APPLICATION OF MULTICRITERIA METHODS TO PLANNING OF INVESTMENT PROJECTS IN THE FIELD OF CIVIL ENGINEERING

In this paper we present a new approach for the planning of an investment project in the field of building construction. The focus is in shaping of the concept that serves to facilitate decision making about investments through providing support to investors’ when they are dealing with a problem - selection of a solution for investment project. The concept is based on the combined use of several different criteria, conventional methods for the evaluation of investment projects and multicriteria methods (PROMETHEE and AHP). Selected criteria encompassed economic, social-administrative, construction-functional and spatial aspects of selection problem. The determining criteria weights process is carried out in two steps. In the first step the AHP method is used to obtain the primary weights. The second step allows interactive involvement of investor in the process of solution selection (an investor-subjective approach to the problem). Presented approach is tested on the planning of nvestment project in Croatia

Cysteine Peptidases, Secreted by Trichomonas gallinae, Are Involved in the Cytopathogenic Effects on a Permanent Chicken Liver Cell Culture

Trichomonas gallinae, the aetiological agent of avian trichomonosis, was shown to secrete soluble factors involved in cytopathogenic effect on a permanent chicken liver (LMH) cell culture. The present study focused on the characterization of these molecules. The addition of specific peptidase inhibitors to the cell-free filtrate partially inhibited the monolayer destruction, which implied the presence of peptidases in the filtrate and their involvement in the cytopathogenic effect. One-dimensional substrate (gelatin) SDS-PAGE confirmed the proteolytic character of the filtrate by demonstrating the proteolytic activity within the molecular weight range from 38 to 110 kDa. In addition, the proteolytic activity was specifically inhibited by addition of TLCK and E-64 cysteine peptidase inhibitors implying their cysteine peptidase nature. Furthermore, variations in the intensity and the number of proteolytic bands were observed between cell-free filtrates of low and high passages of the same T. gallinae clonal culture. Two-dimensional substrate gel electrophoresis of concentrated T. gallinae cell-free filtrate identified at least six proteolytic spots. The mass spectrometric analysis of spots from 2-D gels identified the presence of at least two different Clan CA, family C1, cathepsin L-like cysteine peptidases in the cell-free filtrate of T. gallinae. In parallel, a PCR approach using degenerated primers based on the conserved amino acid sequence region of cysteine peptidases from Trichomonas vaginalis identified the coding sequences for four different Clan CA, family C1, cathepsin L-like cysteine peptidases. Finally, this is the first report analyzing molecules secreted by T. gallinae and demonstrating the ubiquity of peptidases secreted by this protozoon

Establishment of a novel probe-based RT-qPCR approach for detection and quantification of tight junctions reveals age-related changes in the gut barriers of broiler chickens

Tight junctions (TJs) play a dominant role in gut barrier formation, therefore, resolving the structures of TJs in any animal species is crucial but of major importance in fast growing broilers. They are regulated in molecular composition, ultrastructure and function by intracellular proteins and the cytoskeleton. TJ proteins are classified according to their function into barrier-forming, scaffolding and pore-forming types with deductible consequences for permeability. In spite of their importance for gut health and its integrity limited studies have investigated the TJs in chickens, including the comprehensive evaluation of TJs molecular composition and function in the chicken gut. In the actual study sequence-specific probes to target different TJ genes (claudin 1, 3, 5, 7, 10, 19, zonula occludens 1 (ZO1), occludin (OCLN) and tricellulin (MD2)) were designed and probe-based RT-qPCRs were newly developed. Claudin (CLDN) 1, 5, ZO1 and CLDN 3, 7, MD2 were engulfed in multiplex RT-qPCRs, minimizing the number of separate reactions and enabling robust testing of many samples. All RT-qPCRs were standardized for chicken jejunum and caecum samples, which enabled specific detection and quantification of the gene expression. Furthermore, the newly established protocols were used to investigate the age developmental changes in the TJs of broiler chickens from 1-35 days of age in the same organ samples. Results revealed a significant increase in mRNA expression between 14 and 21days of age of all tested TJs in jejunum. However, in caecum, mRNA expression of some TJs decreased after 1 day of age whereas some TJs mRNA remained constant till 35 days of age. Taken together, determining the segment-specific changes in the expression of TJ- proteins by RT-qPCR provides a deeper understanding of the molecular mechanisms underpinning pathophysiological changes in the gut of broiler chickens with various etiologies

INTERSTITIAL 14q31.3-q32.13 DELETION: THE ROLE OF MOLECULAR KARYOTYPING IN CLARIFYING THE ETIOLOGY OF DEVELOPMENTAL DELAY

Aim: With the exception of ring chromosome 14 or translocations, interstitial deletions of the long arm of chromosome 14 are very rare. All patients with these deletions share common phenotypic characteristics, primarily mild dysmorphia and developmental delay. Molecular karyotyping (array CGH) enabled the precise breakpoint determination and improved the analysis of genotype-phenotype correlations. Case presentation: In a 7-year-old girl, array CGH was performed due to developmental delay. The array CGH study showed 8.3Mb de novo interstitial deletion of the 14q31.3–q32.13 region. Conclusions: Comparison of our patient´s phenotype with previously reported chromosome 14q interstitial deletion cases confirmed the presence of common clinical features and highlights the utility of array CGH as a diagnostic tool in clarifying the developmental delay etiology

Identification of a new reovirus causing tendinitis in broilers in France

Clinical cases of tendinitis have appear sporadically and then more regularly in broilers flocks among different regions in France since 2010. These tendinitis have been identified as a consequence of Reovirus infection despite the vaccination of breeders stock with vaccines containing different strains of Reovirus previously described in the field ; additionnally an horizontal transmission, especially at the time of hatch, has been observed. Virological studies conducted in two laboratories lead to the identification of a new type of Reovirus non described in Europe until now. In addition to genetic differences between this new virus and the vaccine strains used on the field, cross seroneutralisation tests have shown antigenic differences which could explain the inefficacy of the vaccines used in the field. In order to prevent the multiplication of theses viruses it seems usefull to update the composition of vaccines for a better protection of breeders and their progenyDes cas cliniques de tendinite sont apparus sporadiquement, puis plus régulièrement, dans des élevages de poulets de chair dans différentes régions de France depuis 2010. Ces tendinites ont été identifiées comme dues à une réovirose en dépit de la vaccination des poules parentales avec des vaccins contenant différentes valences de réovirus précédemment décrits comme présents sur le terrain ; en outre une transmission horizontale notamment lors de l’éclosion a été observée. Les études virologiques conduites dans deux laboratoires aboutissent à l’identification d’un nouveau réovirus jusqu’à présent non décrit en Europe. Outre des différences au plan génétique entre ce nouveau virus et les souches vaccinales utilisées sur le terrain, des tests de séroneutralisation croisée ont montré des différences antigéniques, ce qui peut expliquer l’inefficacité des vaccins observée sur le terrain. Pour prévenir la multiplication de ces virus il apparait donc utile de réactualiser la composition des vaccins de manière à protéger les poules reproductrices et leur descendanc

The NEI/NCBI dbGAP database: Genotypes and haplotypes that may specifically predispose to risk of neovascular age-related macular degeneration

<p>Abstract</p> <p>Background</p> <p>To examine if the significantly associated SNPs derived from the genome wide allelic association study on the AREDS cohort at the NEI (dbGAP) specifically confer risk for neovascular age-related macular degeneration (AMD). We ascertained 134 unrelated patients with AMD who had one sibling with an AREDS classification 1 or less and was past the age at which the affected sibling was diagnosed (268 subjects). Genotyping was performed by both direct sequencing and Sequenom iPLEX system technology. Single SNP analyses were conducted with McNemar's Test (both 2 × 2 and 3 × 3 tests) and likelihood ratio tests (LRT). Conditional logistic regression was used to determine significant gene-gene interactions. LRT was used to determine the best fit for each genotypic model tested (additive, dominant or recessive).</p> <p>Results</p> <p>Before release of individual data, <it>p</it>-value information was obtained directly from the AREDS dbGAP website. Of the 35 variants with <it>P </it>< 10^-6examined, 23 significantly modified risk of neovascular AMD. Many variants located in tandem on 1q32-q22 including those in <it>CFH</it>, <it>CFHR4</it>, <it>CFHR2</it>, <it>CFHR5</it>, <it>F13B</it>, <it>ASPM </it>and <it>ZBTB </it>were significantly associated with AMD risk. Of these variants, single SNP analysis revealed that <it>CFH </it>rs572515 was the most significantly associated with AMD risk (P < 10^-6). Haplotype analysis supported our findings of single SNP association, demonstrating that the most significant haplotype, GATAGTTCTC, spanning <it>CFH</it>, <it>CFHR4</it>, and <it>CFHR2 </it>was associated with the greatest risk of developing neovascular AMD (<it>P </it>< 10^-6). Other than variants on 1q32-q22, only two SNPs, rs9288410 (<it>MAP2</it>) on 2q34-q35 and rs2014307 (<it>PLEKHA1</it>/<it>HTRA1</it>) on 10q26 were significantly associated with AMD status (<it>P </it>= .03 and <it>P </it>< 10^-6respectively). After controlling for smoking history, gender and age, the most significant gene-gene interaction appears to be between rs10801575 (<it>CFH</it>) and rs2014307 (<it>PLEKHA1</it>/<it>HTRA1</it>) (<it>P </it>< 10^-11). The best genotypic fit for rs10801575 and rs2014307 was an additive model based on LRT. After applying a Bonferonni correction, no other significant interactions were identified between any other SNPs.</p> <p>Conclusion</p> <p>This is the first replication study on the NEI dbGAP SNPs, demonstrating that alleles on 1q, 2q and 10q may predispose an individual to AMD.</p

Phylogeny of Parasitic Parabasalia and Free-Living Relatives Inferred from Conventional Markers vs. Rpb1, a Single-Copy Gene

Parabasalia are single-celled eukaryotes (protists) that are mainly comprised of endosymbionts of termites and wood roaches, intestinal commensals, human or veterinary parasites, and free-living species. Phylogenetic comparisons of parabasalids are typically based upon morphological characters and 18S ribosomal RNA gene sequence data (rDNA), while biochemical or molecular studies of parabasalids are limited to a few axenically cultivable parasites. These previous analyses and other studies based on PCR amplification of duplicated protein-coding genes are unable to fully resolve the evolutionary relationships of parabasalids. As a result, genetic studies of Parabasalia lag behind other organisms.Comparing parabasalid EF1α, α-tubulin, enolase and MDH protein-coding genes with information from the Trichomonas vaginalis genome reveals difficulty in resolving the history of species or isolates apart from duplicated genes. A conserved single-copy gene encodes the largest subunit of RNA polymerase II (Rpb1) in T. vaginalis and other eukaryotes. Here we directly sequenced Rpb1 degenerate PCR products from 10 parabasalid genera, including several T. vaginalis isolates and avian isolates, and compared these data by phylogenetic analyses. Rpb1 genes from parabasalids, diplomonads, Parabodo, Diplonema and Percolomonas were all intronless, unlike intron-rich homologs in Naegleria, Jakoba and Malawimonas.The phylogeny of Rpb1 from parasitic and free-living parabasalids, and conserved Rpb1 insertions, support Trichomonadea, Tritrichomonadea, and Hypotrichomonadea as monophyletic groups. These results are consistent with prior analyses of rDNA and GAPDH sequences and ultrastructural data. The Rpb1 phylogenetic tree also resolves species- and isolate-level relationships. These findings, together with the relative ease of Rpb1 isolation, make it an attractive tool for evaluating more extensive relationships within Parabasalia

APPLICATION OF MULTICRITERIA METHODS TO PLANNING OF INVESTMENT PROJECTS IN THE FIELD OF CIVIL ENGINEERING

In this paper we present a new approach for the planning of an investment project in the field of building construction. The focus is in shaping of the concept that serves to facilitate decision making about investments through providing support to investors’ when they are dealing with a problem - selection of a solution for investment project. The concept is based on the combined use of several different criteria, conventional methods for the evaluation of investment projects and multicriteria methods (PROMETHEE and AHP). Selected criteria encompassed economic, social-administrative, construction-functional and spatial aspects of selection problem. The determining criteria weights process is carried out in two steps. In the first step the AHP method is used to obtain the primary weights. The second step allows interactive involvement of investor in the process of solution selection (an investor-subjective approach to the problem). Presented approach is tested on the planning of nvestment project in Croatia

O croata como língua de herança na Argentina: entre a vulnerabilidade e a manutenção linguística

En el contexto sudamericano, la población migrante representa una porción numerosa que, en la mayoría de los casos, deja de usar su lengua de herencia. El propósito de este trabajo es presentar los resultados de un estudio que indagó por la situación de la lengua croata como lengua de herencia en el contexto sudamericano, atendiendo a la tensión entre desplazamiento y mantenimiento lingüístico. El análisis se centra en Argentina, donde se estima que entre 250 000 y 500 000 personas son descendientes de migrantes del pueblo croata. El estudio se sustenta en un corpus de habla croata de migrantes de segunda y tercera generaciones, así como en documentos oficiales y no oficiales, reunidos por los autores. Los resultados sugieren que existen signos de erosión lingüística en los niveles fonológico, morfosintáctico y léxico. A su vez, la identidad y el fortalecimiento de la conciencia lingüística cumplen un rol relevante a favor del mantenimiento de la lengua ancestral en el contexto migratorio. El trabajo aporta al estudio de las lenguas minoritarias y amenazadas, y contribuye a la comprensión de los procesos dinámicos que atraviesa la vitalidad de las lenguas de herencia.In the South American context, migrant population makes up for a large portion of the population who, in most cases, stop using their heritage language. This work aims to present the results of a study that inquired about the status of the Croatian language as a heritage language in South America, and analized the tensions between linguistic displacement and maintenance. The analysis is focused on Argentina, where an estimated 250 000 to 500 000 people are from Croatian descent. The study is based upon a corpus of Croatian speech gathered from second and third-generation migrants and from official and non-official records, gathered by the authors. Findings suggest that there are signs of linguistic attrition at the phonological, morphosyntactic, and lexical levels. They also suggest that identity and language awareness strengthening play a significant role in the maintenance of the ancestral language in this migrating context. The study contributes to the field of minority and endangered languages, and helps understand the dynamic processes influencing the vitality of heritage languages.Dans le contexte sudaméricain, la population migrante représente une grande portion qui, dans la plupart des cas, cesse d’utiliser sa langue de patrimoine. Ce travail présente les résultats d'une étude sur la situation de la langue croate en tant que langue d’héritage dans le contexte sud-américain, en tenant compte de la tension entre déplacement et maintien linguistique. Cette analyse s'est concentrée sur l’Argentine, où l’on estime qu’entre 250 000 et 500 000 personnes sont des descendants de migrants issus du peuple croate. L’étude s’est appuyée sur un corpus croate de migrants de deuxième et troisième générations, ainsi que sur des documents officiels et non officiels, recueillis par les auteurs. Les résultats suggèrent qu’il existe des signes d’attrition linguistique aux niveaux phonologique, morphosyntaxique et lexical. À leur tour, l’identité et le renforcement de la conscience linguistique jouent un rôle pertinent en faveur du maintien de la langue ancestrale dans le contexte migratoire. L’ouvrage contribue à l’étude des langues minoritaires et menacées, et contribue à la compréhension des processus dynamiques que traverse la vitalité des langues patrimoniales.No contexto sul-americano, a população migrante representa uma grande parcela que, na maioria das vezes, deixa de usar sua língua de herança. O objetivo deste trabalho é apresentar um estudo da situação da língua croata como língua de herança no contexto sul-americano, levando em consideração a tensão entre deslocamento e manutenção linguística. A análise tem se concentrado na Argentina, onde se estima que entre 250.000 e 500.000 pessoas sejam descendentes de migrantes do povo croata. O estudo tem se baseado em um corpus croata de migrantes de segunda e terceira geração, bem como em documentos oficiais e não oficiais, coletados pelos autores. Os resultados sugerem que há indícios de erosão linguística nos níveis fonológico, morfossintático e lexical. Por sua vez, a identidade e o reforço da consciência linguística desempenham um papel relevante a favor da manutenção da língua ancestral no contexto migratório. A obra contribui para o estudo das línguas minoritárias e ameaçadas, e contribui para a compreensão dos processos dinâmicos pelos quais passa a vitalidade das línguas de herança.Fil: Bilic, Josip Bruno. Universidad de Buenos Aires. Facultad de Filosofía y Letras; ArgentinaFil: Cúneo, Paola. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Filosofía y Letras. Instituto de Lingüística; ArgentinaFil: Franic, Ivana. Universidad de Zagreb; Croaci

Vaccination against the Protozoan Parasite Histomonas meleagridis Primes the Activation of Toll-like Receptors in Turkeys and Chickens Determined by a Set of Newly Developed Multiplex RT-qPCRs

Histomonosis in turkeys and chickens is caused by the extracellular parasite Histomonas meleagridis, but the outcome of the disease varies depending on the host species. So far, studies on the immune response against histomonosis focus mainly on different traits of the adaptive immune system. Activation of toll like receptors (TLR) leads to the interplay between cells of innate and adaptive immunity with consequences on B and T cell clonal expansion. Therefore, the present investigation focused on the interaction of virulent and/or attenuated histomonads with the innate immune system of turkeys and chickens at 4, 10, 21 days post inoculation. The expression of TLRs (TLR1A, 1B, 2A, 2B, 3, 4, 5, 6(Tu), 7, 13(Tu) and 21(Ch)) and pro-inflammatory cytokines (IL1β and IL6) were analysed in caecum and spleen samples by RT-qPCR. Most frequent significant changes in expression levels of TLRs were observed in the caecum following infection with virulent parasites, an effect noticed to a lower degree in tissue samples from birds vaccinated with attenuated parasites. TLR1B, 2B and 4 showed a continuous up-regulation in the caecum of both species during infection or vaccination, followed by challenge with virulent parasites. Vaccinated birds of both species showed a significant earlier change in TLR expression following challenge than birds kept non-vaccinated but challenged. Expression of TLRs and pro-inflammatory cytokines were associated with severe inflammation of diseased birds in the local organ caecum. In the spleen, changes in TLRs and pro-inflammatory cytokines were less prominent and mainly observed in turkey samples. In conclusion, a detailed comparison of TLRs and pro-inflammatory cytokines of the innate immune system following inoculation with attenuated and/or virulent H. meleagridis of two avian host species provides an insight into regulative mechanisms of TLRs in the development of protection and limitation of the disease