Novel tear film supplements:a potential application for synthetic protein-phospholipid complexes

Purpose: Surfactant proteins A, B, C and D complex with (phospho)lipids to produce surfactants which provide low interfacial tensions. It is likely that similar complexation occurs in the tear film and contributes to its low surface tension. Synthetic protein-phospholipid complexes, with styrene maleic anhydrides (SMAs) as the protein analogue, have been shown to have similarly low surface tensions. This study investigates the potential of modified SMAs and/or SMA-phospholipid complexes, which form under physiological conditions, to supplement natural tear film surfactants. Method: SMAs were modified to provide structural variants which can form complexes under varying conditions. Infrared spectroscopy and Nuclear Magnetic Resonance were used to confirm SMA structure. Interfacial behaviour of the SMA and SMA-phospholipid complexes was studied using Langmuir trough, du Nûoy ring and pulsating bubblemethods. Factors which affect SMA-phospholipid complex formation, such as temperature and pH, were also investigated. Results: Structural manipulation of SMAs allows control over complex formation, including under physiological conditions (e.g. partial SMAesterfication allowed complexation with dimyristoylphosphatidylcholine, at pH7). The low surface tensions of the SMAs (42mN/m for static (du Nûoy ring) and 34mN/m for dynamic (Langmuir) techniques) demonstrate their surface activity at the air-aqueous interface. SMA-phospholipid complexes provide even lower surface tensions (~2 mN/m), approaching that of lung surfactant, as measured by the pulsating bubblemethod. Conclusions: Design of the molecular architecture of SMAs allows control over their surfactant properties. These SMAs could be used as novel tear films supplements, either alone to complex with native tear film phospholipids or delivered as synthetic protein-phospholipid complexes.

The effect of PEO length on the self-assembly of poly(ethylene oxide)−tetrapeptide conjugates prepared by “click” chemistry

Two series of poly(ethylene oxide)-tetrapeptide conjugates have been prepared using a “Click” reaction between an alkyne-modified tetra(phenylalanine) or tetra(valine) and various azide-terminated poly(ethylene oxide) (PEO) oligomers. Three different PEO precursors were used to prepare these conjugates, with number-average molecular weights of 350, 1200, and 1800 Da. Assembly of mPEO-F4-OEt and mPEO-V4-OEt conjugates was achieved by dialysis of a THF solution of the conjugate against water or by direct aqueous rehydration of a thin film. The PEO length has a profound effect on the outcome of the self-assembly, with the F4 conjugates giving rise to nanotubes, fibers, and wormlike micelles, respectively, as the length of the PEO block is increased. For the V4 series, the propensity to form ß-sheets dominates, and hence, the self-assembled structures are reminiscent of those formed by peptides alone, even at the longer PEO lengths. Thus, this systematic study demonstrates that the self-assembly of PEO-peptides depends on both the nature of the peptides and the relative PEO block length.

The development of a new compiler-compiler front-end

The increasing cost of developing complex software systems has created a need for tools which aid software construction. One area in which significant progress has been made is with the so-called Compiler Writing Tools (CWTs); these aim at automated generation of various components of a compiler and hence at expediting the construction of complete programming language translators. A number of CWTs are already in quite general use, but investigation reveals significant drawbacks with current CWTs, such as lex and yacc. The effective use of a CWT typically requires a detailed technical understanding of its operation and involves tedious and error-prone input preparation. Moreover, CWTs such as lex and yacc address only a limited aspect of the compilation process; for example, actions necessary to perform lexical symbol valuation and abstract syntax tree construction must be explicitly coded by the user. This thesis presents a new CWT called CORGI (COmpiler-compiler from Reference Grammar Input) which deals with the entire `front-end' component of a compiler; this includes the provision of necessary data structures and routines to manipulate them, both generated from a single input specification. Compared with earlier CWTs, CORGI has a higher-level and hence more convenient user interface, operating on a specification derived directly from a `reference manual' grammar for the source language. Rather than developing a compiler-compiler from first principles, CORGI has been implemented by building a further shell around two existing compiler construction tools, namely lex and yacc. CORGI has been demonstrated to perform efficiently in realistic tests, both in terms of speed and the effectiveness of its user interface and error-recovery mechanisms.

Transglutaminase 2 interaction with small heat shock proteins mediate cell survival upon excitotoxic stress

Transglutaminase 2 has been postulated to be involved in the pathogenesis of central nervous system neurodegenerative disorders. However, its role in neuronal cell death remains to be elucidated. Excitotoxicity is a common event underlying neurodegeneration. We aimed to evaluate the protein targets for transglutaminase 2 in cell response to NMDA-induced excitotoxic stress, using SH-SY5Y neuroblastoma cells which express high tranglutaminase 2 levels upon retinoic acid-driven differentiation toward neurons. NMDA-evoked calcium increase led to transglutaminase 2 activation that mediated cell survival, as at first suggested by the exacerbation of NMDA toxicity in the presence of R283, a synthetic competitive inhibitor of transglutaminase active site. Assays of R283-mediated transglutaminase inhibition showed the involvement of enzyme activity in NMDA-induced reduction in protein basal levels of pro-apoptotic caspase-3 and the stress protein Hsp20. However, this occurred in a way different from protein cross-linking, given that macromolecular assemblies were not observed in our experimental conditions for both proteins. Co-immunoprecipitation experiments provided evidence for the interaction, in basal conditions, between transglutaminase 2 and Hsp20, as well as between Hsp20 and Hsp27, a major anti-apoptotic protein promoting caspase-3 inactivation and degradation. NMDA treatment disrupted both these interactions that were restored upon transglutaminase 2 inhibition with R283. These results suggest that transglutaminase 2 might be protective against NMDA-evoked excitotoxic insult in neuronal-like SH-SY5Y cells in a way, independent from transamidation that likely involves its interaction with the complex Hsp20/Hsp27 playing a pro-survival role. © 2011 Springer-Verlag.

Abstract Subdifferential Calculus and Semi-Convex Functions

∗ The work is partially supported by NSFR Grant No MM 409/94.

Lack of effect of aspirin in primary CV prevention in type 2 diabetic patients with nephropathy:results from 8 years follow-up of NID-2 study

The risk-to-benefit ratio for the use of low dose of aspirin in primary cardiovascular (CV) prevention in patients with diabetes mellitus remains to be clarified. We assessed the effect of aspirin on risk of CV events in type 2 diabetic patients with nephropathy, in order to verify the usefulness of Guidelines in clinical practice. We carried out a prospective multicentric study in 564 patients with type 2 diabetic nephropathy free of CV disease attending outpatient diabetes clinics. A total of 242 patients received antiplatelet treatment with aspirin 100 mg/day (group A), and 322 were not treated with antiplatelet drugs (group B). Primary end point was the occurrence of total major adverse cardio-vascular events (MACE). Secondary end points were the relative occurrence of fatal MACE. The average follow-up was 8 years. Total MACE occurred in 49 patients from group A and in 52 patients from group B. Fatal MACE occurred in 22 patients from group A and in 20 from group B; nonfatal MACE occurred in 27 patients from group A and in 32 patients from group B. Kaplan-Meier analysis did not show a statistically significant difference of cumulative MACE between the two groups. A not statistically significant difference in the incidence of both fatal (p = 0.225) and nonfatal CV events (p = 0.573) between the two groups was observed. These results were confirmed after adjustment for confounders (HR for MACE 1.11, 95 % CI 0.91-1.35). These findings suggest that low dose of aspirin is ineffective in primary prevention for patients with nephropathy. © 2014 Springer-Verlag Italia.

Biomimetic synthetic lubricants (Biosurfactants)

Poly(styrene-co-maleic anhydride) (PSMA) based copolymers are known to undergo conformational transition in response to environmental stimuli. This smart behaviour makes it possible to mimic the behaviour of native apoproteins. The primary aim of this study was to develop a better understanding of the structure-property relationships of various PSMA-based copolymers sought. The work undertaken in this thesis has revealed that the responsive behaviour of PSMA-based copolymers can be tailored by varying the molecular weight, hydrophobic (styrene) and hydrophilic (maleic acid) balance, and more so in the presence of additional hydrophobic, mono-partial ester moieties. Novel hydrophilic and hydrophobic synthetic surfactant protein analogues have successfully been prepared. These novel lipid solubilising agents possess a broad range of HLB (hydrophilic-lipophilic balance) values that have been estimated. NMR spectroscopy was utilised to confirm the structures for PSMA-based copolymers sought and proved useful in furthering understanding of the structure-property relationships of PSMA-based copolymers. The association of PSMA with the polar phospholipid, 2-dilauryl-sn-glycero-3- phosphocholine (DLPC) produces polymer-lipid complexes analogous to lipoprotein assemblies present in the blood plasma. NMR analysis reveals that the PSMA-based copolymers are not perfectly alternating. Regio-irregular structures, atactic and random monomer sequence distribution have been identified for all materials studied. Novel lipid solubilising agents (polyanionic surfactants) have successfully been synthesised from a broad range of PSMA-based copolymers with desired estimated HLB values that interact with polar phospholipids (DLPC/DPPC) uniquely. Very low static and dynamic surface tensions have been observed via the du Noϋy ring method and Langmuir techniques and correlate well with the estimated HLB values. Synthetic protein-lipid analogues have been successfully synthesised, that mimic the unique surface properties of native biological lubricants without the use of solvents. The novel PSMA-DLPC complexes have successfully been combined with hyaluronan (hyaluronic acid, HA). Today, the employment of HA is economically feasible, because it is readily available from bacterial fermentation processes in a thermally stable form - HyaCare®. The work undertaken in this thesis highlights the usage of HA in biolubrication applications and how this can be optimised and thus justified by carefully selecting the biological source, concentration, molecular weight, purity and most importantly by combining it with compatible boundary lubricating agents (polar phospholipids). Experimental evidence supports the belief that the combined HA and PSMA-DLPC complexes provide a balance of rheological, biotribological and surface properties that are composition dependent, and show competitive advantage as novel synthetic biological lubricants (biosurfactants).

Statistical Inference for Processes Depending on Environments and Application in Regenerative Processes

We build the Conditional Least Squares Estimator of 0 based on the observation of a single trajectory of {Zk,Ck}k, and give conditions ensuring its strong consistency. The particular case of general linear models according to 0=( 0, 0) and among them, regenerative processes, are studied more particularly. In this frame, we may also prove the consistency of the estimator of 0 although it belongs to an asymptotic negligible part of the model, and the asymptotic law of the estimator may also be calculated.

Estimation of the Offspring Mean in a General Single-Type Size-Dependent Branching Process

2000 Mathematics Subject Classification: 60J80, 62F12, 62P10

Implementação de uma arquitetura fuzzy neural em hardware com treinamento online

Computational Intelligence Methods have been expanding to industrial applications motivated by their ability to solve problems in engineering. Therefore, the embedded systems follow the same idea of using computational intelligence tools embedded on machines. There are several works in the area of embedded systems and intelligent systems. However, there are a few papers that have joined both areas. The aim of this study was to implement an adaptive fuzzy neural hardware with online training embedded on Field Programmable Gate Array – FPGA. The system adaptation can occur during the execution of a given application, aiming online performance improvement. The proposed system architecture is modular, allowing different configurations of fuzzy neural network topologies with online training. The proposed system was applied to: mathematical function interpolation, pattern classification and selfcompensation of industrial sensors. The proposed system achieves satisfactory performance in both tasks. The experiments results shows the advantages and disadvantages of online training in hardware when performed in parallel and sequentially ways. The sequentially training method provides economy in FPGA area, however, increases the complexity of architecture actions. The parallel training method achieves high performance and reduced processing time, the pipeline technique is used to increase the proposed architecture performance. The study development was based on available tools for FPGA circuits.

Minicontos: uma prática de letramento emergente na escola

Like numerous other mini ficçion genres, the Flash fiction is configured as a concise and brief genre, but it can still preserve some elements of the traditional tale, such as: narrator, space-temporal unit, characters and plot. For some authors, the emergence of this genre is strictly connected to the speed and diversity of forms of communication in the current social context, that require the reader other skills in addition to reading and writing. Considering the importance of these skills in a literate society and recognizing this genre as potentially effective tool for the development of literacy, this research has as objective to describe the development and implications of didactic sequence directed the literacy practices with the Flash fiction genre. Thus, the research becomes effective in an elementary school vespertine class in 9th grade from Escola Estadual Dr. José Gonçalves de Medeiros, located in the city of Acari-RN. As theoretical assumptions, it will be used fundamentals from Literacy Studies as social practice, especially that ones discussed by the writings Kleiman (1995), Hamilton (2000), Soares (2012), Mortatti (2004) and Rojo (2012). To subsidize the literary literacy, it will be used the theoretical contibutions of Borges (2000), Candido (2004), Pinheiro (2001) and Silva e Silveira (2013). In what relates to the study about the storytellers, it was taken by support the postulates of Gotlib (2006), Bosi (1997), Cortázar (1999), Talbot (2002), Poe (s/d), Luzia de Maria (2004), as well as studies by researchers and writers as Spalding (2007; 2008; 2011), George (2007), Freire (2004) and Trevisan (1994), among others who are dedicated to study Flash fiction. In the methodological field, the proposed perspective anchors itself in an action-researching guidelines (MOREIRA; CALEFFE, 2006) and in the approach of a qualitative data (BOGDAN; BIKLEN, 1994), since it talks to interpretation of diagnosed situation, implementation of intervention actions organized in didactic sequence (SCHNEUWLY; DOLZ, 2004; BARROS; RIVERS-registry, 2014), including description and evaluation of results achieved from the implementation of such actions. The discussions generated indicate that the work with Flash ficition, developed by means of didactic sequences, contributes to the reflection on teaching strategies of Portuguese, used to prepare the students to meet the demands of reading and writing of contemporary society.

On the Mediterranean conveyor belt system

The Mediterranean Sea is a semi-enclosed sea, connected to the Atlantic Ocean through the Gibraltar Strait and subdivided in two different regions by the Sicily Strait. The geographical extension of the basin, the surface heat flux, and the water inflow from the Gibraltar Strait are some of the basic factors determining its horizontal and vertical circulation. In the Mediterranean strong salinity and temperature zonal gradients contribute to maintain the zonal-vertical circulation, while meridional-vertical cells are equally forced by winds and deep water mass formation in three regions, the Gulf of Lyon, the southern Adriatic and the Cretan Sea areas. The objective of this thesis is to study how these cells combine together to form the Mediterranean conveyor belt system. This has never been attempted before so the conclusions are necessarily preliminary. In the first part we discuss the vertical zonal and meridional circulation by reconstructing the Wust Mediterranean vertical salinity and temperature structures in an attempt to evaluate the water mass structure consistent with modern data. Our results confirm that Wust depicted vertical circulation from scarce data is reproduced by the past 27 years observations. The structure of both meridional and zonal circulations was discussed using velocity vertical streamfunctions with two different methods. The first one, eulerian, allowed us to observe vertical structures that were already reported in the literature. Recent studies in the Atlantic Ocean have shown that gyres and eddies have an important influence in the isopycnal vertical circulation. This is called the residual circulation which was computed in this study for the first time. A possible interpretation of horizontal connection between the meridional and zonal cells was discussed using horizontal streamfunction. In the last part of the thesis we have been developing an idealized numerical model to study the vertical circulation in the Mediterranean.

2003 Habitat Maps derived from Grey Nurse Shark (GNS) Project of Wolf Rock, Double Island Point, Queensland, Australia

Surveying habitats critical to the survival of grey nurse sharks in South-East Queensland has mapped critical habitats, gathered species inventories and developed protocols for ecological monitoring of critical habitats in southern Queensland. This information has assisted stakeholders with habitat definition and effective management. In 2002 members of UniDive applied successfully for World Wide Fund for Nature, Threatened Species Network funds to map the critical Grey Nurse Shark Habitats in south east Queensland. UniDive members used the funding to survey, from the boats of local dive operators, Wolf Rock at Double Island Point, Gotham, Cherub's Cave, Henderson's Rock and China Wall at North Moreton and Flat Rock at Point Look Out during 2002 and 2003. These sites are situated along the south east Queensland coast and are known to be key Grey Nurse Shark aggregation sites. During the project UniDive members were trained in mapping and survey techniques that include identification of fish, invertebrates and substrate types. Training was conducted by experts from the University of Queensland (Centre of Marine Studies, Biophysical Remote Sensing) and the Queensland Parks and Wildlife Service who are also UniDive members. The monitoring methods (see methods) are based upon results of the UniDive Coastcare project from 2002, the international established Reef Check program and research conducted by Biophysical Remote Sensing and the Centre of Marine Studies. Habitats were mapped using a combination of towed GPS photo transects, aerial photography, bathymetry surveys and expert knowledge. This data provides georeferenced information regarding the major features of each of Sites mapped including Wolf Rock