Academic literature on the topic 'Vertebrate DNA'
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Journal articles on the topic "Vertebrate DNA"
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Ahmad, Hafiz Ishfaq, Gulnaz Afzal, Sehrish Sadia, Ghulam Haider, Shakeel Ahmed, Saba Saeed, and Jinping Chen. "Structural and Evolutionary Adaptations of Nei-Like DNA Glycosylases Proteins Involved in Base Excision Repair of Oxidative DNA Damage in Vertebrates." Oxidative Medicine and Cellular Longevity 2022 (April 4, 2022): 1–20. http://dx.doi.org/10.1155/2022/1144387.
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Oxidative stress is a type of stress that damages DNA and can occur from both endogenous and exogenous sources. Damage to DNA caused by oxidative stress can result in base modifications that promote replication errors and the formation of sites of base loss, which pose unique challenges to the preservation of genomic integrity. However, the adaptive evolution of the DNA repair mechanism is poorly understood in vertebrates. This research aimed to explore the evolutionary relationships, physicochemical characteristics, and comparative genomic analysis of the Nei-like glycosylase gene family involved in DNA base repair in the vertebrates. The genomic sequences of NEIL1, NEIL2, and NEIL3 genes were aligned to observe selection constraints in the genes, which were relatively low conserved across vertebrate species. The positive selection signals were identified in these genes across the vertebrate lineages. We identified that only about 2.7% of codons in these genes were subjected to positive selection. We also revealed that positive selection pressure was increased in the Fapy-DNA-glyco and H2TH domain, which are involved in the base excision repair of DNA that has been damaged by oxidative stress. Gene structure, motif, and conserved domain analysis indicated that the Nei-like glycosylase genes in mammals and avians are evolutionarily low conserved compared to other glycosylase genes in other “vertebrates” species. This study revealed that adaptive selection played a critical role in the evolution of Nei-like glycosylase in vertebrate species. Systematic comparative genome analyses will give key insights to elucidate the links between DNA repair and the development of lifespan in various organisms as more diverse vertebrate genome sequences become accessible.
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Carvalho, C. B. V. "DNA Barcoding in Forensic Vertebrate Species Identification." Brazilian Journal of Forensic Sciences, Medical Law and Bioethics 4, no. 1 (2014): 12–23. http://dx.doi.org/10.17063/bjfs4(1)y201412.
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Ortega-Recalde, Oscar, and Timothy Alexander Hore. "DNA methylation in the vertebrate germline: balancing memory and erasure." Essays in Biochemistry 63, no. 6 (November 22, 2019): 649–61. http://dx.doi.org/10.1042/ebc20190038.
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Abstract Cytosine methylation is a DNA modification that is critical for vertebrate development and provides a plastic yet stable information module in addition to the DNA code. DNA methylation memory establishment, maintenance and erasure is carefully balanced by molecular machinery highly conserved among vertebrates. In mammals, extensive erasure of epigenetic marks, including 5-methylcytosine (5mC), is a hallmark of early embryo and germline development. Conversely, global cytosine methylation patterns are preserved in at least some non-mammalian vertebrates over comparable developmental windows. The evolutionary mechanisms which drove this divergence are unknown, nevertheless a direct consequence of retaining epigenetic memory in the form of 5mC is the enhanced potential for transgenerational epigenetic inheritance (TEI). Given that DNA methylation dynamics remains underexplored in most vertebrate lineages, the extent of information transferred to offspring by epigenetic modification might be underestimated.
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Ross, Samuel E., Allegra Angeloni, Fan-Suo Geng, Alex de Mendoza, and Ozren Bogdanovic. "Developmental remodelling of non-CG methylation at satellite DNA repeats." Nucleic Acids Research 48, no. 22 (December 4, 2020): 12675–88. http://dx.doi.org/10.1093/nar/gkaa1135.
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Abstract In vertebrates, DNA methylation predominantly occurs at CG dinucleotides however, widespread non-CG methylation (mCH) has been reported in mammalian embryonic stem cells and in the brain. In mammals, mCH is found at CAC trinucleotides in the nervous system, where it is associated with transcriptional repression, and at CAG trinucleotides in embryonic stem cells, where it positively correlates with transcription. Moreover, CAC methylation appears to be a conserved feature of adult vertebrate brains. Unlike any of those methylation signatures, here we describe a novel form of mCH that occurs in the TGCT context within zebrafish mosaic satellite repeats. TGCT methylation is inherited from both male and female gametes, remodelled during mid-blastula transition, and re-established during gastrulation in all embryonic layers. Moreover, we identify DNA methyltransferase 3ba (Dnmt3ba) as the primary enzyme responsible for the deposition of this mCH mark. Finally, we observe that TGCT-methylated repeats are specifically associated with H3K9me3-marked heterochromatin suggestive of a functional interplay between these two gene-regulatory marks. Altogether, this work provides insight into a novel form of vertebrate mCH and highlights the substrate diversity of vertebrate DNA methyltransferases.
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Egeter, Bastian, Sara Peixoto, José C. Brito, Simon Jarman, Pamela Puppo, and Guillermo Velo-Antón. "Challenges for assessing vertebrate diversity in turbid Saharan water-bodies using environmental DNA." Genome 61, no. 11 (November 2018): 807–14. http://dx.doi.org/10.1139/gen-2018-0071.
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The Sahara desert is the largest warm desert in the world and a poorly explored area. Small water-bodies occur across the desert and are crucial habitats for vertebrate biodiversity. Environmental DNA (eDNA) is a powerful tool for species detection and is being increasingly used to conduct biodiversity assessments. However, there are a number of difficulties with sampling eDNA from such turbid water-bodies and it is often not feasible to rely on electrical tools in remote desert environments. We trialled a manually powered filtering method in Mauritania, using pre-filtration to circumvent problems posed by turbid water in remote arid areas. From nine vertebrate species expected in the water-bodies, four were detected visually, two via metabarcoding, and one via both methods. Difficulties filtering turbid water led to severe constraints, limiting the sampling protocol to only one sampling point per study site, which alone may largely explain why many of the expected vertebrate species were not detected. The amplification of human DNA using general vertebrate primers is also likely to have contributed to the low number of taxa identified. Here we highlight a number of challenges that need to be overcome to successfully conduct metabarcoding eDNA studies for vertebrates in desert environments in Africa.
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Varriale, Annalisa. "DNA Methylation, Epigenetics, and Evolution in Vertebrates: Facts and Challenges." International Journal of Evolutionary Biology 2014 (January 16, 2014): 1–7. http://dx.doi.org/10.1155/2014/475981.
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DNA methylation is a key epigenetic modification in the vertebrate genomes known to be involved in biological processes such as regulation of gene expression, DNA structure and control of transposable elements. Despite increasing knowledge about DNA methylation, we still lack a complete understanding of its specific functions and correlation with environment and gene expression in diverse organisms. To understand how global DNA methylation levels changed under environmental influence during vertebrate evolution, we analyzed its distribution pattern along the whole genome in mammals, reptiles and fishes showing that it is correlated with temperature, independently on phylogenetic inheritance. Other studies in mammals and plants have evidenced that environmental stimuli can promote epigenetic changes that, in turn, might generate localized changes in DNA sequence resulting in phenotypic effects. All these observations suggest that environment can affect the epigenome of vertebrates by generating hugely different methylation patterns that could, possibly, reflect in phenotypic differences. We are at the first steps towards the understanding of mechanisms that underlie the role of environment in molding the entire genome over evolutionary times. The next challenge will be to map similarities and differences of DNA methylation in vertebrates and to associate them with environmental adaptation and evolution.
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Carone, A., S. B. Malladi, M. Attimonelli, and C. Saccone. "Vertebrate MitBASE: a specialised database on vertebrate mitochondrial DNA sequences." Nucleic Acids Research 27, no. 1 (January 1, 1999): 150–52. http://dx.doi.org/10.1093/nar/27.1.150.
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Tweedie, S., J. Charlton, V. Clark, and A. Bird. "Methylation of genomes and genes at the invertebrate-vertebrate boundary." Molecular and Cellular Biology 17, no. 3 (March 1997): 1469–75. http://dx.doi.org/10.1128/mcb.17.3.1469.
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Patterns of DNA methylation in animal genomes are known to vary from an apparent absence of modified bases, via methylation of a minor fraction of the genome, to genome-wide methylation. Representative genomes from 10 invertebrate phyla comprise predominantly nonmethylated DNA and (usually but not always) a minor fraction of methylated DNA. In contrast, all 27 vertebrate genomes that have been examined display genome-wide methylation. Our studies of chordate genomes suggest that the transition from fractional to global methylation occurred close to the origin of vertebrates, as amphioxus has a typically invertebrate methylation pattern whereas primitive vertebrates (hagfish and lamprey) have patterns that are typical of vertebrates. Surprisingly, methylation of genes preceded this transition, as many invertebrate genes have turned out to be heavily methylated. Methylation does not preferentially affect genes whose expression is highly regulated, as several housekeeping genes are found in the heavily methylated fraction whereas several genes expressed in a tissue-specific manner are in the nonmethylated fraction.
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GRANTHAM, RICHARD. "CG doublet difficulties in vertebrate DNA." Nature 313, no. 6002 (February 1985): 437. http://dx.doi.org/10.1038/313437a0.
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MAX, EDWARD E. "CG doublet difficulties in vertebrate DNA." Nature 313, no. 6002 (February 1985): 437–38. http://dx.doi.org/10.1038/313437b0.
Full textDissertations / Theses on the topic "Vertebrate DNA"
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Elango, Navin. "Evolutionary impacts of DNA methylation on vertebrate genomes." Diss., Atlanta, Ga. : Georgia Institute of Technology, 2008. http://hdl.handle.net/1853/26691.
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Thesis (Ph.D)--Biology, Georgia Institute of Technology, 2009.Committee Chair: Dr. Soojin Yi; Committee Member: Dr. Eric Vigoda; Committee Member: Dr. James Thomas; Committee Member: Dr. John McDonald; Committee Member: Dr. Kirill Lobachev; Committee Member: Dr. Michael Goodisman. Part of the SMARTech Electronic Thesis and Dissertation Collection.
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Schofield, Julian Paul. "Studies on the structure of vertebrate genes." Thesis, University of Cambridge, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.386858.
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Long, Hannah Katherine. "Evolutionary usage and developmental roles of vertebrate non-methylated DNA." Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:78b14c1d-1fa3-46f1-815f-a8ba55579c43.
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Vertebrate genomes exhibit global methylation of cytosine residues where they occur in a cytosine-guanine dinucleotide (CpG) context and this epigenetic mark is generally thought to be repressive to transcription. Punctuating this pervasive DNA methylation landscape are short, contiguous regions of non-methylated DNA which are found at two thirds of mammalian gene promoters. These non-methylated regions exhibit CpG content close to expected levels as they escape the depletion of CpGs observed across the methylated fraction of the genome. The unique nucleotide properties of these CpG island (CGI) regions enable their identification by computational prediction in mammalian genomes. Owing to a lack of high-resolution genome-wide DNA methylation profiles in non-mammalian species, these CGI predictions have often been used as a proxy for non-methylated DNA in these organisms. In contrast to mammals, CGI predictions in cold-blooded vertebrates rarely coincide with gene promoters, leading to the belief that CGls are significantly divergent between vertebrate species, and that unique promoter-associated features may have been acquired during warmblooded vertebrate evolution. This thesis is primarily concerned with the location, establishment and biological function of non-methylated islands of DNA in vertebrate genomes. To experimentally determine genome-wide profiles of non-methylated DNA, a novel biochemical technique was established called biotinylated ZF-CxxC affinity purification (Bio-CAP), and development of this method is discussed in Chapter 3. Experimental analysis of non-methylated DNA profiles in this thesis initially addresses two main questions: (1) 'How does the non-methylated DNA landscape compare genome-wide for seven vertebrates considering distinct tissue types and developmental stages?' (2) 'How are vertebrate non-methylated regions of DNA defined and interpreted in the nuclear environment?' To address the first question, non-methylated DNA was profiled by Bio-CAP sequencing across the genomes of seven diverse vertebrate species, representing all major branch points of vertebrate evolution, and the results are discussed in Chapters 4 and S. Contrary to previously held dogma, experimentally determined nonmethylated islands of DNA (NMls) constitute an ancient epigenetic feature of vertebrate gene regulatory elements. However, despite having numerous high-resolution maps of vertebrate non-methylated DNA, the means by which NMls are identified and maintained in the nuclear environment remains poorly understood. To address the second question and identify features which determine the methylation state of DNA, exogenous DNA sequences were introduced into mouse embryonic stem (ES) c~.II~. Non-methylated DNA was profiled by Bio-CAP sequencing to investigate how different features, such as sequence-specific binding motifs, chromatin architecture and nucleotide composition of a given DNA sequence impact local DNA methylation patterns. Interestingly, the majority of exogenous promoters were appropriately non-methylated in mouse ES cells, germline and somatic cells suggesting that gene promoters have retained strong signals for the nonmethylated state across millions of years of evolution (discussed in Chapter 6). During mouse embryogenesis, genome-scale DNA demethylation and remethylation events occur to remodel the epigenetic landscape and loss of DNA methylation during this time leads to embryonic lethality. To investigate the biological function of non-methylated DNA, the third question addressed in this thesis is (3) 'What is the developmental importance of non-methylated islands of DNA during vertebrate embryogenesis?' To investigate this, members of the ZF-CxxC domain-containing family of chromatin modifiers were ablated in zebrafish embryos to perturb the chromatin landscape at NMls, and therefore interfere with their function during early development (Chapter 7). Early embryonic development and patterning was disrupted in knockdown embryos, suggesting that interpretation of non-methylated DNA and placement of chromatin modifications at NMls is essential for normal zebrafish embryogenesis. Together this work sheds light on the evolutionary origins of NMls, the mechanisms involved in the recognition and establishment of nonmethylated loci and provides an insight into the function of non-methylated DNA during early embryonic development.
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Chua, Shijia Joy. "Establishing the role of RNF4 in the vertebrate DNA damage response." Thesis, University of Dundee, 2012. https://discovery.dundee.ac.uk/en/studentTheses/20679582-0300-40ac-8d30-e733210c76b4.
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RNF4 belongs to the family of SUMO-targeted ubiquitin E3 ligases (STUbLs). The role of STUbLs in maintaining genomic stability was first discovered in yeast. Theyeast STUbL mutants displayed genomic instability, elevated mutation rates, sensitivity to DNA damaging agents and also demonstrated synthetic lethality with other DNA repair genes. Although the role of vertebrate RNF4 in the DNA damage response was not yet established, it could rescue the Schizosaccaromyces pombe STUbL mutant phenotypes, showing that RNF4 is a functional homologue of the yeast STUbL proteins,and that it might be implicated in the vertebrate DNA damage response.A homozygous knockout of RNF4 in the DT40 chicken lymphocyte cell line was generated to investigate the involvement of vertebrate RNF4 in protecting cells against DNA damage. Although the complete loss of RNF4 did not affect cell proliferation or cell cycle distribution, the RNF4 -/- cells exhibited a selective hypersensitivity to some S-phase specific DNA damaging agents. This hypersensitivity could be rescued by introducing an ortholog of RNF4 from another vertebrate species, and this was dependent on a functional ubiquitin E3 ligase activity of RNF4.To explore the physiological function of RNF4 in the context of a wholeorganism, Danio rerio was chosen as an in vivo model. Danio rerio RNF4 sharedsimilar in vitro biochemical characteristics as RNF4 from other vertebrates – it was able to autoubiquitylate itself and also ubiquitylate SUMO2 chains. In Danio rerio, RNF4 is a maternally provided gene and is highly expressed in the adult gonads. In the ovaries, RNF4 expression was restricted to the early stage oocytes, suggesting a possible role in oocyte development. Loss-of-function studies in Danio rerio were performed using morpholino knockdown and zinc-finger knockout technologies, and the depletion of RNF4 in zebrafish did not affect early embryonic development or viability of the animal.The results presented in this thesis suggests that while vertebrate RNF4 is notlikely to be an essential gene in some vertebrates, it plays a role in the DNA damage response and might be implicated in gonad development in Danio rerio. The zinc-finger knockout model has just been established and a more in-depth analysis is necessary to shed more light on the in vivo functions of RNF4.
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Barr, Alexis. "Characterising the function of CDK5RAP2 in the vertebrate centrosome." Thesis, University of Cambridge, 2010. https://www.repository.cam.ac.uk/handle/1810/228639.
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The centrosome is the major microtubule organising centre in vertebrate cells. CDK5RAP2 is a human protein that localises to the centrosome. At the start of this thesis work, the function of CDK5RAP2 was uncharacterised. Significantly, cdk5rap2 is one of several centrosomal genes that are mutated in the developmental disorder Primary Microcephaly, where affected individuals have smaller brains than expected for the age- and sex-adjusted mean. Orthologues of CDK5RAP2 in the fruit fly (Centrosomin/Cnn) and in fission yeast (Mod20p) have been well characterised and are known to have important roles in maintaining centrosome structure and in regulating microtubule nucleation. CDK5RAP2 shares two evolutionarily conserved domains with Cnn, known as CNN motif 1 and 2. Using the chicken B-cell line, DT40, I have used gene-targeting methods to disrupt both of these domains in CDK5RAP2. This revealed a function for CDK5RAP2 in attaching centrosomes to mitotic spindle poles. Centrosome attachment to spindle poles is mediated by a binding partner of CDK5RAP2, AKAP450. AKAP450 also localises to centrosomes and provides anchorage sites for spindle poles in the centrosome. Disruption of the CNN1 and CNN2 domains of CDK5RAP2 causes mislocalisation of AKAP450 from the centrosome and detachment of centrosomes from spindle poles. My studies in DT40 and in human cell lines revealed that CDK5RAP2 and AKAP450 also cooperate during interphase to maintain the two centrioles in the centrosome as a pair. In addition to a structural role in the centrosome, I also find that CNN motif 1 of CDK5RAP2 plays a role in the cellular response to DNA damage. In the absence of CNN motif 1, cells no longer efficiently arrest the cell cycle in response to damage. Centrosome-mediated mitotic spindle alignment and the DNA damage response have both been implicated in microcephaly. Therefore, defects in these functions of CDK5RAP2 may explain how mutations in cdk5rap2 may lead to microcephaly.
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Veeman, Michael Terrence. "Zebrafish prickle : non-canonical Wnt/PCP functions in vertebrate gastrulation /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/4999.
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Gohm, Birgit [Verfasser]. "The Role of DNA Repair in the Evolution of Vertebrate Longevity / Birgit Gohm." Konstanz : Bibliothek der Universität Konstanz, 2016. http://d-nb.info/1171131860/34.
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Yoshimura, Michio. "Vertebrate POLQ and POLβ cooperate in base excision repair of oxidative DNA damage." Kyoto University, 2007. http://hdl.handle.net/2433/135652.
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Parker, Hugo. "The role of highly conserved non-coding DNA sequences in vertebrate development and evolution." Thesis, Queen Mary, University of London, 2010. http://qmro.qmul.ac.uk/xmlui/handle/123456789/1267.
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Comparisons between vertebrate genome sequences, from mammals to fishes, have revealed thousands of conserved non-coding elements (CNEs) that are associated with developmental genes. Interestingly, the vast majority of these CNEs cannot be found in invertebrate genomes by sequence homology. As many CNEs have been demonstrated to act as enhancers in-vivo, it has been postulated that CNEs represent gene regulatory elements with crucial roles in aspects of development that are shared between vertebrates. To trace the evolution of CNE sequences in vertebrates, a preliminary search for CNEs in the lamprey genome was conducted using the draft lamprey genome sequence. This thesis documents how the CNEs identified in lamprey have been used as a guide to ask questions about the function and evolution of CNEs in the vertebrate lineage. Through the combined use of comparative genomics and developmental biology techniques, including a newly developed reporter assay for sea lamprey embryos, crucial first steps have been taken toward systematically de-coding these ancient gene regulatory elements. Special attention is paid toward utilising the low sequence identity of lamprey CNEs for „phylogenetic footprinting‟, an approach which uncovers striking enrichment of CNEs for a set of motifs that are characteristic of Hox-regulated elements. These findings help to establish CNEs within a developmental and evolutionary context.
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Bednar, Theresa [Verfasser], George [Akademischer Betreuer] Iliakis, and Jürgen [Akademischer Betreuer] Thomale. "Efficient support of DNA replication functions by DNA ligase 3 in vertebrate cells / Theresa Bednar. Gutachter: Jürgen Thomale. Betreuer: George Iliakis." Duisburg, 2013. http://d-nb.info/1042373493/34.
Full textBooks on the topic "Vertebrate DNA"
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IARC Working Group on the Evaluation of Cancer-Preventive Strategies. Cervix cancer screening. Lyon: IARC Press, 2005.
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Kurstak, Edouard, and Christine Kurstak. Vertebrate Animal and Related Viruses: DNA Viruses. Elsevier Science & Technology Books, 2013.
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Kurstak, Edouard. Comparative Diagnosis of Viral Diseases: Vertebrate Animal and Related Viruses Part B--DNA Viruses. Elsevier Science & Technology Books, 2012.
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Bizelli, José Luís, Thaís Conte Vargas, and José Anderson Santos Cruz. Educação Escolar em Ibero-América: 15 anos de trajetória. Editora Ibero-Americana de Educação, 2020. http://dx.doi.org/10.47519/eie.978-65-86839-04-3.
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Muitos são os temas que vertebram toda a atuação do grupo, mas eles podem ser sintetizados em 7: 1. Políticas Públicas e Gestão da Educação; 2. Tecnologias de Informação e Comunicação em Educação; 3. Formação do Educador, Trabalho Docente e Práticas Pedagógicas; 4. Educação Sexual, Sexualidade e Gênero; 5. Educação Especial; 6. Educação Superior; 7. Internacionalização. Este livro reúne 18 capítulos. Na sua construção é possível perceber que alguns dos temas trabalhados pelo grupo estão contemplados. É o caso das Tecnologias de Informação e Comunicação em Educação.
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Penerapan strategi pemetaan konsep dalam pembelajaran untuk mengungkapkan dan memperbaiki kesalahan konsepsi mahasiswa vertebrata di program studi S-1 pendidikan biologi: Laporan penelitian. Mataram: Fakultas Keguruan dan Ilmu Pendidikan Universitas Mataram, 2003.
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Wray, Britt. Rise of the Necrofauna: The Science, Ethics, and Risks of De-Extinction. Greystone Books, 2019.
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Church, George M. (George McDonald), author of foreword and David Suzuki Institute, eds. Rise of the necrofauna: The science, ethics, and risks of de-extinction. Greystone Books, 2017.
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Martínez-Domínguez, Luis Manuel. Una Pedagogía del Nosotros. FERSE, 2020. http://dx.doi.org/10.52154/ferse0001.
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Modelo pedagógico que propone la educación en conciencia, como auténtica, creativa y plena apertura de sí en el Nosotros para autorrealizarse de forma apoteósica. Cuando no se da este desarrollo nosicéntrico, desde donde se habita con felicidad, los sujetos se encierran en egocentrismo o alocentrismo inmaduro que es causa y motivo de violencia. La Pedagogía del Nosotros, desde la fenomenología y el refuerzo de la neurociencia y otras disciplinas, describe desde la Sociedad, la Familia y la Educación, lo que civilizaciones de todos los tiempos y lugares, han procurado desarrollar desde sus diferentes cosmovisiones: el Ágape de la Grecia Clásica, el Ren del confucionismo, la Caritas del cristianismo, el Ubuntu africano, El Iithar (إيثار) del Islam, la Fraternité de la Ilustración…, y tantas otras comunidades humanas que la han inculturado, es decir, que la han vivido desde su cultura, a su manera. Aquí se presenta en lo esencial, como columna vertebral de lo admisible por todos, para educar con un fundamento compartido.
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IARC Handbooks on Cancer Prevention: Cervix Cancer Screening (IARC Handbooks of Cancer Prevention). World Health Organization, 2005.
Find full textBook chapters on the topic "Vertebrate DNA"
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Fukagawa, Tatsuo. "Centromeric Chromatin and Kinetochore Assembly in Vertebrate Cells." In DNA Replication, Recombination, and Repair, 365–87. Tokyo: Springer Japan, 2016. http://dx.doi.org/10.1007/978-4-431-55873-6_14.
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Ehrlich, Melanie, and Kenneth C. Ehrlich. "Effect of DNA methylation on the binding of vertebrate and plant proteins to DNA." In DNA Methylation, 145–68. Basel: Birkhäuser Basel, 1993. http://dx.doi.org/10.1007/978-3-0348-9118-9_7.
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Matkarimov, Bakhyt T., and Murat K. Saparbaev. "DNA Repair and Mutagenesis in Vertebrate Mitochondria: Evidence for Asymmetric DNA Strand Inheritance." In Advances in Experimental Medicine and Biology, 77–100. Cham: Springer International Publishing, 2020. http://dx.doi.org/10.1007/978-3-030-41283-8_6.
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Miklos, George L. Gabor. "Localized Highly Repetitive DNA Sequences in Vertebrate and Invertebrate Genomes." In Molecular Evolutionary Genetics, 241–321. Boston, MA: Springer US, 1985. http://dx.doi.org/10.1007/978-1-4684-4988-4_4.
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Temin, Howard M. "Retrovirus Vectors for Gene Transfer: Efficient Integration into and Expression of Exogenous DNA in Vertebrate Cell Genomes." In Gene Transfer, 149–87. Boston, MA: Springer US, 1986. http://dx.doi.org/10.1007/978-1-4684-5167-2_6.
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Goorha, R. M. "Frog Virus 3 DNA Replication." In Viruses of Lower Vertebrates, 30–38. Berlin, Heidelberg: Springer Berlin Heidelberg, 1989. http://dx.doi.org/10.1007/978-3-642-83727-2_3.
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Ross, Samuel E., and Ozren Bogdanovic. "Generation and Molecular Characterization of Transient tet1/2/3 Knockouts." In Methods in Molecular Biology, 281–318. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1294-1_17.
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Abstract5-methylcytosine (5mC) is a gene-regulatory mark associated with transcriptional repression. 5mC can be erased through the catalytic action of Ten-eleven translocation (TET) methylcytosine dioxygenases (TET1, TET2, TET3), which oxidize 5mC resulting in its removal from the genome. In vertebrates, TET enzymes facilitate DNA demethylation of regulatory regions linked to genes involved in developmental processes. Consequently, TET ablation leads to severe morphological defects and developmental arrest. Here we describe a system that can facilitate the study of relationships between TET enzymes, 5mC, and embryo development. We provide detailed descriptions for the generation of F0 zebrafish tet1/2/3 knockouts using CRISPR/Cas9 technology and elaborate on the strategies to assess the impact of TET loss by reduced representation bisulfite sequencing (RRBS).
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Wu, Shi-Bei, Yu-Ting Wu, Yi-Shing Ma, and Yau-Huei Wei. "Oxidative Stress and its Biochemical Consequences in Mitochondrial DNA Mutation-Associated Diseases: Implications of Redox Therapy for Mitochondrial Diseases." In Oxidative Stress in Vertebrates and Invertebrates, 33–49. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2011. http://dx.doi.org/10.1002/9781118148143.ch3.
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Schmid, Carl W., and Che-Kun James Shen. "The Evolution of Interspersed Repetitive DNA Sequences in Mammals and Other Vertebrates." In Molecular Evolutionary Genetics, 323–58. Boston, MA: Springer US, 1985. http://dx.doi.org/10.1007/978-1-4684-4988-4_5.
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Capriglione, Teresa. "Repetitive DNA as a tool to study the phylogeny of cold-blooded vertebrates." In Chromosomes Today, 183–94. Basel: Birkhäuser Basel, 2000. http://dx.doi.org/10.1007/978-3-0348-8484-6_14.
Full textConference papers on the topic "Vertebrate DNA"
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Hutchinson, Gordon B., and Michael R. Hayden. "SORFIND: A COMPUTER PROGRAM THAT PREDICTS EXONS IN VERTEBRATE GENOMIC DNA." In Proceedings of the 2nd International Conference. WORLD SCIENTIFIC, 1993. http://dx.doi.org/10.1142/9789814503655_0043.
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Scherr, Thomas F., Gerald Knapp, Terrence Tiersch, W. Todd Monroe, and Krishnaswamy Nandakumar. "The Activation of Zebrafish Sperm Cells in a Micromixer." In ASME 2013 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/sbc2013-14734.
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The freshwater fish, Danio rerio (zebrafish), have become widely used as a model organism for vertebrate development, DNA mutation, and human disease studies [1]. Maintaining live colonies of the numerous developed strains of zebrafish under investigation can be prohibitively costly. As such, there is a growing need to catalog their reproductive cells and have them available on demand [2]. Thus cryopreservation of model strain gametes has become an important endeavor, where evaluation of freezing and thawing techniques is currently a bottleneck to these procedures.
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Suharyogi, Ifan Yoga Pratama, Agustina Djafar, Rahajeng Ayu Permana Sari, and Paradita Kenyo Arum Dewantoro. "Geological Museum Innovations to Dealing with Covid-19 Pandemic | Inovasi Museum Geologi dalam Menghadapi Pandemi Covid-19." In The SEAMEO SPAFA International Conference on Southeast Asian Archaeology and Fine Arts (SPAFACON2021). SEAMEO SPAFA, 2021. http://dx.doi.org/10.26721/spafa.pqcnu8815a-34.
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Bandung Geological Museum as the thematic earth museum in Indonesia has been established on 16 May 1929. This museum has 417,882 collections, there are mineral and rock collections, vertebrate, invertebrate, paleobotanical fossils, and artifacts. As a government museum, the Geological Museum has a duty to disseminating geological information. This article aims to identify the Geological Museum’s activities during the Covid-19 pandemic. After the temporary closure in March 2020, the museum activities were carried out virtually, including Collection Talk, Day and Night at the Museum, virtual tours, Bincang Museum, virtual geoscience socialization, and introduce the collections by social media. Museum Geologi Bandung sebagai museum kebumian di Indonesia telah berdiri sejak 16 Mei 1929. Museum ini memiliki 417.882 koleksi, berupa koleksi mineral dan batuan, fosil vertebrata, fosil invertebrata, fosil paleobotani dan artefak. Sebagai instansi yang bertugas menyebarluaskan informasi kegelogian, dimasa pandemi Covid-19, Museum Geologi berinovasi melakukan kegiatan-kegiatan edukasi dalam bentuk virtual. Tujuan penulisan artikel ini adalah melakukan identifikasi kegiatan dilakukan Museum Geologi selama pandemi Covid-19. Pasca penutupan sementara Museum Geologi pada bulan Maret 2020, kegiatan yang dilakukan berupa kegiatan virtual diantaranya: Collection Talk, Day and Night at the Museum, virtual tour, Bincang Museum, sosialisasi kebumian secara virtual, dan pengenalan koleksi melalui sosial media.
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Leitão, Juliana Rodrigues, Marcelo Candido Portilho Gouveia, Bruno Watanabe Minto, and Luis Gustavo Gosuen Gonçalves Dias. "RUPTURA DE BEXIGA URINÁRIA E FRATURA VERTEBRAL EM L5 DECORRENTES DE TRAUMA EM CÃO: RELATO DE CASO." In I Congresso On-line Nacional de Clínica Veterinária de Pequenos Animais. Revista Multidisciplinar em Saúde, 2021. http://dx.doi.org/10.51161/rems/1825.
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Introdução: As fraturas da coluna vertebral são causas potenciais de injúrias da medula espinhal em pequenos animais. Frequentemente, o paciente com trauma toracolombar têm lesões associadas em outros órgãos. Objetivos: Objetivou-se relatar um caso de ruptura da bexiga urinária e fratura completa oblíqua na 5° vértebra lombar e descrever as técnicas cirúrgicas utilizadas. Materiais e métodos: Uma cadela, Shih Tzu, com 5 anos de idade, 5 kg, foi atendido com histórico de trauma há 4 horas pela queda de um botijão de gás cheio na região toracolombar. Paraparesia, disúria, hematúria e dor abdominal eram os sinais clínicos. Ao exame neurológico, a paciente demonstrou dor acentuada na região lombar e propriocepção ausente. O paciente foi submetido ao exame radiográfico da coluna vertebral o qual indicou fratura completa oblíqua na 5° vértebra lombar. Na ultrassonografia abdominal, constatou-se líquido livre na cavidade abdominal. Realizou-se uretrocistografia retrógrada de contraste positivo, confirmando-se a ruptura da vesícula urinária. Optou-se primariamente pela cistorrafia com fio absorvível monofilamentado, padrão contínuo, invaginante Cushing e em 2 planos. Instituiu-se o tratamento clínico para fratura vertebral que incluiu confinamento estrito, imobilização externa e controle da dor (dipirona 25mg/kg SID e tramadol 2 mg/kg SID). Após dois dias, realizou-se laminectomia e estabilização lombar. A fratura em L5 foi reduzida com auxílio de pinça ponta-ponta. Dois parafusos foram colocados tanto na vértebra cranial (L4) quanto caudal (L6) à fratura, com ângulo de 45° de inserção. O cimento ósseo Cimento Ortopédico Veterinário – C-VET foi colocado em torno dos implantes. Efetuou-se o desgaste do processo espinhoso das vértebras L4, L5 e L6. Resultados: No pós-operatório imediato, a paciente foi encaminhada para internação. Foi orientado repouso absoluto por 15 dias. No sétimo dia, o animal deambulava sem dor e urinava normalmente. Após 30 dias, a paciente não apresentava alterações clínicas neurológicas, recebendo alta. Uma intervenção terapêutica rápida limita a extensão da lesão neuronal, favorecendo a recuperação do paciente. Em cães com preservação da percepção de dor profunda, o prognóstico é favorável. Conclusão: Conclui-se que em casos com histórico conhecido de traumatismo vertebral deve-se avaliar o paciente quanto a lesões concorrentes para um resultado pós-cirúrgico satisfatório.
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Borges, Victória Maria Luz, Marília Francisca da Silva Pereira, Isadora Maria de Carvalho Marques, Flávio Luis dos Santos Sousa, Lia Cruz Vaz da Costa Damásio, and Marcelo Barbosa Ribeiro. "Metástase óssea em câncer de colo uterino: um relato de caso." In 45º Congresso da SGORJ XXIV Trocando Ideias. Zeppelini Editorial e Comunicação, 2021. http://dx.doi.org/10.5327/jbg-0368-1416-20211311055.
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Introdução: O câncer de colo de útero é o terceiro câncer mais comum e a quarta causa de morte relacionada a câncer no mundo. Ocorre, na maior parte dos casos, entre a terceira e a quinta décadas de vida e o principal fator de risco relacionado é a presença do papilomavírus humano, com seus subtipos oncogênicos. Metástase óssea raramente ocorre em câncer de colo de útero, com a incidência de 0,8 a 2,3%, e alguns fatores de risco estão associados ao seu desenvolvimento, entre os quais: idade avançada, maiores estágios T e N, tipos histológicos não adenocarcinoma e não escamoso, tumores pouco diferenciados e presença de outras metástases (pulmões, fígado e cérebro). Relato de caso: Paciente de 54 anos, menarca aos 18 e sexarca aos 20. Procurou o atendimento no ambulatório de ginecologia em 2017, relatando queixa de dor em baixo ventre. Ao exame especular, visualizou-se moderada ectopia e colo sangrante. O exame colpocitológico revelou lesão intraepitelial de alto grau. A paciente foi encaminhada à colposcopia, que mostrou NIC III, e foi submetida a tratamento cirúrgico e radioterapia posterior. O exame histopatológico evidenciou carcinoma escamoso GIII, com invasão de 100% da margem comprometida. A paciente evolui com dor em região lombar e em membros inferiores após três anos de seguimento clínico, com presença de lesão lítica em L4 e lesão com aspecto de hemangiomastose no corpo vertebral de L3 à radiografia. A ressonância magnética e a cintilografia óssea exibiram lesão lítica comprometendo o corpo cervical de L4 à direita, o que não foi observado em exames anteriores. Realizou-se biópsia e foi confirmada a neoplasia metastática. Conclusão: A neoplasia de colo uterino surge entre a terceira e a quinta décadas de vida, sendo o tipo histológico carcinoma escamoso o mais comum e condizente com o caso descrito. Apesar de rara, a metástase óssea ocorre em média dois anos após o diagnóstico, sendo o principal sítio a coluna vertebral (36,36%) e destacando-se a topografia lombar (51,9%). Em contrapartida, uma metanálise publicada em 2019 evidenciou que, entre os casos de câncer de colo uterino em estágio avançado, a metástase óssea ocorreu em 23%, perdendo apenas para a metástase pulmonar (59%). Embora não sejam elucidados os mecanismos de metástase, sugere-se relação com o comportamento do tumor primário, o suprimento vascular, o sistema imune e o ambiente ósseo. São necessários maiores estudos para elucidar a oncogênese da metástase óssea. O prognóstico de pacientes com metástases ósseas é bem restrito entre seis e 12 meses, sendo proposto um tratamento paliativo com abordagem multiprofissional, visando aliviar o quadro álgico de elevada intensidade, evitar fraturas patológicas e prevenir incapacidades. O principal objetivo é ter melhor qualidade de vida. O grande avanço preconizado pela Federação Internacional de Ginecologia e Obstetrícia e Federação Brasileira das Sociedades de Ginecologia e Obstetrícia é o rastreamento universal e o diagnóstico precoce de lesões precursoras do câncer de colo uterino, sobretudo por tratar-se de um método de baixo custo e de fácil reprodução.
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Guedes, Samara Mirelly dos Santos, Greifus Greigor Benites, Denise Cristina Moz Vaz Oliane, Gabriel Rodrigues Anacleto, Fausto da Silva Gonçalves, and Carolina Buck. "Gemelaridade imperfeita do tipo parapagus." In 44° Congresso da SGORJ - XXIII Trocando Ideias. Zeppelini Editorial e Comunicação, 2020. http://dx.doi.org/10.5327/jbg-0368-1416-2020130246.
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Introdução: A gemelaridade imperfeita caracteriza-se pela fusão dos fetos em gestações monocoriônicas e monoamnióticas em decorrência da divisão tardia do blastocisto com incidência estimada de 1,5 por 100 mil nascimentos. A gemelaridade imperfeita é classificada de acordo com o local da fusão e seu prognóstico depende do compartilhamento de órgãos, malformações associadas e a possiblidade de separação dos fetos. Objetivo: Relatar o caso de gemelaridade imperfeita do tipo “parapagus”, caracterizada pela união lateral extensa dos fetos com compartilhamento de órgãos, representando um desafio no manejo e no aconselhamento pré e pós-natal. Material e Métodos: Paciente de 21 anos, secundigesta, encaminhada ao serviço de Medicina Fetal em razão de ultrassonografia de 24 semanas apresentando gestação com fetos unidos a partir do tórax (gêmeos siameses). Na avaliação ecográfica realizada em nosso serviço, foram visualizados dois polos cefálicos separados de aspecto habitual, tórax e abdome únicos, com duas colunas vertebrais, dois estômagos e um fígado. Dois membros superiores e dois membros inferiores de aspectos habituais. Em região cardíaca, coração à esquerda aparentemente com quatro câmaras de aspecto habitual e, adjacente a este, à direita, coração aparentemente com duas câmaras; ambos fundidos na linha média. Foram realizados avaliação multidisciplinar e aconselhamento ao casal da impossibilidade da separação cirúrgica pós-natal. Antecipação terapêutica do parto foi solicitada e negada. Paciente evoluiu com trabalho de parto com 36 semanas e 5 dias, sendo realizada cesariana com recém-nascido de 3590 g, Apgar 3/7. Mantém-se vivo aguardando exames complementares para posterior seguimento clínico e conduta. Resultado e Conclusões: O diagnóstico precoce, o conhecimento da correta determinação do tipo de gemelaridade imperfeita e a extensão da fusão podem auxiliar no aconselhamento ao casal, permitindo acompanhamento em centro de Medicina Fetal com equipe multidisciplinar.
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León Vivanco, María Fernanda. "Hecho de fragmentos: reinterpretación de los componentes de un eje urbano en Barcelona." In Seminario Internacional de Investigación en Urbanismo. Barcelona: Maestría en Planeación Urbana y Regional. Pontificia Universidad Javeriana de Bogotá, 2014. http://dx.doi.org/10.5821/siiu.6091.
Full textAbstract:
Cuando el tejido urbano existente es una disposición de fragmentos que se unen entre sí por su cercanía o
vecindad, el eje que los vertebra permite, a través de él, realizar una lectura eficaz de todos sus
componentes. En Barcelona, al mismo tiempo que se empieza a construir el Ensanche, se distinguen sobre
el territorio algunos núcleos periféricos que forman una corona que lo rodea. Hacia el oeste, los pueblos de
Hostafrancs, Sants, Les Corts y Sarrià componen un eje imaginario que, al ir edificándose la ciudad,
también se va consolidando. El presente trabajo pretende realizar un estudio de la configuración del eje,
reconociendo en los fragmentos que le dan forma, aquellas características físicas que permiten su
consolidación; analizándolas no sólo como entidades aisladas sino como elementos que integran un todo,
que se relacionan y articulan entre sí, y, destacar aquellos atributos esenciales de cada pieza urbana que
facultan una reinterpretación de su composición. When the existing urban fabric is an arrangement of fragments that are joined together by their proximity or
neighborhood, the axis that joins them, allows, through it, an efficient reading of all its components. In
Barcelona, at the same time that the city started to build the “Eixample”, there were on the territory some
peripheral towns that were forming a crown around the planned city. To the west, the towns of Hostafrancs,
Sants, Les Corts and Sarrià made up an imaginary axis that at the same time that the city was built it would
also be consolidated. This paper aims to conduct a study of the axis configuration, recognizing in their
fragments, the morphological conditions that shapes it, analyzing them not only as isolated entities but as
elements that make up a whole, that relate and articulate between them, and highlight those essential
attributes of each urban piece that allows a reinterpretation of the axis composition.
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Magadan, Eduarda Druck, Eduarda Kotlinsky Weber, Esthela Rodegheri Trevisan, Luiza Fernandes Xavier, and Virgínia Tafas Da Nóbrega. "TROMBOFILIA EM CRIANÇA PORTADORA DE SÍNDROME DE DOWN: UM RELATO DE CASO." In II Congresso Brasileiro de Hematologia Clínico-laboratorial On-line. Revista Multidisciplinar em Saúde, 2022. http://dx.doi.org/10.51161/hematoclil/127.
Full textAbstract:
Introdução: A trombose arterial é um evento raro na população pediátrica. A chance dele ocorrer é potencializada se associada a mutações ou deficiência de proteínas responsáveis pela formação ou bloqueio do coágulo. Algumas mutações em heterozigose são muito frequentes, como a mutação metilenotetrahidrofolato redutase (MTHFR). Quando isoladas, essas mutações não causam qualquer manifestação clínica; todavia, quando combinadas com outras mutações ou fatores de hipercoagulabilidade, podem desencadear tromboses. Em pacientes com síndrome de Down, não há incidência relatada de trombofilia e eventos tromboembólicos aparentam ser incomuns. Objetivo: Relatar o caso de um paciente pediátrico, portador de síndrome de Down, com AVC isquêmico e trombofilia. Metodologia: Revisão de prontuário, com coleta de dados clínicos e laboratoriais. Resultados/Descrição do caso: Paciente masculino, de 5 anos e 6 meses, portador de síndrome de Down e hipotireoidismo, com AVC prévio, admitido no Hospital da Criança Santo Antônio por quadro de paresia no membro superior direito, desvio da boca para a direita e marcha hemiparética por menos de 24h, com resolução espontânea. Realizou uma tomografia craniana que não apontou sangramentos. À angioressonância magnética, efetuada no dia subsequente, observou-se trombose dos segmentos inclusos da artéria vertebral direita, como também sinais de isquemia aguda no hemisfério cerebelar direito (território da artéria cerebelar póstero-inferior) e no tálamo esquerdo. Ademais, constatou-se encefalomalácia no esplênio do corpo caloso à direita. Os resultados dos exames laboratoriais foram os seguintes: fator V - 127%; fator VIII > 200%; anti-trombina III - 101%; PTN S - 102%; PTN C - 134%; homocisteína - 6,97 µmol/l; anticardiolipinas IgA e IgG - não reagentes; anticardiolipinas IgM - reagente fraco. Detectou-se duas mutações em heterozigose: uma, no gene MTHFR; outra, no gene da protrombina. Fator V de Leiden estava ausente. Atualmente, o paciente encontra-se com anticoagulação profilática para evitar novos eventos trombóticos graves. Conclusão: As mutações em heterozigose no gene da protrombina e no gene MTHFR, associadas à elevação do fator VIII, provocaram um estado de hipercoagulabilidade neste paciente, culminando com trombose em artérias intra e extracerebrais. Portanto, este caso mostra a importância da investigação genética em todos pacientes pediátricos com trombose venosa sem fator desencadeante ou com trombose arterial.
Reports on the topic "Vertebrate DNA"
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Savedoff, Bill, Diana Pinto, and Laura Goyeneche. Convivir con el coronavirus: ¿Cómo pueden las pruebas y el rastreo de contactos frenar la pandemia? Inter-American Development Bank, February 2021. http://dx.doi.org/10.18235/0003047.
Full textAbstract:
Las pruebas y el rastreo de contactos son la columna vertebral de una respuesta eficaz del sistema de salud ante una epidemia. Sin embargo, en el caso del COVID-19, aunque la comunidad científica desarrolló pruebas de diagnóstico tan pronto se reconoció la amenaza pandémica, su producción se retrasó y la mayoría de los países no implementaron pruebas en cantidades suficientes o dentro de una estrategia nacional coherente. El objetivo del presente documento es identificar las estrategias exitosas en el uso de pruebas y el rastreo de contactos para informar a las políticas que puedan implementar los países de América Latina y el Caribe para desacelerar y controlar la pandemia. Se dan énfasis en las políticas de menor costo y las que toman en cuenta restricciones de recursos o en la oferta de insumos diagnósticos.
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Hulata, Gideon, Thomas D. Kocher, and Micha Ron. Elucidating the molecular pathway of sex determination in cultured Tilapias and use of genetic markers for creating monosex populations. United States Department of Agriculture, January 2007. http://dx.doi.org/10.32747/2007.7695855.bard.
Full textAbstract:
The objectives of this project were to: 1) Identify genetic markers linked to sex-determining genes in various experimental and commercial stocks of O. niloticusand O. aureus, as well as red tilapias; 2) Develop additional markers tightly linked to these sex determiners, and develop practical, non-destructive genetic tests for identifying genotypic sex in young tilapia; A third aim, to map sex modifier loci, was removed during budget negotiations at the start of the project. Background to the topic. A major obstacle to profitable farming of tilapia is the tendency of females to reproduce at a small size during the production cycle, diverting feed and other resources to a large population of small, unmarketable fish. Several approaches for producing all-male fingerlings have been tried, including interspecific hybridization, hormonal masculinization, and the use of YY-supermale broodstock. Each method has disadvantages that could be overcome with a better understanding of the genetic basis of sex determination in tilapia. The lack of sex-linked markers has been a major impediment in research and development of efficient monosex populations for tilapia culture. Major conclusions, solutions, achievements. We identified DNA markers linked to sex determining genes in six closely related species of tilapiine fishes. The mode of sex determination differed among species. In Oreochromis karongaeand Tilapia mariaethe sex-determining locus is on linkage group (LG) 3 and the female is heterogametic (WZ-ZZ system). In O. niloticusand T. zilliithe sex-determining locus is on LG1 and the male is heterogametic (XX-XY system). We have nearly identified the series of BAC clones that completely span the region. A more complex pattern was observed in O. aureus and O. mossambicus, in which markers on both LG1 and LG3 were associated with sex. We found evidence for sex-linked lethal effects on LG1, as well as interactions between loci in the two linkage groups. Comparison of genetic and physical maps demonstrated a broad region of recombination suppression harboring the sex-determining locus on LG3. We also mapped 29 genes that are considered putative regulators of sex determination. Amhand Dmrta2 mapped to separate QTL for sex determination on LG23. The other 27 genes mapped to various linkage groups, but none of them mapped to QTL for sex determination, so they were excluded as candidates for sex determination in these tilapia species. Implications, both scientific and agricultural. Phylogenetic analysis suggests that at least two transitions in the mode of sex determination have occurred in the evolution of tilapia species. This variation makes tilapias an excellent model system for studying the evolution of sex chromosomes in vertebrates. The genetic markers we have identified on LG1 in O. niloticusaccurately diagnose the phenotypic sex and are being used to develop monosex populations of tilapia, and eliminate the tedious steps of progeny testing to verify the genetic sex of broodstock animals.
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Funkenstein, Bruria, and Cunming Duan. GH-IGF Axis in Sparus aurata: Possible Applications to Genetic Selection. United States Department of Agriculture, November 2000. http://dx.doi.org/10.32747/2000.7580665.bard.
Full textAbstract:
Many factors affect growth rate in fish: environmental, nutritional, genetics and endogenous (physiological) factors. Endogenous control of growth is very complex and many hormone systems are involved. Nevertheless, it is well accepted that growth hormone (GH) plays a major role in stimulating somatic growth. Although it is now clear that most, if not all, components of the GH-IGF axis exist in fish, we are still far from understanding how fish grow. In our project we used as the experimental system a marine fish, the gilthead sea bream (Sparus aurata), which inhabits lagoons along the Mediterranean and Atlantic coasts of Europe, and represents one of the most important fish species used in the mariculture industry in the Mediterranean region, including Israel. Production of Sparus is rapidly growing, however, in order for this production to stay competitive, the farming of this fish species has to intensify and become more efficient. One drawback, still, in Sparus extensive culture is that it grows relatively slow. In addition, it is now clear that growth and reproduction are physiological interrelated processes that affect each other. In particular sexual maturation (puberty) is known to be closely related to growth rate in fish as it is in mammals, indicating interactions between the somatotropic and gonadotropic axes. The goal of our project was to try to identify the rate-limiting components(s) in Sparus aurata GH-IGF system which might explain its slow growth by studying the ontogeny of growth-related genes: GH, GH receptor, IGF-I, IGF-II, IGF receptor, IGF-binding proteins (IGFBPs) and Pit-1 during early stages of development of Sparus aurata larvae from slow and fast growing lines. Our project was a continuation of a previous BARD project and could be divided into five major parts: i) obtaining additional tools to those obtained in the previous project that are necessary to carry out the developmental study; ii) the developmental expression of growth-related genes and their cellular localization; iii) tissue-specific expression and effect of GH on expression of growth-related genes; iv) possible relationship between GH gene structure, growth rate and genetic selection; v) the possible role of the IGF system in gonadal development. The major findings of our research can be summarized as follows: 1) The cDNAs (complete or partial) coding for Sparus IGFBP-2, GH receptor and Pit-1 were cloned. Sequence comparison reveals that the primary structure of IGFBP-2 protein is 43-49% identical to that of zebrafish and other vertebrates. Intensive efforts resulted in cloning a fragment of 138 nucleotides, coding for 46 amino acids in the proximal end of the intracellular domain of GH receptor. This is the first fish GH receptor cDNA that had been cloned to date. The cloned fragment will enable us to complete the GH - receptor cloning. 2) IGF-I, IGF-II, IGFBP-2, and IGF receptor transcripts were detected by RT-PCR method throughout development in unfertilized eggs, embryos, and larvae suggesting that these mRNAs are products of both the maternal and the embryonic genomes. Preliminary RT-PCR analysis suggest that GH receptor transcript is present in post-hatching larvae already on day 1. 3) IGF-1R transcripts were detected in all tissues tested by RT-PCR with highest levels in gill cartilage, skin, kidney, heart, pyloric caeca, and brain. Northern blot analysis detected IGF receptor only in gonads, brain and gill cartilage but not in muscle; GH increased slightly brain and gill cartilage IGF-1R mRNA levels. 4) IGFBP-2 transcript were detected only in liver and gonads, when analyzed by Northern blots; RT-PCR analysis revealed expression in all tissues studied, with the highest levels found in liver, skin, gonad and pyloric caeca. 5) Expression of IGF-I, IGF-II, IGF-1R and IGFBP-2 was analyzed during gonadal development. High levels of IGF-I and IGFBP-2 expression were found in bisexual young gonads, which decreased during gonadal development. Regardless of maturational stage, IGF-II levels were higher than those of IGF-L 6) The GH gene was cloned and its structure was characterized. It contains minisatellites of tandem repeats in the first and third introns that result in high level of genetic polymorphism. 7) Analysis of the presence of IGF-I and two types of IGF receptor by immunohistochemistry revealed tissue- and stage-specific expression during larval development. Immunohistochemistry also showed that IGF-I and its receptors are present in both testicular and ovarian cells. Although at this stage we are not able to pinpoint which is the rate-limiting step causing the slow growth of Sparus aurata, our project (together with the previous BARD) yielded a great number of experimental tools both DNA probes and antibodies that will enable further studies on the factors regulating growth in Sparus aurata. Our expression studies and cellular localization shed new light on the tissue and developmental expression of growth-related genes in fish.
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Altstein, Miriam, and Ronald J. Nachman. Rational Design of Insect Control Agent Prototypes Based on Pyrokinin/PBAN Neuropeptide Antagonists. United States Department of Agriculture, August 2013. http://dx.doi.org/10.32747/2013.7593398.bard.
Full textAbstract:
The general objective of this study was to develop rationally designed mimetic antagonists (and agonists) of the PK/PBAN Np class with enhanced bio-stability and bioavailability as prototypes for effective and environmentally friendly pest insect management agents. The PK/PBAN family is a multifunctional group of Nps that mediates key functions in insects (sex pheromone biosynthesis, cuticular melanization, myotropic activity, diapause and pupal development) and is, therefore, of high scientific and applied interest. The objectives of the current study were: (i) to identify an antagonist biophores (ii) to develop an arsenal of amphiphilic topically active PK/PBAN antagonists with an array of different time-release profiles based on the previously developed prototype analog; (iii) to develop rationally designed non-peptide SMLs based on the antagonist biophore determined in (i) and evaluate them in cloned receptor microplate binding assays and by pheromonotropic, melanotropic and pupariation in vivo assays. (iv) to clone PK/PBAN receptors (PK/PBAN-Rs) for further understanding of receptor-ligand interactions; (v) to develop microplate binding assays for screening the above SMLs. In the course of the granting period A series of amphiphilic PK/PBAN analogs based on a linear lead antagonist from the previous BARD grant was synthesized that incorporated a diverse array of hydrophobic groups (HR-Suc-A[dF]PRLa). Others were synthesized via the attachment of polyethylene glycol (PEG) polymers. A hydrophobic, biostablePK/PBAN/DH analog DH-2Abf-K prevented the onset of the protective state of diapause in H. zea pupae [EC50=7 pmol/larva] following injection into the preceding larval stage. It effectively induces the crop pest to commit a form of ‘ecological suicide’. Evaluation of a set of amphiphilic PK analogs with a diverse array of hydrophobic groups of the formula HR-Suc-FTPRLa led to the identification of analog T-63 (HR=Decyl) that increased the extent of diapause termination by a factor of 70% when applied topically to newly emerged pupae. Another biostablePK analog PK-Oic-1 featured anti-feedant and aphicidal properties that matched the potency of some commercial aphicides. Native PK showed no significant activity. The aphicidal effects were blocked by a new PEGylated PK antagonist analog PK-dF-PEG4, suggesting that the activity is mediated by a PK/PBAN receptor and therefore indicative of a novel and selective mode-of-action. Using a novel transPro mimetic motif (dihydroimidazole; ‘Jones’) developed in previous BARD-sponsored work, the first antagonist for the diapause hormone (DH), DH-Jo, was developed and shown to block over 50% of H. zea pupal diapause termination activity of native DH. This novel antagonist development strategy may be applicable to other invertebrate and vertebrate hormones that feature a transPro in the active core. The research identifies a critical component of the antagonist biophore for this PK/PBAN receptor subtype, i.e. a trans-oriented Pro. Additional work led to the molecular cloning and functional characterization of the DH receptor from H. zea, allowing for the discovery of three other DH antagonist analogs: Drosophila ETH, a β-AA analog, and a dF analog. The receptor experiments identified an agonist (DH-2Abf-dA) with a maximal response greater than native DH. ‘Deconvolution’ of a rationally-designed nonpeptide heterocyclic combinatorial library with a cyclic bis-guanidino (BG) scaffold led to discovery of several members that elicited activity in a pupariation acceleration assay, and one that also showed activity in an H. zea diapause termination assay, eliciting a maximal response of 90%. Molecular cloning and functional characterization of a CAP2b antidiuretic receptor from the kissing bug (R. prolixus) as well as the first CAP2b and PK receptors from a tick was also achieved. Notably, the PK/PBAN-like receptor from the cattle fever tick is unique among known PK/PBAN and CAP2b receptors in that it can interact with both ligand types, providing further evidence for an evolutionary relationship between these two NP families. In the course of the granting period we also managed to clone the PK/PBAN-R of H. peltigera, to express it and the S. littoralis-R Sf-9 cells and to evaluate their interaction with a variety of PK/PBAN ligands. In addition, three functional microplate assays in a HTS format have been developed: a cell-membrane competitive ligand binding assay; a Ca flux assay and a whole cell cAMP ELISA. The Ca flux assay has been used for receptor characterization due to its extremely high sensitivity. Computer homology studies were carried out to predict both receptor’s SAR and based on this analysis 8 mutants have been generated. The bioavailability of small linear antagonistic peptides has been evaluated and was found to be highly effective as sex pheromone biosynthesis inhibitors. The activity of 11 new amphiphilic analogs has also been evaluated. Unfortunately, due to a problem with the Heliothis moth colony we were unable to select those with pheromonotropic antagonistic activity and further check their bioavailability. Six peptides exhibited some melanotropic antagonistic activity but due to the low inhibitory effect the peptides were not further tested for bioavailability in S. littoralis larvae. Despite the fact that no new antagonistic peptides were discovered in the course of this granting period the results contribute to a better understanding of the interaction of the PK/PBAN family of Nps with their receptors, provided several HT assays for screening of libraries of various origin for presence of PK/PBAN-Ragonists and antagonists and provided important practical information for the further design of new, peptide-based insecticide prototypes aimed at the disruption of key neuroendocrine physiological functions in pest insects.