CO, and solar radiation had positive correlation with PAHs concentrations, while O3, temperature, relative humidity, and wind speed were negatively correlated.

Demo: automatic personal identification system for security in critical services: a case study

The demonstration proposal moves from the capabilities of a wireless biometric badge [4], which integrates a localization and tracking service along with an automatic personal identification mechanism, to show how a full system architecture is devised to enable the control of physical accesses to restricted areas. The system leverages on the availability of a novel IEEE 802.15.4/Zigbee Cluster Tree network model, on enhanced security levels and on the respect of all the users' privacy issues.

Modeling of the lung impedance using a fractional order ladder network with constant phase elements

The self similar branching arrangement of the airways makes the respiratory system an ideal candidate for the application of fractional calculus theory. The fractal geometry is typically characterized by a recurrent structure. This study investigates the identification of a model for the respiratory tree by means of its electrical equivalent based on intrinsic morphology. Measurements were obtained from seven volunteers, in terms of their respiratory impedance by means of its complex representation for frequencies below 5 Hz. A parametric modeling is then applied to the complex valued data points. Since at low-frequency range the inertance is negligible, each airway branch is modeled by using gamma cell resistance and capacitance, the latter having a fractional-order constant phase element (CPE), which is identified from measurements. In addition, the complex impedance is also approximated by means of a model consisting of a lumped series resistance and a lumped fractional-order capacitance. The results reveal that both models characterize the data well, whereas the averaged CPE values are supraunitary and subunitary for the ladder network and the lumped model, respectively.

Study of Proposed Identification of Non-Technical Losses of Electricity in Brazil

The electric utilities have large revenue losses annually due to commercial losses, which are caused mainly by fraud on the part of consumers and faulty meters. Automatic detection of such losses where there is a complex problem, given the large number of consumers and the high cost of each inspection, not to mention the wear of the relationship between company and consumer. Given the above, this paper aims to briefly present some methodologies applied by utilities to identify consumer frauds.

Multidimensional Scaling Visualization using Parametric Similarity Indices

In this paper, we apply multidimensional scaling (MDS) and parametric similarity indices (PSI) in the analysis of complex systems (CS). Each CS is viewed as a dynamical system, exhibiting an output time-series to be interpreted as a manifestation of its behavior. We start by adopting a sliding window to sample the original data into several consecutive time periods. Second, we define a given PSI for tracking pieces of data. We then compare the windows for different values of the parameter, and we generate the corresponding MDS maps of ���points���. Third, we use Procrustes analysis to linearly transform the MDS charts for maximum superposition and to build a global MDS map of ���shapes���. This final plot captures the time evolution of the phenomena and is sensitive to the PSI adopted. The generalized correlation, the Minkowski distance and four entropy-based indices are tested. The proposed approach is applied to the Dow Jones Industrial Average stock market index and the Europe Brent Spot Price FOB time-series.