Isolation and characterisation of EfeM, a periplasmic component of the putative EfeUOBM iron transporter of Pseudomonas syringae pv. syringae

The EfeM protein is a component of the putative EfeUOBM iron-transporter of Pseudomonas syringae pathovar syringae and is thought to act as a periplasmic, ferrous-iron binding protein. It contains a signal peptide of 34 amino acid residues and a C-terminal 'Peptidase_M75' domain of 251 residues. The C-terminal domain contains a highly conserved 'HXXE' motif thought to act as part of a divalent cation-binding site. In this work, the gene (efeM or 'Psyr_3370') encoding EfeM was cloned and over-expressed in Escherichia coli, and the mature protein was purified from the periplasm. Mass spectrometry confirmed the identity of the protein (M(W) 27,772Da). Circular dichroism spectroscopy of EfeM indicated a mainly alpha-helical structure, consistent with bioinformatic predictions. Purified EfeM was crystallised by hanging-drop vapor diffusion to give needle-shaped crystals that diffracted to a resolution of 1.6A. This is the first molecular study of a peptidase M75 domain with a presumed iron transport role.

Response to zinc deficiency of two rice lines with contrasting tolerance is determined by root growth maintenance and organic acid exudation rates, and not by zinc-transporter activity

Zinc (Zn)-deficient soils constrain rice (Oryza sativa) production and cause Zn malnutrition. The identification of Zn-deficiency-tolerant rice lines indicates that breeding might overcome these constraints. Here, we seek to identify processes underlying Zn-deficiency tolerance in rice at the physiological and transcriptional levels. A Zn-deficiency-tolerant line RIL46 acquires Zn more efficiently and produces more biomass than its nontolerant maternal line (IR74) at low Zn(ext) under field conditions. We tested if this was the result of increased expression of Zn(2+) transporters; increased root exudation of deoxymugineic acid (DMA) or low-molecular-weight organic acids (LMWOAs); and/or increased root production. Experiments were performed in field and controlled environment conditions. There was little genotypic variation in transcript abundance of Zn-responsive root Zn(2+)-transporters between the RIL46 and IR74. However, root exudation of DMA and LMWOA was greater in RIL46, coinciding with increased root expression of putative ligand-efflux genes. Adventitious root production was maintained in RIL46 at low Zn(ext), correlating with altered expression of root-specific auxin-responsive genes. Zinc-deficiency tolerance in RIL46 is most likely the result of maintenance of root growth, increased efflux of Zn ligands, and increased uptake of Zn-ligand complexes at low Zn(ext); these traits are potential breeding targets.

Biometrics in ABC: counter-spoofing research

Automated border control (ABC) is concerned with fast and secure processing for intelligence-led identification. The FastPass project aims to build a harmonised, modular reference system for future European ABC. When biometrics is taken on board as identity, spoofing attacks become a concern. This paper presents current research in algorithm development for counter-spoofing attacks in biometrics. Focussing on three biometric traits, face, fingerprint, and iris, it examines possible types of spoofing attacks, and reviews existing algorithms reported in relevant academic papers in the area of countering measures to biometric spoofing attacks. It indicates that the new developing trend is fusion of multiple biometrics against spoofing attacks.

Genetical and comparative genomics of Brassica under altered Ca supply identifies Arabidopsis Ca-transporter orthologs

Although Ca transport in plants is highly complex, the overexpression of vacuolar Ca2+ transporters in crops is a promising new technology to improve dietary Ca supplies through biofortification. Here, we sought to identify novel targets for increasing plant Ca accumulation using genetical and comparative genomics. Expression quantitative trait locus (eQTL) mapping to 1895 cis- and 8015 trans-loci were identified in shoots of an inbred mapping population of Brassica rapa (IMB211 × R500); 23 cis- and 948 trans-eQTLs responded specifically to altered Ca supply. eQTLs were screened for functional significance using a large database of shoot Ca concentration phenotypes of Arabidopsis thaliana. From 31 Arabidopsis gene identifiers tagged to robust shoot Ca concentration phenotypes, 21 mapped to 27 B. rapa eQTLs, including orthologs of the Ca2+ transporters At-CAX1 and At-ACA8. Two of three independent missense mutants of BraA.cax1a, isolated previously by targeting induced local lesions in genomes, have allele-specific shoot Ca concentration phenotypes compared with their segregating wild types. BraA.CAX1a is a promising target for altering the Ca composition of Brassica, consistent with prior knowledge from Arabidopsis. We conclude that multiple-environment eQTL analysis of complex crop genomes combined with comparative genomics is a powerful technique for novel gene identification/prioritization.

Serotonin transporter [corrected] methylation and response to cognitive behaviour therapy in children with anxiety disorders

Anxiety disorders that are the most commonly occurring psychiatric disorders in childhood, are associated with a range of social and educational impairments and often continue into adulthood. Cognitive behaviour therapy (CBT) is an effective treatment option for the majority of cases, although up to 35-45% of children do not achieve remission. Recent research suggests that some genetic variants may be associated with a more beneficial response to psychological therapy. Epigenetic mechanisms such as DNA methylation work at the interface between genetic and environmental influences. Furthermore, epigenetic alterations at the serotonin transporter (SERT) promoter region have been associated with environmental influences such as stressful life experiences. In this study, we measured DNA methylation upstream of SERT in 116 children with an anxiety disorder, before and after receiving CBT. Change during treatment in percentage DNA methylation was significantly different in treatment responders vs nonresponders. This effect was driven by one CpG site in particular, at which responders increased in methylation, whereas nonresponders showed a decrease in DNA methylation. This is the first study to demonstrate differences in SERT methylation change in association with response to a purely psychological therapy. These findings confirm that biological changes occur alongside changes in symptomatology following a psychological therapy such as CBT.

PKC stimulated by glucagon decreases UT-A1 urea transporter expression in rat IMCD

It is well-known that glucagon increases fractional excretion of urea in rats after a protein intravenous infusion. This effect was investigated by using: (a) in vitro microperfusion technique to measure [(14)C]-urea permeability (Pu x 10(-5) cm/s) in inner medullary collecting ducts (IMCD) from normal rats in the presence of 10(-7) M of glucagon and in the absence of vasopressin and (b) immunoblot techniques to determine urea transporter expression in tubule suspension incubated with the same glucagon concentration. Seven groups of IMCDs (n = 47) were studied. Our results revealed that: (a) glucagon decreased urea reabsorption dose-dependently; (b) the glucagon antagonist des-His(1)-[Glu(9)], blocked the glucagon action but not vasopressin action; (c) the phorbol myristate acetate, decreased urea reabsorption but (d) staurosporin, restored its effect; e) staurosporin decreased glucagon action, and finally, (f) glucagon decreased UT-A1 expression. We can conclude that glucagon reduces UT-A1 expression via a glucagon receptor by stimulating PKC.

Na+-glucose transporter-2 messenger ribonucleic acid expression in kidney of diabetic rats correlates with glycemic levels: Involvement of hepatocyte nuclear factor-1 alpha expression and activity

Mutations in Na+-glucose transporters (SGLT)-2 and hepatocyte nuclear factor (HNF)-1 alpha genes have been related to renal glycosuria and maturity-onset diabetes of the young 3, respectively. However, the expression of these genes have not been investigated in type 1 and type 2 diabetes. Here in kidney of diabetic rats, we tested the hypotheses that SGLT2 mRNA expression is altered; HNF-1 alpha is involved in this regulation; and glycemic homeostasis is a related mechanism. The in vivo binding of HNF-1 alpha into the SGLT2 promoter region in renal cortex was confirmed by chromatin immunoprecipitation assay. SGLT2 and HNF-1 alpha mRNA expression (by Northern and RT-PCR analysis) and HNF-1 binding activity of nuclear proteins (by EMSA) were investigated in diabetic rats and treated or not with insulin or phlorizin (an inhibitor of SGLT2). Results showed that diabetes increases SGLT2 and HNF-1 alpha mRNA expression (similar to 50%) and binding of nuclear proteins to a HNF-1 consensus motif (similar to 100%). Six days of insulin or phlorizin treatment restores these parameters to nondiabetic-rat levels. Moreover, both treatments similarly reduced glycemia, despite the differences in plasma insulin and urinary glucose concentrations, highlighting the plasma glucose levels as involved in the observed modulations. This study shows that SGLT2 mRNA expression and HNF-1 alpha expression and activity correlate positively in kidney of diabetic rats. It also shows that diabetes-induced changes are reversed by lowering glycemia, independently of insulinemia. Our demonstration that HNF-1 alpha binds DNA that encodes SGLT2 supports the hypothesis that HNF-1 alpha, as a modulator of SGLT2 expression, may be involved in diabetic kidney disease.

Crystallographic structure and substrate-binding interactions of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

In Xanthomonas axonopodis pv. citri (Xac or X citri), the modA gene codes for a periplasmic protein (ModA) that is capable of binding molybdate and tungstate as part of the ABC-type transporter required for the uptake of micronutrients. In this study, we report the crystallographic structure of the Xac ModA protein with bound molybdate. The Xac ModA structure is similar to orthologs with known three-dimensional structures and consists of two nearly symmetrical domains separated by a hinge region where the oxyanion-binding site lies. Phylogenetic analysis of different ModA orthologs based on sequence alignments revealed three groups of molybdate-binding proteins: bacterial phytopathogens, enterobacteria and soil bacteria. Even though the ModA orthologs are segregated into different groups, the ligand-binding hydrogen bonds are mostly conserved, except for Archaeglobus fulgidus ModA. A detailed discussion of hydrophobic interactions in the active site is presented and two new residues, Ala(38) and Ser(151), are shown to be part of the ligand-binding pocket. (c) 2007 Elsevier B.V All rights reserved.

Implementera en guide för allmänna transporter med buss på mobiltelefoner med Java 2 Micro Edition

Detta examensarbete går ut på att ta fram en applikation, avsedd för mobiltelefoner, där man på ett enkelt sätt kan få reda på avgångarna för Dalarnas kollektivtrafik. Det stigande bensinpriset och bilåkningens negativa miljöpåverkan ligger som grund till att Etex AB och Balanz AB vill undersöka om det går att få människor att välja bussen framför personbilen. Om man genom denna applikation kan underlätta planeringen av människors dagliga resor, hur påverkas då valet av fortskaffningsmedel?Resultatet blev en applikation där man, via sin mobiltelefon, bland annat kan ta reda på avgångar för utvalda linjer av Dalatrafiks bussar, hantera linjekartor och synkronisera avgångar med telefonens kalender. Alla tidtabeller sparas i telefonen för att minska nätverkstrafiken och för att informationen enkelt ska kunna återanvändas.Dalatrafik är det företag som ansvarar för kollektivtrafiken i Dalarna. Det finns i dagsläget ingen möjlighet att få reda på avgångarna för deras bussar om man inte har tillgång till en tidtabell eller ringer till deras kundservice. Vissa kommuner har redan lösningar för tidtabellshantering i mobiltelefonen och nu vill Etex AB och Balanz AB, som är två företag som bl.a. arbetar med trådlösa applikationer, att även Dalarna ska få ta del av detta praktiska hjälpmedel.De flesta mobiltelefoner som säljs idag har stöd för att köra applikationer, eller MIDlets som de också kallas, gjorda i J2ME (Java 2 Platform, Micro Edition). Denna teknik har tagits fram av Sun för att göra applikationer som kan köras på mindre mobila enheter såsom mobiltelefoner och handdatorer. Kombinerar man J2ME med den ökande prestandan i dagens mobiltelefoner kan avancerade applikationer t.ex. spel med 3D-grafik, webbläsare och andra nyttiga program tillverkas.

Pesquisa socioeconômica do ABC 2009

A Pesquisa Socioeconômica do ABC é realizada semestralmente para o conjunto regional que incorpora os municípios de Santo André, São Bernardo do Campo e São Caetano do Sul, pelo Instituto de Pesquisas da USCS, a partir de um levantamento por amostragem probabilística dos domicílios, desde 1983. Recentemente, no mês de setembro de 2009, expandiu a coleta também para os municípios de Diadema, Mauá, Ribeirão Pires e Rio Grande da Serra com previsão de novas coletas anuais. Os entrevistados no domicílio com idade mínima 18 anos, são sorteados a partir de um processo aleatório e segundo cotas estabelecidas para a variável gênero (50% de entrevistados do gênero masculino e 50% do gênero feminino). Os dados são coletados mediante aplicação de questionário estruturado, a partir de entrevista pessoal, com duração média de 60 minutos. A margem de erro estimada, para a região do ABC, em cada levantamento para os resultados obtidos a partir do total de casos é de 4,0% para 95,5% de confiança e para a região do Grande ABC, é de 3,0% para 95,5% de confiança. Para os resultados obtidos mediante um número menor de casos válidos, a margem de erro diferencia-se da anteriormente apresentada no seguinte sentido: “à medida que diminui o número de casos válidos aumenta o erro estatístico”, devendo esse erro ser estimado para cada variável investigada.

As mudanças na organização da produção na indústria gáfica do Grande ABC: um estudo sobre o segmento off set

O presente trabalho tem por objetivo caracterizar a indústria gráfica do ponto de vista da gestão, tecnologia, inovação e competição. A economia ao longo de sua história foi marcante por alguma situação peculiar que a caracterizasse. Em particular a partir da década de 1960 acompanhamos o crescimento em larga escala na região do Grande ABC onde, a indústria automobilística e seus fornecedores necessitaram de maior estrutura e apoio para as suas operações, provocando alterações nos modos de atuação das empresas industriais no país. Nessa mesma direção pelas necessidades criadas à época, a indústria gráfica na região do ABC também teve seu crescimento para atender a demanda e, criando o caráter da regionalidade e se fortalecendo economicamente. Entretanto a partir da década de 1980 com as crises econômicas e a abertura de mercado, houve uma redução nos postos de trabalho, mas as empresas também alteraram as formas de produzir. O trabalho de campo foi apoiado num referencial teórico baseado na revisão bibliográfica e um roteiro de entrevistas que orientou a coleta de dados. Na presente pesquisa foram realizadas entrevistas com os sujeitos relacionados e verificação de documentos. A análise realizada buscou confrontar os dados coletados e sistematizados com o quadro conceitual utilizado para a elaboração da pesquisa, possibilitando apurar as principais transformações verificadas no segmento estudado, bem como oferecendo embasamento para a tomada de decisão dos atores econômicos envolvidos com o desenvolvimento da indústria gráfica no Grande ABC.