Relevant bibliographies by topics / Ligase IV

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Academic literature on the topic 'Ligase IV'

Author: Grafiati
Published: 4 June 2021
Last updated: 6 September 2023

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Journal articles on the topic "Ligase IV"

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Nick McElhinny, Stephanie A., Carey M. Snowden, Joseph McCarville, and Dale A. Ramsden. "Ku Recruits the XRCC4-Ligase IV Complex to DNA Ends." Molecular and Cellular Biology 20, no. 9 (May 1, 2000): 2996–3003. http://dx.doi.org/10.1128/mcb.20.9.2996-3003.2000.

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ABSTRACT Genetic experiments have determined that Ku, XRCC4, and ligase IV are required for repair of double-strand breaks by the end-joining pathway. The last two factors form a tight complex in cells. However, ligase IV is only one of three known mammalian ligases and is intrinsically the least active in intermolecular ligation; thus, the biochemical basis for requiring this ligase has been unclear. We demonstrate here a direct physical interaction between the XRCC4-ligase IV complex and Ku. This interaction is stimulated once Ku binds to DNA ends. Since XRCC4-ligase IV alone has very low DNA binding activity, Ku is required for effective recruitment of this ligase to DNA ends. We further show that this recruitment is critical for efficient end-joining activity in vitro. Preformation of a complex containing Ku and XRCC4-ligase IV increases the initial ligation rate 20-fold, indicating that recruitment of the ligase is an important limiting step in intermolecular ligation. Recruitment by Ku also allows XRCC4-ligase IV to use Ku's high affinity for DNA ends to rapidly locate and ligate ends in an excess of unbroken DNA, a necessity for end joining in cells. These properties are conferred only on ligase IV, because Ku does not similarly interact with the other mammalian ligases. We have therefore defined cell-free conditions that reflect the genetic requirement for ligase IV in cellular end joining and consequently can explain in molecular terms why this factor is required.
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Malashetty, Vidyasagar, Audrey Au, Jose Chavez, Mary Hanna, Jennifer Chu, Jesse Penna, and Patricia Cortes. "The DNA binding domain and the C-terminal region of DNA Ligase IV specify its role in V(D)J recombination." PLOS ONE 18, no. 2 (February 24, 2023): e0282236. http://dx.doi.org/10.1371/journal.pone.0282236.

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DNA Ligase IV is responsible for the repair of DNA double-strand breaks (DSB), including DSBs that are generated during V(D)J recombination. Like other DNA ligases, Ligase IV contains a catalytic core with three subdomains—the DNA binding (DBD), the nucleotidyltransferase (NTD), and the oligonucleotide/oligosaccharide-fold subdomain (OBD). Ligase IV also has a unique C-terminal region that includes two BRCT domains, a nuclear localization signal sequence and a stretch of amino acid that participate in its interaction with XRCC4. Out of the three mammalian ligases, Ligase IV is the only ligase that participates in and is required for V(D)J recombination. Identification of the minimal domains within DNA Ligase IV that contribute to V(D)J recombination has remained unresolved. The interaction of the Ligase IV DNA binding domain with Artemis, and the interaction of its C-terminal region with XRCC4, suggest that both of these regions that also interact with the Ku70/80 heterodimer are important and might be sufficient for mediating participation of DNA Ligase IV in V(D)J recombination. This hypothesis was investigated by generating chimeric ligase proteins by swapping domains, and testing their ability to rescue V(D)J recombination in Ligase IV-deficient cells. We demonstrate that a fusion protein containing Ligase I NTD and OBDs flanked by DNA Ligase IV DBD and C-terminal region is sufficient to support V(D)J recombination. This chimeric protein, which we named Ligase 37, complemented formation of coding and signal joints. Coding joints generated with Ligase 37 were shorter than those observed with wild type DNA Ligase IV. The shorter length was due to increased nucleotide deletions and decreased nucleotide insertions. Additionally, overexpression of Ligase 37 in a mouse pro-B cell line supported a shift towards shorter coding joints. Our findings demonstrate that the ability of DNA Ligase IV to participate in V(D)J recombination is in large part mediated by its DBD and C-terminal region.
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Chistiakov, Dimitry A., Natalia V. Voronova, and Alexander P. Chistiakov. "Ligase IV syndrome." European Journal of Medical Genetics 52, no. 6 (November 2009): 373–78. http://dx.doi.org/10.1016/j.ejmg.2009.05.009.

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Wei, Y. F., P. Robins, K. Carter, K. Caldecott, D. J. Pappin, G. L. Yu, R. P. Wang, B. K. Shell, R. A. Nash, and P. Schär. "Molecular cloning and expression of human cDNAs encoding a novel DNA ligase IV and DNA ligase III, an enzyme active in DNA repair and recombination." Molecular and Cellular Biology 15, no. 6 (June 1995): 3206–16. http://dx.doi.org/10.1128/mcb.15.6.3206.

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Three distinct DNA ligases, I to III, have been found previously in mammalian cells, but a cloned cDNA has been identified only for DNA ligase I, an essential enzyme active in DNA replication. A short peptide sequence conserved close to the C terminus of all known eukaryotic DNA ligases was used to search for additional homologous sequences in human cDNA libraries. Two different incomplete cDNA clones that showed partial homology to the conserved peptide were identified. Full-length cDNAs were obtained and expressed by in vitro transcription and translation. The 103-kDa product of one cDNA clone formed a characteristic complex with the XRCC1 DNA repair protein and was identical with the previously described DNA ligase III. DNA ligase III appears closely related to the smaller DNA ligase II. The 96-kDa in vitro translation product of the second cDNA clone was also shown to be an ATP-dependent DNA ligase. A fourth DNA ligase (DNA ligase IV) has been purified from human cells and shown to be identical to the 96-kDa DNA ligase by unique agreement between mass spectrometry data on tryptic peptides from the purified enzyme and the predicted open reading frame of the cloned cDNA. The amino acid sequences of DNA ligases III and IV share a related active-site motif and several short regions of homology with DNA ligase I, other DNA ligases, and RNA capping enzymes. DNA ligases III and IV are encoded by distinct genes located on human chromosomes 17q11.2-12 and 13q33-34, respectively.
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Zhao, Bailin, Tasmin Naila, Michael R. Lieber, and Alan E. Tomkinson. "NAD+ is not utilized as a co-factor for DNA ligation by human DNA ligase IV." Nucleic Acids Research 48, no. 22 (December 2, 2020): 12746–50. http://dx.doi.org/10.1093/nar/gkaa1118.

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Abstract As nucleotidyl transferases, formation of a covalent enzyme-adenylate intermediate is a common first step of all DNA ligases. While it has been shown that eukaryotic DNA ligases utilize ATP as the adenylation donor, it was recently reported that human DNA ligase IV can also utilize NAD+ and, to a lesser extent ADP-ribose, as the source of the adenylate group and that NAD+, unlike ATP, enhances ligation by supporting multiple catalytic cycles. Since this unexpected finding has significant implications for our understanding of the mechanisms and regulation of DNA double strand break repair, we attempted to confirm that NAD+ and ADP-ribose can be used as co-factors by human DNA ligase IV. Here, we provide evidence that NAD+ does not enhance ligation by pre-adenylated DNA ligase IV, indicating that this co-factor is not utilized for re-adenylation and subsequent cycles of ligation. Moreover, we find that ligation by de-adenylated DNA ligase IV is dependent upon ATP not NAD+ or ADP-ribose. Thus, we conclude that human DNA ligase IV cannot use either NAD+ or ADP-ribose as adenylation donor for ligation.
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Rusché, Laura N., Catherine E. Huang, Kenneth J. Piller, Michael Hemann, Elizabeth Wirtz, and Barbara Sollner-Webb. "The Two RNA Ligases of the Trypanosoma brucei RNA Editing Complex: Cloning the Essential Band IV Gene and Identifying the Band V Gene." Molecular and Cellular Biology 21, no. 4 (February 15, 2001): 979–89. http://dx.doi.org/10.1128/mcb.21.4.979-989.2001.

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ABSTRACT Kinetoplastid RNA editing is a posttranscriptional insertion and deletion of U residues in mitochondrial transcripts that involves RNA ligase. A complex of seven different polypeptides purified fromTrypanosoma brucei mitochondria that catalyzes accurate RNA editing contains RNA ligases of ∼57 kDa (band IV) and ∼50 kDa (band V). From a partial amino acid sequence, cDNA and genomic clones of band IV were isolated, making it the first cloned component of the minimal RNA editing complex. It is indeed an RNA ligase, for when expressed inEscherichia coli, the protein autoadenylylates and catalyzes RNA joining. Overexpression studies revealed that T. brucei can regulate of total band IV protein at the level of translation or protein stability, even upon massively increased mRNA levels. The protein's mitochondrial targeting was confirmed by its location, size when expressed in T. brucei and E. coli, and N-terminal sequence. Importantly, genetic knockout studies demonstrated that the gene for band IV is essential in procyclic trypanosomes. The band IV and band V RNA ligases of the RNA editing complex therefore serve different functions. We also identified the gene for band V RNA ligase, a protein much more homologous to band IV than to other known ligases.
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Tomkinson, Alan E., Tasmin Naila, and Seema Khattri Bhandari. "Altered DNA ligase activity in human disease." Mutagenesis 35, no. 1 (October 20, 2019): 51–60. http://dx.doi.org/10.1093/mutage/gez026.

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Abstract The joining of interruptions in the phosphodiester backbone of DNA is critical to maintain genome stability. These breaks, which are generated as part of normal DNA transactions, such as DNA replication, V(D)J recombination and meiotic recombination as well as directly by DNA damage or due to DNA damage removal, are ultimately sealed by one of three human DNA ligases. DNA ligases I, III and IV each function in the nucleus whereas DNA ligase III is the sole enzyme in mitochondria. While the identification of specific protein partners and the phenotypes caused either by genetic or chemical inactivation have provided insights into the cellular functions of the DNA ligases and evidence for significant functional overlap in nuclear DNA replication and repair, different results have been obtained with mouse and human cells, indicating species-specific differences in the relative contributions of the DNA ligases. Inherited mutations in the human LIG1 and LIG4 genes that result in the generation of polypeptides with partial activity have been identified as the causative factors in rare DNA ligase deficiency syndromes that share a common clinical symptom, immunodeficiency. In the case of DNA ligase IV, the immunodeficiency is due to a defect in V(D)J recombination whereas the cause of the immunodeficiency due to DNA ligase I deficiency is not known. Overexpression of each of the DNA ligases has been observed in cancers. For DNA ligase I, this reflects increased proliferation. Elevated levels of DNA ligase III indicate an increased dependence on an alternative non-homologous end-joining pathway for the repair of DNA double-strand breaks whereas elevated level of DNA ligase IV confer radioresistance due to increased repair of DNA double-strand breaks by the major non-homologous end-joining pathway. Efforts to determine the potential of DNA ligase inhibitors as cancer therapeutics are on-going in preclinical cancer models.
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Przewloka, Marcin R., Paige E. Pardington, Steven M. Yannone, David J. Chen, and Robert B. Cary. "In Vitro and In Vivo Interactions of DNA Ligase IV with a Subunit of the Condensin Complex." Molecular Biology of the Cell 14, no. 2 (February 2003): 685–97. http://dx.doi.org/10.1091/mbc.e01-11-0117.

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Several findings have revealed a likely role for DNA ligase IV, and interacting protein XRCC4, in the final steps of mammalian DNA double-strand break repair. Recent evidence suggests that the human DNA ligase IV protein plays a critical role in the maintenance of genomic stability. To identify protein–protein interactions that may shed further light on the molecular mechanisms of DSB repair and the biological roles of human DNA ligase IV, we have used the yeast two-hybrid system in conjunction with traditional biochemical methods. These efforts have resulted in the identification of a physical association between the DNA ligase IV polypeptide and the human condensin subunit known as hCAP-E. The hCAP-E polypeptide, a member of the Structural Maintenance of Chromosomes (SMC) super-family of proteins, coimmunoprecipitates from cell extracts with DNA ligase IV. Immunofluorescence studies reveal colocalization of DNA ligase IV and hCAP-E in the interphase nucleus, whereas mitotic cells display colocalization of both polypeptides on mitotic chromosomes. Strikingly, the XRCC4 protein is excluded from the area of mitotic chromosomes, suggesting the formation of specialized DNA ligase IV complexes subject to cell cycle regulation. We discuss our findings in light of known and hypothesized roles for ligase IV and the condensin complex.
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Wang, Yu, Brandon J. Lamarche, and Ming-Daw Tsai. "Human DNA Ligase IV and the Ligase IV/XRCC4 Complex: Analysis of Nick Ligation Fidelity†." Biochemistry 46, no. 17 (May 2007): 4962–76. http://dx.doi.org/10.1021/bi0621516.

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Liu, Sicheng, Xunyue Liu, Radhika Pankaj Kamdar, Rujira Wanotayan, Mukesh Kumar Sharma, Noritaka Adachi, and Yoshihisa Matsumoto. "C-terminal region of DNA ligase IV drives XRCC4/DNA ligase IV complex to chromatin." Biochemical and Biophysical Research Communications 439, no. 2 (September 2013): 173–78. http://dx.doi.org/10.1016/j.bbrc.2013.08.068.

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Dissertations / Theses on the topic "Ligase IV"

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De, Melo Abinadabe Jackson. "Molecular basis for the structural role of human DNA ligase IV." Thesis, Aix-Marseille, 2016. http://www.theses.fr/2016AIXM4040.

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Les défauts dans la réparation des cassures double-brin de l'ADN (DSBs) peuvent avoir d'importantes conséquences pouvant entrainer une instabilité génomique et conduire à la mort cellulaire ou au développement de cancers. Dans la plupart des cellules mammifères, le mécanisme de Jonction des Extrémités Non Homologues (NHEJ) est le principal mécanisme de réparation des DSBs. L'ADN Ligase IV (LigIV) est une protéine unique dans sa capacité à promouvoir la NHEJ classique. Elle s'associe avec deux autres protéines structuralement similaires, XRCC4 et XLF (ou Cernunnos). LigIV interagit directement avec XRCC4 pour former un complexe stable, tandis que l'interaction entre XLF et ce complexe est médiée par XRCC4. XLF stimule fortement l'activité de ligation du complexe LigIV/XRCC4 par un mécanisme encore indéterminé. Récemment, un rôle structurel non catalytique a été attribué à LigIV (Cottarel et al., 2013). Dans le travail de thèse présenté ici, nous avons reconstitué l'étape de ligation de la NHEJ en utilisant des protéines recombinantes produites dans des bactéries afin d’une part, d'explorer les bases moléculaires du rôle structural de LigIV, d’autre part de comprendre le mécanisme par lequel XLF stimule le complexe de ligation, et enfin de mieux comprendre comment ces trois protéines coopèrent au cours de la NHEJ. Nos analyses biochimiques suggèrent que XLF via son interaction avec XRCC4 lié à LigIV, pourrait induire un changement conformationnel dans la LigIV. Ce réarrangement de la ligase exposerait son interface de liaison à l'ADN ce qui lui permettrait alors de ponter deux molécules indépendantes d'ADN, une capacité indépendante de l'activité catalytique de LigIV
Failure to repair DNA double-strand breaks (DSBs) may have deleterious consequences inducing genomic instability and even cell death. In most mammalian cells, Non-Homologous End Joining (NHEJ) is a prominent DSB repair pathway. DNA ligase IV (LigIV) is unique in its ability to promote classical NHEJ. It associates with two structurally related proteins called XRCC4 and XLF (aka Cernunnos). LigIV directly interacts with XRCC4 forming a stable complex while the XLF interaction with this complex is mediated by XRCC4. XLF strongly stimulates the ligation activity of the LigIV/XRCC4 complex by an unknown mechanism. Recently, a structural noncatalytic role of LigIV has been uncovered (Cottarel et al., 2013). Here, we have reconstituted the end joining ligation step using recombinant proteins produced in bacteria to explore not only the molecular basis for the structural role of LigIV, but also to understand the mechanism by which XLF stimulates the ligation complex, and how these three proteins work together during NHEJ. Our biochemical analysis suggests that XLF, through interactions with LigIV/XRCC4 complex, could induce a conformational change in LigIV. Rearrangement of the LigIV would expose its DNA binding interface that is able to bridge two independent DNA molecules. This bridging ability is fully independent of LigIV’s catalytic activity. We have mutated this interface in order to attempt to disrupt the newly identified DNA bridging ability. In vitro analysis of this LigIV mutant will be presented as well as a preliminary in vivo analysis
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Nijnik, A. "ENU mouse mutant with a hypomorphic mutation in DNA ligase IV." Thesis, University of Oxford, 2006. http://ora.ox.ac.uk/objects/uuid:5a8ae151-6261-4ad4-9541-d5c59ba654a7.

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Stiff, Thomas. "Characterisation of proteins that stimulate the DNA ligase IV/Xrcc4 complex." Thesis, University of Sussex, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.249402.

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Wang, Yu. "Mismatch ligation during non-homologous end joining pathway kinetic characterization of human DNA ligase IV/XRCC4 complex /." Columbus, Ohio : Ohio State University, 2007. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1179947467.

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Amram, Jérémy. "Etude structurale et fonctionnelle des complexes multi-protéiques de la voie de réparation NHEJ chez l’homme." Thesis, Paris 11, 2015. http://www.theses.fr/2015PA114822/document.

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La voie de réparation NHEJ (Non-Homologous End-Joining) est une voie majeure de réparation des cassures double-brin chez l’homme. Les protéines de cette voie interagissent et forment des complexes dynamiques dont les mécanismes moléculaires sont encore largement méconnus. Nous avons dans un premier temps mis au point des protocoles de production à l’échelle de plusieurs milligrammes des protéines cœur de la voie NHEJ en cellules d’insecte à l’aide du système MultiBac. Nous avons ainsi purifié les complexes Ku70/Ku80 et Ligase4/XRCC4 et les protéines Cernunnos et Artemis à homogénéité. Des essais de cristallisation, des études par SAXS et des analyses par microscopie électronique ont été réalisés sur différents complexes formés par ces protéines cœur du NHEJ. Nous avons également caractérisé par chromatographie d’exclusion de taille et calorimétrie, les interactions effectuées entre les protéines de la voie NHEJ. L’ensemble de ces travaux a permis d’établir des bases biochimiques solides en vue des études structurales et fonctionnelles de la voie NHEJ chez l’homme
Human DNA repair pathway NHEJ (Non-Homologous End-Joining) is a major pathway of double-strand breaks repair. The proteins involved in this pathway interact and form dynamic complexes whose molecular mechanisms are largely unknown. Firstly, we established protocols to be able to purify milligrams of those NHEJ pathway core proteins using MultiBac insect cells system. We then purified Ku70/Ku80 and Ligase4/XRCC4 complexes, Artemis and Cernunnos to homogeneity. Crystallogenesis assays, SAXS experiments and Transmission Electronic Microscopy experiments have been performed on several complexes formed by these core NHEJ proteins. We also characterized the interactions between these proteins by Size Exclusion Chromatography and Isothermal Calorimetry. These experiments have led to biochemical results sufficient to establish a solid basis to initiate the structural and functional study of the Human NHEJ Pathway
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Ochi, Takashi. "Non-homologous end-joining DNA double strand break repair : structural studies of the DNA ligase IV/XRCC4 complex." Thesis, University of Cambridge, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.609793.

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PATRIZI, LAURA. "ANALYSIS OF B LYMPHOCYTES IN MOUSE MODEL LIGASE IV WITH HYPOMORPHIC MUTATION IN VDJ RECOMBINATION ASSOCIATED WITH GROWTH DEFECT." Doctoral thesis, Università degli Studi di Milano, 2010. http://hdl.handle.net/2434/150193.

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The major mechanism for the repair of DNA doublestrand breaks (DSBs) in mammalian cells is non-homologous end-joining (NHEJ), a process that involves the DNA-dependent protein kinase , XRCC4 and DNA ligase IV. Rodent cells and mice defective in these components are radiation-sensitive and defective in V(D)J-recombination, showing that NHEJ also functions to rejoin DSBs introduced during lymphocyte development. We have generated a knock-in mouse model with a homozygous Lig4 arginine to histidine (R278H) mutation that corresponds to the mutation identified in the first LIG4-deficient patient, who developed T cell leukemia associated with increased cellular radiosensitivity. The clinical presentation of the syndrome is complex and heterogeneous and may include varying degrees of lymphopenia, growth retardation and microcephaly. The phenotypic effects of the impaired repair of non programmed DNA damage are more diverse and difficult to study. Although such defects in cell survival and proliferation are likely to have an impact on the immune system, their contribution to the immunodeficiency of the LigIV syndrome remains unknown.
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Menchon, Grégory. "Criblage virtuel et fonctionnel sur le complexe XRCC4/ADN ligase IV/Cer-XLF de ligature des cassures double-brin de l'ADN : application en radiosensibilisation tumorale." Thesis, Toulouse 3, 2015. http://www.theses.fr/2015TOU30395.

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En cancérologie, la radiothérapie est une des armes essentielles pour éradiquer les cellules tumorales. Les cassures des deux brins de l'ADN dites "double-brin" qu'elle induit sont particulièrement toxiques et constituent la principale cause de mort cellulaire. La NHEJ (Jonction d'Extrémités Non-Homologues) est la voie métabolique majeure de réparation de ces cassures double-brin de l'ADN et par ce mécanisme, les cellules humaines adoptent une résistance à la radiothérapie. Ce mécanisme de réparation constitue donc une cible de choix pour un traitement anticancéreux combiné en vue d'augmenter la sensibilité des cellules cancéreuses aux rayons ionisants (radiosensibilisation). Au cours du mécanisme NHEJ, la ligature finale des extrémités d'ADN est assurée par le complexe protéique tripartite: XRCC4/ADN Ligase IV/Cernunnos-XLF. Les interfaces protéiques concernées représentent toutes des cibles potentielles dans une stratégie rationnelle d'isolement de molécules inhibitrices, guidée par les structures tridimensionnelles de chaque protéine. A travers des expériences de criblage virtuel et de validation à la fois biophysique et biochimique, nous avons isolé les premières molécules capable de prévenir in vitro les interactions protéine-protéine pour les complexes XRCC4/Lig4 et XRCC4/Cer-XLF, respectivement. Ces composés sont des points de départ pour l'élaboration d'inhibiteurs potentiels de plus haute affinité grâce à l'apport de la biologie structurale, en vue d'un effet radiosensibilisant cellulaire
Radiotherapy is a major weapon used against cancer. Radio-induced DNA double strand breaks (DSB) are the main lesions responsible for cell death. Non-homologous end-joining (NHEJ) is a predominant DSB repair mechanism which contributes to cancer cells resistance to radiotherapy. NHEJ is thus a good target for strategies which aim at increasing the radio-sensitivity of tumors. Through in silico screening and biophysical and biochemical assays, our objective was to find specific ligands for the XRCC4/Lig4 and XRCC4/Cer-XLF protein-protein interactions involved in NHEJ. Here, we isolated the first compounds able to prevent their interaction in vitro. These early stage inhibitors are promising tools for cancer therapy with the hope to develop more specific compounds for cellular assays through the 3D structure of the protein/inhibitor complexes
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Tatavarthi, Haritha. "Action of Tyrosyl DNA Phosphodiesterase on 3'-Phosphoglycolate Terminated DNA Strand Breaks." VCU Scholars Compass, 2006. http://hdl.handle.net/10156/1799.

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Costa, Vânia Aparecida da. "Efeitos termoelétricos em ligas e nanoestruturas de semicondutores IV-VI." Instituto Nacional de Pesquisas Espaciais (INPE), 2015. http://urlib.net/sid.inpe.br/mtc-m21b/2015/01.30.11.47.

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Este trabalho apresenta um estudo teórico das propriedades termoelétricas de ligase nanoestruturas de semicondures IV-VI. Estes materiais têm sido muito estudados por possuírem altos valores de figura de mérito e por suas nanoestruturas (superredes, fios e poços quânticos) apresentarem possibilidades na melhora da resposta termoelétrica. Dentre os modelos teóricos desenvolvidos até o momento, pode-se observar que ainda não foi possível modelar quantitativamente os coeficientes mensurados e nem determinar o principal mecanismo responsável pela resposta termoelétrica. Para contribuir nessa área, foram estudados os coeficientes de transporte de ligas \emph{Pb$_{1-x}$Sn$_{x}$Te}, fios quânticos e super-redes de \emph{PbTe}. Partindo do modelo \emph{k . p}, conhecido como modelo de Dimmock, foi calculada a estrutura de bandas incluindo efeitos de não parabolicidade, anisotropia e múltiplos vales. Os coeficientes de transporte do \emph{bulk} e super-rede foram calculados dentro do formalismo semiclássico que utiliza a solução da equação de Boltzmann na aproximação do tempo de relaxação. No caso do fio quântico, foi utilizado o formalismo de Landauer, onde os coeficientes de transporte são dados em termos da probabilidade de transmissão de elétrons em movimento balístico entre contatos representados por reservatórios em equilíbrio. Com base na análise das variações de temperatura, concentração de portadores, percentual de estanho do \emph{bulk}, geometria e parâmetros das nanoestrururas, foram discutidos os mecanismos para o aumento da eficiência termoelétrica destes materiais. Os resultados indicam que o modelo utilizado para o \emph{bulk} possui um bom acordo com a experiência e que o aumento de estanho em ligas \emph{Pb$_{1-x}$Sn$_{x}$Te} melhoraos valores de figura de mérito. Um fator de ganho foi introduzido para avaliar a resposta termoelétrica devido à alteração na dimensionalidade e empacotamento de nanoestruturas no interior de um \emph{bulk} 3D finito. Alterações na eficiência termoelétrica de super-redes IV-VI podem ser obtidas explorando a sua anisotropia o que implica em novas possilibidades de aplicação em dispositivos termoelétricos.
This work presents a theoretical study of thermoelectric properties of IV-VI semiconductor alloys and nanostructures. These materials have a been widely studied because they have high figure of merit and their nanostructures (superlattices, quantum wires and quantum wells) are able to improving the thermoelectric response. The theoretical models, developed until now, either have not been able to model quantitatively the measured coefficients and neither to determine the main mechanism responsible for thermoelectric performance. To contribute in this area, the transport coefficients of \emph{Pb$_{1-x}$Sn$_{x}$Te} alloys, quantum wires and \emph{PbT} e superlattices were studied. From \emph{k . p} model, known as Dimmock model, it was calculated the band structure including non parabolicity, anisotropy and multiple valleys effects. The bulk transport coefficients and superlattice were calculated within the semiclassical formalism which uses the Boltzmann equation solution in the relaxation time approximation. In the quantum wire case, it was used the Landauer formalism, where the transport coefficients are given in terms of the transmission probability of electrons in the ballistic regime, between contacts represented by reservoirs in equilibrium. Based on the analysis of temperature variations, carrier concentration, percentage of tin percentage in bulk, geometry and parameters of nanostructures, were discussed the mechanism to increase the thermoelectric efficiency of these materials. The results indicate that the model used for the bulk has a good agreement with the experiment and the increase of tin in \emph{Pb$_{1-x}$Sn$_{x}$Te} alloys increase the figures of merit values. A gain factor has been introduced to study the thermoelectric performance due to changes in dimensionality and nanostructures packaging within a finite-3D bulk. Changes in the thermoelectric efficiency of IV -VI superlattices can be obtained exploiting its anisotropy, this implies new possilibidades of application in thermoelectric devices.
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Books on the topic "Ligase IV"

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Brûlart, Nicolas. Journal d'un ligueur parisien: Des barricades à la levée du siège de Paris par Henri IV, 1588-1590. Genève: Droz, 1999.

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2

Un prêtre anglais contre Henri IV: Archéologie d'une haine religieuse. Paris: Harmattan, 2011.

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Płoski, Marcin. Liga Polskich Rodzin jako aktor społeczny. Wrocław: Wydawnictwo Instytutu Socjologii Uniwersytetu Wrocławskiego, 2016. https://www.repozytorium.uni.wroc.pl/dlibra/publication/80114.

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This book is a monograph, on the political party, that shows the birth, rise and fall of the League of Polish Families. In the chapter I the division was made into individual and collective actors. Chapter II is an attempt to describe the birth and functioning of LPF in the context of a broader national movement. Chapter III focuses on the fields of activities of the national party. Chapter IV is an attempt to recreate the role of LPF on the political scene. Chapter V is an attempt to play the role of LPF on the European political scene. Chapter VI is the description of a research taken on the party groups, surveyed with the usage of a questionnaire technique. Chapter VII is an empirical reflection of the views of LPF deputies and senators about political transformation and Polish integration with European structures. The tool for this method was mainly based on an open interview. Chapter VIII is an attempt to present the Polish society in the perspective of political transformation, and the reflection on the degree of civility and democracy and finaly presents the fall of LPF. \n\nNiniejsza książka ma charakter monografii poświęconej partii politycznej, która ukazuje narodziny, rozkwit i upadek Ligi Polskich Rodzin. W rozdziale I dokonany został podział na aktorów zbiorowych i indywidualnych. Rozdział II jest próbą opisu narodzin i funkcjonowania LPR w ramach szerszego ruchu narodowego. Rozdział III koncentruje się na polach aktywności partii narodowej. Rozdział IV jest próbą odtworzenia roli LPR-u na scenie politycznej. Rozdział V jest próbą odtworzenia roli LPR na europejskiej scenie politycznej. Rozdział VI stanowi badawczy portret kół partyjnych, przeprowadzony za pomocą metody ankietowej z zastosowaniem kwestionariusza ankiety. Rozdział VII jest empirycznym odzwierciedleniem poglądów posłów i senatorów LPR na temat transformacji ustrojowej i integracji Polski ze strukturami europejskimi. Narzędziem tej metody był wywiad swobodny otwarty. Rozdział VIII jest próbą przedstawienia społeczeństwa polskiego w perspektywie transformacji ustrojowej, refleksji nad stopniem obywatelskości i demokracji oraz upadkiem LPR.
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Stiff, Thomas. Characterisation of proteins that stimulate the DNA Ligase IV/Xrcc4 complex. 2002.

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The Paris of Henry of Navarre as Seen by Pierre de l'Estoile. Harvard University Press, 2014.

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IV Міжнародна студентська наукова конференція «ДІДЖИТАЛІЗАЦІЯ НАУКИ ЯК ВИКЛИК СЬОГОДЕННЯ». Європейська наукова платформа, 2023. http://dx.doi.org/10.36074/liga-inter-26.05.2023.

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Викладено матеріали учасників IV Міжнародної мультидисциплінарної студентської наукової конференції «Діджиталізація науки як виклик сьогодення», яка відбулася 26 травня 2023 року у місті Луцьк, Україна.
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IV Міжнародна студентська наукова конференція «СУЧАСНІ АСПЕКТИ ТА ПЕРСПЕКТИВНІ НАПРЯМКИ РОЗВИТКУ НАУКИ». Європейська наукова платформа, 2022. http://dx.doi.org/10.36074/liga-inter-09.12.2022.

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Викладено матеріали учасників IV Міжнародної мультидисциплінарної студентської наукової конференції «Сучасні аспекти та перспективні напрямки розвитку науки», яка відбулася 9 грудня 2022 року у місті Ужгород, Україна.
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IV Міжнародна студентська наукова конференція «АКТУАЛЬНІ ПИТАННЯ ТА ПЕРСПЕКТИВИ ПРОВЕДЕННЯ НАУКОВИХ ДОСЛІДЖЕНЬ». Європейська наукова платформа, 2023. http://dx.doi.org/10.36074/liga-inter-17.03.2023.

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Викладено матеріали учасників IV Міжнародної мультидисциплінарної студентської наукової конференції «Актуальні питання та перспективи проведення наукових досліджень», яка відбулася 17 березня 2023 року у місті Полтава, Україна.
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IV Міжнародна студентська наукова конференція «ТЕОРЕТИЧНЕ ТА ПРАКТИЧНЕ ЗАСТОСУВАННЯ РЕЗУЛЬТАТІВ СУЧАСНОЇ НАУКИ». Європейська наукова платформа, 2023. http://dx.doi.org/10.36074/liga-inter-07.04.2023.

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Викладено матеріали учасників IV Міжнародної мультидисциплінарної студентської наукової конференції «Теоретичне та практичне застосування результатів сучасної науки», яка відбулася 7 квітня 2023 року у місті Одеса, Україна.
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IV Міжнародна студентська наукова конференція «ЦИФРОВІЗАЦІЯ НАУКИ ТА СУЧАСНІ ТРЕНДИ ЇЇ РОЗВИТКУ». Європейська наукова платформа, 2023. http://dx.doi.org/10.36074/liga-inter-05.05.2023.

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Викладено матеріали учасників IV Міжнародної мультидисциплінарної студентської наукової конференції «Цифровізація науки та сучасні тренди її розвитку», яка відбулася 5 травня 2023 року у місті Вінниця, Україна.
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Book chapters on the topic "Ligase IV"

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Chistiakov, Dimitry A. "Ligase IV Syndrome." In Advances in Experimental Medicine and Biology, 175–85. New York, NY: Springer New York, 2010. http://dx.doi.org/10.1007/978-1-4419-6448-9_16.

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Dückers, Gregor, and Tim Niehues. "DNA Ligase IV Deficiency." In Genetic Syndromes, 1–2. Cham: Springer International Publishing, 2022. http://dx.doi.org/10.1007/978-3-319-66816-1_67-1.

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Markaverich, Barry M., Brian S. Middleditch, and James Clark. "Inhibition of Cell Proliferation by a Nuclear Type II Ligand: Methyl P-Hydroxyphenylactate." In Gene Regulation by Steroid Hormones IV, 213–19. New York, NY: Springer New York, 1989. http://dx.doi.org/10.1007/978-1-4612-3666-5_13.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of oxovanadium(IV) complex with acetylacetone ligand." In Magnetic Properties of Paramagnetic Compounds, 189. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_100.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of oxovanadium(IV) complex with macrocyclic ligand." In Magnetic Properties of Paramagnetic Compounds, 177–78. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_94.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of oxovanadium(IV) complex with macrocyclic ligand." In Magnetic Properties of Paramagnetic Compounds, 179–80. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_95.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of oxovanadium(IV) complex with macrocyclic ligand." In Magnetic Properties of Paramagnetic Compounds, 181–82. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_96.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of oxovanadium(IV) complex with macrocyclic ligand." In Magnetic Properties of Paramagnetic Compounds, 183–84. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_97.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of oxovanadium(IV) complex with macrocyclic ligand." In Magnetic Properties of Paramagnetic Compounds, 185–86. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_98.

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Pardasani, R. T., and P. Pardasani. "Magnetic properties of μ-cyanide bridged vanadium(IV)-molybdenum(IV) complex with substituted amido ligand." In Magnetic Properties of Paramagnetic Compounds, 127–28. Berlin, Heidelberg: Springer Berlin Heidelberg, 2017. http://dx.doi.org/10.1007/978-3-662-54228-6_68.

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Conference papers on the topic "Ligase IV"

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Hooks, Grace, Yasmin Anchondo, Neelam Sharma, Jac Nickoloff, and Amanda Ashley. "Abstract LB-164: DNA ligase IV modulates the cellular response to DNA replication stress." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-lb-164.

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Lin, Tingting, Zhilian Zhou, Lifeng Zhu, Yandan Fan, Xiaofen Ding, and Yingming Sun. "Abstract 3066: DNA ligase IV inhibitor and X-ray exert a synthetic lethal in loss-of-function p53 cells." In Proceedings: AACR Annual Meeting 2021; April 10-15, 2021 and May 17-21, 2021; Philadelphia, PA. American Association for Cancer Research, 2021. http://dx.doi.org/10.1158/1538-7445.am2021-3066.

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Tseng, Hui-Min, David Shum, Hakim Djaballah, and David Scheinberg. "Abstract 3689: Identification of DNA ligase IV inhibitors as possible drug and probe candidates for enhancement of radiation treatment and chemotherapy." In Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-3689.

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Silva, Alessandro Márcio Hakme Da, Alessandro Fraga Farah, Abimael Caleb Ribeiro, Alexandre José Ribeiro, and Jonas De Carvalho. "MODELO COMPUTACIONAL DE ANÁLISE DO COMPORTAMENTO MECÂNICO DA LIGA TI-6AL-4V EM COMPARAÇÃO A LIGA Ti-20Nb." In IV Simpósio de Tecnologias da Fatec de Sertãozinho (SITEFA/Stz). Fatec Sertãozinho, 2022. http://dx.doi.org/10.33635/sitefa.v4i1.163.

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O presente trabalho apresenta um modelo computacional com análise pelo método dos elementos finitos aplicado a uma liga Titânio-Nióbio Ti-20Nb comparativamente a liga de titânio TI-6AL-4V. Os dois modelos de compressão mecânica foram criados e simulados no Software Ansys. Os objetivos do trabalho foram analisar comparativamente, através de um modelo computacional, as ligas a fim de se interpretar as características mecânicas de tensões e deformações sob carregamento estático, buscando compreender e verificar se os comportamentos das ligas são similares, em vista que o nióbio poderá ser usado como material para próteses ortodônticas e ortopédicas em um cenário futuro. Foi utilizado o Método dos Elementos Finitos (MEF), que é um método numérico usado para resolver, de forma aproximada, equações diferenciais que governam o comportamento mecânico dos materiais das ligas consideradas no estudo. Foi utilizado o software de desenho 3D SpaceClaim, para a criação dos modelos simplificados TiM1 (titânio malha 1) e NbM1 (Nióbio malha 1). A análise comparativa entre os resultados computacionais e experimentais foi realizada em termos qualitativos e quantitativos. Na comparação dos comportamentos mecânicos das ligas, Ti-6Al-4V e Ti-20Nb, as tensões se concentraram na porção onde foi feita a vinculação do modelo e os maiores valores de tensão encontrados diferiram entre os modelos considerados da ordem de 7%. As tensões médias encontradas ficaram bem próximas do modelo numérico para escalas em tecidos ósseos com implantes, e as deformações totais foram pequenas comparadas com modelos para ligas de titânio mais utilizadas em medicina, o que corrobora que a liga Ti-Nb é bastante promissora para aplicações na área de saúde.
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Oliveira, Lucas Fernandes de, Amanda Lilian Monteiro de Almeida, Claudieli dos Santos Moreira, Maria Alix Leite Araújo, Ana Fátima Braga Rocha, Ana Karinne Dantas de Oliveira, Marilene Alves Oliveira Guanabara, Simone Paes de Melo, Valéria Lima de Barros, and Aline Sales Nunes Félix. "Estudo da epidemiologia como ferramenta de ensino na extensão universitária: um relato de experiência." In XIII Congresso da Sociedade Brasileira de DST - IX Congresso Brasileiro de AIDS - IV Congresso Latino Americano de IST/HIV/AIDS. Zeppelini Editorial e Comunicação, 2021. http://dx.doi.org/10.5327/dst-2177-8264-202133p048.

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Introdução: Capaz de transcender ampla influência social, agindo diretamente na sociedade, tendo como características histórico-sociais, mostrando ter relevância no processo de melhoria na sociedade, as ligas acadêmicas mostram-se fundamentais no processo de formação profissional, social, pois busca fazer com que os discentes participantes alcancem o trabalho pensando no tripé saúde, comunidade e universidade, tornando-se para o aluno peça fundamental para formação. Objetivo: Relatar a vivência dos discentes participantes da Liga Acadêmica de Gênero e Infecções Sexualmente Transmissíveis no processo de entendimento da epidemiologia. Métodos: Trata-se de um relato de experiência desenvolvido pela Liga Acadêmica de Gênero e Infecções Sexualmente Transmissíveis, sendo composta de membros de cursos que o Centro de Ciências da Saúde engloba em maio de 2021. O estudo fundamenta-se pela discussão acerca da metodologia de saúde e doença na comunidade e a utilização de conceitos epidemiológicos que auxiliem na execução de atividades que a liga desenvolve. Resultados: A utilização de ferramentas para compreensão dos problemas de saúde e seu ciclo é complexo. Para isso, a liga estuda por bases de dados de domínio público, a fim de entender melhor o quadro de epidemias e as endemias das doenças sexualmente transmissíveis mais prevalentes na comunidade. Os dados são coletados em sistemas virtuais, assim podendo ser discutidos em reunião em busca do desenvolvimento de atividades educativas em saúde para disseminar medidas de promoção em saúde na comunidade. Conclusão: Evidencia-se que a conscientização é a melhor ferramenta de suporte para promover os cuidados necessários para prevenção e, diante da epidemiologia, o grupo pode destacar os principais focos de assistência e proporcionar suporte para os serviços de saúde, tornando-se um modelo de ensino aprendizado essencial no processo de formação.
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Akbar, Himyan, Salma Habib, Mohammed Mahroof Tahir, and Lakshmaiah Sreerama. "Synthesis and Characterization of Vanadium (IV)-Flavonoid Complexes and its Antioxidant ability toward Superoxide and Radical Scavenging." In Qatar University Annual Research Forum & Exhibition. Qatar University Press, 2020. http://dx.doi.org/10.29117/quarfe.2020.0109.

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In this project Vanadium complex -Vanadium (IV) - flavone was synthesized using vanadium (IV) acetylacetonate (VO(acac)2) complex and 3-hydroxy-6-methyl flavone ligand. The complex stability was checked using FTIR and UV-vis spectroscopies. Peackes around 990 cm-1 conforms the formation of (V=O) in the complex, as well as (V-O) around 790 cm-1. In UV-Vis spectrum peak around 400-450 nm was noticed, which conforms the formation of the vanadium complex that correspond to the ligand to metal charge transfer (LMCT) transition. The radical scavenging abilities of vanadium complex were investigated using DPPH. The anti-oxidant activity using (BHA) as a standard reference, the complex synthesized displayed strong DPPH antioxidant radical scavenging activity compared to VO(acac)2 and BHA, with IC50 value of (105, 95 and 96) mM respectively. The absorbance in which the reducing power occurred were found to be (0.397, 0.825 and 0.228) for the complex, VO(acac)2 and BHA.
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Lima, Lorena Santos, Paula Rosane Sousa Andrade, Amanda Almeida Mendonça, Anne Hellen Brito Leite, Cleiton Mateus Santos Siqueira, Dayanne Cristina Barreto da Paixão, Maria de Lara Lucilla de Carvalho, Maria Rafaella Carvalho de Jesus, Flávia Luryane Ferreira Sandes, and Brenda Evelin Barreto da Silva. "Liga acadêmica de sexualidade e infecções sexualmente transmissíveis no contexto da formação de enfermeiros em Sergipe." In XIII Congresso da Sociedade Brasileira de DST - IX Congresso Brasileiro de AIDS - IV Congresso Latino Americano de IST/HIV/AIDS. Zeppelini Editorial e Comunicação, 2021. http://dx.doi.org/10.5327/dst-2177-8264-202133p295.

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Introdução: As Ligas Acadêmicas visam à aproximação dos alunos à concepção do tripé de ensino, pesquisa e extensão e complementam o conhecimento/atuação em áreas não contempladas pelo currículo da graduação. Objetivo: Apresentar a importância da Liga Acadêmica de Sexualidade e Infecções Sexualmente Transmissíveis da Universidade Federal de Sergipe na formação acadêmica em Enfermagem. Métodos: Há 12 anos, a Liga Acadêmica de Sexualidade e Infecções Sexualmente Transmissíveis atua dentro do curso de Enfermagem da Universidade Federal de Sergipe. É composta de discentes da graduação do segundo ao décimo período, docentes do curso e discentes de mestrado e doutorado. Cada membro possui função especificada em regimento próprio. Resultados: A liga oferece diversas oportunidades científicas e assistenciais que são importantes para a formação acadêmica e profissional, podendo-se destacar: a) reuniões quinzenais: planejamento e organização de ações, discussão de artigos científicos e de casos clínicos; b) participação e apresentação de trabalhos em eventos científicos; c) estímulo e preparo para serem divulgadores e multiplicadores de conhecimento das atividades da liga aos demais alunos e comunidade, inclusive por meio de redes sociais; d) capacitação e realização de campanhas de testes rápidos, ações de educação sexual e preventiva; e) acompanhamento de profissionais em ambulatórios de referência de infecções sexualmente transmissíveis/vírus da imunodeficiência humana/aids; f) colaboração em ações/eventos do calendário dos programas estadual e municipal de infecções sexualmente transmissíveis/vírus da imunodeficiência humana/ aids. Assim, a relevância deste projeto está na capacitação de futuros enfermeiros sobre sexualidade, prevenção e controle das infecções sexualmente transmissíveis e assistência de enfermagem a pessoas com infecções sexualmente transmissíveis ou vivendo com vírus da imunodeficiência humana. A participação na Liga Acadêmica de Sexualidade e Infecções Sexualmente Transmissíveis garante experiências inovadoras para desenvolvimento de raciocínio crítico, tomada de decisão e ampliação do conhecimento sobre a temática. Conclusão: A Liga Acadêmica de Sexualidade e Infecções Sexualmente Transmissíveis contribui para a formação de enfermeiros em relação à sexualidade e infecções sexualmente transmissíveis por meio de atividades de ensino, pesquisa e extensão, a fim de melhor atender as necessidades de saúde para as diversidades e vulnerabilidades da sociedade sergipana.
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"Synthesis, Characterization and Antibacterial Activity of Organotin (IV) Complexes with Benzoylacetone Benzhydrazone Ligand." In 3rd International Conference on Biological, Chemical and Environmental Sciences. International Institute of Chemical, Biological & Environmental Engineering, 2015. http://dx.doi.org/10.15242/iicbe.c0915063.

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Wei, Meng, Qian’ge He, Xuegang Liu, and Jing Chen. "N,N,N′,N′-Tetra-Methyl-3-Oxy-Pentane-1,5-Diamide (TMPDA): A Promising Back Extractant for Ln(III) and Zr(IV)." In 18th International Conference on Nuclear Engineering. ASMEDC, 2010. http://dx.doi.org/10.1115/icone18-29358.

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Water-soluble oxa-diamide ligand, N,N,N′,N′-tetra-methyl-3-oxy-pentane-1,5-diamid (TMPDA) has been synthesized and purified. Its crystal structure, melting point, decomposition temperature, solubilities in aqueous phase and organic phase, distribution ratio between aqueous and organic phase, etc. are reported. The effect of TMPDA concentration in aqueous phase and HNO3 concentration in the equilibrium aqueous phase on the extraction efficiency of La(III), Ce(III), Pr(III), Nd(III), Zr(IV), Fe(III), Y(III), Mo(VI), Ru(III) and Pd(II) by 30% TRPO/kerosene have been studied. The results indicate that TMPDA dissolve well in aqueous phase but almost insoluble in kerosene or 30%TRPO/kerosene in the bi-phase system. It can effectively reduce the extraction of Ln(III), Y(III) and Zr(IV) into 30%TRPO/kerosene at a moderate acid system (0.24mol/L∼0.27mol/L HNO3). TMPDA is a promising stripping agent for Ln(III), Y(III) and Zr(IV) from loaded TRPO.
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Alves Luzia da Silva, Rafaela, and Viviane Lopes dos Santos. "LIGA DA CIÊNCIA: APROXIMANDO O LETRAMENTO RACIAL DO ENSINO FUNDAMENTAL I." In IV Arvorecer Negro. ,: Even3, 2022. http://dx.doi.org/10.29327/ivarvorecernegro_2021.423385.

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Reports on the topic "Ligase IV"

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Szigethy, Geza. Rational Ligand Design for U(VI) and Pu(IV). Office of Scientific and Technical Information (OSTI), August 2009. http://dx.doi.org/10.2172/972716.

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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.

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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.
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3

Altstein, Miriam, and Ronald Nachman. Rationally designed insect neuropeptide agonists and antagonists: application for the characterization of the pyrokinin/Pban mechanisms of action in insects. United States Department of Agriculture, October 2006. http://dx.doi.org/10.32747/2006.7587235.bard.

Full text
Abstract:
The general objective of this BARD project focused on rationally designed insect neuropeptide (NP) agonists and antagonists, their application for the characterization of the mechanisms of action of the pyrokinin/PBAN (PK-PBAN) family and the development of biostable, bioavailable versions that can provide the basis for development of novel, environmentally-friendly pest insect control agents. The specific objectives of the study, as originally proposed, were to: (i) Test stimulatory potencies of rationally designed backbone cyclic (BBC) peptides on pheromonotropic, melanotropic, myotropic and pupariation activities; (ii) Test the inhibitory potencies of the BBC compounds on the above activities evoked either by synthetic peptides (PBAN, LPK, myotropin and pheromonotropin) or by the natural endogenous mechanism; (iii) Determine the bioavailability of the most potent BBC compounds that will be found in (ii); (iv) Design, synthesize and examine novel PK/PBAN analogs with enhanced bioavailability and receptor binding; (v) Design and synthesize ‘magic bullet’ analogs and examine their ability to selectively kill cells expressing the PK/PBAN receptor. To achieve these goals the agonistic and antagonistic activities/properties of rationally designed linear and BBC neuropeptide (NP) were thoroughly studied and the information obtained was further used for the design and synthesis of improved compounds toward the design of an insecticide prototype. The study revealed important information on the structure activity relationship (SAR) of agonistic/antagonistic peptides, including definitive identification of the orientation of the Pro residue as trans for agonist activity in 4 PK/PBANbioassays (pheromonotropic, pupariation, melanotropic, & hindgut contractile) and a PK-related CAP₂b bioassay (diuretic); indications that led to the identification of a novel scaffold to develop biostbiostable, bioavailable peptidomimetic PK/PBANagonists/antagonists. The work led to the development of an arsenal of PK/PBAN antagonists with a variety of selectivity profiles; whether between different PKbioassays, or within the same bioassay between different natural elicitors. Examples include selective and non-selective BBC and novel amphiphilic PK pheromonotropic and melanotropic antagonists some of which are capable of penetrating the moth cuticle in efficacious quantities. One of the latter analog group demonstrated unprecedented versatility in its ability to antagonize a broad spectrum of pheromonotropic elicitors. A novel, transPro mimetic motif was proposed & used to develop a strong, selective PK agonist of the melanotropic bioassay in moths. The first antagonist (pure) of PK-related CAP₂b diuresis in flies was developed using a cisPro mimetic motif; an indication that while a transPro orientation is associated with receptor agonism, a cisPro orientation is linked with an antagonist interaction. A novel, biostablePK analog, incorporating β-amino acids at key peptidase-susceptible sites, exhibited in vivo pheromonotropic activity that by far exceeded that of PBAN when applied topically. Direct analysis of neural tissue by state-of-the-art MALDI-TOF/TOF mass spectrometry was used to identify specific PK/PK-related peptides native to eight arthropod pest species [house (M. domestica), stable (S. calcitrans), horn (H. irritans) & flesh (N. bullata) flies; Southern cattle fever tick (B. microplus), European tick (I. ricinus), yellow fever mosquito (A. aegypti), & Southern Green Stink Bug (N. viridula)]; including the unprecedented identification of mass-identical Leu/Ile residues and the first identification of NPs from a tick or the CNS of Hemiptera. Evidence was obtained for the selection of Neb-PK-2 as the primary pupariation factor of the flesh fly (N. bullata) among native PK/PK-related candidates. The peptidomic techniques were also used to map the location of PK/PK-related NP in the nervous system of the model fly D. melanogaster. Knowledge of specific PK sequences can aid in the future design of species specific (or non-specific) NP agonists/antagonists. In addition, the study led to the first cloning of a PK/PBAN receptor from insect larvae (S. littoralis), providing the basis for SAR analysis for the future design of 2ⁿᵈgeneration selective and/or nonselective agonists/antagonists. Development of a microplate ligand binding assay using the PK/PBAN pheromone gland receptor was also carried out. The assay will enable screening, including high throughput, of various libraries (chemical, molecular & natural product) for the discovery of receptor specific agonists/antagonists. In summary, the body of work achieves several key milestones and brings us significantly closer to the development of novel, environmentally friendly pest insect management agents based on insect PK/PBANNPs capable of disrupting critical NP-regulated functions.
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