Phosphorylated and unphosphorylated forms of human single-stranded DNA-binding protein are equally active in simian virus 40 DNA replication and in nucleotide excision repair.

The trimeric human single-stranded DNA-binding protein (HSSB; also called RP-A) plays an essential role in DNA replication, nucleotide excision repair, and homologous DNA recombination. The p34 subunit of HSSB is phosphorylated at the G1/S boundary of the cell cycle or upon exposure of cells to DNA damage-inducing agents including ionizing and UV radiation. We have previously shown that the phosphorylation of p34 is catalyzed by both cyclin-dependent kinase-cyclin A complex and DNA-dependent protein kinase. In this study, we investigated the effect of phosphorylation of p34 by these kinases on the replication and repair function of HSSB. We observed no significant difference with the unphosphorylated and phosphorylated forms of HSSB in the simian virus 40 DNA replication or nucleotide excision repair systems reconstituted with purified proteins. The phosphorylation status of the p34 subunit of HSSB was unchanged during the reactions. We suggest that the phosphorylated HSSB has no direct effect on the basic mechanism of DNA replication and nucleotide excision repair reactions in vitro, although we cannot exclude a role of p34 phosphorylation in modulating HSSB function in vivo through a yet poorly understood control pathway in the cellular response to DNA damage and replication.

Resistência à fadiga de tubo API 5L X65 cladeado e soldado circunferencialmente com eletrodos de Inconel® 625

As recentes descobertas de petróleo e gás na camada do Pré-sal representam um enorme potencial exploratório no Brasil, entretanto, os desafios tecnológicos para a exploração desses recursos minerais são imensos e, consequentemente, têm motivado o desenvolvimento de estudos voltados a métodos e materiais eficientes para suas produções. Os tubos condutores de petróleo e gás são denominados de elevadores catenários ou do inglês \"risers\", e são elementos que necessariamente são soldados e possuem fundamental importância nessa cadeia produtiva, pois transportam petróleo e gás natural do fundo do mar à plataforma, estando sujeitos a carregamentos dinâmicos (fadiga) durante sua operação. Adicionalmente, um dos problemas centrais à produção de óleo e gás das reservas do Pré-Sal está diretamente associado a meios altamente corrosivos, tais como H2S e CO2. Uma forma mais barata de proteção dos tubos é a aplicação de uma camada de um material metálico resistente à corrosão na parte interna desses tubos (clad). Assim, a união entre esses tubos para formação dos \"risers\" deve ser realizada pelo emprego de soldas circunferenciais de ligas igualmente resistentes à corrosão. Nesse contexto, como os elementos soldados são considerados possuir defeitos do tipo trinca, para a garantia de sua integridade estrutural quando submetidos a carregamentos cíclicos, é necessário o conhecimento das taxas de propagação de trinca por fadiga da solda circunferencial. Assim, neste trabalho, foram realizados ensaios de propagação de trinca por fadiga na região da solda circunferencial de Inconel® 625 realizada em tubo de aço API 5L X65 cladeado, utilizando corpos de prova do tipo SEN(B) (Single Edge Notch Bending) com relações entre espessura e largura (B/W) iguais a 0,5, 1 e 2. O propósito central deste trabalho foi de obter a curva da taxa de propagação de trinca por fadiga (da/dN) versus a variação do fator de intensidade de tensão (ΔK) para o metal de solda por meio de ensaios normatizados, utilizando diferentes técnicas de acompanhamento e medição da trinca. A monitoração de crescimento da trinca foi feita por três técnicas: variação da flexibilidade elástica (VFE), queda de potencial elétrico (QPE) e análise de imagem (Ai). Os resultados mostraram que as diferentes relações B/W utilizadas no estudo não alteraram significantemente as taxas de propagação de trinca por fadiga, respeitado que a propagação aconteceu em condições de escoamento em pequena escala na frente da trinca. Os resultados de propagação de trinca por fadiga permitiram a obtenção das regiões I e II da curva da/dN versus ΔK para o metal de solda. O valor de ΔKlim obtido para o mesmo foi em torno de 11,8 MPa.m1/2 e os valores encontrados das constantes experimentais C e m da equação de Paris-Erdogan foram respectivamente iguais a 1,55 x10-10 [(mm/ciclo)/(MPa.m1/2)m] e 4,15. A propagação de trinca no metal de solda deu-se por deformação plástica, com a formação de estrias de fadiga.

A relação inconsciente-consciente na produção do pensamento consciente

Tese de mestrado, Psicologia (Ciência Cognitiva), Universidade de Lisboa, Faculdade de Psicologia, Faculdade de Letras, Faculdade de Ciências, Faculdade de Medicina, 2016

GFP-targeting allows visualization of the apicoplast throughout the life cycle of live malaria parasites.

BACKGROUND INFORMATION The Plasmodium parasite, during its life cycle, undergoes three phases of asexual reproduction, these being repeated rounds of erythrocytic schizogony, sporogony within oocysts on the mosquito midgut wall and exo-erythrocytic schizogony within the hepatocyte. During each phase of asexual reproduction, the parasite must ensure that every new daughter cell contains an apicoplast, as this organelle cannot be formed de novo and is essential for parasite survival. To date, studies visualizing the apicoplast in live Plasmodium parasites have been restricted to the blood stages of Plasmodium falciparum. RESULTS In the present study, we have generated Plasmodium berghei parasites in which GFP (green fluorescent protein) is targeted to the apicoplast using the specific targeting sequence of ACP (acyl carrier protein), which has allowed us to visualize this organelle in live Plasmodium parasites. During each phase of asexual reproduction, the apicoplast becomes highly branched, but remains as a single organelle until the completion of nuclear division, whereupon it divides and is rapidly segregated into newly forming daughter cells. We have shown that the antimicrobial agents azithromycin, clindamycin and doxycycline block development of the apicoplast during exo-erythrocytic schizogony in vitro, leading to impaired parasite maturation. CONCLUSIONS Using a range of powerful live microscopy techniques, we show for the first time the development of a Plasmodium organelle through the entire life cycle of the parasite. Evidence is provided that interference with the development of the Plasmodium apicoplast results in the failure to produce red-blood-cell-infective merozoites.

The Plasmodium serine-type SERA proteases display distinct expression patterns and non-essential in vivo roles during life cycle progression of the malaria parasite

Parasite proteases play key roles in several fundamental steps of the Plasmodium life cycle, including haemoglobin degradation, host cell invasion and parasite egress. Plasmodium exit from infected host cells appears to be mediated by a class of papain-like cysteine proteases called 'serine repeat antigens' (SERAs). A SERA subfamily, represented by Plasmodium falciparum SERA5, contains an atypical active site serine residue instead of a catalytic cysteine. Members of this SERAser subfamily are abundantly expressed in asexual blood stages, rendering them attractive drug and vaccine targets. In this study, we show by antibody localization and in vivo fluorescent tagging with the red fluorescent protein mCherry that the two P. berghei serine-type family members, PbSERA1 and PbSERA2, display differential expression towards the final stages of merozoite formation. Via targeted gene replacement, we generated single and double gene knockouts of the P. berghei SERAser genes. These loss-of-function lines progressed normally through the parasite life cycle, suggesting a specialized, non-vital role for serine-type SERAs in vivo. Parasites lacking PbSERAser showed increased expression of the cysteine-type PbSERA3. Compensatory mechanisms between distinct SERA subfamilies may thus explain the absence of phenotypical defect in SERAser disruptants, and challenge the suitability to develop potent antimalarial drugs based on specific inhibitors of Plasmodium serine-type SERAs.

Global database of surface ocean particulate organic carbon export fluxes diagnosed from the 234Th technique

Through the processes of the biological pump, carbon is exported to the deep ocean in the form of dissolved and particulate organic matter. There are several ways by which downward export fluxes can be estimated. The great attraction of the 234Th technique is that its fundamental operation allows a downward flux rate to be determined from a single water column profile of thorium coupled to an estimate of POC/234Th ratio in sinking matter. We present a database of 723 estimates of organic carbon export from the surface ocean derived from the 234Th technique. Data were collected from tables in papers published between 1985 and 2013 only. We also present sampling dates, publication dates and sampling areas. Most of the open ocean Longhurst provinces are represented by several measurements. However, the Western Pacific, the Atlantic Arctic, South Pacific and the South Indian Ocean are not well represented. There is a variety of integration depths ranging from surface to 220m. Globally the fluxes ranged from -22 to 125 mmol of C/m**2/d. We believe that this database is important for providing new global estimate of the magnitude of the biological carbon pump.

The critical role of tryptophan-116 in the catalytic cycle of dimethylsulfoxide reductase from Rhodobacter capsulatus

In dimethylsulfoxide reductase of Rhodobacter capsulatus tryptophan-116 forms a hydrogen bond with a single oxo ligand bound to the molybdenum ion. Mutation of this residue to phenylalanine affected the UV/visible spectrum of the purified Mo-VI form of dimethylsulfoxide reductase resulting in the loss of the characteristic transition at 720 nm. Results of steady-state kinetic analysis and electrochemical studies suggest that tryptophan 116 plays a critical role in stabilizing the hexacoordinate monooxo Mo-VI form of the enzyme and prevents the formation of a dioxo pentacoordinate Mo-VI species, generated as a consequence of the dissociation of one of the dithiolene ligands of the molybdopterin cofactor from the Mo ion. (C) 2004 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.

Optimal control and operation of drum granulation processes

Process optimisation and optimal control of batch and continuous drum granulation processes are studied in this paper. The main focus of the current research has been: (i) construction of optimisation and control relevant, population balance models through the incorporation of moisture content, drum rotation rate and bed depth into the coalescence kernels; (ii) investigation of optimal operational conditions using constrained optimisation techniques; (iii) development of optimal control algorithms based on discretized population balance equations; and (iv) comprehensive simulation studies on optimal control of both batch and continuous granulation processes. The objective of steady state optimisation is to minimise the recycle rate with minimum cost for continuous processes. It has been identified that the drum rotation-rate, bed depth (material charge), and moisture content of solids are practical decision (design) parameters for system optimisation. The objective for the optimal control of batch granulation processes is to maximize the mass of product-sized particles with minimum time and binder consumption. The objective for the optimal control of the continuous process is to drive the process from one steady state to another in a minimum time with minimum binder consumption, which is also known as the state-driving problem. It has been known for some time that the binder spray-rate is the most effective control (manipulative) variable. Although other possible manipulative variables, such as feed flow-rate and additional powder flow-rate have been investigated in the complete research project, only the single input problem with the binder spray rate as the manipulative variable is addressed in the paper to demonstrate the methodology. It can be shown from simulation results that the proposed models are suitable for control and optimisation studies, and the optimisation algorithms connected with either steady state or dynamic models are successful for the determination of optimal operational conditions and dynamic trajectories with good convergence properties. (c) 2005 Elsevier Ltd. All rights reserved.

Photonic crystal fibers design for 2 μm wavelenght operation

New photonic crystal fiber designs are presented and numerically investigated in order to improve the state of art of high power fiber lasers. The focus of this work is targeted on the region of 2 μm laser emission, which is of high interest due to its eye-safe nature and due to the large amount of applications permitted. Thulium doped fiber amplifiers are suitable for emitting in this region. Different fiber designs have been proposed, both flexible and rod-type, with the aim to enlarge mode area while maintaining robust single mode operation. The analysis of thermal effects, caused by the high thulium quantum defect, have been taken in consideration. Solutions to counteract issues derived by detrimental thermal effects have been implemented.

Simultaneous demultiplexing and clock recovery using a single electroabsorption modulator in a novel bi-directional configuration

A single electroabsorption modulator was used to demultiplex a 10 Gbit/s channel from a 40 Gbit/s OTDM data stream, whilst simultaneously recovering the 10 GHz electrical clock. This was achieved using a new bi-directional operation of the EA modulator, combined with a simple phase-locked loop feedback circuit. Excellent system performance was achieved, indicating that operation up to and beyond 100 Gbit/s is possible using current technology.

Mathematical modelling and optimisation of a water heat pump

The purpose of the work described here has been to seek methods of narrowing the present gap between currently realised heat pump performance and the theoretical limit. The single most important pre-requisite to this objective is the identification and quantitative assessment of the various non-idealities and degradative phenomena responsible for the present shortfall. The use of availability analysis has been introduced as a diagnostic tool, and applied to a few very simple, highly idealised Rankine cycle optimisation problems. From this work, it has been demonstrated that the scope for improvement through optimisation is small in comparison with the extensive potential for improvement by reducing the compressor's losses. A fully instrumented heat pump was assembled and extensively tested. This furnished performance data, and led to an improved understanding of the systems behaviour. From a very simple analysis of the resulting compressor performance data, confirmation of the compressor's low efficiency was obtained. In addition, in order to obtain experimental data concerning specific details of the heat pump's operation, several novel experiments were performed. The experimental work was concluded with a set of tests which attempted to obtain definitive performance data for a small set of discrete operating conditions. These tests included an investigation of the effect of two compressor modifications. The resulting performance data was analysed by a sophisticated calculation which used that measurements to quantify each dagradative phenomenon occurring in that compressor, and so indicate where the greatest potential for improvement lies. Finally, in the light of everything that was learnt, specific technical suggestions have been made, to reduce the losses associated with both the refrigerant circuit and the compressor.

Surface acoustic wave devices with low loss and high frequency operation

This thesis describes an industrial research project carried out in collaboration with STC Components, Harlow, Essex. Technical and market trends in the use of surface acoustic wave (SAW) devices are reviewed. As a result, three areas not previously addressed by STC were identified: lower insertion loss designs, higher operating frequencies and improved temperature dependent stability. A review of the temperature performance of alternative lower insertion loss designs,shows that greater use could be made of the on-site quartz growing plant. Data is presented for quartz cuts in the ST-AT range. This data is used to modify the temperature performance of a SAW filter. Several recently identified quartz orientations have been tested. These are SST, LST and X33. Problems associated with each cut are described and devices demonstrated. LST quartz, although sensitive to accuracy of cut, is shown to have an improved temperature coefficient over the normal ST orientation. Results show that its use is restricted due to insertion loss variations with temperature. Effects associated with split-finger transducers on LST-quartz are described. Two low-loss options are studied, coupled resonator filters for very narrow bandwidth applications and single phase unidirectional transducers (SPUDT) for fractional bandwidths up to about 1%. Both designs can be implemented with one quarter wavelength transducer geometries at operating frequencies up to 1GHz. The SPUDT design utilised an existing impulse response model to provide analysis of ladder or rung transducers. A coupled resonator filter at 400MHz is demonstrated with a matched insertion loss of less than 3.5dB and bandwidth of 0.05%. A SPUDT device is designed as a re-timing filter for timing extraction in a long haul PCM transmission system. Filters operating at 565MHz are demonstrated with insertion losses of less than 6dB. This basic SPUDT design is extended to a maximally distributed version and demonstrated at 450MHz with 9.8dB insertion loss.

Single- and dual-wavelength switchable erbium-doped fiber ring laser based on intracavity polarization selective tilted fiber gratings

We propose and demonstrate a single- and dual-wavelength switchable erbium-doped fiber laser (EDFL) by utilizing intracavity polarization selective filters based on tilted fiber gratings (TFGs). In the cavity, one 45° TFG functions as an in-fiber polarizer and the other 77° TFG is used as a fiber polarization dependent loss (PDL) filter. The combined polarization effect from these two TFGs enables the laser to switch between the single- and the dual-wavelength operation with a single-polarization state at room temperature. The laser output at each wavelength shows an optical signal-to-noise ratio (OSNR) of >60 dB, a side mode suppression ratio (SMSR) of >50 dB, and a polarization extinction ratio of ~35 dB. The proposed EDFL can give stable output under laboratory conditions.

Single polarisation fibre ring laser by utilising intracavity 45° tilted fibre Bragg grating

Single polarisation operation of fibre ring laser has been realised by employing an intracavity 45deg-tilted fibre Bragg grating (45deg-TFBG). The degree of polarisation up to 99.94% of the laser was demonstrated with good stability.

Realisation of single polarisation state of fibre ring laser by utilising intracavity 45° tilted fibre Bragg grating

Single polarisation operation of fibre ring laser has been realised by employing an intracavity 45°tilted fibre Bragg gratings (45° TFBGs). The degree of polarisation of 99.94% of the laser was demonstrated with good stability.