Evaluation of the mechanical properties of cements with fillers derived from the CO2 reduction of cement plants

This work introduces a novel method for the development of CO2 recovery systems derived from the production process of cement in order to obtain CaCO3 nanofiller in cement-based composites. Research was carried out in collaboration between the Department of Applied Science and Technology (DISAT) and the Department of Structural, Construction and Geotechnical Engineering (DISEG) of Politecnico di Torino. The objective of this method was dual. Firstly, it aimed to obtain a precipitated calcium carbonate - nanoCaCO3 - with a high degree of purity. Secondly, it aimed to optimize the characteristics of these nanoparticles e.g. additional percentages, morphology, particle size distribution or crystal phase, according to their use in cement-based composites. The synthesized nanoCaCO3 particles were subsequently added into the cementitious composites in different percentages according to the weight of the cement, in order to understand their behaviour within the cement matrix. The mechanical properties were also evaluated, both at 7 and 28 days, through three point bending and compression tests. The results of the mechanical tests showed a promising improvement in strength and toughness. This study is a first step towards developing a CO2 circular economy

Identification of New Genes Related to Virulence of Xanthomonas axonopodis Pv. Citri during Citrus Host Interactions

A mutant library of the bacterium Xanthomonas citri subsp. citri strain 306 pathotype A (Xac), the causative agent of most aggressive Asiatic type A citrus canker, was screened regarding altered canker symptoms after inoculations into Citrus sinensis and Citrus limonia host leaves. Twenty-six mutants have shown phenotypic virulence changes and have respectively knocked out gene identified by sequencing. In vivo growth curves were obtained for nine mutants to quantify how the mutations could affect pathogen's adaptability to growth inside and attack host plant infected tissue. Among identified genes in mutated strains, we could find those that until now had not been reported as being involved in Xac adaptation and/or virulence, such as predicted to encode for xylose repressor-like protein (XAC Delta xylR), Fe-S oxidoredutase (XAC Delta aslB), helicase IV (XAC Delta helD), ubiquinol cytochrome c oxidoreductase iron-sulfur subunit (XAC Delta petA), chromosome partitioning protein (XAC Delta parB) and cell division protein FtsB (XAC Delta ftsB), in addition to genes predicted to encode for hypothetical proteins. The new genes found in this study as being relevant to adaptation and virulence, improve the understanding of Xac fitness during citrus plant attack and canker symptoms development.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo-FAPESPCoordenacao de Aperfeicoamento de Pessoal de Nivel Superior-CAPESCNPqFundo de Defesa da Citricultura (FUNDECITRUS)CAPESFUNDECITRUSFAPESPSao Paulo State Univ UNESP, Fac Agr Sci & Vet, Dept Technol, Jaboticabal, SP, BrazilCitriculture Res Ctr Sylvio Moreira IAC Campinas, Campinas, SP, BrazilUniv Fed Ouro Preto, Inst Exact & Biol Sci, Dept Biol Sci, Ouro Preto, MG, BrazilUniv Fed Ouro Preto, Res Ctr Biol Sci NUPEB, Inst Exact & Biol Sci, Dept Biol Sci, Ouro Preto, MG, BrazilSao Paulo State Univ UNESP, Fac Agr Sci & Vet, Plant Hlth Dept, Jaboticabal, SP, BrazilFed Univ Sao Paulo UNIFESP, Dept Biol Sci, Diadema, SP, BrazilFed Univ Sao Paulo UNIFESP, Dept Biol Sci, Diadema, SP, BrazilFAPESP: 04/02006-7Web of Scienc

Selection of endogenous genes for gene expression studies in Eucalyptus under biotic (Puccinia psidii) and abiotic (acibenzolar-S-methyl) stresses using RT-qPCR

<p>Abstract</p> <p>Background</p> <p>Rust caused by <it>Puccinia psidii </it>Winter has been limiting for the establishment of new <it>Eucalyptus </it>plantations, as well as for resprouting of susceptible genetic materials. Identifying host genes involved in defense responses is important to elucidate resistance mechanisms. Reverse transcription-quantitative PCR is the most common method of mRNA quantitation for gene expression analysis. This method generally employs a reference gene as an internal control to normalize results. A good endogenous control transcript shows minimal variation due to experimental conditions.</p> <p>Findings</p> <p>We analyzed the expression of 13 genes to identify transcripts with minimal variation in leaves of 60-day-old clonal seedlings of two <it>Eucalyptus </it>clones (rust-resistant and susceptible) subjected to biotic (<it>P. psidii</it>) and abiotic (acibenzolar-S-methyl, ASM) stresses.</p> <p>Conclusions</p> <p>For tissue samples of clones that did not receive any stimulus, a combination of the <it>eEF2 </it>and <it>EglDH </it>genes was the best control for normalization. When pathogen-inoculated and uninoculated plant samples were compared, <it>eEF2 </it>and <it>UBQ </it>together were more appropriate as normalizers. In ASM-treated and untreated leaves of both clones, transcripts of the <it>CYP </it>and <it>elF4B </it>genes combined were the ones with minimal variation. Finally, when comparing expression in both clones for ASM-treated leaves, <it>P. psidii</it>-inoculated leaves, ASM-treated plus <it>P. psidii</it>-inoculated leaves, and their respective controls, the genes with the most stable expression were <it>EgIDH </it>and <it>UBQ</it>. The chitinase gene, which is highly expressed in studies on plant resistance to phytopathogens, was used to confirm variation in gene expression due to the treatments.</p

New genes of Xanthomonas citri subsp. citri involved in pathogenesis and adaptation revealed by a transposon-based mutant library

<p>Abstract</p> <p>Background</p> <p>Citrus canker is a disease caused by the phytopathogens <it>Xanthomonas citri </it>subsp. <it>citri</it>, <it>Xanthomonas fuscans </it>subsp. <it>aurantifolli </it>and <it>Xanthomonas alfalfae </it>subsp. <it>citrumelonis</it>. The first of the three species, which causes citrus bacterial canker type A, is the most widely spread and severe, attacking all citrus species. In Brazil, this species is the most important, being found in practically all areas where citrus canker has been detected. Like most phytobacterioses, there is no efficient way to control citrus canker. Considering the importance of the disease worldwide, investigation is needed to accurately detect which genes are related to the pathogen-host adaptation process and which are associated with pathogenesis.</p> <p>Results</p> <p>Through transposon insertion mutagenesis, 10,000 mutants of <it>Xanthomonas citri </it>subsp. <it>citri </it>strain 306 (Xcc) were obtained, and 3,300 were inoculated in Rangpur lime (<it>Citrus limonia</it>) leaves. Their ability to cause citrus canker was analyzed every 3 days until 21 days after inoculation; a set of 44 mutants showed altered virulence, with 8 presenting a complete loss of causing citrus canker symptoms. Sequencing of the insertion site in all 44 mutants revealed that 35 different ORFs were hit, since some ORFs were hit in more than one mutant, with mutants for the same ORF presenting the same phenotype. An analysis of these ORFs showed that some encoded genes were previously known as related to pathogenicity in phytobacteria and, more interestingly, revealed new genes never implicated with <it>Xanthomonas </it>pathogenicity before, including hypothetical ORFs. Among the 8 mutants with no canker symptoms are the <it>hrpB4 </it>and <it>hrpX </it>genes, two genes that belong to type III secretion system (TTSS), two hypothetical ORFS and, surprisingly, the <it>htrA </it>gene, a gene reported as involved with the virulence process in animal-pathogenic bacteria but not described as involved in phytobacteria virulence. Nucleic acid hybridization using labeled cDNA probes showed that some of the mutated genes are differentially expressed when the bacterium is grown in citrus leaves. Finally, comparative genomic analysis revealed that 5 mutated ORFs are in new putative pathogenicity islands.</p> <p>Conclusion</p> <p>The identification of these new genes related with Xcc infection and virulence is a great step towards the understanding of plant-pathogen interactions and could allow the development of strategies to control citrus canker.</p

Proteome of the phytopathogen Xanthomonas citri subsp. citri: a global expression profile

<p>Abstract</p> <p>Background</p> <p>Citrus canker is a disease caused by <it>Xantomonas citri </it>subsp.<it>citri (Xac)</it>, and has emerged as one of the major threats to the worldwide citrus crop because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in citrus growing areas. In this work, the first proteome of <it>Xac </it>was analyzed using two methodologies, two-dimensional liquid chromatography (2D LC) and tandem mass spectrometry (MS/MS).</p> <p>Results</p> <p>In order to gain insight into the metabolism of <it>Xac</it>, cells were grown on two different media (<b>NB </b>- Nutrient Broth and <b>TSE </b>- Tryptone Sucrose broth enriched with glutamic acid), and proteins were proteolyzed with trypsin and examined by 2D LC-MS/MS. Approximately 39% of all predicted proteins by annotation of <it>Xac </it>were identified with their component peptides unambiguously assigned to tandem mass spectra. The proteins, about 1,100, were distributed in all annotated functional categories.</p> <p>Conclusions</p> <p>This is the first proteomic reference map for the most aggressive strain of <it>Xanthomonas </it>pathogen of all orange varieties. The compilation of metabolic pathways involved with bacterial growth showed that <it>Xac </it>expresses a complete central and intermediary metabolism, replication, transcription and translation machineries and regulation factors, distinct membrane transporters (ABC, MFS and pumps) and receptors (MCP, TonB dependent and metabolites acquisition), two-component systems (sensor and regulatory components) and response regulators. These data corroborate the growth curve <it>in vitro </it>and are the first reports indicating that many of these genome annotated genes are translated into operative in <it>Xac</it>. This proteomic analysis also provided information regarding the influence of culture medium on growth and protein expression of <it>Xac</it>.</p

Proteomics-based identification of differentially abundant proteins reveals adaptation mechanisms of Xanthomonas citri subsp citri during Citrus sinensis infection

Background: Xanthomonas citri subsp. citri (Xac) is the causal agent of citrus canker. A proteomic analysis under in planta infectious and non-infectious conditions was conducted in order to increase our knowledge about the adaptive process of Xac during infection. Results: For that, a 2D-based proteomic analysis of Xac at 1, 3 and 5 days after inoculation, in comparison to Xac growth in NB media was carried out and followed by MALDI-TOF-TOF identification of 124 unique differentially abundant proteins. Among them, 79 correspond to up-regulated proteins in at least one of the three stages of infection. Our results indicate an important role of proteins related to biofilm synthesis, lipopolysaccharides biosynthesis, and iron uptake and metabolism as possible modulators of plant innate immunity, and revealed an intricate network of proteins involved in reactive oxygen species adaptation during Plants'Oxidative Burst response. We also identified proteins previously unknown to be involved in Xac-Citrus interaction, including the hypothetical protein XAC3981. A mutant strain for this gene has proved to be non-pathogenic in respect to classical symptoms of citrus canker induced in compatible plants. Conclusions: This is the first time that a protein repertoire is shown to be active and working in an integrated manner during the infection process in a compatible host, pointing to an elaborate mechanism for adaptation of Xac once inside the plant.Fundacao de Amparo a Pesquisa do Estado de Sao PauloFundacao de Amparo a Pesquisa do Estado de Minas GeraisBIGA grant-CAPESUniv Fed Ouro Preto, Inst Ciencias Exatas & Biol, Dept Ciencias Biol DECBI, Ouro Preto, MG, BrazilUniv Fed Ouro Preto, Nucleo Pesquisas Ciencias Biol NUPEB, Ouro Preto, MG, BrazilUniv Fed Rio de Janeiro, Inst Quim, Dept Bioquim DBq, Rio De Janeiro, RJ, BrazilUniv Estadual Paulista, UNESP, Dept Tecnol, Fac Ciencias Agr & Vet Jaboticabal, Jaboticabal, SP, BrazilUniv Estadual Campinas, UNICAMP, Inst Quim, Campinas, SP, BrazilUniv Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo, SP, BrazilUniv Fed Sao Paulo UNIFESP, Dept Ciencias Biol, Diadema, SP, BrazilVirginia Tech, Biocomplex Inst, Blacksburg, VA USAUniv Sao Paulo, Inst Quim, Dept Bioquim, Sao Paulo, SP, BrazilUniv Fed Sao Paulo UNIFESP, Dept Ciencias Biol, Diadema, SP, BrazilCAPESFAPESP: 04/02006-7Fundacao de Amparo a Pesquisa do Estado de Minas Gerais: CBB-APQ-04425-10BIGA grant-CAPES: 3385/2013Web of Scienc

TOIMINNANOHJAUSJÄRJESTELMÄN KÄYTTÖÖNOTON VAIKUTUS

Tässä tutkielmassa tutkitaan kuinka toiminnanohjausjärjestelmän käyttöönotto on vaikuttanut yrityksen tulokseen ja suorituskykyyn suomalaisissa yrityksissä 1999-2015. Tavoitteena on testata empiirisesti aiheuttaako toiminnanohjausjärjestelmän käyttöönotto merkittäviä säästöjä, yrityk-sen toiminnan tehokkuuden parannuksia ja muutoksia yrityksen kannattavuudessa. Toiminnanohjaus- eli ERP-järjestelmät integroivat yrityksen eri toimintoja esimerkiksi logistiikkaa, tuotantoa, kirjanpitoa, varastointia ja laskutusta yhteen järjestelmään ja näin ollen tehostavat yrityksen toimintaa useiden eri järjestelmien käytön sijaan. Onkin tärkeää selvittää kuinka toiminnanohjausjärjestelmien käyttöönotto on vaikuttanut yritysten kannattavuuteen ja voidaanko investointi nähdä tuottavana. Aikaisempien tutkimusten perusteella tulokset ovat ristiriitaisia; tutkimukset osoittavat, että vaikka ERP- järjestelmien käyttöönotto tutkitusti vaikuttaa ei-rahamääräisiin mittareihin, vaikutusta rahamääräisiin mittareihin ei aina pystytä todistamaan. Yritysten kannalta mielenkiintoista onkin ovatko kalliit ERP-järjestelmät, sekä niiden implementointiprosessit todella kannattavia investointeja. Tutkimuksen empiirisessä osiossa selvitettiin toiminnanohjausjärjestelmien käyttöönoton vaiku-tuksia suorituskykyyn suomalaisissa yhtiöissä, jotka ovat implementoineet toiminnanohjausjärjestelmän vuosina 1999-2015. Aineistona on käytetty julkisuudessa ilmoitettuja järjestelmähank-keita sekä tilinpäätöstietoja implementointiprosessin jälkeisiltä sekä edeltäviltä vuosilta. Suorituskyvyn muutosta on seurattu vertailemalla suorituskyvyn ja kannattavuuden muutosta implementoinnin jälkeen. Tutkimus on suoritettu määrällisenä tutkimuksena ja tilastollisina menetelminä on käytetty toistomittausten varianssianalyysiä sekä t-testiä parivertailulle. Tutkimuksessa havaittiin, että näyttäisi siltä, ettei toiminnanohjausjärjestelmien käyttöönotolla ei ole ollut vaikutusta kohdeyhtiöiden rahamääräiseen suorituskykyyn.fi=Opinnäytetyö kokotekstinä PDF-muodossa.|en=Thesis fulltext in PDF format.|sv=Lärdomsprov tillgängligt som fulltext i PDF-format

Detection and identification of Xanthomonas pathotypes associated with citrus diseases using comparative genomics and multiplex PCR

Background. In Citrus cultures, three species of Xanthomonas are known to cause distinct diseases. X. citri subsp. citri patothype A, X. fuscans subsp. aurantifolii pathotypes B and C, and X. alfalfae subsp. citrumelonis, are the causative agents of cancrosis A, B, C, and citrus bacterial spots, respectively. Although these species exhibit different levels of virulence and aggressiveness, only limited alternatives are currently available for proper and early detection of these diseases in the fields. The present study aimed to develop a new molecular diagnostic method based on genomic sequences derived from the four species of Xanthomonas. Results. Using comparative genomics approaches, primers were synthesized for the identification of the four causative agents of citrus diseases. These primers were validated for their specificity to their target DNA by both conventional and multiplex PCR. Upon evaluation, their sensitivity was found to be 0.02 ng/mu l in vitro and 1.5 x 10(4) CFU ml(-1) in infected leaves. Additionally, none of the primers were able to generate amplicons in 19 other genomes of Xanthomonas not associated with Citrus and one species of Xylella, the causal agent of citrus variegated chlorosis (CVC). This denotes strong specificity of the primers for the different species of Xanthomonas investigated in this study. Conclusions. We demonstrated that these markers can be used as potential candidates for performing in vivo molecular diagnosis exclusively for citrus-associated Xanthomonas. The bioinformatics pipeline developed in this study to design specific genomic regions is capable of generating specific primers. It is freely available and can be utilized for any other model organism.7CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAIS - FAPEMIG481226/2013-3CFP 51/2013; 3385/2013APQ-02387-1