Natural and amyloid self-assembly of S100 proteins: structural basis of functional diversity

The S100 proteins are 10-12 kDa EF-hand proteins that act as central regulators in a multitude of cellular processes including cell survival, proliferation, differentiation and motility. Consequently, many S100 proteins are implicated and display marked changes in their expression levels in many types of cancer, neurodegenerative disorders, inflammatory and autoimmune diseases. The structure and function of S100 proteins are modulated by metal ions via Ca2+ binding through EF-hand motifs and binding of Zn2+ and Cu2+ at additional sites, usually at the homodimer interfaces. Ca2+ binding modulates S100 conformational opening and thus promotes and affects the interaction with p53, the receptor for advanced glycation endproducts and Toll-like receptor 4, among many others. Structural plasticity also occurs at the quaternary level, where several S100 proteins self-assemble into multiple oligomeric states, many being functionally relevant. Recently, we have found that the S100A8/A9 proteins are involved in amyloidogenic processes in corpora amylacea of prostate cancer patients, and undergo metal-mediated amyloid oligomerization and fibrillation in vitro. Here we review the unique chemical and structural properties of S100 proteins that underlie the conformational changes resulting in their oligomerization upon metal ion binding and ultimately in functional control. The possibility that S100 proteins have intrinsic amyloid-forming capacity is also addressed, as well as the hypothesis that amyloid self-assemblies may, under particular physiological conditions, affect the S100 functions within the cellular milieu.

The impact of pharmaceutical policy measures: a structural-break approach

Pharmaceutical spending in many other countries has had a steep increase in the last decade. The Portuguese Government has adopted several measures to reduce pharmaceutical expenditure growth, ranging from increased co-payments to price decreases determined administratively. Promotion of generic consumption has also ranked high in political priorities. We assess the overall impact of the several policy measures on total pharmaceutical spending, using monthly data over the period January 1995 – August 2008. Endogenous structural breaks (time-series) methods were employed. Our findings suggest that policy measures aimed at controlling pharmaceutical expenditure have been, in general, unsuccessful. Two breaks were identified. Both coincide with administratively determined price decreases. Measures aimed at increasing competition in the market had no visible effect on the dynamics of Government spending in pharmaceutical products. In particular, the introduction of reference pricing had only a transitory effect of less than one year, with historical growth resuming quickly. The consequence of it is a transfer of financial burden from the Government to the patients, with no apparent effect on the dynamics of pharmaceutical spending. This strongly suggests that pharmaceutical companies have been able to adjust to policy measures, in order to sustain their sales. It remains a challenge for the future to identify firms’ strategies that supported continued growth of sales, despite the several policy measures adop

Robustness of corroded reinforced concrete structures. A structural performance approach

Structure and Infrastructure Engineering, 1-17

Acetyl-L-Carnitine Prevents Methamphetamine-Induced Structural Damage on Endothelial Cells via ILK-Related MMP-9 Activity

Methamphetamine (METH) is a potent psychostimulant highly used worldwide. Recent studies evidenced the involvement of METH in the breakdown of the blood-brain-barrier (BBB) integrity leading to compromised function. The involvement of the matrix metalloproteinases (MMPs) in the degradation of the neurovascular matrix components and tight junctions (TJs) is one of the most recent findings in METH-induced toxicity. As BBB dysfunction is a pathological feature of many neurological conditions, unveiling new protective agents in this field is of major relevance. AcetylL-carnitine (ALC) has been described to protect the BBB function in different paradigms, but the mechanisms underling its action remain mostly unknown. Here, the immortalized bEnd.3 cell line was used to evaluate the neuroprotective features of ALC in METH-induced damage. Cells were exposed to ranging concentrations of METH, and the protective effect of ALC 1 mM was assessed 24 h after treatment. F-actin rearrangement, TJ expression and distribution, and MMPs activity were evaluated. Integrin-linked kinase (ILK) knockdown cells were used to assess role of ALC in ILK mediated METHtriggered MMPs’ activity. Our results show that METH led to disruption of the actin filaments concomitant with claudin-5 translocation to the cytoplasm. These events were mediated by MMP-9 activation in association with ILK overexpression. Pretreatment with ALC prevented METH-induced activation of MMP-9, preserving claudin-5 location and the structural arrangement of the actin filaments. The present results support the potential of ALC in preserving BBB integrity, highlighting ILK as a new target for the ALC therapeutic use.

Influence of structural features on Portuguese toddler child care quality

Whereas child care quality has been extensively studied in the U.S., there is much less information about the quality of child care in other countries.With one of the highest maternal employment rates in Europe, it is important to examine child care in Portugal. Thirty toddler classrooms in child care centers were observed. The purpose of this studywas to determine whether structural features account for overall toddler child care quality. Results showed younger and better-paid teachers provided better toddler child care quality. Space available per childwas not a statistically significant predictor of toddler child care quality.Overall quality results suggest some issues to be addressed by early education policy makers and indicate the need to promote quality in Portuguese toddler child care programs.

Functional and structural characterization of the RNase II-family of enzymes

Dissertation presented to obtain a Doctoral Degree in Biology by Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa

The effect of adhesive thickness on the mechanical behavior of a structural polyurethane adhesive

One parameter that influences the adhesively bonded joints performance is the adhesive layer thickness. Hence, its effect has to be investigated experimentally and should be taken into consideration in the design of adhesive joints. Most of the results from literature are for typical structural epoxy adhesives which are generally formulated to perform in thin sections. However, polyurethane adhesives are designed to perform in thicker sections and might have a different behavior as a function of adhesive thickness. In this study, the effect of adhesive thickness on the mechanical behavior of a structural polyurethane adhesive was investigated. The mode I fracture toughness of the adhesive was measured using double-cantilever beam (DCB) tests with various thicknesses of the adhesive layer ranging from 0.2 to 2 mm. In addition, single lap joints (SLJs) were fabricated and tested to assess the influence of adhesive thickness on the lap-shear strength of the adhesive. An increasing fracture toughness with increasing adhesive thickness was found. The lap-shear strength decreases as the adhesive layer gets thicker, but in contrast to joints with brittle adhesives the decrease trend was less pronounced.

Structural adhesives modified with thermally expandable particles

In this study, the behaviour of two structural adhesives modified with thermally expandable particles (TEPs) was investigated as a preliminary study for further investigations on the potential of TEPs in adhesive joints. Tensile bulk tests were performed to get the tensile properties of the adhesives and TEPs-modified adhesives. In order to determine the expansion temperature of the particles while encapsulated in these particular adhesive systems, the variation of the volume of adhesive samples modified with different TEPs concentration as a function of temperature was measured. Further, the possibility of any chemical interactions between TEPs and adhesives matrix in the TEPs-modified specimens was verified by a Fourier transform infrared spectroscopy analysis. Finally, the fracture surfaces of the unmodified and TEPs-modified specimens, as well as the dispersion and the morphology of the particles, were examined by a scanning electron microscopy analysis. It was found that the stiffness of the TEPs-modified adhesives is not affected by incorporation of TEPs in the adhesives matrix, while the tensile yield strength decreased by increasing the wt% TEPs content. In applications of such particular materials (TEPs-modified adhesives), the temperature should be controlled to stay between 90°C and 120°C in order to obtain the highest expansion ratio. At a lower temperature, not all the particles will expand, and above, the TEPs will deteriorate and as a result the TEPs-modified adhesives will deteriorate.

Biosynthesis-Assisted Structural Elucidation of the Bartolosides, Chlorinated Aromatic Glycolipids from Cyanobacteria

The isolation of the bartolosides, unprecedented cyanobacterial glycolipids featuring aliphatic chains with chlorine substituents and C-glycosyl moieties, is reported. Their chlorinated dialkylresorcinol (DAR) core presented a major structural-elucidation challenge. To overcome this, we discovered the bartoloside (brt) biosynthetic gene cluster and linked it to the natural products through in vitro characterization of the DAR-forming ketosynthase and aromatase. Bioinformatic analysis also revealed a novel potential halogenase. Knowledge of the bartoloside biosynthesis constrained the DAR core structure by defining key pathway intermediates, ultimately allowing us to determine the full structures of the bartolosides. This work illustrates the power of genomics to enable the use of biosynthetic information for structure elucidation.

Structural change: a brief review

This is a short summary of some aspects of structural change models, and is not intended to be comprehensive survey or review. Many important works have been left out. Also, the presentation was not intended to be rigorous nor general. It’s main purpose is only to focus on some recent developments in this area and motivate the reader to learn more.

In situ measured cross section geometry of old timber structures and its influence on structural safety

Old timber structures may show significant variation in the cross section geometry along the same element, as a result of both construction methods and deterioration. As consequence, the definition of the geometric parameters in situ may be both time consuming and costly. This work presents the results of inspections carried out in different timber structures. Based on the obtained results, different simplified geometric models are proposed in order to efficiently model the geometry variations found. Probabilistic modelling techniques are also used to define safety parameters of existing timber structures, when subjected to dead and live loads, namely self-weight and wind actions. The parameters of the models have been defined as probabilistic variables, and safety of a selected case study was assessed using the Monte Carlo simulation technique. Assuming a target reliability index, a model was defined for both the residual cross section and the time dependent deterioration evolution. As a consequence, it was possible to compute probabilities of failure and reliability indices, as well as, time evolution deterioration curves for this structure. The results obtained provide a proposal for definition of the cross section geometric parameters of existing timber structures with different levels of decay, using a simplified probabilistic geometry model and considering a remaining capacity factor for the decayed areas. This model can be used for assessing the safety of the structure at present and for predicting future performance.

Active bionanoconjugates of laccase and gold nanoparticles: kinetic and structural studies

Thesis for the master degree in Structural and Functional Biochemistry

An NMR structural study of nickel-substituted rubredoxin

J Biol Inorg Chem (2010) 15:409–420 DOI 10.1007/s00775-009-0613-6

Kinetic, Structural, and EPR Studies Reveal That Aldehyde Oxidoreductase from Desulfovibrio gigas Does Not Need a Sulfido Ligand for Catalysis and Give Evidence for a Direct Mo-C Interaction in a Biological System

J. Am. Chem. Soc., 2009, 131 (23), pp 7990–7998 DOI: 10.1021/ja809448r

Correlating EPR and X-ray structural analysis of arsenite-inhibited forms of aldehyde oxidoreductase

J Biol Inorg Chem (2007) 12:353–366 DOI 10.1007/s00775-006-0191-9