Metallothioneins in the clam Ruditapes decussatus: an overview

The clam Ruditapes decussatus is a suspension-feeding bivalve mollusc widely distributed in European waters and in the Mediterranean. Due to is economic importance it is heavily harvested in many countries, and particularly in Portugal. Its ability to accumulate high metal concentrations along with its economic importance was the main reason for its selection as a bioindicator. Metallothionein (MT) concentrations in the clams R. decussatus followed by gel filtration chromatography, differential pulse polarography and SDS-PAGE, after Cd exposure, revealed that MT is induced in different tissues (whole soft tissues, gills, digestive gland and remaining tissues) but the level of MT induction is tissue dependent. MT from the gills and the digestive gland give a more sensitive response to assess the effects of metal exposure directly from the water or from the food than in the whole soft tissues. MT levels were also measured in the gills, digestive gland and remaining tissues of R. decussatus collected in the Ria Formosa lagoon (Portugal) from areas of different metal load and during the period of sexual differentiation of the clam. Data revealed that there were significant differences of MT concentrations among sites and season but not among sex. Purification of MT from the digestive gland of R. decussatus revealed four MT isoforms. The molecular weight of one of these isoforms, determined by SDS-PAGE, was of the same order of magnitude as that of MT from other bivalve species. Similarly the amino acid sequence of the beta domain of the MT of the digestive gland of the clam also shows some degree of similarity with the similar MT sequence from mussels and oysters. It is, therefore suggested that there is some degree of similarity in the MT structure among these species.info:eu-repo/semantics/publishedVersio

Quantitative image analysis of polyhydroxyalkanoates inclusions from microbial mixed cultures under different SBR operation strategies

Polyhydroxyalkanoates (PHAs) produced from mixed microbial cultures (MMC), regarded as potential substitutes of petrochemical plastics, can be found as intracellular granules in various microorganisms under limited nutrient conditions and excess of carbon source. PHA is traditionally quantified by laborious and time-consuming chromatography analysis, and a simpler and faster method to assess PHA contents from MMC, such as quantitative image analysis (QIA), is of great interest. The main purpose of the present work was to upgrade a previously developed QIA methodology (Mesquita et al., 2013a, 2015) for MMC intracellular PHA contents quantification, increase the studied intracellular PHA concentration range and extend to different sequencing batch reactor (SBR) operation strategies. Therefore, the operation of a new aerobic dynamic feeding (ADF) SBR allowed further extending the studied operating conditions, dataset, and range of the MMC intracellular PHA contents from the previously reported anaerobic/aerobic cycle SBR. Nile Blue A (NBA) staining was employed for epifluorescence microscope visualization and image acquisition, further fed to a custom developed QIA. Data from each of the feast and famine cycles of both SBR were individually processed using chemometrics analysis, obtaining the correspondent partial least squares (PLS) models. The PHA concentrations determined from PLS models were further plotted against the results obtained in the standard chromatographic method. For both SBR the predicted ability was higher at the end of the feast stage than for the famine stage. Indeed, an independent feast and famine QIA data treatment was found to be fundamental to obtain the best prediction abilities. Furthermore, a promising overall correlation (R2 of 0.83) could be found combining the overall QIA data regarding the PHA prediction up to a concentration of 1785.1 mgL-1 (37.3 wt%). Thus, the results confirm that the presented QIA methodology can be seen as promising for estimating higher intracellular PHA concentrations for a larger reactors operation systems and further extending the prediction range of previous studies.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE01-0145-FEDER-000004) funded by European Regional Development Fundunder the scope ofNorte2020 - ProgramaOperacional Regional do Norte.The authors also acknowledge the financial support to Cristiano S. Leal (PTDC/EBB-EBI/103147/2008, FCOMP-01-0124-FEDER009704) and Daniela P. Mesquita through the FCT postdoctoral grant (SFRH/BPD/82558/2011).info:eu-repo/semantics/publishedVersio

Determination of βS haplotypes in patients with sickle-cell anemia in the state of Rio Grande do Norte, Brazil

βS haplotypes were studied in 47 non-related patients with sickle-cell anemia from the state of Rio Grande do Norte, Brazil. Molecular analysis was conducted by PCR/RFLP using restriction endonucleases XmnI, HindIII, HincII and HinfI to analyze six polymorphic sites from the beta cluster. Twenty-seven patients (57.5%) were identified with genotype CAR/CAR, 9 (19.1%) CAR/BEN, 6 (12.8%) CAR/CAM, 1 (2.1%) BEN/BEN, 2 (4.3%) CAR/Atp, 1 (2.1%) BEN/Atp and 1 (2.1%) with genotype Atp/Atp. The greater frequency of Cameroon haplotypes compared to other Brazilian states suggests the existence of a peculiarity of African origin in the state of Rio Grande do Norte

Autoimmune Disease Classification by Inverse Association with SNP Alleles

With multiple genome-wide association studies (GWAS) performed across autoimmune diseases, there is a great opportunity to study the homogeneity of genetic architectures across autoimmune disease. Previous approaches have been limited in the scope of their analysis and have failed to properly incorporate the direction of allele-specific disease associations for SNPs. In this work, we refine the notion of a genetic variation profile for a given disease to capture strength of association with multiple SNPs in an allele-specific fashion. We apply this method to compare genetic variation profiles of six autoimmune diseases: multiple sclerosis (MS), ankylosing spondylitis (AS), autoimmune thyroid disease (ATD), rheumatoid arthritis (RA), Crohn's disease (CD), and type 1 diabetes (T1D), as well as five non-autoimmune diseases. We quantify pair-wise relationships between these diseases and find two broad clusters of autoimmune disease where SNPs that make an individual susceptible to one class of autoimmune disease also protect from diseases in the other autoimmune class. We find that RA and AS form one such class, and MS and ATD another. We identify specific SNPs and genes with opposite risk profiles for these two classes. We furthermore explore individual SNPs that play an important role in defining similarities and differences between disease pairs. We present a novel, systematic, cross-platform approach to identify allele-specific relationships between disease pairs based on genetic variation as well as the individual SNPs which drive the relationships. While recognizing similarities between diseases might lead to identifying novel treatment options, detecting differences between diseases previously thought to be similar may point to key novel disease-specific genes and pathways

Sequencing of \u3ci\u3eAspergillus nidulans\u3c/i\u3e and comparative analysis with \u3ci\u3eA. fumigatus\u3c/i\u3e and \u3ci\u3eA. oryzae\u3c/i\u3e

The aspergilli comprise a diverse group of filamentous fungi spanning over 200 million years of evolution. Here we report the genome sequence of the model organism Aspergillus nidulans, and a comparative study with Aspergillus fumigatus, a serious human pathogen, and Aspergillus oryzae, used in the production of sake, miso, and soy sauce. Our analysis of genome structure provided a quantitative evaluation of forces driving long-term eukaryotic genome evolution. It also led to an experimentally validated model of mating-type locus evolution, suggesting the potential for sexual reproduction in A. fumigatus and A. oryzae. Our analysis of sequence conservation revealed over 5,000 non-coding regions actively conserved across all three species. Within these regions, we identified potential functional elements including a previously uncharacterized TPP riboswitch and motifs suggesting regulation in filamentous fungi by Puf family genes. We further obtained comparative and experimental evidence indicating widespread translational regulation by upstream open reading frames. These results enhance our understanding of these widely studied fungi as well as provide new insight into eukaryotic genome evolution and gene regulation. Document includes all supplementary information (820 pages). Supplementary files are also attached below as Related files. THERE IS NO SUPPLEMENTARY FILE #7. PDF file size (with supplementary files included) is 10 Mbytes. An optimized version of the ARTICLE ONLY is attached as a Related File and is 1.9 Mbytes

Sequencing of \u3ci\u3eAspergillus nidulans\u3c/i\u3e and comparative analysis with \u3ci\u3eA. fumigatus\u3c/i\u3e and \u3ci\u3eA. oryzae\u3c/i\u3e

The aspergilli comprise a diverse group of filamentous fungi spanning over 200 million years of evolution. Here we report the genome sequence of the model organism Aspergillus nidulans, and a comparative study with Aspergillus fumigatus, a serious human pathogen, and Aspergillus oryzae, used in the production of sake, miso, and soy sauce. Our analysis of genome structure provided a quantitative evaluation of forces driving long-term eukaryotic genome evolution. It also led to an experimentally validated model of mating-type locus evolution, suggesting the potential for sexual reproduction in A. fumigatus and A. oryzae. Our analysis of sequence conservation revealed over 5,000 non-coding regions actively conserved across all three species. Within these regions, we identified potential functional elements including a previously uncharacterized TPP riboswitch and motifs suggesting regulation in filamentous fungi by Puf family genes. We further obtained comparative and experimental evidence indicating widespread translational regulation by upstream open reading frames. These results enhance our understanding of these widely studied fungi as well as provide new insight into eukaryotic genome evolution and gene regulation. Document includes all supplementary information (820 pages). Supplementary files are also attached below as Related files. THERE IS NO SUPPLEMENTARY FILE #7. PDF file size (with supplementary files included) is 10 Mbytes. An optimized version of the ARTICLE ONLY is attached as a Related File and is 1.9 Mbytes