APP RERMS enriched matrix as an adhesion substrate for neuritic outgrowth

Specific domains can determine protein structural functional relationships. For the Alzheimer’s Amyloid Precursor Protein (APP) several domains have been described, both in its intracellular and extracellular fragments. Many functions have been attributed to APP including an important role in cell adhesion and cell to cell recognition. This places APP at key biological responses, including synaptic transmission. To fulfil these functions, extracellular domains take on added significance. The APP extracellular domain RERMS is in fact a likely candidate to be involved in the aforementioned physiological processes. A multidisciplinary approach was employed to address the role of RERMS. The peptide RERMS was crosslinked to PEG (Polyethylene glycol) and the reaction validated by FTIR (Fourier transform infrared spectrometry). FTIR proved to be the most efficient at validating this reaction because it requires only a drop of sample, and it gives information about the reactions occurred in a mixture. The data obtained consist in an infrared spectra of the sample, where peaks positions give information about the structure of the molecules, and the intensity of peaks is related to the concentration of the molecules. Subsequently substrates of PEG impregnated with RERMS were prepared and SH-SY5Y (human neuroblastoma cell line) cells were plated and differentiated on the latter. Several morphological alterations were clearly evident. The RERMS peptide provoked cells to take on a flatter appearance and the cytoskeletal architecture changed, with the appearance of stress fibres, a clear indicator of actin reorganization. Given that focal adhesions play a key role in determining cellular structure the latter were directly investigated. Focal adhesion kinase (FAK) is one of the most highly expressed proteins in the CNS (central nervous system) during development. It has been described to be crucial for radial migration of neurons. FAK can be localized in growth cones and mediated the response to attractive and repulsive cues during migration. One of the mechanisms by which FAK becomes active is by auto phosphorylation at tyrosine 397. It became clearly evident that in the presence of the RERMS peptide pFAK staining at focal adhesions intensified and more focal adhesions became apparent. Furthermore speckled structures in the nucleus, putatively corresponding to increased expression activity, also increased with RERMS. Taken together these results indicate that the RERMS domain in APP plays a critical role in determining cellular physiological responses. Here is suggested a model by which RERMS domain is recognized by integrins and mediate intracellular responses involving FAK, talin, actin filaments and vinculin. This mechanism probably is responsible for mediating cell adhesion and neurite outgrowth on neurons.

Extraction and purification of immunoglobulin G with aqueous biphasic systems

The immune system is able to produce antibodies, which have the capacity to recognize and to bind to foreign molecules or pathogenic organisms. Currently, there are a diversity of diseases that can be treated with antibodies, like immunoglobulins G (IgG). Thereby, the development of cost-efficient processes for their extraction and purification is an area of main interest in biotechnology. Aqueous biphasic systems (ABS) have been investigated for this purpose, once they allow the reduction of costs and the number of steps involved in the process, when compared with conventional methods. Nevertheless, typical ABS have not showed to be selective, resulting in low purification factors and yields. In this context, the addition of ionic liquids (ILs) as adjuvants can be a viable and potential alternative to tailor the selectivity of these systems. In this work, ABS composed of polyethylene glycol (PEG) of different molecular weight, and a biodegradable salt (potassium citrate) using ILs as adjuvants (5 wt%), were studied for the extraction and purification of IgG from a rabbit source. Initially, it was tested the extraction time, the effect on the molecular weight of PEG in a buffer solution of K3C6H5O7/C6H8O7 at pH≈7, and the effect of pH (59) on the yield (YIgG) and extraction efficiency (EEIgG%) of IgG. The best results regarding EEIgG% were achieved with a centrifugation step at 1000 rpm, during 10 min, in order to promote the separation of phases followed by 120 min of equilibrium. This procedure was then applied to the remaining experiments. The results obtained in the study of PEGs with different molecular weights, revealed a high affinity of IgG for the PEG-rich phase, and particularly for PEGs of lower molecular weight (EEIgG% of 96 % with PEG 400). On the other hand, the variation of pH in the buffer solution did not show a significant effect on the EEIgG%. Finally, it was evaluated the influence of the addition of different ILs (5% wt) on the IgG extraction in ABS composed of PEG 400 at pH≈7. In these studies, it was possible to obtain EEIgG% of 100% with the ILs composed of the anions [TOS]-, [CH3CO2]-and Cl-, although the obtained YIgG% were lower than 40%. On the other hand, the ILs composed of the anions Br-, as well as of the cation [C10mim]+, although not leading to EEIgG% of 100%, provide an increase in the YIgG%. ABS composed of PEG, a biodegradable organic salt and ILs as adjuvants, revealed to be an alternative and promising method to purify IgG. However, additional studies are still required in order to reduce the loss of IgG.

Nanotubos de carbono de parede simples funcionalizados com polietilenoglicol: avaliação de parâmetros in vivo e in vitro

Os nanomateriais apresentam uma escala na qual ao menos uma das dimensões varia entre 1 e 100 nm e possuem propriedades químicas, físicas ou biológicas dependentes da nanoestrutura e que lhes confere características funcionais de interesse para fins comerciais ou aplicações na área médica. Dentre os nanomateriais mais estudados e utilizados, destacam-se os de carbono, que incluem os fulerenos e os nanotubos de carbono (NT). Uma potencial utilização dos nanomateriais de carbono é na área biomédica, já que estes podem interagir com os sistemas biológicos em nível molecular e supramolecular com alto grau de especificidade. Em contrapartida, é importante considerar que os nanotubos de carbono podem exercer efeitos tóxicos, tendo como possível mecanismo o estresse oxidativo. Sendo assim, o objetivo desse trabalho foi investigar a ação dos nanotubos de carbono de parede única funcionalizados com polietilenoglicol (SWNT-PEG) em Danio rerio “zebrafish” (Teleostei, Cyprinidae). Avaliaram-se parâmetros bioquímicos, histológicos, comportamentais e de biodistribuição para entender como esse material se comporta in vitro e in vivo. Foi observado que o tipo de funcionalização é determinante para a ação desse material em meio biológico. No experimento in vitro o SWNT-PEG não mostrou efeito pró-oxidante nas avaliações de peroxidação lipídica, capacidade antioxidante total, conteúdo de GSH e atividade de GCL. Na exposição intraperitoneal em zebrafish constatou-se a agregação e geração de processo inflamatório, o que sugere que a cadeia de PEG utilizada para a funcionalização dos NT possui um tamanho inadequado e/ou uma funcionalização ineficiente para manter a estabilidade do material em meio biológico e evitar uma resposta inflamatória por parte do organismo exposto. Possivelmente devido a esta característica do nanomaterial, nas análises de biodistribuição, através de espectroscopia Raman, não se observou distribuição de SWNT-PEG no sistema nervoso central de zebrafish. No entanto, através da análise histológica foi observado processo inflamatório no tecido nervoso central, bem como alterações comportamentais avaliadas na tarefa de campo aberto.

Badanie metodą EPR funkcjonalizowanych nanocząstek magnetytu w surowicy i pełnej krwi ludzkiej

Wydział Fizyki

Does Epoetin Beta Still Have a Place in Peginterferon Alpha-2a Plus Ribavirin Treatment Strategies for Chronic Hepatitis C?

International audience

Interventions for dysphagia in acute stroke (Review)

Background It is unclear how dysphagic patients should be fed and treated after acute stroke. Objectives The objective of this review was to assess the effect of different management strategies for dysphagic stroke patients, in particular how and when to feed, whether to supplement nutritional intake, and how and whether to treat dysphagia. Search strategy We searched the Cochrane Stroke Group trials register, Medline, Embase, ISI, and existing review articles.We contacted researchers in the field and equipment manufacturers. Date of the most recent searches: March 1999. Selection criteria Unconfounded truly or quasi randomised controlled trials in dysphagic patients with acute/subacute (within 3 months) stroke. Data collection and analysis Three reviewers independently applied the trial inclusion criteria. Two reviewers assessed trial quality and extracted the data. Main results Percutaneous endoscopic gastrostomy (PEG) versus nasogastric tube (NGT) feeding: two trials (49 patients) suggest that PEG reduces end-of-trial case fatality (Peto Odds Ratio, OR 0.28, 95% CI 0.09 to 0.89) and treatment failures (OR 0.10, 95% CI 0.02 to 0.52), and improves nutritional status, assessed as weight (Weighted Men Difference, WMD +4.1 kg, 95% CI -4.3 to +12.5), mid-arm circumference (WMD +2.2 cm, 95% CI -0.5 to +4.9) or serum albumin (WMD + 7.0 g/l, 95% CI +4.9 to +9.1) as compared with NGT feeding; two larger studies are ongoing. Timing of feeding: no completed trials; one large study is ongoing. Swallowing therapy for dysphagia: two trials (85 patients) suggest that formal swallowing therapy does not significantly reduce end-of-trial dysphagia rates (OR 0.55, 95%CI 0.18 to 1.66). Drug therapy for dysphagia: one trial (17 patients); nifedipine did not alter end-of-trial case fatality or the frequency of dysphagia. Nutritional supplementation: one trial (42 patients) found a non-significant trend to a lower case fatality, and significantly increased energy and protein intake; one large trial is ongoing and data is awaited from two other studies. Fluid supplementation: one trial (20 patients) found that supplementation did not alter the time to resolution of dysphagia. Authors’ conclusions Too few studies have been performed, and these have involved too few patients. PEG feeding may improve outcome and nutrition as compared with NGT feeding. Further research is required to assess how and when patients are fed, and the effect of swallowing or drug therapy on dysphagia.

Factores determinantes de la pérdida de valor accionario en octubre de 2008: aplicación a las empresas que integran el índice de precios y cotizaciones de la Bolsa Mexicana de Valores

La crisis financiera global de 2008, golpeó fuertemente a todas las economías del mundo y puso de manifiesto la debilidad con la que se mueve el mudo de las finanzas. La velocidad con la que se mueve la información a través de nuevas y mejoradas tecnologías de información; productos financieros más sofisticados y hasta cierto punto, difíciles de interpretar; instituciones financieras cuyo principal objetivo es ganar ofreciendo productos financieros a empresas, que en la gran mayoría de las veces, no son los que necesitan. Durante el período de la crisis financiera, la economía mexicana no fue la excepción, la mayoría de las empresas en México, experimentaron pérdidas importantes que ocasionó incluso, el cierre de operacione en algunos casos. En esta Tesis, se investigó el comportamiento accionario de las empresas que integraron el IPyC de la Bolsa Mexicana de Valores en el segundo semestre de 2008, se observó que tomando el punto más alto que alcanzó el índice en el 2008 comparado con el momento en que la crisis pegó en la economía global (Octubre 2008), todas las empresas sin excepción, mostraron pérdidas en sus valores accionarios. Continuando con la observación del índice accionario mexicano, durante el 2009, al momento en el que el Índice recupera el mejor nivel que se observó en el 2008, se podría asumir que todas las empresas tendrían la misma recuperación, sin embargo esto no sucedió así, hubo empresas que habiendo tenido un buen desempeño accionario en el 2008, continuaron presentando pérdidas aún de que el Índice se había recuperado en su totalidad. A raíz de esta observación se investigaron las causas que ocasionaron este desfase en la recuperación del valor accionario de las empresas que integran el Índice. Para cada compañía, se analizaron diversas variables que a través de una análisis de correlación y de la aplicación de las variables en las regresiones, se seleccionaron solo tres variables que fueron las más representativas: Los movimientos del tipo de cambio, la utilidad operativa más la depreciación y amortización (UAFIRDA) y las variaciones en el capital contable. Aunado a lo anterior, se analizaron los resultados obtenidos por la gestión del personal de las áreas financieras, esto para analizar el impacto que sus decisiones pudieron haber contribuido en la pérdida o en la recuperación del valor accionario en las diferentes organizaciones. En la investigación se utilizó el modelo de datos panel estimando los efectos aleatorios en las unidades (empresas) con un análisis transversal en el momento en que detona la crisis y longitudinal para el análisis de 32 períodos trimestrales. Los resultados obtenidos muestran evidencia significativa de que la pérdida accionaria está estrechamente relacionada a la toma de decisiones de la administración sobre la utilización de productos financieros y a la alta exposición a las variaciones del tipo de cambio. Por lo tanto, se rechaza la hipótesis nula planteada y se concluye que la falta de conocimiento y habilidades técnicas de los administradores de las empresas que integran el IPyC de la BMV, provocaron la no recuperación con el mismo dinamismo que se recuperó el IPyC.

"Os Conflitos ambientais em Portugal (1974-2015): uma breve retrospectiva"

In this chapter, the authors made a survey of the research undertaken by social scientists and their reflections on environmental conflicts in Portugal. In these, a critical discourse has emerged concerning, on the one hand, the weak public environmental awareness, and, on the other, the progressive obliteration of environmental movements and their institutionalization throughout the creation of different environmental groups and the incorporation of the “environment” in legislation and in political discourse. In a brief retrospective, we review several conflicts taking into account different relevant factors: level of mobilization, media attention received, organization, impact, and ideological reference they have had in Portugal since the seventies. We have particularly highlighted the movement against nuclear energy and the construction of dams, the pollution caused by intensive breeding, the expansion of eucalyptus plantations, the conflicts against “wild” forms of mining, the business of toxic waste, the expansion of the economy of the concrete, the installation of landfills, and the defense of the natural heritage. This survey has considered three periods: the 1970s, marked by the emergence and performance of ecological movements of different ideological extraction; the second half of the 1980s, marked by the institutionalization of the environment and the imposition of a new legal framework with impact on environmental policies resulting from the integration into the European Economic Community; and finally, a third period, from the 1990s to present, marked by increasing environmental media coverage, with a particular emphasis on environmental conflicts in a context of an increasing liberalization of economic activities and the expansion of extraction and of the concrete economy. This is also the period where the environment emerges in disputes over the uses of the territory as economic and asset value. Most of these conflicts arise from the activities of local agents or national environmental groups that quickly gain strong local roots and sometimes even have some success. However, their impact on the national and Community legislation seems to be less relevant.

Conflitos ambientais e progresso técnico na indústria mineira e metalúrgica em Portugal (1858-1938)

This chapter explores the relationship between environmental conflicts and technical progress, trying to understand, in the case of large mines of the Iberian Pyrite Belt, in Alentejo, how emerging environmental problems conditioned the performance or led to the search for alternative technical solutions, taking as chronological limit for this observation the beginning of World War II. In the absence of the archives of the companies, the research was based on existing administrative documents in the state archives (mining engineers reports, the licensing of mining activities), on reports and documents published in specialized mining press, in particular, the Bulletin of the Ministry of Public Works, Trade and Industry, the Journal of Public Works, Trade and Industry (both in Portuguese), and finally in the local press. Despite that limitation, the information available shows that in global competition markets, the success of the British enterprise in Santo Domingo had the active search for new technical solutions for the creation and adaptation of existing knowledge to local problems in order to maximize the mineral resources available. The early development of the hydrometallurgical processes for the treatment of poor ores, named ‘natural cementation’, can be explained as the way these companies tried to solve problems of competitiveness, boosting economies of scale. Thus, they transferred the environmental costs previously limited to agriculture to more fragile social groups, the poor fishermen of Guadiana River and of Vila Real de Santo António. Therefore, the hydrometallurgy of pyrites was developed locally, pioneered in Santo Domingo that allowed the survival and expansion of the British company from the late 1870s, that is, at a time when most small mines shut since they were not able to compete globally. Through different consented and regulated processes (judicial), through conflict or parliamentary mediation, the State imposed exceptionally additional costs to companies, either for compensation, the imposing the application of remediation measures to reduce the environmental damage in some cases, thus contributing to derail some projects. These cases suggest that the interaction between local conflicts, corporate behavior and technological progress proves to be complex. This article aims to contribute to the debate on economic and social history between the environment and technological progress, arguing that the fixed costs and economic imponderable social risks were factors that encouraged the companies to search for new solutions and to introduce innovations since that would allow the expansion of their activity. In this process the companies sometimes faced environmental dilemmas and unforeseen costs with consequences on the economy of firms. The nature of the knowledge needed to address the environmental problems they created, however, is of a very different nature from that knowledge needed to face the environmental burdens that were inherent to the development of its activity.

O passado e o presente nos conflitos ambientais na indústria mineira e metalúrgica

This chapter reflects on several forthcoming key topics on environmental social conflits in historical perspective, namely, the relationship between the expansion of extractivism and the reactive and preemptive conflicts, the role of scientists and scientific discourses on social mobilization, the environmental movements and the empowerment of the poor, the strategic response behavior of large mining corporations. The conclusive analysis looks for continuities and contrasts between contemporary environmental conflicts and the conflicts of the past, often identified as 'peasant conflicts' over industrial polution.

TRANSFORMATIONS OF TASK-DEPENDENT PLASTICITY FROM A1 TO HIGHER-ORDER AUDITORY CORTEX

Everyday, humans and animals navigate complex acoustic environments, where multiple sound sources overlap. Somehow, they effortlessly perform an acoustic scene analysis and extract relevant signals from background noise. Constant updating of the behavioral relevance of ambient sounds requires the representation and integration of incoming acoustical information with internal representations such as behavioral goals, expectations and memories of previous sound-meaning associations. Rapid plasticity of auditory representations may contribute to our ability to attend and focus on relevant sounds. In order to better understand how auditory representations are transformed in the brain to incorporate behavioral contextual information, we explored task-dependent plasticity in neural responses recorded at four levels of the auditory cortical processing hierarchy of ferrets: the primary auditory cortex (A1), two higher-order auditory areas (dorsal PEG and ventral-anterior PEG) and dorso-lateral frontal cortex. In one study we explored the laminar profile of rapid-task related plasticity in A1 and found that plasticity occurred at all depths, but was greatest in supragranular layers. This result suggests that rapid task-related plasticity in A1 derives primarily from intracortical modulation of neural selectivity. In two other studies we explored task-dependent plasticity in two higher-order areas of the ferret auditory cortex that may correspond to belt (secondary) and parabelt (tertiary) auditory areas. We found that representations of behaviorally-relevant sounds are progressively enhanced during performance of auditory tasks. These selective enhancement effects became progressively larger as you ascend the auditory cortical hierarchy. We also observed neuronal responses to non-auditory, task-related information (reward timing, expectations) in the parabelt area that were very similar to responses previously described in frontal cortex. These results suggests that auditory representations in the brain are transformed from the more veridical spectrotemporal information encoded in earlier auditory stages to a more abstract representation encoding sound behavioral meaning in higher-order auditory areas and dorso-lateral frontal cortex.

Porous self-protonating spiropyran-based NIPAAm gels with improved reswelling kinetics

The final publication is available at Springer via http://dx.doi.org/[10.1007/s10853-015-9458-2]

REGULATION OF SHAPE DYNAMICS AND ACTIN POLYMERIZATION DURING COLLECTIVE CELL MIGRATION

This thesis aims to understand how cells coordinate their motion during collective migration. As previously shown, the motion of individually migrating cells is governed by wave-like cell shape dynamics. The mechanisms that regulate these dynamic behaviors in response to extracellular environment remain largely unclear. I applied shape dynamics analysis to Dictyostelium cells migrating in pairs and in multicellular streams and found that wave-like membrane protrusions are highly coupled between touching cells. I further characterized cell motion by using principle component analysis (PCA) to decompose complex cell shape changes into a serial shape change modes, from which I found that streaming cells exhibit localized anterior protrusion, termed front narrowing, to facilitate cell-cell coupling. I next explored cytoskeleton-based mechanisms of cell-cell coupling by measuring the dynamics of actin polymerization. Actin polymerization waves observed in individual cells were significantly suppressed in multicellular streams. Streaming cells exclusively produced F-actin at cell-cell contact regions, especially at cell fronts. I demonstrated that such restricted actin polymerization is associated with cell-cell coupling, as reducing actin polymerization with Latrunculin A leads to the assembly of F-actin at the side of streams, the decrease of front narrowing, and the decoupling of protrusion waves. My studies also suggest that collective migration is guided by cell-surface interactions. I examined the aggregation of Dictyostelim cells under distinct conditions and found that both chemical compositions of surfaces and surface-adhesion defects in cells result in altered collective migration patterns. I also investigated the shape dynamics of cells suspended on PEG-coated surfaces, which showed that coupling of protrusion waves disappears on touching suspended cells. These observations indicate that collective migration requires a balance between cell-cell and cell-surface adhesions. I hypothesized such a balance is reached via the regulation of cytoskeleton. Indeed, I found cells actively regulate cytoskeleton to retain optimal cell-surface adhesions on varying surfaces, and cells lacking the link between actin and surfaces (talin A) could not retain the optimal adhesions. On the other hand, suspended cells exhibited enhanced actin filament assembly on the periphery of cell groups instead of in cell-cell contact regions, which facilitates their aggregation in a clumping fashion.

Differential expression in “rogue” paramutation in peas (Pisum sativum L.)

Dissertação de Mestrado, Engenharia Biológica, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2014

Flixweed vs. Polyethylene Glycol in the Treatment of Childhood Functional Constipation: A Randomized Clinical Trial

Background: Polyethylene glycol (PEG) is often considered as the first-line treatment for functional constipation in children. Descurainia sophia (L.) Webb et Berth (D. sophia) is a safe recommended medicine in Iranian folk and Traditional Persian Medicine for the treatment of constipation. Objectives: To clinically compare D. sophia with PEG 4000 (without electrolyte) in pediatric constipation and to assess its efficacy and side effects. Patients and Methods: 120 patients aged 2 - 12 years with constipation for at least 3 months were included in an 8 weeks lasting randomized controlled trial within two parallel-groups. Children received either PEG, 0.4 g/kg/day, or D. sophia seeds, 2 grams (for children aged 2 - 4 years) and 3 grams (for those aged > 4 years) per day. Results: A total of 109 patients completed the study (56 in D. sophia and 53 in PEG group). At the end of the study, 36 (64.3%) patients in D. sophia group and 29 (54.7%) in PEG group were out of Rome III criteria (P = 0.205). Median weekly stool frequency in 0, 1, 2, 3 weeks of the treatment was found to be 2, 5, 5, 5 in D. sophia and 3, 4, 4, 5 in PEG group (P = 0.139, 0.076, 0.844, 0.294), respectively. The number of patients who suffered flatulence was less (5, 8.9%) in D. sophia group as compared to PEG group (6, 11.3%) at the end of the trial (P = 0.461). D. sophia taste was less tolerated. Conclusions: D. sophia is introduced as a cheap and available medication which can be applied as a safe alternative to conventional PEG in the management of pediatric chronic functional constipation.